WO2015017589A1 - 1,4-disubstituted pyridazine derivatives and their use for treating smn-deficiency-related conditions - Google Patents

1,4-disubstituted pyridazine derivatives and their use for treating smn-deficiency-related conditions Download PDF

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Publication number
WO2015017589A1
WO2015017589A1 PCT/US2014/048984 US2014048984W WO2015017589A1 WO 2015017589 A1 WO2015017589 A1 WO 2015017589A1 US 2014048984 W US2014048984 W US 2014048984W WO 2015017589 A1 WO2015017589 A1 WO 2015017589A1
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Prior art keywords
methyl
pyridazin
amino
tetramethylpiperidin
alkyl
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PCT/US2014/048984
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French (fr)
Inventor
Atwood Cheung
Natalie Dales
Timothy Brian Hurley
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Novartis Ag
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Priority to BR112016001899-0A priority Critical patent/BR112016001899B1/en
Priority to US14/909,052 priority patent/US20160184305A1/en
Application filed by Novartis Ag filed Critical Novartis Ag
Priority to EP22159905.3A priority patent/EP4105208A1/en
Priority to AU2014296255A priority patent/AU2014296255B2/en
Priority to MX2020011652A priority patent/MX2020011652A/en
Priority to CA2918805A priority patent/CA2918805C/en
Priority to JP2016531875A priority patent/JP6461150B2/en
Priority to KR1020217034080A priority patent/KR20210130843A/en
Priority to PL14750931.9T priority patent/PL3027600T3/en
Priority to CN201480054025.8A priority patent/CN105636953B/en
Priority to ES14750931T priority patent/ES2917979T3/en
Priority to KR1020167004818A priority patent/KR102318727B1/en
Priority to EA201690287A priority patent/EA030631B1/en
Priority to MX2016001400A priority patent/MX2016001400A/en
Priority to EP14750931.9A priority patent/EP3027600B1/en
Publication of WO2015017589A1 publication Critical patent/WO2015017589A1/en
Priority to US16/259,441 priority patent/US11672799B2/en
Priority to US18/322,521 priority patent/US20240009189A1/en

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    • 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
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/501—Pyridazines; Hydrogenated pyridazines not condensed and containing further heterocyclic rings
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • 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
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/502—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with carbocyclic ring systems, e.g. cinnoline, phthalazine
    • 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
    • A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
    • 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
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/02—Muscle relaxants, e.g. for tetanus or cramps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • 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
    • 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
    • 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
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/10—Spiro-condensed systems

Definitions

  • Proximal spinal muscular atrophy is an inherited, clinically heterogeneous group of neuromuscular disorders characterized by degeneration of the anterior horn cells of the spinal cord. Patients suffer from symmetrical weakness of trunk and limb muscles, the legs being more affected than the arms and the proximal muscles weaker than the distal ones; diaphragm, facial and ocular muscles are spared.
  • SMA spinal muscular atrophy
  • Type I (Werdnig-Hoffmann disease) is the most acute and severe form, with onset before six months and death usually before two years; children are never able to sit without support.
  • Symptoms of the disease can be present in utero, as reduction of fetal movements; at birth; or more often, within the first four months of life.
  • Affected children are particularly floppy, experience feeding difficulties and diaphragmatic breathing, and are characterized by a general weakness in the intercostals and accessory respiratory muscles. Affected children never sit or stand and usually die before the age of 2; death is generally due to respiratory insufficiency.
  • Type II intermediate, chronic form
  • muscular fasciculations are common, and tendon reflexes progressively reduce.
  • Children are unable to stand or walk without aid.
  • Feeding and swallowing problems are not usually present in Type II SMA, although in some patients a feeding tube may become necessary.
  • Most patients generally develop a progressive muscular scoliosis which can require surgical correction.
  • clearing of tracheal secretions and coughing might become difficult because of poor bulbar function and weak intercostal muscles.
  • These patients have profound hypotonia, symmetrical flaccid paralysis, and no control of head movement.
  • Type III Kergelberg-Welander disease, or Juvenile Spinal Muscular Atrophy
  • motor milestones achievement is normal, and deambulation can be preserved until variable ages.
  • These patients often develop scoliosis, and symptoms of joint overuse, generally caused by weakness, are frequently seen. Life expectancy is almost normal but quality of life is markedly compromised.
  • Types I, II and III progress over time, accompanied by deterioration of the patient's condition.
  • Type IV SMA is characterized by weakness in the second or third decade of life, with mild motor impairment not accompanied by respiratory or nutritional problems.
  • Adult SMA is characterized by insidious onset and very slow progression. The bulbar muscles are rarely affected in Type IV. It is not clear that Type IV SMA is etiologically related to the Type l-lll forms.
  • spinal muscular atrophy examples include X-linked disease, spinal muscular atrophy with respiratory distress (SMARD), spinal and bulbar muscular atrophy (Kennedy's disease, or Bulbo-Spinal Muscular Atrophy), and distal spinal muscular atrophy.
  • SMARD spinal muscular atrophy with respiratory distress
  • Kennedy's disease spinal and bulbar muscular atrophy
  • Bulbo-Spinal Muscular Atrophy spinal muscular atrophy
  • SMA is due to mutations in the Survival of Motor Neuron (SMN) gene, which exists in two forms in humans (SMN1 and SMN2). Loss of SMN is deleterious to motor neurons and results in neuromuscular insufficiency, a hallmark of the disease. From a genetic point of view, SMA is an autosomal recessive condition, caused by disruption of SMN1 gene, located in 5q13 (Lefebvre S., et al. (1995) Cell 80: 155-165). More than 98% of patients with spinal muscular atrophy have a homozygous disruption of SMN1 by deletion, rearrangement, or mutation. All these patients, however, retain at least one copy of SMN2.
  • the mRNA transcribed from the SMN1 gene is generally a full-length mRNA with only a small fraction of its transcripts spliced to remove exon 3, 5, or 7 (Gennarelli et al. (1995) Biochem. Biophys. Res. Commun. 213:342-348; Jong et al. (2000) J. Neurol. Sci. 173:147-153). All SMA subjects have at least one, and generally two to four copies of the SMN2 gene, which encodes the same protein as SMN1 ; however, the SMN2 gene produces only low levels of full- length SMN protein.
  • the SMNA7 protein is non-functional and thought to be rapidly degraded. About 10% of
  • SMN2 pre-mRNA is properly spliced and subsequently translated into full length SMN protein (FL- SMN), and the rest being the SMNA7 copy.
  • FL- SMN full length SMN protein
  • the efficiency of SMN2 splicing might be dependent on severity of disease, and production of a full length transcript of SMN2 could range from 10% to 50%.
  • presence or absence of the SMN1 gene roughly 90% of which becomes the FL-SMN gene product and protein, influences the severity of SMA by whether or not it can compensate for the truncated SMNA7 copies.
  • a low level of SMN protein allows embryonic development, but is not sufficient to sustain the survival of motor neurons of the spinal cord.
  • SMN protein function of SMN protein is still partially unknown, and studies indicate that it can be involved in mRNA metabolism (Meister G, et al. (2002). Trends Cell Biol. 12:472-8; Pellizzoni L, et al. (2002). Science. 298: 1775-9), and probably in transport of proteins/mRNA to neuromuscular junctions (Ci-fuentes-Diaz C, et al. (2002) Hum Mol. Genet. 1 1 : 1439-47; Chan Y B, et al. (2003) Hum Mol. Genet. 12:1367-76; McWhorter M L, et al. (2003) J. Cell Biol. 162:919-31 ; Rossoll W, et al. (2003) J. Cell Biol. 163:801-812).
  • Congenital AMC congenital AMC
  • ALS amyotrophic lateral sclerosis
  • the invention provides compounds, salts thereof, pharmaceutical formulations thereof and
  • the invention further provides methods of treating, preventing, or ameliorating Spinal Muscular Atrophy, comprising administering to a subject in need thereof an effective amount of an SMN modulator (e.g., a compound of the invention).
  • an SMN modulator e.g., a compound of the invention.
  • SMN modulators provided herein are compounds of Formula IA and salts thereof:
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound according to the definition of formula (I) or subformulae thereof and one or more pharmaceutically acceptable carriers.
  • the invention provides a combination, in particular a pharmaceutical combination, comprising a therapeutically effective amount of the compound according to the definition of formula (I) or subformulae thereof and one or more therapeutically active.
  • One embodiment of the invention is to provide a method for treating, preventing, or ameliorating an SMN-deficiency-related condition, comprising administering to a subject in need thereof an effective amount of an SMN modulator, or a pharmaceutical composition comprising the same.
  • Another embodiment of the invention is a method of modulating SMN protein through the administration of an SMN modulator.
  • said SMN modulator is capable of increasing one or more of FL-SMN or SMNA7 levels.
  • said SMN modulator is capable of preventing exon 7 from being spliced from the SMN transcript.
  • the present invention is based on the discovery that the SMN modulators of the invention (e.g., compounds of formula (I) and/or compounds of formula (l-A) are capable of modulating SMN proteins, e.g., through SMN promoter activation, splicing modulation (e.g., preventing exon7 from being spliced out of the SMN gene), and/or SMN protein stability modulation.
  • the SMN modulators of the invention e.g., compounds of formula (I) and/or compounds of formula (l-A) are capable of modulating SMN proteins, e.g., through SMN promoter activation, splicing modulation (e.g., preventing exon7 from being spliced out of the SMN gene), and/or SMN protein stability modulation.
  • the present invention provides compounds that modulate SMN activity. Such compounds may be used in vitro or in vivo to modulate (preferably increase) SMN production and activity in a variety of contexts.
  • the invention provides compounds of Formula (IA) and
  • A is bicyclic heteroaryl or heterocyle having 9 or 10 ring atoms and 1 or 2 ring N atoms and 0 or 1 O atoms, which bicyclic heteroaryl or heterocycle is substituted with 0, 1 , 2, 3, 4 or 5 substituents independently selected from -C(0)NH 2 ,-C(0)0-Ci-C 4 alkyl, aryl, oxo, cyano, halogen, hydroxy, Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, d-C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl CrC 4 alkyl, CrC 4 alkyl aryl, CrC 4 alkyl heterocyclyl, CrC 4 alkyl heteroaryl, CrC 4 alkoxy aryl, Ci-C 4 alkoxy heterocyclyl, Ci-C 4 alkoxy heterocyclyl, Ci-C 4
  • R, R-i, R 2 , R3, and R 4 are independently selected from the group consisting of hydrogen, CrC 4 alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C 4 akylamino;
  • R 5 and R 6 are independently selected from hydrogen and fluorine; or
  • R and R 3 taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S;
  • Ri and R 3 taken in combination form a CrC 3 alkylene group;
  • Ri and R 5 taken in combination form a Ci-C 3 alkylene group;
  • R 3 and R 4 taken in combination with the carbon atom to which they attach, form a spirocyclicC 3 -C 6 cycloalkyl;
  • X is CR A RB, O, N R 7 or a bond;
  • R 7 is hydrogen, or CrC 4 alkyl
  • the invention is a the compound, or salt thereof, according to the first embodiment wherein the compound is of Formula (I):
  • A is bicyclic heteroaryl having 10 ring atoms and 1 or 2 ring N atoms, which bicyclic heteroaryl is substituted with 0, 1 , or 2 substituents independently selected from oxo, cyano, halogen, hydroxy, CrC 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, CrC 4 alkoxy, C 3 -C 7 cycloalkyl,
  • heterocyclyl heteroaryl, heterocyclyl CrC 4 alkyl, CrC 4 alkyl aryl, CrC 4 alkyl heterocyclyl, Ci-C 4 alkyl heteroaryl, CrC 4 alkoxy aryl, CrC 4 alkoxy heterocyclyl, CrC 4 alkoxy heteroaryl, Ci-C 4 alkoxy substituted with hydroxy, Ci-C 4 alkoxy, amino and mono-and di-Ci-C 4 alkylamino; and B is a group of the formula:
  • R, R-i, R 2 , R3, and R 4 are independently selected from the group consisting of hydrogen, d-C 4 alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C 4 akylamino;
  • R 5 and R 6 are independently selected from hydrogen and fluorine; or
  • R and R 3 taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S;
  • R-i and R 3 taken in combination form a C-
  • R-i and R 5 taken in combination form a C-
  • R 3 and R 4 taken in combination with the carbon atom to which they attach, form a spirocyclicC 3 -C 6 cycloalkyl;
  • X is CR A RB, O, N R 7 or a bond
  • R 9 and Ri 3 are independently selected from hydrogen and CrC 4 alkyl
  • R 10 and R14 are independently selected from hydrogen, amino, mono- and di-Ci-C 4 akylamino and Ci-C 4 alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C 4 akylamino
  • Rn is hydrogen, Ci-C 4 alkyl, amino or mono- and di-Ci-C 4 akylamino
  • R 12 is hydrogen or CrC 4 alkyl; or R 9 and Rn, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1-3 C 1 -C 4 alkyl groups; or R ⁇ and R 12 , taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 C 1 -C 4 alkyl groups.
  • the invention is the compound according to any one the first or second embodiments, or salt thereof, wherein A is selected from:
  • u and v are each, independently, 0, 1 , 2 or 3; and each R a and R are, independently, selected from , cyano, halogen, hydroxy, C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 alkoxy, C 3 - C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C 1 -C 4 alkyl, C 1 -C 4 alkyl aryl, C 1 -C 4 alkyl heterocyclyl, C 1 -C 4 alkyl heteroaryl, C 1 -C 4 alkoxy aryl, C 1 -C 4 alkoxy heterocyclyl, C 1 -C 4 alkoxy heteroaryl, and CrC 4 alkoxy substituted with hydroxy, CrC 4 alkoxy, amino and mono-and di-Cr C 4 alkylamino.
  • the invention is the compound of any combination of any combination of
  • u and v are each, independently, 0, 1 , 2 or 3; and each R a and R are, independently, selected from , cyano, halogen, hydroxy, C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 alkoxy, C 3 - C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C 1 -C 4 alkyl, C 1 -C 4 alkyl aryl, C 1 -C 4 alkyl heterocyclyl, C-i-C 4 alkyl heteroaryl, CrC 4 alkoxy aryl, Ci-C 4 alkoxy heterocyclyl, Ci-C 4 alkoxy heteroaryl, and CrC 4 alkoxy substituted with hydroxy, CrC 4 alkoxy, amino and mono-and di-Cr C 4 alkylamino.
  • the invention is the compound according to any one of the first through fourth embodiments, wherein A is substituted in the ortho position with a hydroxyl group.
  • the inveniotn is the compound according to any one of the first through fourth embodiments, or salt thereof, wherein A is selected from:
  • the invneiotn is the compound of any one of the first through fifth embodiments, or a salt thereof, wherein A has a single N atom
  • the invention is the compound, according to any one of the first through sixth emebodiments, or salt thereof, wherein the compound is of formula II:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C 1 -C 4 alkyl, C 1 -C 4 alkyl aryl, C 1 -C 4 alkyl heterocyclyl, C 1 -C 4 alkyl heteroaryl, C 1 -C 4 alkoxy aryl, CrC 4 alkoxy heterocyclyl, CrC 4 alkoxy heteroaryl, CrC 4 alkoxy substituted with hydroxy, C C 4 alkoxy, amino and mono-and di-Ci-C 4 alkylamino.
  • the invention is the compound, or salt thereof, according to any one of the first through sixth embodiments, wherein the compound is of formula III:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C 1 -C 4 alkyl, C 1 -C 4 alkyl aryl, C 1 -C 4 alkyl heterocyclyl, C 1 -C 4 alkyl heteroaryl, C 1 -C 4 alkoxy aryl, C 1 -C 4 alkoxy heterocyclyl, C 1 -C 4 alkoxy heteroaryl, C 1 -C 4 alkoxy substituted with hydroxy, C C 4 alkoxy, amino and mono-and di-Ci-C 4 alkylamino.
  • the invention is the compound, according to any one of the first through sixth embodiments, or salt thereof, wherein the compound is of formula IV:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, d-C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C 4 alkyl, Ci-C 4 alkyl aryl, CrC 4 alkyl heterocyclyl, CrC 4 alkyl heteroaryl, Ci-C 4 alkoxy aryl, CrC 4 alkoxy heterocyclyl, CrC 4 alkoxy heteroaryl, CrC 4 alkoxy substituted with hydroxy, C
  • the invention is the compound, according to any one of the first through sixth emebodiments, or salt thereof, wherein the compound is of formula V:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C 4 alkyl, d-C 4 alkyl aryl, CrC 4 alkyl heterocyclyl, CrC 4 alkyl heteroaryl, Ci-C 4 alkoxy aryl, CrC 4 alkoxy heterocyclyl, CrC 4 alkoxy heteroaryl, CrC 4 alkoxy substituted with hydroxy, C 4 alkoxy, amino and mono-and di-Ci-C 4 alkylamino.
  • the invention is the compound, according to any one of cthe first through sixth emebodiments, or salt thereof, wherein the compound is of formula VI:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C 1 -C 4 alkyl, C 1 -C 4 alkyl aryl, C 1 -C 4 alkyl heterocyclyl, C 1 -C 4 alkyl heteroaryl, C 1 -C 4 alkoxy aryl, CrC 4 alkoxy heterocyclyl, CrC 4 alkoxy heteroaryl, CrC 4 alkoxy substituted with hydroxy, C C 4 alkoxy, amino and mono-and di-Ci-C 4 alkylamino.
  • the invention is the compound, according to any one of the first through sixth embodiments, or salt thereof, wherein the compound is of formula VII:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C 4 alkyl, Ci-C 4 alkyl aryl, CrC 4 alkyl heterocyclyl, CrC 4 alkyl heteroaryl, Ci-C 4 alkoxy aryl, C 1 -C 4 alkoxy heterocyclyl, C 1 -C 4 alkoxy heteroaryl, C 1 -C 4 alkoxy substituted with hydroxy, C C 4 alkoxy, amino and mono-and di-C 1 -C 4 alkylamino.
  • the invention is the compound, according to any one of the first through sixth embodiments, or salt thereof, wherein the compound is of formula VIII:
  • R c and R d are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, d-C 4 alkoxy, C 3 -C 7 cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C 4 alkyl, Ci-C 4 alkyl aryl, CrC 4 alkyl heterocyclyl, CrC 4 alkyl heteroaryl, Ci-C 4 alkoxy aryl, C 1 -C 4 alkoxy heterocyclyl, C 1 -C 4 alkoxy heteroaryl, C 1 -C 4 alkoxy substituted with hydroxy, C C 4 alkoxy, amino and mono-and di-C 1 -C 4 alkylamino.
  • the invention is the compound according to any one of the first through thirteenth embodiments, or a salt thereof, wherein B is a group of the formula:
  • R, R-i, R 2 , R3, and R 4 are independently selected from the group consisting of hydrogen, CrC 4 alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C 4 akylamino;
  • R 5 and R 6 are hydrogen; or
  • R and R 3 taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S;
  • Ri and R 3 taken in combination form a CrC 3 alkylene group;
  • Ri and R 5 taken in combination form a Ci-C 3 alkylene group;
  • R 3 and R 4 taken in combination with the carbon atom to which they attach, form a spirocyclicC 3 -C 6 cycloalkyl;
  • X is CR A RB, O, N R 7 or a bond;
  • R A and R B are independently selected from hydrogen and CrC 4 alkyl
  • the invention is the compound according to any one of the first through thirteenth embodiments, or a salt thereof, wherein B is a group of the formula:
  • R 9 and Ri 3 are independently selected from hydrogen and CrC 4 alkyl
  • R 10 and R14 are independently selected from hydrogen, amino, mono- and di-Ci-C 4 akylamino and Ci-C 4 alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C 4 akylamino
  • Rn is hydrogen, d-C 4 alkyl, amino or mono- and di-Ci-C 4 akylamino
  • R 12 is hydrogen or CrC 4 alkyl; or R 9 and Rn, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1-3 C 1 -C 4 alkyl groups; or R ⁇ and R 12 , taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 C 1 -C 4 alkyl groups.
  • the invention is the compound of any one of the first through fifteenth embodiments, wherein B is selected from the group consisting of:
  • X is O or N(Me) or NH; and Ri 7 is hydrogen or methyl.
  • the invention is the compound, or salt thereof, according to any one of the first through sixteenth embodiments, wherein B is:
  • the invention is the compound, or salt thereof, according to any one of the first through seventeenth embodiments, wherein X is -0-.
  • the invention is the compound, or salt thereof, according to any one of the first through eighteenth embodiments, wherein X is N(Me).
  • theinvention is a compound, or salt thereof, selected from the group consisting of:
  • the invention is a compound, or salt thereof, selected from the group consisting of:
  • the invention is a compound, or salt thereof, selected from the group consisting of:
  • the inventiuon is a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of the first through twentythird embodiments, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.
  • the invention is a combination comprising a therapeutically effective amount of a compound according to any one of the first through twentysecond
  • the invention is a method to treat, prevent or ameliorate an
  • SMN-deficiency-related condition comprising administering to a subject in need thereof an effective amount of a compound or salt thereof of any one of the first through twentysecond embodiments.
  • the invention is the method of the twentyfifth embodiment, wherein said SMN-deficiency-related condition is Spinal Muscular Atrophy.
  • the invention is a compound according to any one of the first through twentysecond embodiments, or a pharmaceutically acceptable salt thereof, for use as a medicament.
  • the invention is a compound according to any one of the first through twentysecond embodiments, or a pharmaceutically acceptable salt thereof, for use in the treatment of an SMN-deficiency-related condition.
  • the invention is the compound according to the twentyeighth embodiment, or pharmaceutically acceptable salt thereof, for use in the treatment of spinal muscular atrophy.
  • the invention is the use of a compound according to any one of the first through twentysecond embodiments, or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of spinal muscular atrophy.
  • SMSN modulator includes agents, such as the compounds of the invention, which possess the ability to modulate, e.g., increase, SMN protein levels by at least one of multiple possible mechanisms.
  • a non-limiting set of mechanisms includes SMN promoter activation, splicing modulation (e.g., preventing exon7 from being spliced out of the SMN gene), and SMN protein stability modulation.
  • SMN modulators can modulate, e.g., increase FL-SMN and/or SMNA7 levels via any of said mechanisms, and/or can prevent SMNA7 from being degraded.
  • SMA Spinal Muscular Atrophy
  • ALS amyotrophic lateral sclerosis
  • SMA Spinal Muscular Atrophy
  • Type I Wirednig-Hoffmann disease
  • Type II Intermediate, chronic form
  • Type III Kugelberg-Welander disease, or Juvenile Spinal Muscular Atrophy
  • SMA Spinal Muscular Atrophy
  • SMARD spinal muscular atrophy with respiratory distress
  • SMARD spinal and bulbar muscular atrophy
  • distal spinal muscular atrophy distal spinal muscular atrophy.
  • Ci-i 0 alkyl refers to a fully saturated branched or unbranched hydrocarbon moiety having 1 to 10 carbon atoms.
  • the terms “Ci -6 alkyl” and “Ci -4 alkyl” are to be construed accordingly.
  • Ci-i 0 alkyl include, but are not limited to, methyl, ethyl, n-propyl, / ' so-propyl, n-butyl, sec-butyl, / ' so-butyl, ferf-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2- dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl and n-decyl.
  • Ci-i 0 alkylene refers to divalent alkyl group as defined herein above having 1 to 10 carbon atoms.
  • the terms “Ci -6 alkylene” and “Ci -4 alkylene” are to be construed accordingly.
  • Ci-i 0 alkylene include, but are not limited to, methylene, ethylene, n-propylene, / ' so-propylene, n-butylene, sec-butylene, / ' so-butylene, tert- butylene, n-pentylene, isopentylene, neopentylene, n-hexylene, 3-methylhexylene, 2,2- dimethylpentylene, 2,3-dimethylpentylene, n-heptylene, n-octylene, n-nonylene and n-decylene.
  • haloC 1-10 alkyl refers to a C 1-10 alkyl group as defined herein, wherein at least one of the hydrogen atoms is replaced by a halo atom.
  • the haloC 1-10 alkyl group can be monohaloC 1-10 alkyl, dihaloC 1-10 alkyl or polyhaloC 1-10 alkyl including perhaloC 1-10 alkyl.
  • a monohaloCi-i 0 alkyl can have one iodo, bromo, chloro or fluoro within the alkyl group.
  • DihaloC-i. - l oalkyl and polyhaloCi-i 0 alkyl groups can have two or more of the same halo atoms or a
  • haloCi-i 0 alkyl group contains up to 12, or 10, or 8, or 6, or 4, or 3, or 2 halo groups.
  • Non-limiting examples of haloCi-i 0 alkyl include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl and dichloropropyl.
  • a perhaloCi-i 0 alkyl group refers to an Ci-i 0 alkyl group having all hydrogen atoms replaced with halo atoms.
  • aryl refers to an aromatic hydrocarbon group having 6-20 carbon atoms in the ring portion. Typically, aryl is monocyclic, bicyclic or tricyclic aryl having 6-20 carbon atoms and includes one or more aromatic rings fused to one or more non-aromatic hydrocarbon rings. Non- limiting examples include phenyl, naphthyl or tetrahydronaphthyl.
  • Ci-i 0 alkyl-O- refers to Ci-i 0 alkyl-O-, wherein Ci-i 0 alkyl is defined herein above.
  • Representative examples of Ci-i 0 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, ferf-butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy- and decyloxy-.
  • heterocyclyl or “heterocyclo” refers to a saturated or unsaturated non-aromatic ring or ring system, which is a 4-, 5-, 6-, or 7-membered monocyclic ring containing 1 , 2 or 3 heteroatoms selected from O, S and N, a 7-, 8-, 9-, 10-, 1 1 -, or 12-membered bicyclic ring system containing 1 , 2, 3, 4 or 5 heteroatoms selected from O, S and N, or a 10-, 1 1-, 12-, 13-, 14- or 15-membered tricyclic ring system and containing 1 , 2, 3, 4, 5, 6 or 7 heteroatoms selected from O, S and N, where the N and S can also optionally be oxidized to various oxidation states.
  • the heterocyclic group can be attached via a heteroatom or a carbon atom.
  • the heterocyclyl can include fused or bridged rings as well as spirocyclic rings.
  • heterocycles include tetrahydrofuran (THF), dihydrofuran, 1 , 4-dioxane, morpholine, 1 ,4-dithiane, piperazine, piperidine, 1 ,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1 ,3-dioxane, 1 ,3-dithiane, oxathiane and thiomorpholine.
  • C 3- i 2 cycloalkyl refers to saturated or unsaturated monocyclic, bicyclic or tricyclic hydrocarbon groups of 3-12 carbon atoms.
  • C 3-18 cycloalkyl refers to a fully saturated or unsaturated monocyclic hydrocarbon group of 3-8 carbon atoms.
  • Exemplary monocyclic hydrocarbon groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl and cyclohexenyl.
  • Exemplary bicyclic hydrocarbon groups include bornyl, indyl, hexahydroindyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1 .1 ]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1 .1 ]heptyl, 2,6,6- trimethylbicyclo[3.1.1 ]heptyl, bicyclo[2.2.2]octyl.
  • Exemplary tricyclic hydrocarbon groups include, for example, adamantyl.
  • C 3 12 cycloalklyoxy refers to C 3- i 2 cycloalkyl-0-, wherein C 3- i 2 cycloalkyl is defined herein above.
  • Representative examples of C 3. i 2 cycloalklyoxy include , but are not limited to monocyclic groups such as cyclopropoxy, cyclobutoxy, cyclopentyloxy,
  • hydrocarbon groups include bornyloxy, indyloxy, hexahydroindyloxy, tetrahydronaphthyloxy, decahydronaphthyloxy, bicyclo[2.1 .1]hexyloxy, bicyclo[2.2.1]heptyloxy, bicyclo[2.2.1]heptenyloxy, 6,6-dimethylbicyclo[3.1 .1 ]heptyloxy, 2,6,6-trimethylbicyclo[3.1.1 ]heptyloxy, bicyclo[2.2.2]octyloxy and the like.
  • Exemplary tricyclic hydrocarbon groups include, for example, adamantyloxy.
  • aryloxy refers to both an— O-aryl and an— O-heteroaryl group, wherein aryl and heteroaryl are defined herein.
  • heteroaryl refers to a 5-, 6-, or 7-membered monocyclic aromatic ring containing 1 , 2, 3 or 4 heteroatoms selected from O, S and N, an 8-, 9-, or 10-membered fused bicyclic ring system containing 1 , 2, 3, 4 or 5 heteroatoms selected from O, S and N, or an 1 1-, 12-, 13-, or 14-membered fused tricyclic ring system containing 1 , 2, 3, 4, 5 or 6 heteroatoms selected from O, S and N, wherein at least one of the rings of the bicyclic or tricyclic ring systems is fully aromatic.
  • Typical heteroaryl groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or 5- imidazolyl, 3-, 4-, or 5- pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or 5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or 5-1 ,2,4-triazolyl, 4- or 5-1 ,2, 3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3- or 4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, 2-, 4-, or 5-pyrimidinyl, 1-, 2-, 3-, 5-, 6-, 7-, or 8- indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl
  • halogen refers to fluoro, chloro, bromo, and iodo.
  • the term “isomers” refers to different compounds that have the same molecular formula but differ in arrangement and configuration of the atoms.
  • an optical isomer or “a stereoisomer” refers to any of the various stereo isomeric configurations which may exist for a given compound of the present invention and includes geometric isomers. It is understood that a substituent may be attached at a chiral center of a carbon atom. Therefore, the invention includes enantiomers, diastereomers or racemates of the compound. "Enantiomers” are a pair of stereoisomers that are non- superimposable mirror images of each other.
  • a 1 : 1 mixture of a pair of enantiomers is a "racemic" mixture.
  • the term is used to designate a racemic mixture where appropriate.
  • "Diastereoisomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.
  • the absolute stereochemistry is specified according to the Cahn- Ingold- Prelog R-S system. When a compound is a pure enantiomer the stereochemistry at each chiral carbon may be specified by either R or S.
  • Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) depending on the direction (dextro- or levorotatory) which they rotate plane polarized light at the wavelength of the sodium D line.
  • Certain of the compounds described herein contain one or more asymmetric centers or axes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-.
  • the present invention is meant to include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures.
  • Optically active (R)- and (S)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be E or Z configuration. If the compound contains a disubstituted cycloalkyi, the cycloalkyi substituent may have a cis- or trans-configuration. All tautomeric forms are also intended to be included.
  • salt refers to an acid addition or base addition salt of a compound of the invention.
  • Salts include in particular “pharmaceutical acceptable salts.”
  • pharmaceutically acceptable salts refers to salts that retain the biological effectiveness and properties of the compounds of this invention and, which typically are not biologically or otherwise undesirable.
  • the compounds of the present invention are capable of forming acid and/or base salts by virtue of the presence of amino and/or carboxyl groups or groups similar thereto.
  • Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids, e.g., acetate, aspartate, benzoate, besylate, bromide/hydrobromide,
  • chlortheophyllonate citrate, ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate, laurylsulfate, malate, maleate, malonate, mandelate, mesylate, methylsulphate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, subsalicylate, tartrate, tosylate and trifluoroacetate salts.
  • Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
  • Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like.
  • Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.
  • Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I to XII of the periodic table.
  • the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper;
  • particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.
  • Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like.
  • Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and
  • the pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound, a basic or acidic moiety, by conventional chemical methods.
  • such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate or the like), or by reacting free base forms of these compounds with a stoichiometric amount of the appropriate acid.
  • a stoichiometric amount of the appropriate base such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate or the like
  • Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two.
  • use of non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is desirable, where practicable.
  • any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds.
  • Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number.
  • isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 F 31 P, 32 P, 35 S, 36 CI, 125 l respectively.
  • the invention includes various isotopically labeled compounds as defined herein, for example those into which radioactive isotopes, such as 3 H, 13 C, and 14 C , are present.
  • isotopically labelled compounds are useful in metabolic studies (with 14 C), reaction kinetic studies (with, for example 2 H or 3 H), detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients.
  • PET positron emission tomography
  • SPECT single-photon emission computed tomography
  • an 18 F or labeled compound may be particularly desirable for PET or SPECT studies.
  • Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.
  • isotopic enrichment factor means the ratio between the isotopic abundance and the natural abundance of a specified isotope.
  • a substituent in a compound of this invention is denoted deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).
  • Isotopically-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations using an appropriate isotopically-labeled reagents in place of the non-labeled reagent previously employed.
  • solvates in accordance with the invention include those wherein the solvent of crystallization may be isotopically substituted, e.g. D 2 0, d 6 -acetone, d 6 - DMSO.
  • Compounds of the invention i.e. compounds of formula (I) that contain groups capable of acting as donors and/or acceptors for hydrogen bonds may be capable of forming co-crystals with suitable co-crystal formers.
  • These co-crystals may be prepared from compounds of formula (I) by known co-crystal forming procedures. Such procedures include grinding, heating, co-subliming, co- melting, or contacting in solution compounds of formula (I) with the co-crystal former under crystallization conditions and isolating co-crystals thereby formed.
  • Suitable co-crystal formers include those described in WO 2004/078163.
  • the invention further provides co-crystals comprising a compound of formula (I).
  • a therapeutically effective amount of a compound of the present invention refers to an amount of the compound of the present invention that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc.
  • a therapeutically effective amount refers to the amount of the compound of the present invention that, when administered to a subject, is effective to (1 ) at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease (i) mediated by Survival of Motor Neuron (SMN) gene or gene product, or by SMNA7 degradation, or by the relative levels of FL-SMN and SMNA7 (ii) associated with SMN activity, or (iii) characterized by activity (normal or abnormal) of SMN; or (2) reducing or inhibiting the activity of SMN; or (3) reducing or inhibiting the expression of SMN 1 or SMN2.
  • SSN Motor Neuron
  • a therapeutically effective amount refers to the amount of the compound of the present invention that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reducing or inhibiting the activity of SMN; or at least partially reducing or inhibiting the expression of SMN, in both cases by modulating the relative levels of FL-SMN and SMNA7.
  • terapéuticaally effective amount and "effective amount” are used herein to mean an amount sufficient to reduce by at least about 15 percent, preferably by at least 50 percent, more preferably by at least 90 percent, and most preferably prevent, a clinically significant deficit in the activity, function and response of the host.
  • a therapeutically effective amount is sufficient to cause an improvement in a clinically significant condition/symptom in the host.
  • the effective amount can vary depending on such factors as the size and weight of the subject, the type of illness, or the particular compound of the invention. For example, the choice of the compound of the invention can affect what constitutes an "effective amount.”
  • One of ordinary skill in the art would be able to study the factors contained herein and make the determination regarding the effective amount of the compounds of the invention without undue experimentation.
  • the regimen of administration can affect what constitutes an effective amount.
  • the compound of the invention can be administered to the subject either prior to or after the onset of an SMN-deficiency-related condition. Further, several divided dosages, as well as staggered dosages, can be administered daily or sequentially, or the dose can be continuously infused, or can be a bolus injection. Further, the dosages of the compound(s) of the invention can be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.
  • the term "subject" refers to an animal. Typically the animal is a mammal. A subject also refers to for example, primates (e.g., humans, male or female), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds and the like. In certain embodiments, the subject is a primate. In yet other embodiments, the subject is a human.
  • primates e.g., humans, male or female
  • the subject is a primate.
  • the subject is a human.
  • the term “inhibit”, “inhibition” or “inhibiting” refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.
  • treat refers in one embodiment, to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one of the clinical symptoms thereof).
  • ameliorating i.e., slowing or arresting or reducing the development of the disease or at least one of the clinical symptoms thereof.
  • treat,” “treating,” or “treatment” refers to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the patient.
  • “treat”, “treating” or “treatment” refers to modulating the disease or disorder, either physically (e.g., through stabilization of a discernible symptom), physiologically, (e.g., through stabilization of a physical parameter), or both.
  • “treat,” “treating,” or “treatment” refers to preventing or delaying the onset or development or progression of the disease or disorder.
  • a subject is "in need of” a treatment if such subject would benefit biologically, medically or in quality of life from such treatment.
  • any asymmetric atom (e.g., carbon or the like) of the compound(s) of the present invention can be present in racemic or enantiomerically enriched, for example the (/?)-, (S)- or ⁇ R,S)- configuration.
  • each asymmetric atom has at least 50 % enantiomeric excess, at least 60 % enantiomeric excess, at least 70 % enantiomeric excess, at least 80 % enantiomeric excess, at least 90 % enantiomeric excess, at least 95 % enantiomeric excess, or at least 99 % enantiomeric excess in the (R)- or (S)- configuration.
  • Substituents at atoms with unsaturated bonds may, if possible, be present in cis- (Z)- or trans- (E)- form.
  • a compound of the present invention can be in the form of one of the possible isomers, rotamers, atropisomers, tautomers or mixtures thereof, for example, as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (antipodes), racemates or mixtures thereof.
  • Any resulting mixtures of isomers can be separated on the basis of the physicochemical differences of the constituents, into the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and/or fractional crystallization.
  • any resulting racemates of final products or intermediates can be resolved into the optical antipodes by known methods, e.g., by separation of the diastereomeric salts thereof, obtained with an optically active acid or base, and liberating the optically active acidic or basic compound.
  • a basic moiety may thus be employed to resolve the compounds of the present invention into their optical antipodes, e.g., by fractional crystallization of a salt formed with an optically active acid, e.g., tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-0,0'-p-toluoyl tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid.
  • Racemic products can also be resolved by chiral chromatography, e.g., high pressure liquid chromatography (HPLC) using a chiral adsorbent.
  • HPLC high pressure liquid chromatography
  • the compounds of the present invention may also form internal salts, e.g., zwitterionic molecules.
  • the compounds of the present invention can also be obtained in the form of their hydrates, or include other solvents used for their crystallization.
  • the compounds of the present invention may inherently or by design form solvates with
  • solvate refers to a molecular complex of a compound of the present invention (including pharmaceutically acceptable salts thereof) with one or more solvent molecules.
  • solvent molecules are those commonly used in the pharmaceutical art, which are known to be innocuous to the recipient, e.g., water, ethanol, and the like.
  • hydrate refers to the complex where the solvent molecule is water.
  • the compounds of the present invention including salts, hydrates and solvates thereof, may inherently or by design form polymorphs.
  • the invention further includes any variant of the present processes, in which an
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition can be formulated for particular routes of administration such as oral administration, parenteral administration, and rectal administration, etc.
  • the pharmaceutical compositions of the present invention can be made up in a solid form (including without limitation capsules, tablets, pills, granules, powders or suppositories), or in a liquid form (including without limitation solutions, suspensions or emulsions).
  • compositions can be subjected to conventional pharmaceutical operations such as sterilization and/or can contain conventional inert diluents, lubricating agents, or buffering agents, as well as adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers and buffers, etc.
  • the pharmaceutical compositions are tablets or gelatin capsules comprising the active ingredient together with
  • diluents e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine;
  • lubricants e.g., silica, talcum, stearic acid, its magnesium or calcium salt and/or polyethyleneglycol; for tablets also
  • lubricants e.g., silica, talcum, stearic acid, its magnesium or calcium salt and/or polyethyleneglycol
  • binders e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth,
  • disintegrants e.g., starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and/or
  • Tablets may be either film coated or enteric coated according to methods known in the art.
  • suitable compositions for oral administration include an effective amount of a compound of the invention in the form of tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs.
  • Compositions intended for oral use are prepared according to any method known in the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations.
  • Tablets may contain the active ingredient in admixture with nontoxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets.
  • excipients are, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc.
  • the tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.
  • a time delay material such as glyceryl monostearate or glyceryl distearate can be employed.
  • Formulations for oral use can be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil.
  • an inert solid diluent for example, calcium carbonate, calcium phosphate or kaolin
  • water or an oil medium for example, peanut oil, liquid paraffin or olive oil.
  • compositions are aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fatty emulsions or suspensions.
  • compositions may be sterilized and/or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances.
  • adjuvants such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers.
  • Said compositions are prepared according to conventional mixing, granulating or coating methods, respectively, and contain about 0.1-75%, or contain about 1-50%, of the active ingredient.
  • compositions for transdermal application include an effective amount of a compound of the invention with a suitable carrier.
  • Carriers suitable for transdermal delivery include absorbable pharmacologically acceptable solvents to assist passage through the skin of the host.
  • transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound of the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin.
  • compositions for topical application include aqueous solutions, suspensions, ointments, creams, gels or sprayable formulations, e.g., for delivery by aerosol or the like.
  • topical delivery systems will in particular be appropriate for dermal application, e.g., for the treatment of skin cancer, e.g., for prophylactic use in sun creams, lotions, sprays and the like. They are thus particularly suited for use in topical, including cosmetic, formulations well-known in the art.
  • Such may contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.
  • a topical application may also pertain to an inhalation or to an intranasal application. They may be conveniently delivered in the form of a dry powder (either alone, as a mixture, for example a dry blend with lactose, or a mixed component particle, for example with phospholipids) from a dry powder inhaler or an aerosol spray presentation from a pressurized container, pump, spray, atomizer or nebulizer, with or without the use of a suitable propellant.
  • a dry powder either alone, as a mixture, for example a dry blend with lactose, or a mixed component particle, for example with phospholipids
  • the present invention further provides anhydrous pharmaceutical compositions and dosage forms comprising the compounds of the present invention as active ingredients, since water may facilitate the degradation of certain compounds.
  • Anhydrous pharmaceutical compositions and dosage forms of the invention can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions.
  • An anhydrous pharmaceutical composition may be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions are packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastics, unit dose containers (e. g., vials), blister packs, and strip packs.
  • compositions and dosage forms that comprise one or more agents that reduce the rate by which the compound of the present invention as an active ingredient will decompose.
  • agents which are referred to herein as “stabilizers,” include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers, etc.
  • the compounds of formula I in free form or in salt form exhibit valuable pharmacological properties, e.g. full length SMN protein production modulating properties, e.g. as indicated in in vitro and in vivo tests as provided in the next sections, and are therefore indicated for therapy or for use as research chemicals, e.g. as tool compounds.
  • valuable pharmacological properties e.g. full length SMN protein production modulating properties, e.g. as indicated in in vitro and in vivo tests as provided in the next sections, and are therefore indicated for therapy or for use as research chemicals, e.g. as tool compounds.
  • the present invention provides the use of a compound of formula (I) or a salt thereof in therapy.
  • the therapy is selected from a disease which may be treated by modulating full length SMN protein production.
  • the disease is selected from the afore-mentioned list, suitably spinal muscular atrophy.
  • the invention provides a method of treating a disease which is treated by modulating full length SMN protein production comprising administration of a
  • the disease is selected from the afore-mentioned list, suitably spinal muscular atrophy.
  • the present invention provides the use of a compound of formula (I) or salt thereof for the manufacture of a medicament.
  • the medicament is for treatment of a disease which may be treated by modulation of SMN protein production.
  • the disease is selected from the afore-mentioned list, suitably spinal muscular atrophy.
  • the pharmaceutical composition or combination of the present invention can be in unit dosage of about 0.01-1000 mg of active ingredient(s) for a subject of about .05-70 kg or about 1 -20 kg, or about 1 -500 mg or about 1 -250 mg or about 1 -150 mg or about 0.5-100 mg, or about 0.01-1 mg or about 0.01-0.1 mg or about 1-50 mg of active ingredients.
  • the therapeutically effective dosage of a compound, the pharmaceutical composition, or the combinations thereof, is dependent on the species of the subject, the body weight, age and individual condition, the disorder or disease or the severity thereof being treated. A physician, clinician or veterinarian of ordinary skill can readily determine the effective amount of each of the active ingredients necessary to prevent, treat or inhibit the progress of the disorder or disease.
  • the above-cited dosage properties are demonstrable in vitro and in vivo tests using advantageously mammals, e.g., mice, rats, dogs, monkeys or isolated organs, tissues and preparations thereof.
  • the compounds of the present invention can be applied in vitro in the form of solutions, e.g., aqueous solutions, and in vivo either enterally, parenterally, advantageously intravenously, e.g., as a suspension or in aqueous solution.
  • the dosage in vitro may range between about 10 "3 molar and 10 "9 molar concentrations.
  • a therapeutically effective amount in vivo may range depending on the route of administration, between about 0.1-500 mg/kg, or between about 1-100 mg/kg.
  • the compound of the present invention may be administered either simultaneously with, or before or after, one or more other therapeutic agent.
  • the compound of the present invention may be administered separately, by the same or different route of administration, or together in the same pharmaceutical composition as the other agents.
  • the invention provides a product comprising a compound of formula (I) and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy.
  • the therapy is the treatment of a spinal muscular atrophy.
  • Products provided as a combined preparation include a composition comprising the compound of formula (I) and the other therapeutic agent(s) together in the same pharmaceutical composition, or the compound of formula (I) and the other therapeutic agent(s) in separate form, e.g. in the form of a kit.
  • the invention provides a pharmaceutical composition comprising a compound of formula (I) and another therapeutic agent(s).
  • the pharmaceutical composition may comprise a pharmaceutically acceptable carrier, as described above.
  • the invention provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of formula (I).
  • the kit comprises means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet.
  • An example of such a kit is a blister pack, as typically used for the packaging of tablets, capsules and the like.
  • the kit of the invention may be used for administering different dosage forms, for example, oral and parenteral, for administering the separate compositions at different dosage intervals, or for titrating the separate compositions against one another.
  • the kit of the invention typically comprises directions for administration.
  • the compound of the invention and the other therapeutic agent may be manufactured and/or formulated by the same or different manufacturers. Moreover, the compound of the invention and the other therapeutic may be brought together into a combination therapy: (i) prior to release of the combination product to physicians (e.g. in the case of a kit comprising the compound of the invention and the other therapeutic agent); (ii) by the physician themselves (or under the guidance of the physician) shortly before administration; (iii) in the patient themselves, e.g. during sequential administration of the compound of the invention and the other therapeutic agent.
  • Suitable protecting groups include hydroxy, phenol, amino and carboxylic acid.
  • Suitable protecting groups for hydroxy or phenol include trialkylsilyl or diarylalkylsilyl (e.g., t- butyldimethylsilyl, f-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, substituted benzyl, methyl, and the like.
  • Suitable protecting groups for amino, amidino and guanidino include t- butoxycarbonyl, benzyloxycarbonyl, and the like.
  • Suitable protecting groups for carboxylic acid include alkyl, aryl or arylalkyl esters.
  • Protecting groups may be added or removed in accordance with standard techniques, which are well-known to those skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley.
  • the protecting group may also be a polymer resin, such as a Wang resin or a 2-chlorotrityl-chloride resin.
  • reaction schemes illustrate methods to make compounds of this invention. It is understood that one skilled in the art would be able to make these compounds by similar methods or by methods known to one skilled in the art.
  • starting components and reagents may be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, Strem, other commercial vendors, or synthesized according to sources known to those skilled in the art, or prepared as described in this invention.
  • A, B, X, R, R 1 , R 2 , R 3 , R 4 are defined as in the Specification unless specifically defined.
  • pyridazine compounds of Formula (I) of this invention can be synthesized following the general procedure described in Scheme 1.
  • Di-halopyridazine (1) reacts in a displacement reaction or a metal-mediated cross coupling reaction (Buchwald) with an alcohol or an amine (B) to provide pyridazine intermediate (2).
  • Transition metal-mediated cross coupling reaction such as a Suzuki reaction, between halide compound (2) and a substituted aryl or heteroaryl compound A, such as a boronate acid or boronate ester, provides compound (3) of Formula (I) of the invention.
  • Di-halopyridazine (1) reacts in a transition metal-mediated cross coupling reaction, such as a Suzuki reaction, with a substituted aryl or heteroaryl compound A, such as a boronate acid or ester, to provide pyridazine intermediate (4).
  • Pyridazine intermediate (4) reacts via a displacement reaction with an alcohol or an amine (B) to provide pyridazine (3) of Formula (I) of the invention.
  • Compounds of Formula (I) can also be prepared following the general procedure described in Scheme 3.
  • Di-halopyridazine (1) reacts in a transition metal-mediated cross coupling reaction, such as a Suzuki reaction, with a substituted aryl or heteroaryl compound A, such as a boronate acid or ester, to provide pyridazine intermediate (4).
  • a transition metal-mediated cross coupling reaction such as a Suzuki reaction
  • a substituted aryl or heteroaryl compound A such as a boronate acid or ester
  • Pyridazine intermediate (4) reacts via second metal- mediated cross coupling, such as a Suzuki reaction, to provide pyridazine (3) of Formula (I) of the invention.
  • All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, catalysts and scavengers utilized to synthesize the compounds of the present invention are either commercially available or can be produced by organic synthesis methods known to one of ordinary skill in the art. Further, the compounds of the present invention can be produced by organic synthesis methods known to one of ordinary skill in the art as shown in the following examples.
  • Step 2 (7-Methoxyquinolin-6-yl)boronic acid Butyllithium (1.6 M in heptane, 4.90 mL, 7.90 mmol) was added dropwise to a solution of 6- bromo-7-methoxyquinoline (1 .71 g, 7.18 mmol) cooled to -78 °C. The solution was stirred for 0.5 h after which time trimethyl borate (2.0 mL, 18 mmol) was added in a single portion. The reaction was allowed to warm to room temperature and stir overnight. The crude reaction was concentrated to dryness, then concentrated from heptane (2x).
  • 6-bromo-7-methoxyisoquinoline 400 mg, 1 .68 mmol
  • bis(pinacolato)diboron 853 mg, 3.36 mmol
  • potassium acetate 989 mg, 10.1 mmol
  • Oxalyl chloride (4.18 ml, 47.7 mmol) was added dropwise to a suspension of 4-bromo-5- fluoro-2-nitrobenzoic acid (1 1 .46 g, 43.4 mmol) in DCM (175 mL) and catalytic DMF (100 uL). After 3 h the volatiles were removed via rotary evaporation. The remaining residue was redissolved into DCM (25 mL) and added portion-wise to rapidly stirring methanol (100 mL). The solvent was again removed and the residue concentrated from toluene (2x) to afford methyl 4-bromo-5-fluoro-2- nitrobenzoate as a yellow liquid (12.07 g) which was taken on without purification.
  • Zinc powder (6.24 g, 95.4 mmol) was added portion-wise over 30 min (Caution! exotherm possible) to a solution of methyl 4-bromo-5-methoxy-2-nitrobenzoate (3.076 g, 10.60 mmol) in 3:1 DCM/AcOH (55 mL). The mixture was stirred at room temperature for three days. The mixture was filtered through Celite ® , washing the filter pad with EtOAc, and concentrated. The crude product was redissolved into EtOAc, washed with saturated NaHC0 3 , brine, dried over MgS0 4 and concentrated to a yellow oil.
  • Step 3 (7-Methoxy-2-methyl-4-oxo-1,4-dihydroquinolin-6-yl)boronic acid and 7-Methoxy-2- methyl-6-(4, 4, 5, 5-tetramethyl-1 ,3, 2-dioxaborolan-2-yl)quinolin-4(1H)-one
  • the reaction mixture was sealed, then evacuated and back-filled with nitrogen (4x), and heated via microwave irradiation at 1 10 °C for 1 hour.
  • the mixture was filtered through Celite ® washing with EtOAc.
  • the resulting filtrate was concentrated and acidified to pH 3 by 4 M HCI in 1 ,4-dioxane.
  • Step 5 6-(4-Chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2,2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
  • Step 6 4-Chloro-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino
  • 6-(4-Chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (195 mg, 0.43 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using boron tribromide (1 M solution in DCM, 2.49 mL, 2.49 mmol).
  • Step 1 5-( ((4-Bromo-3-methoxyphenyl)amino)methylene)-2, 2-dimethyl-1 , 3-dioxane-4, 6- dione
  • Step 3 7-Met oxy-6-(4 ! 4 ! 5 ! 5-tetramet yl-1 ! 3 ! 2-dioxaborolan-2-yl)quinolin-4(1H)-one and, (7-methoxy-4-oxo- 1, 4-dihydroquinolin-6-yl)boronic acid
  • 6-bromo-7-methoxyquinolin-4(1 H)-one (0.92 g, 2.72 mmol)
  • dppf 151 mg, 0.272 mmol
  • PdCl2(dppf).CH2Cl2 222 mg, 0.272 mmol
  • KOAc (1 .33 g, 13.6 mmol
  • the reaction mixture was evacuated and backfilled with nitrogen (4x), then heated at 80 °C for 0.5 hours.
  • Bis(pinacolato) diboron (1 .72 g, 6.79 mmol) was then added, and the mixture was heated at 90 °C overnight.
  • the reaction mixture was cooled to room temperature, filtered through Celite ® , washed with 9:1 MeOH/DCM, then 1 :1 MeOH/DCM. The filtrates were concentrated in vacuo.
  • Step 4 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-4( 1H)-one
  • the vessel was evacuated and back-filled with nitrogen (4x), and heated via microwave irradiation at 1 10 °C for 1 .5 hour.
  • the mixture was cooled to room temperature, diluted with MeOH (3 ml_), filtered through Celite ® , and washed with 9:1 MeOH/DCM and then with 1 :1 MeOH/DCM.
  • the resulting filtrate was concentrated and acidified to pH 3 with 1 M HCI in diethyl ether.
  • Step 5 6-(4-Chloro-7-methoxyquinolin-6-yl)-N-methyl-N-(2 ! 2,6fi-tetramethy ⁇
  • Step 6 4-chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino
  • 6-(4-Chloro-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (62 mg, 0.14 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using boron tribromide (1 M solution in DCM , 0.70 mL, 0.70 mmol).
  • Step 1 5-(((3-Chloro-4-methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxane-4,6- dione
  • Triethoxymethane (1 1 mL, 64 mmol) and 2,2-dimethyl-1 ,3-dioxane-4,6-dione (Meldrum's acid, 10.7 g, 74.2 mmol) were added to 3-chloro-4-methoxyaniline (10.0 g, 63.5 mmol) in ethanol (60 mL). The mixture was stirred at reflux overnight. An additional portion of triethoxymethane (2.1 mL, 13 mmol) and 2,2-dimethyl-1 ,3-dioxane-4,6-dione (2.74 g, 19.0 mmol) were added to the reaction and the mixture was stirred at reflux for 3 h.
  • Step 2 7-Chloro-6-methoxyquinolin-4(1H)-one and 5-chloro-6-methoxyquinolin-4(1H)-one 5-(((3-chloro-4-methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxan (12.9 g, 41.4 mmol) in diphenyl ether (100 mL) was heated at 190 °C for 2 hour. The reaction mixture was cooled to room temperature and diluted with diethyl ether (100 mL).
  • Step 4 6-(4-Chloro-6-methoxyquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • Step 5 4-Chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-6-ol
  • a mixture of a chloropyridazine intermediate, such as Intermediate 1 (1.0 equivalents), potassium triphosphate (5.0 equivalents), and 2 nd generation XPhos precatalyst (0.07 equivalents), in DMF (0.3 M) is degassed via vacuum/N 2 purge (3x), and heated at 50 °C.
  • a 50 or 100 mg/mL solution of a boronic acid, such as Intermediate 7 or Intermediate 8 in DMF is added in 1 ml. aliquots every 2-3 h until Intermediate 1 is consumed.
  • the reaction is cooled to room temperature, filtered through Celite ® washing with EtOAc, and partially concentrated.
  • the crude product is purified by SCX (GENERAL METHOD 3-1 ) and silica gel chromatography.
  • a mixture of a chloropyridazine intermediate, such as Intermediate 1 (1.0 equivalent), boronic acid reagent (1.5-2 equivalents), and Na 2 C0 3 (3 equivalents) in 3:1 DME/water (0.12 M) in a microwave vial is degassed with a stream of dry nitrogen.
  • Pd(PPh 3 ) 4 (0.1 equivalents) is added to the reaction mixture and the vial is sealed and heated in a Biotage ® Initiator microwave reactor at 140 °C for 30 min.
  • the mixture is diluted with water and extracted with DCM (4x).
  • the organic extracts are dried over MgS0 4 , filtered, and concentrated to afford the crude product.
  • the crude product is purified via silica chromatography as described in GENERAL METHOD 4-1.
  • the methoxy substrate (1 equivalent) and pyridine hydrochloride (20-30 equivalents) are heated at 160-190 °C for 15-120 min in a Biotage ® Initiator microwave reactor.
  • the reaction mixture is solubilized with MeOH/DMSO and purified via preparative reverse-phase HPLC (15 to 45% acetonitrile in water, 5 mM ammonium hydroxide modifier).
  • the methoxy substrate (1 equivalent) and pyridine hydrochloride (20-25 equivalents) are heated at 160-190 °C for 15-120 min in a Biotage ® Initiator microwave reactor.
  • the reaction mixture is solubilized with MeOH and the solution loaded onto a mixture of solid sodium bicarbonate (30 equivalents) and silica gel ( ⁇ 6 g/mmol methoxy substrate), and concentrated to dryness.
  • Column chromatography as described in GENERAL METHOD 4-1 provided the product.
  • the crude product (either neat, dissolved into DCM, or loaded onto a solid support such as silica gel or Celite ® ) is subjected to normal phase flash column chromatography using RediSep ® Rf pre-packed silica gel cartridges (Teledyne Isco, Inc.) with an elution gradient of 1-20% ammonia in MeOH (1.5 N, or 3.5 N concentration), in DCM.
  • Residual palladium, and certain other metal contaminants can be scavenged from reaction products by dissolution in an appropriate solvent (e.g., THF, DCM, or acetonitrile) and stirring with SiliaMetS ® Dimercaptotriazine (DMT) resin (Silicycle, Inc.) for 4-24 h using 3-5 equivalents relative to the amount of the palladium catalyst (or other metal) used. Filtration, and washing of the resin, followed by concentration provides the purified product.
  • an appropriate solvent e.g., THF, DCM, or acetonitrile
  • SiliaMetS ® Dimercaptotriazine (DMT) resin Silicycle, Inc.
  • Example 2-1 Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 3-1 Synthesis of 7-(6-(methyl(1 , 2,2,6, 6-pentamethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
  • Step 1 6-( 6-Methoxyisoquinolin-7-yl)-N-methyl-N-( 1, 2, 2, 6, 6-pentamethylpiperidin-4- yl)pyridazin-3-amine
  • Step 2 7-( 6-(Methyl( 1, 2, 2, 6, 6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6- ol
  • the title compound (26 mg) was prepared from 6-(6-methoxyisoquinolin-7-yl)-N-methyl-N- (1 ,2,2,6,6-pentamethylpiperidin-4-yl)pyridazin-3-amine (63 mg, 0.150 mmol) following GENERAL METHOD 2-4 for demethylation using thiophenol.
  • Example 3-2 Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
  • Step 1 6-( 6-Methoxyisoquinolin-7-yl)-N-methyl-N-( 2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • Example 3-3 Synthesis of 7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)isoquinolin-6-ol
  • Step 1 6-Methoxy-7-(6-((2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinolin
  • the title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction from Intermediate 3 (200 mg, 0.741 mmol) and Intermediate 7 (256 mg, 1 .26 mmol).
  • MS (M+1 ) 393.0
  • Example 3-4 Synthesis of 7-(6-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol- 2(1 H)-yl)pyridazin-3-yl)isoquinolin-6-ol
  • Example 3-5 Synthesis of 1 -cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
  • Benzyl bromide (7 mL, 57.8 mmol) was added to a mixture of 3-hydroxy-4-iodobenzoic acid (5.13 g, 18.9 mmol) and K 2 C0 3 (8.0 g, 58 mmol) in acetone (200 mL). The mixture was then heated at reflux overnight. The mixture was cooled to room temperature then filtered through a small Celite ® pad. The filtrate was concentrated in vacuo. MeOH (30 mL) was added to the residue followed by 1 M NaOH (94 mL, 94 mmol). The resulting mixture was heated in a 70 °C oil bath for 1 h.
  • p-Toluenesulfonic acid monohydrate (0.215 g, 1.129 mmol) was added to a solution of (3- (benzyloxy)-4-iodophenyl)methanol (1.92 g, 5.64 mmol) and 3,4-dihydro-2H-pyran (0.64 mL, 6.8 mmol) in DCM (50 mL) at room temperature. After 2 h, the solution was diluted with DCM (30 mL), then washed with saturated aqueous NaHC0 3 solution (2 x 20 mL). The organic layer was washed with brine, dried over MgS0 4 , filtered, and concentrated in vacuo. The resulting oil was stored under high vacuum overnight.
  • the reaction flask was equipped with a reflux condenser, and the flask evacuated and filled with N 2 (2x) then heated in a 100 °C oil bath overnight (18 h).
  • the mixture was cooled to room temperature then filtered through Celite ® with a MeOH wash.
  • the filtrate was acidified to pH 2 using 1 M HCI then heated at 70 °C for 1 h.
  • the mixture was cooled to room temperature then filtered through Celite ® washing with MeOH.
  • SCX purification (GENERAL
  • Step 9 N-(2-Hydroxy-2-(3-methoxy-4-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)phenyl)ethyl)cyclopropanecarboxamide
  • Step 10 6-( 1 -Cyclopropyl-6-methoxyisoquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
  • Example 3-6 Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 ,6-diol
  • Step 4. 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline- 1, 6- diol
  • Example 3-7 Synthesis of 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 -carbonitrile
  • Example 4-1 Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-7-ol
  • Butyllithium (2.5 M in heptane, 0.42 ml_, 1.1 mmol) was added dropwise to a solution of 7-
  • Step 3 6-(7-(Benzyloxy)quinolin-8-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • Step 4 8-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
  • 6-(7-(benzyloxy)quinolin-8-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine 176 mg, 0.365 mmol
  • EtOAc/MeOH 3.6 ml.
  • Example 6-1 Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
  • Butyllithium (1.6 M in heptane, 1.90 ml_, 3.04 mmol) was added drop-wise over the course of one hour to a solution of 7-bromo-6-methoxyquinoline (0.603 g, 2.53 mmol) and triisopropyl borate (0.823 ml, 3.55 mmol) in THF (12.6 ml.) cooled to -78 °C. After the addition was complete, the cooling bath was removed and the slurry allowed to warm to room temperature. LC/MS analysis shows near quantitative conversion to the boronic acid.
  • Step 4 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol 6-(6-Methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1.
  • Example 6-2 Synthesis of 2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
  • Step 1 7-Bromo-6-methoxy-2-methylquinoline According to the general procedure of Matsugi, M, et al, Tetrahedron Lett, 2000, 41, 8523, to a solution of 3-bromo-4-methoxyaniline (1 .00 g, 4.95 mmol) in 6 M HCI (25 ml.) and toluene (6 ml.) heated to 100 °C was added dropwise crotonaldehyde (0.407 ml_, 4.95 mmol). Stirring was continued for 2 h at 100 °C and the mixture then allowed to cool to room temperature. The toluene layer was removed and the aqueous layer basified to pH 7 by slow addition of 2 M NaOH.
  • the aqueous phase was extracted with DCM (6x) and concentrated to a green residue.
  • the residue was dissolved into 1 M HCI (75 ml.) and zinc chloride (1 .349 g, 9.90 mmol) was added resulting in immediate formation of a dark gummy solid. Sonication and stirring of the mixture transformed the gummy solid to a light brown precipitate.
  • the solids were isolated by filtration, washing with 1 M HCI, 2-propanol, then water. The solids were partitioned between 1 :1
  • Butyllithium (1.6 M in heptane, 0.41 mL, 0.65 mmol) was added dropwise over the course of 1 h to a solution of 7-bromo-6-methoxy-2-methylquinoline (150 mg, 0.60 mmol) and triisopropyl borate (0.180 mL, 0.77 mmol) cooled to -78 °C. After addition was complete, the reaction was stirred at -78 °C for one hour and then allowed to warm to room temperature and stir overnight. LC/MS analysis shows a 9:1 ratio of the title boronic acid to the debrominated side product (6- methoxy-2-methylquinoline).
  • Step 3 6-( 6-Methoxy-2-methylquinolin- 7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • Example 7-1 Synthesis of 3-chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol hydrochloride salt
  • Step 1 6-(3-Chloro- 7-methoxyquinolin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • Example 7-2 Synthesis of 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Step 1 6-(3-Bromo-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • 6-(7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 9, 100 mg, 0.247 mmol), was borylated according to GENERAL METHOD 5-1.
  • To the crude boronic acid solution was added water (800 uL) and CuBr 2 (165 mg, 0.740 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was partitioned between DCM/water. The aqueous phase was extracted with EtOAc (5x), DCM (2x), and the combined organic extracts were concentrated.
  • Example 7-3 Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-3-carbonitrile
  • Example 7-4 Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(1 -methyl-1 H-imidazol-4-yl)quinolin-7-ol
  • Example 7-5 Synthesis of 3-(1 H-imidazol-1 -yl)-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol hydrochloride salt
  • Step 1 6-(3-(1H-lmidazol-1-yl)-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
  • yl)boronic acid (Example 7-4 Step 1 , 0.06 g, 0.14 mmol), imidazole (0.06 g, 0.8 mmol) and copper(ll) nitrate hemi(pentahydrate) (0.04 g, 0.2 mmol) were added to tetramethylethylenediamine (0.03 ml_, 0.20 mmol) in methanol (2 ml_). The reaction mixture was heated at 40 °C overnight. A 28% ammonium hydroxide solution was added to the reaction and the aqueous phase was extracted with DCM (3x). The combined organic phases were dried over anhydrous Na 2 S0 4, filtered and concentrated under reduced pressure.
  • Example 7-6 Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-3,7-diol formate salt.
  • Example 8-1 Synthesis of 3-ethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Benzophenone 0.65 g, 4.20 mmol
  • potassium ferf-butoxide 0.514 g, 4.58 mmol
  • the mixture was cooled to room temperature, diluted with EtOAc, washed with saturated aqueous sodium bicarbonate and brine.
  • the organic phase was extracted with 1 M HCI (3x).
  • the acidic extracts were basified to pH 12 by addition of 2 M NaOH and extracted with EtOAc (3x) and diethyl ether (2x).
  • Step 3 6-(3 ⁇ thyl-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetra ⁇
  • 6-(3-Ethyl-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine (100 mg, 0.231 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-2. The crude material was subjected to silica gel chromatography
  • Example 8-2 Synthesis of 3-isopropyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 9-1 Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one
  • Step 3 (7-Methoxy-2-oxo-1 ,2-dihydroquinolin-6-yl)boronic acid and 7-methoxy-6-(4,4,5,5- tetramethyl-1 , 3, 2-dioxaborolan-2-yl)quinolin-2( 1H)-one
  • Step 4 7-Methoxy-6-(6-(methyl(2,2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-2(1H)-one
  • Step 5 7-Hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-2( 1H)-one
  • Example 9-2 Synthesis of 7-hydroxy-1 -methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one hydrochloride salt
  • Methyltrioxorhenium (0.03 g, 0.12 mmol) was added to 6-chloro-7-methoxyquinoline (1 .69 g, 8.73 mmol) in DCM (40 mL). A solution of hydrogen peroxide (50 wt % in water, 1.0 mL, 17 mmol) was then added at 5 °C and the mixture was stirred at room temperature overnight. An additional portion of methyltrioxorhenium (0.03 g, 0.12 mmol) was added followed by hydrogen peroxide (50 wt % in water, 1.0 mL, 17 mmol) and the reaction was stirred for 4 h.
  • Trifluoroacetic anhydride (6.0 mL, 42 mmol) was added dropwise to 6-chloro-7- methoxyquinoline 1-oxide (1.18 g, 5.63 mmol) in DMF (8.0 mL) at 5 °C.
  • the reaction mixture was heated in a Biotage ® Initiator microwave reactor at 100 °C for 30 min. Water (20 mL) was added to the reaction and the resulting solid was collected by filtration. The solid was triturated with hot acetonitrile and filtered to afford 6-chloro-7-methoxyquinolin-2(1 H)-one (0.39 g).
  • MS (M+1 ) 210.1.
  • Potassium bis(trimethylsilyl)amide (0.5 M in toluene, 2.7 mL, 1.4 mmol) was added to 6- chloro-7-methoxyquinolin-2(1 H)-one (0.20 g, 0.95 mmol) in toluene (5 mL) and the mixture was stirred for 10 min at room temperature, lodomethane (0.07 mL, 1 mmol) was added to the reaction and the mixture was stirred 2 h at 50 °C.
  • the mixture was stirred at 80 °C for 2 h, then a 4% solution of Na 2 C0 3 was added to the reaction and the aqueous phase was extracted with DCM (3x).
  • the combined organic phases were extracted with 2 M HCI (3x).
  • the combined acidic aqueous phases were basified to pH 1 1 with 6 M NaOH then extracted with DCM (3x).
  • the combined organic phases were dried over anhydrous Na 2 S0 4, filtered and concentrated under reduced pressure.
  • Step 6 7-Hydroxy- 1 -methyl-6-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinolin-2(1H)-one hydrochloride salt.
  • Example 10-1 Synthesis of 4-methoxy-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 10-2 Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(pyrrolidin-1 -yl)quinolin-7-ol
  • Example 10-1 or Example 10-2 The following example compounds were prepared from the Intermediate 11 and the appropriate sodium alkoxide or amine according to the preparations of Example 10-1 or Example 10-2.
  • Example 11 -1 Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(1 -methyl-1 H-pyrazol-4-yl)quinolin-7-ol
  • Step 1 6-(7-Methoxy-2-methyl-4-( 1 -methyl- 1 H-pyrazol-4-yl)quinolin-6-yl)-N-methyl-N-
  • Step 2 2-Methyl-6-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-( methyl- 1H-pyrazol-4-yl)quinolin-7-ol
  • Example 12-1 Synthesis of 4-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol formate salt
  • Example 13-1 Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoxalin-6-ol hydrochloride salt
  • Step 3 6-(7-Ethoxyquinoxalin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4-yl)pyridazin- 3-amine
  • the reaction mixture was cooled to room temperature and the volatiles were removed under vacuum. 2 M aqueous HCI was added to the crude material and the aqueous phase was washed with DCM and basified to pH 1 1 with 6 M aqueous NaOH. The precipitate was collected by filtration and the filtrate was extracted with 9:1 DCM/MeOH (3x). The organic extracts and the collected precipitate were mixed and volatiles were removed under vacuum.
  • the crude material was purified by silica gel chromatography (2-15% 7 M ammonia in MeOH gradient, in DCM) followed by a second purification via reverse-phase preparative HPLC (25-50% acetonitrile in water, 5 mM NH 4 OH as modifier).
  • Step 4 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol hydrochloride salt.
  • Step 1 2-Hydroxy-4-methoxy-5-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)benzaldehyde
  • the title compound (3.5 g, 8.78 mmol, 53%) was prepared following GENERAL METHOD 9-1 for Suzuki reaction of Intermediate 1 (4.7 g, 16.62 mmol) and Intermediate 15 (7.86 g, 28.3 mmol).
  • MS (M+1 ) 399.2.
  • Step 1 2-Hydroxy-4-methoxy-5-( 6-((2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)benzaldehyde
  • MP-carbonate a resin-bound base
  • Biotage part number 800267 for 10 grams.
  • the chemical name for MP-carbonate is macroporous
  • Example 14-1 Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(tetrahydro-2H-pyran-4-yl)quinolin-7-ol
  • Example 15-1 Synthesis of 3-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
  • Example 15-2 Synthesis of 3-bromo-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
  • Example 15-3 Synthesis of 3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
  • Example 16-1 Synthesis of 5-bromo-3-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol
  • Example 17-1 Synthesis of 6-hydroxy-1 -methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-4(1H)-one
  • Step 1 6-methoxy-1-methyl-7-(6-(methyl(2,2, 6, 6-tetramethylpiperidin-4-yl)amino)pyrM 3-yl)quinolin-4(1H)-one
  • Step 2 6-Hydroxy- 1 -methyl- 7-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinolin-4(1H)-one
  • Step 3 6-(7-Ethoxy-2, 3-dimethylquinoxalin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
  • the reaction mixture was sealed and heated in a Biotage ® Initiator microwave reactor at 130 °C for 1 h.
  • a solution of NaHC0 3 was added to the reaction and the aqueous phase was extracted with DCM (3x).
  • the combined organic phases were dried over anhydrous Na 2 S0 4 filtered, and concentrated under reduced pressure.
  • the crude material was purified by silica gel chromatography (1 -15% 7 M ammonia in MeOH gradient, in DCM) followed by a second purification via reverse-phase preparative HPLC (5-20% acetonitrile in water, 7.5% formic acid modifier).
  • Step 4 2, 3-Dimethyl-7-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol
  • Examples 18-2 and 18-3 Synthesis of 2-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol and 3-methyl-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol
  • Step 1 6-Bromo-7-ethoxy-2-methylquinoxaline and 7-bromo-6-ethoxy-2-methylquinoxaline 2-Oxopropanal (8.26 g, 45.8 mmol) was added dropwise to 4-bromo-5-ethoxybenzene-1 ,2- diamine (Example 13-1 Step 1) (8.41 g, 36.4 mmol) in THF (360 mL) and the reaction was stirred overnight.
  • Step 2 6-Ethoxy-2-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoxaline, (7- ethoxy-3-methylquinoxalin-6-yl)boronic acid, and 7-ethoxy-2-methyl-6-(4, 4, 5, 5-tetramethyl- 1, 3, 2- dioxaborolan-2-yl)quinoxaline and (7-ethoxy-2-methylquinoxalin-6-yl)boronic acid
  • Step 3 6-(7-Ethoxy-3-methylquinoxalin-6-yl)-N-methyl-N-(2,2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine, and 6-(7-ethoxy-2-methylquinoxalin-6-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
  • Step 4 2-Methyl-7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol, and 3-methyl- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol
  • Example 19-1 Synthesis of 4-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 20-1 Synthesis of 4-(azetidin-1 -yl)-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 20-2 Synthesis of 7-hydroxy-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-4-carbonitrile
  • Example 20-3 Synthesis of 4-cyclopropyl-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Step 1 6-(4-Cyclopropyl-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
  • Step 2 4-Cyclopropyl-2-met yl-6-(6-(met yl(2,2, 6, 6-tetramet ylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 20-4 Synthesis of 4-(3,6-dihydro-2H-pyran-4-yl)-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
  • Example 20-5 Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)quinolin-7-ol formate salt
  • Example 20-4 A mixture of Example 20-4 (14 mg, 0.029 mmol), Pd-C (3 mg, 10 wt % on carbon, 3 ⁇ ), Pd(OH) 2 (2.0 mg, 20 wt % on carbon, 3 ⁇ ) and one drop of concentrated HCI aqueous solution in methanol (10 mL) was evacuated, filled with H 2 (4x) and shaken under H 2 (50 psi) on a Parr shaker hydrogenator at room temperature overnight. The reaction mixture was filtered through Celite®, washed with MeOH, concentrated, and subjected to SCX purification (GENERAL
  • Example 20-6 Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(oxetan-3-yl)quinolin-7-ol
  • Example 21 -1 Synthesis of 4-(dimethylamino)-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol di-formate salt
  • Step 3 7-Methoxy-6-(4, 4, 5, 5-tetramethyl-1 ,3, 2-dioxaborolan-2-yl)quinazolin-4( 1H)-one
  • Step 4 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazolin-4( 1H)-one 7-Methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazolin-4(1 H)-one (305 mg, -61 % pure, 0.62 mmol ), 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 1 , 145 mg, 0.51 mmol), Pd(PPh 3 ) 4 (47 mg, 0.041 mmol), aqueous Na 2 C0 3 (2.0 M, 0.64 ml_, 1.3 mmol) and 1 ,4-dioxane (2.4 ml.) were combined in a microwave vessel.
  • the vessel was evacuated and back-filled with nitrogen (4x), and heated via microwave irradiation at 1 10 °C for 1 h.
  • the mixture was cooled to room temperature and acidified to pH 2 with 4 M HCI in 1 ,4- dioxane.
  • SCX purification (GENERAL METHOD 3-1 ) followed by silica gel chromatography (0-20% MeOH in DCM) provided 7-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinazolin-4(1 H)-one (137 mg) as a light brown solid.
  • MS (M+1 ) 423.1. 1 H NMR
  • Step 5 7-Hydroxy-6-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazolin-4( 1H)-one

Abstract

The present invention provides a compound of formula IA or a pharmaceutically acceptable salt thereof; 5 (IA) a method for manufacturing the compounds of the invention, and its therapeutic uses. The present invention further provides a combination of pharmacologically active agents and a pharmaceutical composition.

Description

1 ,4-DISUBSTITUTED PYRIDAZINE DERIVATIVES AND THEIR USE FOR TREATING
SMN-DEFICIENCY-RELATED CONDITIONS
BACKGROUND OF THE INVENTION
Proximal spinal muscular atrophy (SMA) is an inherited, clinically heterogeneous group of neuromuscular disorders characterized by degeneration of the anterior horn cells of the spinal cord. Patients suffer from symmetrical weakness of trunk and limb muscles, the legs being more affected than the arms and the proximal muscles weaker than the distal ones; diaphragm, facial and ocular muscles are spared. There are three forms of childhood-onset SMA (types I, II and III), and a relatively recently categorized adult-onset form IV, all of which can be distinguished on the basis of age of onset and severity of the clinical course assessed by clinical examination, muscle biopsy and electromyography (EMG)(Munsat T L, Davies K E (1992)).
Type I (Werdnig-Hoffmann disease) is the most acute and severe form, with onset before six months and death usually before two years; children are never able to sit without support.
Symptoms of the disease can be present in utero, as reduction of fetal movements; at birth; or more often, within the first four months of life. Affected children are particularly floppy, experience feeding difficulties and diaphragmatic breathing, and are characterized by a general weakness in the intercostals and accessory respiratory muscles. Affected children never sit or stand and usually die before the age of 2; death is generally due to respiratory insufficiency.
Type II (intermediate, chronic form) has onset between six and eighteen months of age; muscular fasciculations are common, and tendon reflexes progressively reduce. Children are unable to stand or walk without aid. Feeding and swallowing problems are not usually present in Type II SMA, although in some patients a feeding tube may become necessary. Most patients generally develop a progressive muscular scoliosis which can require surgical correction. Like patients with type I disease, clearing of tracheal secretions and coughing might become difficult because of poor bulbar function and weak intercostal muscles. These patients have profound hypotonia, symmetrical flaccid paralysis, and no control of head movement.
Type III (Kugelberg-Welander disease, or Juvenile Spinal Muscular Atrophy) is a mild, chronic form, with onset after the age of 18 months; motor milestones achievement is normal, and deambulation can be preserved until variable ages. These patients often develop scoliosis, and symptoms of joint overuse, generally caused by weakness, are frequently seen. Life expectancy is almost normal but quality of life is markedly compromised.
Types I, II and III progress over time, accompanied by deterioration of the patient's condition.
Adult-onset type IV is characterized by weakness in the second or third decade of life, with mild motor impairment not accompanied by respiratory or nutritional problems. Adult SMA is characterized by insidious onset and very slow progression. The bulbar muscles are rarely affected in Type IV. It is not clear that Type IV SMA is etiologically related to the Type l-lll forms.
Other forms of spinal muscular atrophy include X-linked disease, spinal muscular atrophy with respiratory distress (SMARD), spinal and bulbar muscular atrophy (Kennedy's disease, or Bulbo-Spinal Muscular Atrophy), and distal spinal muscular atrophy.
SMA is due to mutations in the Survival of Motor Neuron (SMN) gene, which exists in two forms in humans (SMN1 and SMN2). Loss of SMN is deleterious to motor neurons and results in neuromuscular insufficiency, a hallmark of the disease. From a genetic point of view, SMA is an autosomal recessive condition, caused by disruption of SMN1 gene, located in 5q13 (Lefebvre S., et al. (1995) Cell 80: 155-165). More than 98% of patients with spinal muscular atrophy have a homozygous disruption of SMN1 by deletion, rearrangement, or mutation. All these patients, however, retain at least one copy of SMN2.
At the genomic level, only five nucleotides have been found that differentiate the SMN1 gene from the SMN2 gene. Furthermore, the two genes produce identical mRNAs, except for a silent nucleotide change in exon 7, i.e., a C→T change six base pairs inside exon 7 in SMN2. This mutation modulates the activity of an exon splicing enhancer (Lorson and Androphy (2000) Hum. Mol. Genet. 9:259-265). The result of this and the other nucleotide changes in the intronic and promoter regions is that most SMN2 are alternatively spliced, and their transcripts lack exons 3, 5, or 7. In contrast, the mRNA transcribed from the SMN1 gene is generally a full-length mRNA with only a small fraction of its transcripts spliced to remove exon 3, 5, or 7 (Gennarelli et al. (1995) Biochem. Biophys. Res. Commun. 213:342-348; Jong et al. (2000) J. Neurol. Sci. 173:147-153). All SMA subjects have at least one, and generally two to four copies of the SMN2 gene, which encodes the same protein as SMN1 ; however, the SMN2 gene produces only low levels of full- length SMN protein.
The SMNA7 protein is non-functional and thought to be rapidly degraded. About 10% of
SMN2 pre-mRNA is properly spliced and subsequently translated into full length SMN protein (FL- SMN), and the rest being the SMNA7 copy. The efficiency of SMN2 splicing might be dependent on severity of disease, and production of a full length transcript of SMN2 could range from 10% to 50%. Furthermore, presence or absence of the SMN1 gene, roughly 90% of which becomes the FL-SMN gene product and protein, influences the severity of SMA by whether or not it can compensate for the truncated SMNA7 copies. A low level of SMN protein allows embryonic development, but is not sufficient to sustain the survival of motor neurons of the spinal cord.
The clinical severity of SMA patients inversely correlates with the number of SMN2 genes and with the level of functional SMN protein produced (Lorson C L, et al. (1999) PNAS; 96:6307- 631 1 )(Vitali T. et al. (1999) Hum Mol Genet; 8:2525-2532)(Brahe C. (2000) Neuromusc. Disord.; 10:274-275)(Feldkotter M, et al. (2002) Am J Hum Genet; 70:358-368)(Lefebvre S, et al. (1997) Nature Genet; 16:265-269)(Coovert D D, et al. (1997) Hum Mol Genet; 6:1205-1214)(Patrizi A L, et al. (1999) Eur J Hum Genet; 7:301-309).
Current therapeutic strategies for SMA are mostly centered on elevating full length (wild type) SMN protein levels, modulating splicing towards exon 7 inclusion, stabilizing the wild type protein, and to a lesser extent, on restoring muscle function in SMA by providing trophic support or by inhibiting skeletal muscle atrophy.
The mechanism leading to motorneuron loss and to muscular atrophy still remains obscure, although the availability of animal models of the disease is rapidly increasing knowledge in this field
(Frugier T, et al. (2000) Hum Mol. Genet. 9:849-58; Monani U R, et al. (2000) Hum Mol Genet 9:333-9; Hsieh-Li H M, et al. (2000) Nat Genet 24:66-70; Jablonka S, et al. (2000) Hum Mol. Genet.
9:341-6). Also the function of SMN protein is still partially unknown, and studies indicate that it can be involved in mRNA metabolism (Meister G, et al. (2002). Trends Cell Biol. 12:472-8; Pellizzoni L, et al. (2002). Science. 298: 1775-9), and probably in transport of proteins/mRNA to neuromuscular junctions (Ci-fuentes-Diaz C, et al. (2002) Hum Mol. Genet. 1 1 : 1439-47; Chan Y B, et al. (2003) Hum Mol. Genet. 12:1367-76; McWhorter M L, et al. (2003) J. Cell Biol. 162:919-31 ; Rossoll W, et al. (2003) J. Cell Biol. 163:801-812).
In addition to the SMAs, a subclass of neurogenic-type arthrogryposis multiplex congenita
(congenital AMC) has separately been reported to involve SMN1 gene deletion, suggesting that some degree of pathology in those afflicted is likely due to low levels of motor neuron SMN. (L. Burgien et al., (1996) J. Clin. Invest. 98(5):1 130-32. Congenital AMC affects humans and animals, e.g., horses, cattle, sheep, goats, pigs, dogs, and cats. (M. Longeri et al., (2003) Genet. Sel. Evol.
35:S167-S175). Also, the risk of development or the severity of amyotrophic lateral sclerosis (ALS) has been found to be correlated with low levels of motor neuron SMN.
There is no cure for SMA available to date and therefore it would be advantageous to provide novel methods for modulating SMN in order to treat those afflicted with SMA, with neurogenic congenital AMC, ALS, or with other SMN-deficiency-related conditions. It would further be advantageous to provide novel drug targets that could be used as a basis for developing effective therapeutics or diagnostics for such neuronal conditions. SUMMARY OF THE INVENTION
There remains a need for new treatments and therapies for Spinal Muscular Atrophy. The invention provides compounds, salts thereof, pharmaceutical formulations thereof and
combinations thereof which compounds are Spinal Muscular Atrophy modulators. The invention further provides methods of treating, preventing, or ameliorating Spinal Muscular Atrophy, comprising administering to a subject in need thereof an effective amount of an SMN modulator (e.g., a compound of the invention). Various embodiments of the invention are described herein. It will be recognized that features specified in each embodiment may be combined with other specified features to provide further embodiments.
Within certain aspects, SMN modulators provided herein are compounds of Formula IA and salts thereof:
Figure imgf000005_0001
(IA)
In another embodiment, the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound according to the definition of formula (I) or subformulae thereof and one or more pharmaceutically acceptable carriers.
In another embodiment, the invention provides a combination, in particular a pharmaceutical combination, comprising a therapeutically effective amount of the compound according to the definition of formula (I) or subformulae thereof and one or more therapeutically active.
One embodiment of the invention is to provide a method for treating, preventing, or ameliorating an SMN-deficiency-related condition, comprising administering to a subject in need thereof an effective amount of an SMN modulator, or a pharmaceutical composition comprising the same.
Another embodiment of the invention is a method of modulating SMN protein through the administration of an SMN modulator. In another embodiment, said SMN modulator is capable of increasing one or more of FL-SMN or SMNA7 levels. In still another embodiment, said SMN modulator is capable of preventing exon 7 from being spliced from the SMN transcript.
The present invention is based on the discovery that the SMN modulators of the invention (e.g., compounds of formula (I) and/or compounds of formula (l-A) are capable of modulating SMN proteins, e.g., through SMN promoter activation, splicing modulation (e.g., preventing exon7 from being spliced out of the SMN gene), and/or SMN protein stability modulation.
DETAILED DESCRIPTION OF THE INVENTION
As noted above, the present invention provides compounds that modulate SMN activity. Such compounds may be used in vitro or in vivo to modulate (preferably increase) SMN production and activity in a variety of contexts.
In a first embodiment, the invention provides compounds of Formula (IA) and
pharmaceutically acceptable salts thereof, which modulate SMN activity. Compounds of Formula I are represented by the structure (IA):
Figure imgf000006_0001
(IA)
wherein A is bicyclic heteroaryl or heterocyle having 9 or 10 ring atoms and 1 or 2 ring N atoms and 0 or 1 O atoms, which bicyclic heteroaryl or heterocycle is substituted with 0, 1 , 2, 3, 4 or 5 substituents independently selected from -C(0)NH2,-C(0)0-Ci-C4alkyl, aryl, oxo, cyano, halogen, hydroxy, Ci-C4alkyl, C2-C4alkenyl, C2-C4alkynyl, d-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl CrC4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, CrC4alkyl heteroaryl, CrC4alkoxy aryl, Ci-C4alkoxy heterocyclyl, Ci-C4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, Ci-C4alkoxy, amino and mono-and di-Ci-C4alkylamino; and B is a group of the formula:
Figure imgf000006_0002
wherein m, n and p are independently selected from 0 or 1 ; R, R-i, R2, R3, and R4 are independently selected from the group consisting of hydrogen, CrC4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino; R5 and R6 are independently selected from hydrogen and fluorine; or R and R3, taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S; Ri and R3, taken in combination form a CrC3alkylene group; Ri and R5, taken in combination form a Ci-C3alkylene group; R3 and R4, taken in combination with the carbon atom to which they attach, form a spirocyclicC3-C6cycloalkyl; X is CRARB, O, N R7 or a bond; R7 is hydrogen, or CrC4alkyl; RA and RB are independently selected from hydrogen and CrC4alkyl, or RA and RB, taken in combination, form a divalent C2-C5alkylene group; Z is CR8 or N; when Z is N, X is a bond; R8 is hydrogen or taken in combination with R6 form a double bond; or B is a group of the formula:
Figure imgf000007_0001
wherein p and q are independently selected from the group consisting of 0, 1 , and 2; R9 and R13 are independently selected from hydrogen and C1-C4alkyl; R10 and R14 are independently selected from hydrogen, amino, mono- and di-C1-C4akylamino and C1-C4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-C C4akylamino; Rn is hydrogen, d-C4alkyl, amino or mono- and di-Ci-C4akylamino; Ri2 is hydrogen or CrC4alkyl; or R9 and Rn, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1-3 Ci-C4alkyl groups; or R-n and Ri2, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 CrC4alkyl groups.
In a second embodiment the invention is a the compound, or salt thereof, according to the first embodiment wherein the compound is of Formula (I):
Figure imgf000007_0002
(I)
wherein A is bicyclic heteroaryl having 10 ring atoms and 1 or 2 ring N atoms, which bicyclic heteroaryl is substituted with 0, 1 , or 2 substituents independently selected from oxo, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, CrC4alkoxy, C3-C7cycloalkyl,
heterocyclyl, heteroaryl, heterocyclyl CrC4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, Ci-C4alkoxy substituted with hydroxy, Ci-C4alkoxy, amino and mono-and di-Ci-C4alkylamino; and B is a group of the formula:
Figure imgf000007_0003
wherein m, n and p are independently selected from 0 or 1 ; R, R-i, R2, R3, and R4 are independently selected from the group consisting of hydrogen, d-C4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino; R5 and R6 are independently selected from hydrogen and fluorine; or R and R3, taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S; R-i and R3, taken in combination form a C-|-C3alkylene group; R-i and R5, taken in combination form a C-|-C3alkylene group; R3 and R4, taken in combination with the carbon atom to which they attach, form a spirocyclicC3-C6cycloalkyl; X is CRARB, O, N R7 or a bond; R7 is hydrogen, or CrC4alkyl; RA and RB are independently selected from hydrogen and CrC4alkyl, or RA and RB, taken in combination, form a divalent C2-C5alkylene group; Z is CR8 or N; when Z is N, X is a bond; R8 is hydrogen or taken in combination with R6 form a double bond; or B is a group of the formula:
Figure imgf000008_0001
wherein p and q are independently selected from the group consisting of 0, 1 , and 2; R9 and Ri3 are independently selected from hydrogen and CrC4alkyl; R10 and R14 are independently selected from hydrogen, amino, mono- and di-Ci-C4akylamino and Ci-C4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino; Rn is hydrogen, Ci-C4alkyl, amino or mono- and di-Ci-C4akylamino; R12 is hydrogen or CrC4alkyl; or R9 and Rn, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1-3 C1-C4alkyl groups; or R^ and R12, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 C1-C4alkyl groups.
In a third embodiment, the invention is the compound according to any one the first or second embodiments, or salt thereof, wherein A is selected from:
Figure imgf000009_0001
Figure imgf000009_0002
Figure imgf000009_0003
Figure imgf000009_0004
Figure imgf000009_0005
Figure imgf000009_0006
wherein u and v are each, independently, 0, 1 , 2 or 3; and each Ra and R are, independently, selected from , cyano, halogen, hydroxy, C1-C4alkyl, C2-C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3- C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C1-C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C1-C4alkyl heteroaryl, C1-C4alkoxy aryl, C1-C4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, and CrC4alkoxy substituted with hydroxy, CrC4alkoxy, amino and mono-and di-Cr C4alkylamino. In a fourth embodiment, the invention is the compound of any one of the first through third embodiments, or salt thereof, wherein A is selected from:
Figure imgf000010_0001
and
wherein u and v are each, independently, 0, 1 , 2 or 3; and each Ra and R are, independently, selected from , cyano, halogen, hydroxy, C1-C4alkyl, C2-C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3- C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C1-C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C-i-C4alkyl heteroaryl, CrC4alkoxy aryl, Ci-C4alkoxy heterocyclyl, Ci-C4alkoxy heteroaryl, and CrC4alkoxy substituted with hydroxy, CrC4alkoxy, amino and mono-and di-Cr C4alkylamino.
In another embodiment, the invention is the compound according to any one of the first through fourth embodiments, wherein A is substituted in the ortho position with a hydroxyl group.
In a fifth embodiment, the inveniotn is the compound according to any one of the first through fourth embodiments, or salt thereof, wherein A is selected from:
Figure imgf000011_0001
Figure imgf000011_0002
Figure imgf000011_0003
Figure imgf000011_0004
and
Figure imgf000011_0005
In a sixth embodiment, the invneiotn is the compound of any one of the first through fifth embodiments, or a salt thereof, wherein A has a single N atom
In a seventh embodiment, the invention is the compound, according to any one of the first through sixth emebodiments, or salt thereof, wherein the compound is of formula II:
Figure imgf000011_0006
(II)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C1-C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C1-C4alkyl heteroaryl, C1-C4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, C C4alkoxy, amino and mono-and di-Ci-C4alkylamino.
In an eighth embodiment, the invention is the compound, or salt thereof, according to any one of the first through sixth embodiments, wherein the compound is of formula III:
Figure imgf000012_0001
(III)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C1-C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C1-C4alkyl heteroaryl, C1-C4alkoxy aryl, C1-C4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, C1-C4alkoxy substituted with hydroxy, C C4alkoxy, amino and mono-and di-Ci-C4alkylamino.
In a ninth embodiment, the invention is the compound, according to any one of the first through sixth embodiments, or salt thereof, wherein the compound is of formula IV:
Figure imgf000012_0002
(IV)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, d-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C4alkyl, Ci-C4alkyl aryl, CrC4alkyl heterocyclyl, CrC4alkyl heteroaryl, Ci-C4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, C
C4alkoxy, amino and mono-and di-Ci-C4alkylamino.
In a tenth embodiment, the invention is the compound, according to any one of the first through sixth emebodiments, or salt thereof, wherein the compound is of formula V:
Figure imgf000012_0003
(V)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, Ci-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C4alkyl, d-C4alkyl aryl, CrC4alkyl heterocyclyl, CrC4alkyl heteroaryl, Ci-C4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, C C4alkoxy, amino and mono-and di-Ci-C4alkylamino.
In an eleventh embodiment, the invention is the compound, according to any one of cthe first through sixth emebodiments, or salt thereof, wherein the compound is of formula VI:
Figure imgf000013_0001
(VI)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl C1-C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C1-C4alkyl heteroaryl, C1-C4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, C C4alkoxy, amino and mono-and di-Ci-C4alkylamino.
In a twelfth embodiment, the invention is the compound, according to any one of the first through sixth embodiments, or salt thereof, wherein the compound is of formula VII:
Figure imgf000013_0002
(VII)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, Ci-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C4alkyl, Ci-C4alkyl aryl, CrC4alkyl heterocyclyl, CrC4alkyl heteroaryl, Ci-C4alkoxy aryl, C1-C4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, C1-C4alkoxy substituted with hydroxy, C C4alkoxy, amino and mono-and di-C1-C4alkylamino.
In a thirteenth embodiment, the invention is the compound, according to any one of the first through sixth embodiments, or salt thereof, wherein the compound is of formula VIII:
Figure imgf000013_0003
(VIII) wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, C C4alkyl, C2-C4alkenyl, C2-C4alkynyl, d-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-C4alkyl, Ci-C4alkyl aryl, CrC4alkyl heterocyclyl, CrC4alkyl heteroaryl, Ci-C4alkoxy aryl, C1-C4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, C1-C4alkoxy substituted with hydroxy, C C4alkoxy, amino and mono-and di-C1-C4alkylamino.
In a fourteenth embodiment, the invention is the compound according to any one of the first through thirteenth embodiments, or a salt thereof, wherein B is a group of the formula:
Figure imgf000014_0001
wherein m, n and p are independently selected from 0 or 1 ; R, R-i, R2, R3, and R4 are independently selected from the group consisting of hydrogen, CrC4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino; R5 and R6 are hydrogen; or R and R3, taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S; Ri and R3, taken in combination form a CrC3alkylene group; Ri and R5, taken in combination form a Ci-C3alkylene group; R3 and R4, taken in combination with the carbon atom to which they attach, form a spirocyclicC3-C6cycloalkyl; X is CRARB, O, N R7 or a bond; RA and RB are independently selected from hydrogen and CrC4alkyl, or RA and RB, taken in combination, form a divalent C2-C5alkylene group; Z is CR8 or N; when Z is N, X is a bond; R8 is hydrogen or taken in combination with R6 form a double bond.
In a fifteenth embodiment, the invention is the compound according to any one of the first through thirteenth embodiments, or a salt thereof, wherein B is a group of the formula:
Figure imgf000014_0002
wherein p and q are independently selected from the group consisting of 0, 1 , and 2; R9 and Ri3 are independently selected from hydrogen and CrC4alkyl; R10 and R14 are independently selected from hydrogen, amino, mono- and di-Ci-C4akylamino and Ci-C4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino; Rn is hydrogen, d-C4alkyl, amino or mono- and di-Ci-C4akylamino; R12 is hydrogen or CrC4alkyl; or R9 and Rn, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1-3 C1-C4alkyl groups; or R^ and R12, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 C1-C4alkyl groups.
In a sixteenth embodiment, the invention is the compound of any one of the first through fifteenth embodiments, wherein B is selected from the group consisting of:
Figure imgf000015_0001
wherein X is O or N(Me) or NH; and Ri7 is hydrogen or methyl.
In a seventeenth embodiment, the invention is the compound, or salt thereof, according to any one of the first through sixteenth embodiments, wherein B is:
Figure imgf000015_0002
In an eighteenth embodiment, the invention is the compound, or salt thereof, according to any one of the first through seventeenth embodiments, wherein X is -0-.
In a nineteenth emebodiment, the invention is the compound, or salt thereof, according to any one of the first through eighteenth embodiments, wherein X is N(Me).
In a twentieth embodiment, theinvention is a compound, or salt thereof, selected from the group consisting of:
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one; 6- (6-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1 H)-yl)pyridazin-3-yl)quinolin-7-ol;
7- hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui 2(1 H)-one;
6- (6-(methyl(1 ,2,2,6,6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-morphoN
7- ol;
4-chloro-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7
3-ethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7
3-(1 H-imidazol-1-yl)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-y ol;
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(1-methyl-1 H-imidazol-4- yl)quinolin-7-ol;
3- isopropyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3J-diol;
7- hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoli carbonitrile;
6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
4- (dimethylamino)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyn yl)quinolin-7-ol;
3- chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperid^
4- methoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui ol;
6- (3-(benzyloxy)isoquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-am 8-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
7- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline-1 ,6-d
7-(6-(methyl(1 ,2,2,6,6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
1- cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquino 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
6-(1 -(benzyloxy)isoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-am 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol;
2- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(1-meth H- pyrazol-4-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin
4-ethoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui 4- chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperi
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(tetrahydro-2H-p
yl)quinolin-7-ol;
3-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl^^
3-bromo-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6
3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin
5- bromo-3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
6- hydroxy-1-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
4(1 H)-one;
2,3-dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4^
2- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin
3- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin
4- methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoli
4-(azetidin-1-yl)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
yl)quinolin-7-ol;
7- hydroxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qu carbonitrile;
4-cyclopropyl-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-y ^
7- ol;
4-(3,6-dihydro-2H-pyran-4-yl)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(tetrahyd
pyran-4-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)a
yl)quinolin-7-ol;
4-(dimethylamino)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-^
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin H)-one;
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin-7-ol;
7- hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3 dihydroquinolin-2(1 H)-one;
7-hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3 dihydroquinolin-2(1 H)-one;
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinol^
carbonitrile;
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quin
carbonitrile; 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-y^
carbonitrile;
6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidi
carboxamide;
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin carboxamide;
6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
carboxamide;
methyl 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2- carboxylate;
6- hydroxy-7-(6-(piperazin-1 -yl)pyridazin-3-yl)quinoline-2-carbonitrile;
7- hydroxy-6-(6-(piperazin-1 -yl)pyridazin-3-yl)quinoline-2-carbonitrile;
7-(6-(piperazin-1-yl)pyridazin-3-yl)isoquinolin-6-ol;
7-(6-(1 ,2,3,6-tetrahydropyridin-4-yl)pyridazin-3-yl)quinolin-6-ol;
1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-7-ol; 1-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol; 1 ,3-dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin 7-hydroxy-3-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquino 1 -carbonitrile;
1 -amino-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin
7-hydroxy-1 ,3-dimethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazoline-2,4(1 H,3H)-dione;
6-hydroxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)benzo[d]oxazol- 2(3H)-one;
2-methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-2H-indazol-6-ol;
1- methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-1 H-indazol-6-ol;
6- hydroxy-2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoqui 1 (2H)-one;
2- ethyl-6-hydroxy-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinolin-1 (^ 1-ethoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin
7- (6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinoline-1 ,6-diol;
7-(6-(methyl(2,2,6,6-tetramethyl-piperidin-4-yl)amino)-pyridazin-3-yl)-3-phenylisoquinolin
3- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-^^ 3-cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethyl-piperidin-4-yl)amino)pyridazin-3-yl)isoquinolin 3-isopropyl-7-(6-(methyl(2,2,6,6-tetramethyl-piperidin-4-yl)amino)pyridazin-3-yl)isoquinolin 3-propyl-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)-pyridazin-3-yl)isoquinolin-6-ol; 3-isopropyl-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)-pyridazin-3-yl)isoquinoli and
3- methyl-7-(6-(piperazin-1 -yl)pyridazin-3-yl)isoquinolin-6-ol;
In a twentyfirst embodiment, the invention is a compound, or salt thereof, selected from the group consisting of:
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one;
6- (6-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1 H)-yl)pyridazin-3-yl)quinolin-7-ol;
7- hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin- 2(1 H)-one;
6- (6-(methyl(1 ,2,2,6,6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-morpholinoquinolin-
7- ol;
4- chloro-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol; 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
3-ethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
3-(1 H-imidazol-1-yl)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7- ol;
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(1-methyl-1 H-imidazol-4- yl)quinolin-7-ol;
3- isopropyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3,7-diol;
7- hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3- carbonitrile;
6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
4- (dimethylamino)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol;
3- chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
4- methoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7- ol;
6- (3-(benzyloxy)isoquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine; 8-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
7- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline-1 ,6-diol;
7-(6-(methyl(1 ,2,2,6,6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
1-cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol; 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
6-(1 -(benzyloxy)isoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine; 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol; 2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)am H- pyrazol-4-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)^
4-ethoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
and
4-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol.
In a twentysecond embodiment, the invention is a compound, or salt thereof, selected from the group consisting of:
7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
2- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol;
3- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol;
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin-7-ol;
7- hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2- carbonitrile; and
1-amino-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol.
In a twentythird embodiment, the inventiuon is a pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of the first through twentythird embodiments, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.
In a twentyfourth embodiment, the invention is a combination comprising a therapeutically effective amount of a compound according to any one of the first through twentysecond
embodiments, or a pharmaceutically acceptable salt thereof and one or more therapeutically active co-agents.
In a twentyfifth embodiment, the invention is a method to treat, prevent or ameliorate an
SMN-deficiency-related condition, comprising administering to a subject in need thereof an effective amount of a compound or salt thereof of any one of the first through twentysecond embodiments.
In a twentysixth embodiment, the invention is the method of the twentyfifth embodiment, wherein said SMN-deficiency-related condition is Spinal Muscular Atrophy.
In a twentyseventh embodiment, the invention is a compound according to any one of the first through twentysecond embodiments, or a pharmaceutically acceptable salt thereof, for use as a medicament.
In a twentyeighth embodiment, the invention is a compound according to any one of the first through twentysecond embodiments, or a pharmaceutically acceptable salt thereof, for use in the treatment of an SMN-deficiency-related condition. In a twentyninth embodiment, the invention is the compound according to the twentyeighth embodiment, or pharmaceutically acceptable salt thereof, for use in the treatment of spinal muscular atrophy.
In a thirtieth embodiment, the invention is the use of a compound according to any one of the first through twentysecond embodiments, or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of spinal muscular atrophy.
For purposes of interpreting this specification, the following definitions will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa.
As used herein, the term "SMN modulator" includes agents, such as the compounds of the invention, which possess the ability to modulate, e.g., increase, SMN protein levels by at least one of multiple possible mechanisms. A non-limiting set of mechanisms includes SMN promoter activation, splicing modulation (e.g., preventing exon7 from being spliced out of the SMN gene), and SMN protein stability modulation. SMN modulators can modulate, e.g., increase FL-SMN and/or SMNA7 levels via any of said mechanisms, and/or can prevent SMNA7 from being degraded.
As used herein, the term "compounds of the invention" include but are not limited to the compounds of formula (I) and the compounds of formula (l-A)
As used herein, the term "SMN-deficiency-related conditions" includes but is not limited to Spinal Muscular Atrophy (SMA), neurogenic-type arthrogryposis multiplex congenita (congenital AMC), and amyotrophic lateral sclerosis (ALS).
As used herein, the term "Spinal Muscular Atrophy", "SMA," include three forms of childhood-onset SMA: Type I (Werdnig-Hoffmann disease); Type II (intermediate, chronic form), Type III (Kugelberg-Welander disease, or Juvenile Spinal Muscular Atrophy); Adult-onset type IV; as well as other forms of SMA, including X-linked disease, spinal muscular atrophy with respiratory distress (SMARD), spinal and bulbar muscular atrophy (Kennedy's disease, or Bulbo-Spinal Muscular Atrophy), and distal spinal muscular atrophy.
For purposes of interpreting this specification, the following definitions will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa.
As used herein, the term "Ci-i0alkyl" refers to a fully saturated branched or unbranched hydrocarbon moiety having 1 to 10 carbon atoms. The terms "Ci-6alkyl" and "Ci-4alkyl" are to be construed accordingly. Representative examples of Ci-i0alkyl include, but are not limited to, methyl, ethyl, n-propyl, /'so-propyl, n-butyl, sec-butyl, /'so-butyl, ferf-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2- dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl and n-decyl.
As used herein, the term "Ci-i0alkylene" refers to divalent alkyl group as defined herein above having 1 to 10 carbon atoms. The terms "Ci-6alkylene" and "Ci-4alkylene" are to be construed accordingly. Representative examples of Ci-i0alkylene include, but are not limited to, methylene, ethylene, n-propylene, /'so-propylene, n-butylene, sec-butylene, /'so-butylene, tert- butylene, n-pentylene, isopentylene, neopentylene, n-hexylene, 3-methylhexylene, 2,2- dimethylpentylene, 2,3-dimethylpentylene, n-heptylene, n-octylene, n-nonylene and n-decylene.
As used herein, the term "haloC1-10alkyl" refers to a C1-10alkyl group as defined herein, wherein at least one of the hydrogen atoms is replaced by a halo atom. The haloC1-10alkyl group can be monohaloC1-10alkyl, dihaloC1-10alkyl or polyhaloC1-10alkyl including perhaloC1-10alkyl. A monohaloCi-i0alkyl can have one iodo, bromo, chloro or fluoro within the alkyl group. DihaloC-i. -loalkyl and polyhaloCi-i0alkyl groups can have two or more of the same halo atoms or a
combination of different halo groups within the alkyl. Typically the polyhaloCi-i0alkyl group contains up to 12, or 10, or 8, or 6, or 4, or 3, or 2 halo groups. Non-limiting examples of haloCi-i0alkyl include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl and dichloropropyl. A perhaloCi-i0alkyl group refers to an Ci-i0alkyl group having all hydrogen atoms replaced with halo atoms.
The term "aryl" refers to an aromatic hydrocarbon group having 6-20 carbon atoms in the ring portion. Typically, aryl is monocyclic, bicyclic or tricyclic aryl having 6-20 carbon atoms and includes one or more aromatic rings fused to one or more non-aromatic hydrocarbon rings. Non- limiting examples include phenyl, naphthyl or tetrahydronaphthyl.
As used herein, the term "C1-10alkoxy" refers to Ci-i0alkyl-O-, wherein Ci-i0alkyl is defined herein above. Representative examples of Ci-i0alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, ferf-butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy- and decyloxy-.
As used herein, the term "heterocyclyl" or "heterocyclo" refers to a saturated or unsaturated non-aromatic ring or ring system, which is a 4-, 5-, 6-, or 7-membered monocyclic ring containing 1 , 2 or 3 heteroatoms selected from O, S and N, a 7-, 8-, 9-, 10-, 1 1 -, or 12-membered bicyclic ring system containing 1 , 2, 3, 4 or 5 heteroatoms selected from O, S and N, or a 10-, 1 1-, 12-, 13-, 14- or 15-membered tricyclic ring system and containing 1 , 2, 3, 4, 5, 6 or 7 heteroatoms selected from O, S and N, where the N and S can also optionally be oxidized to various oxidation states. The heterocyclic group can be attached via a heteroatom or a carbon atom. The heterocyclyl can include fused or bridged rings as well as spirocyclic rings. Examples of heterocycles include tetrahydrofuran (THF), dihydrofuran, 1 , 4-dioxane, morpholine, 1 ,4-dithiane, piperazine, piperidine, 1 ,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1 ,3-dioxane, 1 ,3-dithiane, oxathiane and thiomorpholine.
As used herein, the term "C3-i2cycloalkyl" refers to saturated or unsaturated monocyclic, bicyclic or tricyclic hydrocarbon groups of 3-12 carbon atoms. The term "C3-18cycloalkyl" refers to a fully saturated or unsaturated monocyclic hydrocarbon group of 3-8 carbon atoms. Exemplary monocyclic hydrocarbon groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl and cyclohexenyl. Exemplary bicyclic hydrocarbon groups include bornyl, indyl, hexahydroindyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1 .1 ]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1 .1 ]heptyl, 2,6,6- trimethylbicyclo[3.1.1 ]heptyl, bicyclo[2.2.2]octyl. Exemplary tricyclic hydrocarbon groups include, for example, adamantyl.
As used herein the term "C3 12cycloalklyoxy" refers to C3-i2cycloalkyl-0-, wherein C3- i2cycloalkyl is defined herein above. Representative examples of C3.i2cycloalklyoxy include , but are not limited to monocyclic groups such as cyclopropoxy, cyclobutoxy, cyclopentyloxy,
cyclopentenyloxy, cyclohexyloxy and cyclohexenyloxy and the like. Exemplary bicyclic
hydrocarbon groups include bornyloxy, indyloxy, hexahydroindyloxy, tetrahydronaphthyloxy, decahydronaphthyloxy, bicyclo[2.1 .1]hexyloxy, bicyclo[2.2.1]heptyloxy, bicyclo[2.2.1]heptenyloxy, 6,6-dimethylbicyclo[3.1 .1 ]heptyloxy, 2,6,6-trimethylbicyclo[3.1.1 ]heptyloxy, bicyclo[2.2.2]octyloxy and the like. Exemplary tricyclic hydrocarbon groups include, for example, adamantyloxy.
As used herein, the term "aryloxy" refers to both an— O-aryl and an— O-heteroaryl group, wherein aryl and heteroaryl are defined herein.
As used herein, the term "heteroaryl" refers to a 5-, 6-, or 7-membered monocyclic aromatic ring containing 1 , 2, 3 or 4 heteroatoms selected from O, S and N, an 8-, 9-, or 10-membered fused bicyclic ring system containing 1 , 2, 3, 4 or 5 heteroatoms selected from O, S and N, or an 1 1-, 12-, 13-, or 14-membered fused tricyclic ring system containing 1 , 2, 3, 4, 5 or 6 heteroatoms selected from O, S and N, wherein at least one of the rings of the bicyclic or tricyclic ring systems is fully aromatic. Typical heteroaryl groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or 5- imidazolyl, 3-, 4-, or 5- pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or 5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or 5-1 ,2,4-triazolyl, 4- or 5-1 ,2, 3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3- or 4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, 2-, 4-, or 5-pyrimidinyl, 1-, 2-, 3-, 5-, 6-, 7-, or 8- indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7- indazolyl, 2-, 4-, 5-, 6-, 7-, or 8- purinyl, 1 -, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinoliyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 1 -, 4-, 5-, 6-, 7-, or 8-phthalazinyl, 2-, 3-, 4-, 5-, or 6-naphthyridinyl, 2-, 3- , 5-, 6-, 7-, or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7-, or 8-cinnolinyl, 2-, 4-, 6-, or
7- pteridinyl, 1 -, 2-, 3-, 4-, 5-, 6-, 7-, or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-carbzaolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl, 1 -, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1- , 2-, 3-, 4-, 5- , 6-, 7-, 8-, or 9-acridinyl, 1 -, 2-, 4-, 5-, 6-, 7-, 8-, or 9-perimidinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9-, or 10- phenathrolinyl, 1 -, 2- , 3-, 4-, 6-, 7-, 8-, or 9-phenazinyl, 1 -, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10- phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenoxazinyl, 2-, 3-, 4-, 5-, 6-, or I-, 3-, 4-, 5-, 6-, 7-,
8- , 9-, or 10- benzisoqinolinyl, 2-, 3-, 4-, or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10 -, or 1 1 - 7H-pyrazino[2,3-c]carbazolyl,2-, 3-, 5-, 6-, or 7-2H- furo[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7-, or 8-5H- pyrido[2,3-d]-o-oxazinyl, 1 -, 3-, or 5-1 H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d] thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or 6- imidazo[2, 1 -b] thiazolyl, 1 -, 3-, 6-, 7-, 8-, or 9-furo[3,4-c]cinnolinyl, 1 -, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10, or 1 1 -4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6-, or 7-imidazo[1 ,2-b][1 ,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5- , 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6-, or 7-benzothiazolyl, 1 -, 2-, 4-, 5-, 6-, 7-, 8-, or 9- benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1 -, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 1 1 -1 H- pyrrolo[1 ,2-b][2]benzazapinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1 -, 3-, 4-, 5-, 6-, 7-, or 8- isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5- , 6-, or 7- benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl.
As used herein, the term "halogen" or "halo" refers to fluoro, chloro, bromo, and iodo.
As used herein, the term "isomers" refers to different compounds that have the same molecular formula but differ in arrangement and configuration of the atoms. Also as used herein, the term "an optical isomer" or "a stereoisomer" refers to any of the various stereo isomeric configurations which may exist for a given compound of the present invention and includes geometric isomers. It is understood that a substituent may be attached at a chiral center of a carbon atom. Therefore, the invention includes enantiomers, diastereomers or racemates of the compound. "Enantiomers" are a pair of stereoisomers that are non- superimposable mirror images of each other. A 1 : 1 mixture of a pair of enantiomers is a "racemic" mixture. The term is used to designate a racemic mixture where appropriate. "Diastereoisomers" are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other. The absolute stereochemistry is specified according to the Cahn- Ingold- Prelog R-S system. When a compound is a pure enantiomer the stereochemistry at each chiral carbon may be specified by either R or S. Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) depending on the direction (dextro- or levorotatory) which they rotate plane polarized light at the wavelength of the sodium D line. Certain of the compounds described herein contain one or more asymmetric centers or axes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-. The present invention is meant to include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R)- and (S)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be E or Z configuration. If the compound contains a disubstituted cycloalkyi, the cycloalkyi substituent may have a cis- or trans-configuration. All tautomeric forms are also intended to be included.
As used herein, the terms "salt" or "salts" refers to an acid addition or base addition salt of a compound of the invention. "Salts" include in particular "pharmaceutical acceptable salts." The term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of this invention and, which typically are not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid and/or base salts by virtue of the presence of amino and/or carboxyl groups or groups similar thereto.
Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids, e.g., acetate, aspartate, benzoate, besylate, bromide/hydrobromide,
bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonate, chloride/hydrochloride,
chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate, laurylsulfate, malate, maleate, malonate, mandelate, mesylate, methylsulphate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, subsalicylate, tartrate, tosylate and trifluoroacetate salts.
Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.
Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I to XII of the periodic table. In certain embodiments, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper;
particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.
Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and
tromethamine.
The pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound, a basic or acidic moiety, by conventional chemical methods. Generally, such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate or the like), or by reacting free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two. Generally, use of non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is desirable, where practicable. Lists of additional suitable salts can be found, e.g., in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).
Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18F 31P, 32P, 35S, 36CI, 125l respectively. The invention includes various isotopically labeled compounds as defined herein, for example those into which radioactive isotopes, such as 3H, 13C, and 14C , are present. Such isotopically labelled compounds are useful in metabolic studies (with 14C), reaction kinetic studies (with, for example 2H or 3H), detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients. In particular, an 18F or labeled compound may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.
Further, substitution with heavier isotopes, particularly deuterium (i.e., 2H or D) may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements or an improvement in therapeutic index. It is understood that deuterium in this context is regarded as a substituent of a compound of the formula (I). The concentration of such a heavier isotope, specifically deuterium, may be defined by the isotopic enrichment factor. The term "isotopic enrichment factor" as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope. If a substituent in a compound of this invention is denoted deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). Isotopically-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations using an appropriate isotopically-labeled reagents in place of the non-labeled reagent previously employed.
Pharmaceutically acceptable solvates in accordance with the invention include those wherein the solvent of crystallization may be isotopically substituted, e.g. D20, d6-acetone, d6- DMSO.
Compounds of the invention, i.e. compounds of formula (I) that contain groups capable of acting as donors and/or acceptors for hydrogen bonds may be capable of forming co-crystals with suitable co-crystal formers. These co-crystals may be prepared from compounds of formula (I) by known co-crystal forming procedures. Such procedures include grinding, heating, co-subliming, co- melting, or contacting in solution compounds of formula (I) with the co-crystal former under crystallization conditions and isolating co-crystals thereby formed. Suitable co-crystal formers include those described in WO 2004/078163. Hence the invention further provides co-crystals comprising a compound of formula (I).
The term "a therapeutically effective amount" of a compound of the present invention refers to an amount of the compound of the present invention that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc. In one non-limiting embodiment, the term "a therapeutically effective amount" refers to the amount of the compound of the present invention that, when administered to a subject, is effective to (1 ) at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease (i) mediated by Survival of Motor Neuron (SMN) gene or gene product, or by SMNA7 degradation, or by the relative levels of FL-SMN and SMNA7 (ii) associated with SMN activity, or (iii) characterized by activity (normal or abnormal) of SMN; or (2) reducing or inhibiting the activity of SMN; or (3) reducing or inhibiting the expression of SMN 1 or SMN2.
In another non-limiting embodiment, the term "a therapeutically effective amount" refers to the amount of the compound of the present invention that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reducing or inhibiting the activity of SMN; or at least partially reducing or inhibiting the expression of SMN, in both cases by modulating the relative levels of FL-SMN and SMNA7.
The phrases "therapeutically effective amount" and "effective amount" are used herein to mean an amount sufficient to reduce by at least about 15 percent, preferably by at least 50 percent, more preferably by at least 90 percent, and most preferably prevent, a clinically significant deficit in the activity, function and response of the host. Alternatively, a therapeutically effective amount is sufficient to cause an improvement in a clinically significant condition/symptom in the host. The effective amount can vary depending on such factors as the size and weight of the subject, the type of illness, or the particular compound of the invention. For example, the choice of the compound of the invention can affect what constitutes an "effective amount." One of ordinary skill in the art would be able to study the factors contained herein and make the determination regarding the effective amount of the compounds of the invention without undue experimentation.
The regimen of administration can affect what constitutes an effective amount. The compound of the invention can be administered to the subject either prior to or after the onset of an SMN-deficiency-related condition. Further, several divided dosages, as well as staggered dosages, can be administered daily or sequentially, or the dose can be continuously infused, or can be a bolus injection. Further, the dosages of the compound(s) of the invention can be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.
As used herein, the term "subject" refers to an animal. Typically the animal is a mammal. A subject also refers to for example, primates (e.g., humans, male or female), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds and the like. In certain embodiments, the subject is a primate. In yet other embodiments, the subject is a human.
As used herein, the term "inhibit", "inhibition" or "inhibiting" refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.
As used herein, the term "treat", "treating" or "treatment" of any disease or disorder refers in one embodiment, to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another
embodiment "treat," "treating," or "treatment" refers to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the patient. In yet another embodiment, "treat", "treating" or "treatment" refers to modulating the disease or disorder, either physically (e.g., through stabilization of a discernible symptom), physiologically, (e.g., through stabilization of a physical parameter), or both. In yet another embodiment, "treat," "treating," or "treatment" refers to preventing or delaying the onset or development or progression of the disease or disorder.
As used herein, a subject is "in need of" a treatment if such subject would benefit biologically, medically or in quality of life from such treatment.
As used herein, the term "a," "an," "the" and similar terms used in the context of the present invention (especially in the context of the claims) are to be construed to cover both the singular and plural unless otherwise indicated herein or clearly contradicted by the context.
All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. "such as") provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed.
Any asymmetric atom (e.g., carbon or the like) of the compound(s) of the present invention can be present in racemic or enantiomerically enriched, for example the (/?)-, (S)- or {R,S)- configuration. In certain embodiments, each asymmetric atom has at least 50 % enantiomeric excess, at least 60 % enantiomeric excess, at least 70 % enantiomeric excess, at least 80 % enantiomeric excess, at least 90 % enantiomeric excess, at least 95 % enantiomeric excess, or at least 99 % enantiomeric excess in the (R)- or (S)- configuration. Substituents at atoms with unsaturated bonds may, if possible, be present in cis- (Z)- or trans- (E)- form.
Accordingly, as used herein a compound of the present invention can be in the form of one of the possible isomers, rotamers, atropisomers, tautomers or mixtures thereof, for example, as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (antipodes), racemates or mixtures thereof.
Any resulting mixtures of isomers can be separated on the basis of the physicochemical differences of the constituents, into the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and/or fractional crystallization.
Any resulting racemates of final products or intermediates can be resolved into the optical antipodes by known methods, e.g., by separation of the diastereomeric salts thereof, obtained with an optically active acid or base, and liberating the optically active acidic or basic compound. In particular, a basic moiety may thus be employed to resolve the compounds of the present invention into their optical antipodes, e.g., by fractional crystallization of a salt formed with an optically active acid, e.g., tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-0,0'-p-toluoyl tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. Racemic products can also be resolved by chiral chromatography, e.g., high pressure liquid chromatography (HPLC) using a chiral adsorbent.
Compounds of the present invention are either obtained in the free form, as a salt thereof, or as prodrug derivatives thereof.
When both a basic group and an acid group are present in the same molecule, the compounds of the present invention may also form internal salts, e.g., zwitterionic molecules.
Furthermore, the compounds of the present invention, including their salts, can also be obtained in the form of their hydrates, or include other solvents used for their crystallization. The compounds of the present invention may inherently or by design form solvates with
pharmaceutically acceptable solvents (including water); therefore, it is intended that the invention embrace both solvated and unsolvated forms. The term "solvate" refers to a molecular complex of a compound of the present invention (including pharmaceutically acceptable salts thereof) with one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical art, which are known to be innocuous to the recipient, e.g., water, ethanol, and the like. The term "hydrate" refers to the complex where the solvent molecule is water.
The compounds of the present invention, including salts, hydrates and solvates thereof, may inherently or by design form polymorphs.
The invention further includes any variant of the present processes, in which an
intermediate product obtainable at any stage thereof is used as starting material and the remaining steps are carried out, or in which the starting materials are formed in situ under the reaction conditions, or in which the reaction components are used in the form of their salts or optically pure material.
Compounds of the invention and intermediates can also be converted into each other according to methods generally known to those skilled in the art.
In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition can be formulated for particular routes of administration such as oral administration, parenteral administration, and rectal administration, etc. In addition, the pharmaceutical compositions of the present invention can be made up in a solid form (including without limitation capsules, tablets, pills, granules, powders or suppositories), or in a liquid form (including without limitation solutions, suspensions or emulsions). The pharmaceutical compositions can be subjected to conventional pharmaceutical operations such as sterilization and/or can contain conventional inert diluents, lubricating agents, or buffering agents, as well as adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers and buffers, etc.
Typically, the pharmaceutical compositions are tablets or gelatin capsules comprising the active ingredient together with
diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine;
lubricants, e.g., silica, talcum, stearic acid, its magnesium or calcium salt and/or polyethyleneglycol; for tablets also
binders, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth,
methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; if desired
disintegrants, e.g., starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and/or
absorbents, colorants, flavors and sweeteners.
Tablets may be either film coated or enteric coated according to methods known in the art. Suitable compositions for oral administration include an effective amount of a compound of the invention in the form of tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use are prepared according to any method known in the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets may contain the active ingredient in admixture with nontoxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients are, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be employed.
Formulations for oral use can be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil.
Certain injectable compositions are aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fatty emulsions or suspensions. Said
compositions may be sterilized and/or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances. Said compositions are prepared according to conventional mixing, granulating or coating methods, respectively, and contain about 0.1-75%, or contain about 1-50%, of the active ingredient.
Suitable compositions for transdermal application include an effective amount of a compound of the invention with a suitable carrier. Carriers suitable for transdermal delivery include absorbable pharmacologically acceptable solvents to assist passage through the skin of the host. For example, transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound of the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin.
Suitable compositions for topical application, e.g., to the skin and eyes, include aqueous solutions, suspensions, ointments, creams, gels or sprayable formulations, e.g., for delivery by aerosol or the like. Such topical delivery systems will in particular be appropriate for dermal application, e.g., for the treatment of skin cancer, e.g., for prophylactic use in sun creams, lotions, sprays and the like. They are thus particularly suited for use in topical, including cosmetic, formulations well-known in the art. Such may contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.
As used herein a topical application may also pertain to an inhalation or to an intranasal application. They may be conveniently delivered in the form of a dry powder (either alone, as a mixture, for example a dry blend with lactose, or a mixed component particle, for example with phospholipids) from a dry powder inhaler or an aerosol spray presentation from a pressurized container, pump, spray, atomizer or nebulizer, with or without the use of a suitable propellant.
The present invention further provides anhydrous pharmaceutical compositions and dosage forms comprising the compounds of the present invention as active ingredients, since water may facilitate the degradation of certain compounds.
Anhydrous pharmaceutical compositions and dosage forms of the invention can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions. An anhydrous pharmaceutical composition may be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions are packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastics, unit dose containers (e. g., vials), blister packs, and strip packs.
The invention further provides pharmaceutical compositions and dosage forms that comprise one or more agents that reduce the rate by which the compound of the present invention as an active ingredient will decompose. Such agents, which are referred to herein as "stabilizers," include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers, etc.
The compounds of formula I in free form or in salt form, exhibit valuable pharmacological properties, e.g. full length SMN protein production modulating properties, e.g. as indicated in in vitro and in vivo tests as provided in the next sections, and are therefore indicated for therapy or for use as research chemicals, e.g. as tool compounds.
Thus, as a further embodiment, the present invention provides the use of a compound of formula (I) or a salt thereof in therapy. In a further embodiment, the therapy is selected from a disease which may be treated by modulating full length SMN protein production. In another embodiment, the disease is selected from the afore-mentioned list, suitably spinal muscular atrophy.
In another embodiment, the invention provides a method of treating a disease which is treated by modulating full length SMN protein production comprising administration of a
therapeutically acceptable amount of a compound of formula (I) or salt thereof to a patient in need of such therapy. In a further embodiment, the disease is selected from the afore-mentioned list, suitably spinal muscular atrophy. Thus, as a further embodiment, the present invention provides the use of a compound of formula (I) or salt thereof for the manufacture of a medicament. In a further embodiment, the medicament is for treatment of a disease which may be treated by modulation of SMN protein production. In another embodiment, the disease is selected from the afore-mentioned list, suitably spinal muscular atrophy.
The pharmaceutical composition or combination of the present invention can be in unit dosage of about 0.01-1000 mg of active ingredient(s) for a subject of about .05-70 kg or about 1 -20 kg, or about 1 -500 mg or about 1 -250 mg or about 1 -150 mg or about 0.5-100 mg, or about 0.01-1 mg or about 0.01-0.1 mg or about 1-50 mg of active ingredients. The therapeutically effective dosage of a compound, the pharmaceutical composition, or the combinations thereof, is dependent on the species of the subject, the body weight, age and individual condition, the disorder or disease or the severity thereof being treated. A physician, clinician or veterinarian of ordinary skill can readily determine the effective amount of each of the active ingredients necessary to prevent, treat or inhibit the progress of the disorder or disease.
The above-cited dosage properties are demonstrable in vitro and in vivo tests using advantageously mammals, e.g., mice, rats, dogs, monkeys or isolated organs, tissues and preparations thereof. The compounds of the present invention can be applied in vitro in the form of solutions, e.g., aqueous solutions, and in vivo either enterally, parenterally, advantageously intravenously, e.g., as a suspension or in aqueous solution. The dosage in vitro may range between about 10"3 molar and 10"9 molar concentrations. A therapeutically effective amount in vivo may range depending on the route of administration, between about 0.1-500 mg/kg, or between about 1-100 mg/kg.
The compound of the present invention may be administered either simultaneously with, or before or after, one or more other therapeutic agent. The compound of the present invention may be administered separately, by the same or different route of administration, or together in the same pharmaceutical composition as the other agents.
In one embodiment, the invention provides a product comprising a compound of formula (I) and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is the treatment of a spinal muscular atrophy. Products provided as a combined preparation include a composition comprising the compound of formula (I) and the other therapeutic agent(s) together in the same pharmaceutical composition, or the compound of formula (I) and the other therapeutic agent(s) in separate form, e.g. in the form of a kit.
In one embodiment, the invention provides a pharmaceutical composition comprising a compound of formula (I) and another therapeutic agent(s). Optionally, the pharmaceutical composition may comprise a pharmaceutically acceptable carrier, as described above. In one embodiment, the invention provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of formula (I). In one embodiment, the kit comprises means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet. An example of such a kit is a blister pack, as typically used for the packaging of tablets, capsules and the like.
The kit of the invention may be used for administering different dosage forms, for example, oral and parenteral, for administering the separate compositions at different dosage intervals, or for titrating the separate compositions against one another. To assist compliance, the kit of the invention typically comprises directions for administration.
In the combination therapies of the invention, the compound of the invention and the other therapeutic agent may be manufactured and/or formulated by the same or different manufacturers. Moreover, the compound of the invention and the other therapeutic may be brought together into a combination therapy: (i) prior to release of the combination product to physicians (e.g. in the case of a kit comprising the compound of the invention and the other therapeutic agent); (ii) by the physician themselves (or under the guidance of the physician) shortly before administration; (iii) in the patient themselves, e.g. during sequential administration of the compound of the invention and the other therapeutic agent.
The following examples are intended to illustrate the invention and are not to be construed as being limitations thereon. Temperatures are given in degrees Celsius. If not mentioned otherwise, all evaporations are performed under reduced pressure, typically between about 15 mm Hg and 100 mm Hg (= 20-133 mbar). The structure of final products, intermediates and starting materials is confirmed by standard analytical methods, e.g., microanalysis and spectroscopic characteristics, e.g., MS, IR, NMR. Abbreviations used are those conventional in the art.
All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts utilized to synthesis the compounds of the present invention are either commercially available or can be produced by organic synthesis methods known to one of ordinary skill in the art (Houben-Weyl 4th Ed. 1952, Methods of Organic Synthesis, Thieme, Volume 21 ). Further, the compounds of the present invention can be produced by organic synthesis methods known to one of ordinary skill in the art as shown in the following examples.
Preparations of Compounds
It is understood that in the following description, combinations of substituents and/or variables of the depicted formulae are permissible only if such contributions result in stable compounds.
It will also be appreciated by those skilled in the art that in the processes described below the functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxy, phenol, amino and carboxylic acid. Suitable protecting groups for hydroxy or phenol include trialkylsilyl or diarylalkylsilyl (e.g., t- butyldimethylsilyl, f-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, substituted benzyl, methyl, and the like. Suitable protecting groups for amino, amidino and guanidino include t- butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl or arylalkyl esters.
Protecting groups may be added or removed in accordance with standard techniques, which are well-known to those skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. The protecting group may also be a polymer resin, such as a Wang resin or a 2-chlorotrityl-chloride resin.
It will also be appreciated by those skilled in the art, although such protected derivatives of compounds of this invention may not possess pharmacological activity as such, they may be administered to a subject and thereafter metabolized in the body to form compounds of the invention which are pharmacologically active. Such derivatives may therefore be described as "prodrugs". All prodrugs of compounds of this invention are included within the scope of the invention.
The following reaction schemes illustrate methods to make compounds of this invention. It is understood that one skilled in the art would be able to make these compounds by similar methods or by methods known to one skilled in the art. In general, starting components and reagents may be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, Strem, other commercial vendors, or synthesized according to sources known to those skilled in the art, or prepared as described in this invention. A, B, X, R, R1, R2, R3, R4, are defined as in the Specification unless specifically defined.
In general, pyridazine compounds of Formula (I) of this invention can be synthesized following the general procedure described in Scheme 1.
Figure imgf000035_0001
(I)
General Scheme 1
Figure imgf000036_0001
Formula I
The starting materials for the above reaction scheme are commercially available or can be prepared according to methods known to one skilled in the art or by methods disclosed herein. In general, the compounds of the invention are prepared in the above reaction Scheme 1 as follows:
Di-halopyridazine (1) reacts in a displacement reaction or a metal-mediated cross coupling reaction (Buchwald) with an alcohol or an amine (B) to provide pyridazine intermediate (2).
Transition metal-mediated cross coupling reaction, such as a Suzuki reaction, between halide compound (2) and a substituted aryl or heteroaryl compound A, such as a boronate acid or boronate ester, provides compound (3) of Formula (I) of the invention.
In a complementary manner, compounds of Formula (I) can be synthesized following the general procedure described in Scheme 2.
General Scheme 2
Figure imgf000036_0002
ormu a
The starting materials for the above reaction scheme are commercially available or can be prepared according to methods known to one skilled in the art or by methods disclosed herein. In general, the compounds of the invention are prepared in the above reaction Scheme 2 as follows:
Di-halopyridazine (1) reacts in a transition metal-mediated cross coupling reaction, such as a Suzuki reaction, with a substituted aryl or heteroaryl compound A, such as a boronate acid or ester, to provide pyridazine intermediate (4). Pyridazine intermediate (4) reacts via a displacement reaction with an alcohol or an amine (B) to provide pyridazine (3) of Formula (I) of the invention. Compounds of Formula (I) can also be prepared following the general procedure described in Scheme 3.
General Scheme 3
Figure imgf000037_0001
ormu a
The starting materials for the above reaction scheme are commercially available or can be prepared according to methods known to one skilled in the art or by methods disclosed herein. In general, the compounds of the invention are prepared in the above reaction Scheme 3 as follows:
Di-halopyridazine (1) reacts in a transition metal-mediated cross coupling reaction, such as a Suzuki reaction, with a substituted aryl or heteroaryl compound A, such as a boronate acid or ester, to provide pyridazine intermediate (4). Pyridazine intermediate (4) reacts via second metal- mediated cross coupling, such as a Suzuki reaction, to provide pyridazine (3) of Formula (I) of the invention.
General Schemes 1 , 2 and 3 can be followed for a variety of aromatic A groups such as substituted phenols, naphthyls, heteroaryls, and the like, and for a variety of amine or alcohol B groups such as substituted aminopiperdines, piperazines, homopiperazines, 4-hydroxy piperidines, and the like, to provide compounds of Formula (I) of the invention. Routine protecting group strategies may be required to achieve final compounds of Formula (I).
All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, catalysts and scavengers utilized to synthesize the compounds of the present invention are either commercially available or can be produced by organic synthesis methods known to one of ordinary skill in the art. Further, the compounds of the present invention can be produced by organic synthesis methods known to one of ordinary skill in the art as shown in the following examples.
The following examples are intended to illustrate the invention and are not to be construed as being limitations thereon. Temperatures are given in degrees centigrade. If not mentioned otherwise, all evaporations are performed under reduced pressure, preferably between about 15 mm Hg and 100 mm Hg (= 20-133 mbar). The structure of final products, intermediates and starting materials is confirmed by standard analytical methods, e.g., microanalysis and spectroscopic characteristics, e.g., LCMS, NMR, CHN. Abbreviations used are those conventional in the art, a list of which is provided at the end of the experimental section.
PREPARATION 1 Intermediate 1 : Synthesis of 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine:
Figure imgf000038_0001
A solution of 3,6-dichloropyridazine (4.00 g, 26.8 mmol) and N, 2,2,6, 6-pentamethylpiperidin- 4-amine (7.32 g, 43.0 mmol) in butan-1 -ol (67 mL) was heated at 120 °C for 72 h. The solvent was removed via rotary evaporation and the residue was partitioned between water and DCM, then the water layer was further extracted with DCM. The combined organic layers were washed with water, brine, dried over MgS04, and concentrated in vacuo. The black crude material was stirred in small amount of EtOAc overnight, and the resulting off-white solid was collected to provide 6-chloro-N- methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 1 (4.18 g). MS (M+1 ) = 283.5. 1H NMR (400 MHz, METHANOL-^) δ 7.40 (d, J=9.60 Hz, 1 H), 7.14 (d, J=9.60 Hz, 1 H), 4.96-5.13 (m, 1 H), 2.93 (s, 3H), 1 .59-1.68 (m, 2H), 1.51 (t, J=12.38 Hz, 2H), 1 .20 (s, 6H), 1 .33 (s, 6H).
PREPARATION 2
Intermediate 2: Synthesis of 6-chloro-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine
Figure imgf000038_0002
A mixture of 3,6-dichloropyridazine (6.26 g, 42.0 mmol) and 2,2,6,6-tetramethylpiperidin-4- amine (14.7 mL, 84 mmol) was stirred at 120 °C for 1 h, neat. To this mixture was added butan-1 -ol (40 mL), and the reaction was stirred at 120 °C for 1 h. The mixture was cooled to room
temperature and partitioned between water and DCM. The organic layer was dried over MgS04, filtered, and concentrated. The crude material was recrystallized from acetonitrile to provide 6- chloro-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 2 (7.3 g) as an off- white solid. MS (M+1 ) = 269.2. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.08 (d, J=9.3 Hz, 1 H), 6.54 (d, J=9.3 Hz, 1 H), 4.53 (d, J=7.6 Hz, 1 H), 4.05-4.26 (m, 1 H), 1 .98 (dd, J=12.6, 3.8 Hz, 2H), 1.22 (s, 6H), 1 .08 (s, 6H), 0.93 (apparent t, J=12.1 Hz, 2H).
PREPARATION 3
Intermediate 3: Synthesis of 3-chloro-6-(2,2,6,6-tetramethylpiperidin-4- yloxy)pyridazine
Figure imgf000039_0001
To a solution of 2,2,6, 6-tetramethylpiperidin-4-ol (106 mg, 0.67 mmol) in DMF (6.7 mL) was added NaH (60 wt %, 35 mg, 0.87 mmol). The solution was stirred at room temperature for 30 min, then 3,6-dichloropyridazine (100 mg, 0.67 mmol) was added and the reaction was stirred for 1 h. The crude reaction mixture was diluted with EtOAc. The organic layer was washed with water (5x), brine, dried over Na2S04, filtered and concentrated under reduced pressure to provide 3-chloro-6- (2,2,6, 6-tetramethylpiperidin-4-yloxy)pyridazine, Intermediate 3 (135 mg). MS (M+1 ) = 270.2. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.53 (s, 1 H), 7.37 (d, J=9.1 Hz, 1 H), 6.91 (d, J=9.1 Hz, 1 H), 5.68-5.78 (m, 1 H), 2.20 (dd, J=12.4, 4.0 Hz, 2H), 1 .32 (s, 6H), 1 .27-1.29 (m, 2H), 1.20 (s, 6H). PREPARATION 4
Intermediate 4: Synthesis of 6-chloro-N-methyl-N-(1 , 2,2,6, 6-pentamethylpiperidin-4- yl)pyridazin-3-amine
Figure imgf000039_0002
To a suspension of 6-chloro-N-(1 ,2,2,6,6-pentamethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 2 (4.0 g, 14.1 mmol) in DMF (140 mL) cooled to 0 °C was added NaH (60 wt %, 735 mg, 18.39 mmol) portion-wise. The reaction was warmed to room temperature and stirred for 60 min. After 60 minutes, methyl iodide (0.88 mL, 14.1 mmol) was added and the reaction was stirred an additional 3 h, then slowly quenched with water at room temperature. The mixture was diluted with EtOAc. The organic layer was washed with water (5x), brine, dried over MgS04, filtered and concentrated under reduced pressure to provide 6-chloro-N-methyl-N-(1 ,2,2,6,6- pentamethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 4 (3.98 g). MS (M+1 ) = 297.0. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.10 (d, J=9.6 Hz, 1 H), 6.69 (d, J=9.6 Hz, 1 H), 4.79-5.00 (m, 1 H), 2.86 (s, 3H), 2.22 (s, 3H), 1 .61 -1.73 (m, 2H), 1.48-1.57 (m, 2H), 1.14 (s, 6H), 1.10 (s, 6H).
PREPARATION 5
Intermediate 5: Synthesis of (3aR,6aS)-2-(6-chloropyridazin-3-yl)-5- methyloctahydropyrrolo[3,4-c]pyrrole
Figure imgf000040_0001
To a solution of 3,6-dichloropyridazine (0.596 g, 4.00 mmol) in butan-1 -ol (10 mL) was added DIPEA (1 .40 mL, 8.00 mmol) and 2-methyloctahydropyrrolo[3,4-c]pyrrole (0.505 g, 4.00 mmol). The solution was heated at 120 °C overnight. The reaction mixture was concentrated and the residue purified by silica gel chromatography (0-25% 2 N ammonia in MeOH gradient, in DCM) to provide (3aR,6aS)-2-(6-chloropyridazin-3-yl)-5-methyloctahydropyrrolo[3,4-c]pyrrole,
Intermediate 5 (0.67 g), as an off-white solid. MS (M+1 ) = 239.1. 1H NMR (400 MHz, METHANOL- d4) δ 7.47 (d, J=9.6 Hz, 1 H), 7.1 1 (d, J=9.6 Hz, 1 H), 3.58-3.70 (m, 4H), 3.47 (dd, J=10.9, 7.3 Hz, 2H), 3.3 (2H, partially obscured by methanol residual solvent peak), 3.15 (d, J=1 1 .1 Hz, 2H), 2.80 (s, 3H).
PREPARATION 6
Intermediate 6: Synthesis of (7-methoxyquinolin-6-yl)boronic acid
Figure imgf000040_0002
Step 1: 6-Bromo-7-methoxyquinoline
In a 100 mL round bottom flask, a solution of sulfuric acid (7.1 mL, 130 mmol) in water (6.5 mL) was treated with 3-nitrobenzenesulfonic acid (7.07 g, 34.8 mmol) and glycerol (8.7 mL, 120 mmol) to give a thick grey suspension. The suspension was heated to 1 10 °C and 4-bromo-3- methoxyaniline (6.7 g, 33 mmol) was added resulting in a slurry. Additional quantities of water (6 mL), glycerol (6 mL), sulfuric acid (6 mL) were added and the temperature increased to 140 °C. After about 3 h the mixture became a homogeneous dark brown solution. The solution was cooled to room temperature, poured onto ice, and the pH adjusted to 8 by addition of concentrated ammonium hydroxide. The mixture was extracted with 1 :1 EtOAc/diethyl ether (5x), dried over Mg2S04 and concentrated to a brown liquid. Silica gel chromatography (0-20% EtOAc in DCM) provided the title compound which was further purified by silica gel chromatography (0-20% MeOH in DCM) to provide 6-bromo-7-methoxyquinoline (3.94 g) as an off-white solid. MS (M+1 ) = 240.1 . 1H NMR (400 MHz, CHLOROFORM-d) δ 8.86 (dd, J=4.0, 1 .5 Hz, 1 H), 8.01 -8.12 (m, 2H), 7.53 (s, 1 H), 7.34 (dd, J=8.1 , 4.5 Hz, 1 H), 4.07 (s, 3H).
Step 2: (7-Methoxyquinolin-6-yl)boronic acid Butyllithium (1.6 M in heptane, 4.90 mL, 7.90 mmol) was added dropwise to a solution of 6- bromo-7-methoxyquinoline (1 .71 g, 7.18 mmol) cooled to -78 °C. The solution was stirred for 0.5 h after which time trimethyl borate (2.0 mL, 18 mmol) was added in a single portion. The reaction was allowed to warm to room temperature and stir overnight. The crude reaction was concentrated to dryness, then concentrated from heptane (2x). The resulting solids were passed through a plug of silica gel (80 mL, dry measure) eluting with 9:1 DCM/MeOH to provide (7-methoxyquinolin-6- yl)boronic acid, Intermediate 6, as an orange solid (778 mg). MS (M+1 ) = 204.1
PREPARATION 7
Intermediate 7: Synthesis of (6-methoxyisoquinolin-7-yl)boronic acid
Figure imgf000041_0001
Step 1. 7-Bromo-6-methoxyisoquinoline
7-Bromo-6-methoxyisoquinoline was prepared as described in WO2007000240. MS (M+1 ) = 240.3. 1 H NMR (400 MHz, CHLOROFORM-d) δ 9.08 (s, 1 H), 8.49 (d, J=5.8 Hz, 1 H), 8.20 (s, 1 H), 7.56 (d, J=5.8 Hz, 1 H), 7.1 1 (s, 1 H), 4.05 (s, 3H).
Step 2. (6-Methoxyisoquinolin-7-yl)boronic acid
To a 50 mL round bottom flask was added 7-bromo-6-methoxyisoquinoline (300 mg, 1 .26 mmol), bis(pinacolato)diboron (640 mg, 2.52 mmol), potassium acetate (742 mg, 7.56 mmol), and PdCI2(dppf).CH2CI2 (103 mg, 0.126 mmol). 1 ,4-Dioxane (0.2 M) was added and the flask was evacuated/purged with N2 (3x). The reaction mixture was stirred at 100 °C for 2 h. The reaction was complete after 3 h as determined by LC/MS analysis; only the boronic acid was observed (no pinacol boronate ester). The mixture was diluted with EtOAc, filtered through Celite® and concentrated to provide (6-methoxyisoquinolin-7-yl)boronic acid, Intermediate 7, which was used without further purification. MS (M+1 ) = 204.4
PREPARATION 8
Intermediate 8: Synthesis of (7-methoxyisoquinolin-6-yl)boronic acid
Figure imgf000041_0002
Step 1: (E)-N-(4-Bromo-3-methoxybenzylidene)-2, 2-dimethoxyethanamine
In a 100 mL round-bottom flask fitted with a Dean-Stark trap and reflux condenser, 4-bromo- 3-methoxybenzaldehyde (3.65 g, 17.0 mmol) and 2,2-dimethoxyethanamine (1.32 mL, 18.6 mmol) were refluxed in benzene (37 mL) overnight. The mixture was concentrated in vacuo to provide the title compound as an amber oil (3.65 g). 1H NMR (400 MHz, CHLOROFORM-d) δ 8.20-8.26 (m, 1 H), 7.58 (d, J=8.0 Hz, 1 H), 7.40 (d, J=1.5 Hz, 1 H), 7.10 (dd, J=1 .76, 8.0 Hz, 1 H), 4.69 (t, J=5.4 Hz, 1 H), 3.96 (s, 3H), 3.78 (dd, J=1 .3, 5.3 Hz, 2H), 3.43 (s, 6H).
Step 2: 6-Bromo-7-methoxyisoquinoline
A 100 mL round-bottom flask was charged with (E)-N-(4-bromo-3-methoxybenzylidene)-2,2- dimethoxyethanamine (3.65 g, 12.1 mmol) and THF (24 mL) to give a tan solution. Under N2, the mixture was cooled to 0 °C, and ethyl chloroformate (1 .16 mL, 12.1 mmol) was added dropwise. After 5 min, triethyl phosphite (2.54 mL, 14.5 mmol) was added dropwise, and the mixture allowed to warm to room temperature and stir overnight. LC/MS analysis indicated consumption of the starting material, and the mixture was concentrated in vacuo. Toluene was added and removed in vacuo (2x). The crude residue was dissolved in chloroform (39 mL) and TiCI4 (5.17 mL, 46.8 mmol) was added to give a brown solution. The solution was refluxed for two days under N2. Product formation was confirmed by LC/MS analysis, and the solution was cooled to room temperature. The reaction mixture was poured into an ice/water bath (300 mL) and stirred for several hours. The pH was adjusted to neutral with aqueous ammonium hydroxide whereupon a brownish precipitate formed. EtOAc (200 mL) was added and the mixture stirred for 1 h, then filtered through Celite® to remove insoluble material. The aqueous layer was extracted with EtOAc (2x 200 mL). The combined organic extracts were dried over MgS04, filtered, and concentrated in vacuo to provide the crude product. Silica gel chromatography (80 g column, 30-80% EtOAc/heptane) provided the title compound (1 .64 g). MS (M+1 ) = 238.3. 1H NMR (400 MHz, CHLOROFORM-d) δ 9.16 (s, 1 H), 8.45 (d, J=5.8 Hz, 1 H), 8.08 (s, 1 H), 7.52 (d, J=5.5 Hz, 1 H), 7.25 (s, 1 H), 4.05 (s, 3H).
Step 3: (7-Methoxyisoquinolin-6-yl)boronic acid
To a 50 mL round bottom flask was added 6-bromo-7-methoxyisoquinoline (400 mg, 1 .68 mmol), bis(pinacolato)diboron (853 mg, 3.36 mmol), potassium acetate (989 mg, 10.1 mmol),
PdCI2(dppf).CH2Cl2 (137 mg, 0.168 mmol), and 1 ,4-dioxane (10 mL). The flask was vacuum/purged with N2 (3x), then stirred at 100 °C for 2 h. The mixture was diluted with EtOAc and filtered through Celite® to provide (7-methoxyisoquinolin-6-yl)boronic acid, Intermediate 8, which was used without further purification. MS (M+1 ) = 204.4
PREPARATION 9
Intermediate 9: Synthesis of 6-(7-methoxyquinolin-6-yl)-N-methyl-N-(2, 2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine
Figure imgf000043_0001
6-Chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 1 , 0.660 g, 2.334 mmol), was coupled with (7-methoxyquinolin-6-yl)boronic acid, (Intermediate 6, 0.771 g, 3.27 mmol), using GENERAL METHOD 1 -2 for Suzuki coupling. The mixture was diluted with water, extracted with DCM (5x), and concentrated. The crude product was purified by silica gel chromatography as described in GENERAL METHOD 4-1 to provide 6-(7-methoxyquinolin-6-yl)-N- methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 9 (0.857 g), as a yellow solid. MS (M+1 ) = 406.2. 1 H NMR (400 MHz, METHANOL-^) δ 8.80 (dd, J=4.3, 1 .8 Hz, 1 H), 8.37 (d, J=7.1 Hz, 1 H), 8.16 (s, 1 H), 7.84 (d, J=9.6 Hz, 1 H), 7.53 (s, 1 H), 7.44 (dd, J=8.1 , 4.5 Hz, 1 H), 7.15 (d, J=10.1 Hz, 1 H), 5.25 (t, J=12.4 Hz, 1 H), 4.03 (s, 3H), 3.01 (s, 3H), 1 .66-1 .74 (m, 2H), 1 .51 -1 .63 (m, 2H), 1 .37 (s, 6H), 1 .23 (s, 6H).
PREPARATION 10
Intermediate 10: Synthesis of (2-amino-4-bromo-5-methoxyphenyl)methanol
Figure imgf000043_0002
Step 1 : 4-Bromo-5-fluoro-2-nitrobenzoic acid
According to the procedure of Beaulieu, P., et al, WO 2010037210, a mixture of 4-bromo-3- fluorobenzoic acid (10.0 g, 45.7 mmol) and concentrated sulfuric acid (75 mL) was stirred until a homogeneous solution formed (-15 min). Potassium nitrate (4.85 g, 47.9 mmol) was added portion- wise over 5 min and the reaction mixture was allowed to stir overnight. The mixture was poured into 500 mL crushed ice. After the ice melted, the mixture was extracted with 1 :1 ethyl acetate/diethyl ether (3x). The extracts were washed with brine, dried over MgS04 and concentrated to provide 4- bromo-5-fluoro-2-nitrobenzoic acid as a dull yellow solid (12.14 g). 1 H NMR (400 MHz,
CHLOROFORM-d) δ ppm 8.18 (d, JH-F =5.6 HZ, 1 H), 7.61 (d, JH-F =7.6 HZ, 1 H).
Step 2: Methyl 4-bromo-5-fluoro-2-nitrobenzoate
Oxalyl chloride (4.18 ml, 47.7 mmol) was added dropwise to a suspension of 4-bromo-5- fluoro-2-nitrobenzoic acid (1 1 .46 g, 43.4 mmol) in DCM (175 mL) and catalytic DMF (100 uL). After 3 h the volatiles were removed via rotary evaporation. The remaining residue was redissolved into DCM (25 mL) and added portion-wise to rapidly stirring methanol (100 mL). The solvent was again removed and the residue concentrated from toluene (2x) to afford methyl 4-bromo-5-fluoro-2- nitrobenzoate as a yellow liquid (12.07 g) which was taken on without purification. 1H NMR (400 MHz, METHANOL-^) δ 8.39 (d, JH-F =5.6 HZ, 1 H), 7.71 (d, JH-F =8.1 HZ, 1 H), 3.91 (s, 3H).
Step 3: Methyl 4-bromo-5-methoxy-2-nitrobenzoate
Sodium methoxide (25 wt % solution in methanol, 10.9 mL, 47.7 mmol) was added to a solution of methyl 4-bromo-5-fluoro-2-nitrobenzoate (12.07 g, 43.4 mmol) in methanol (125 mL) and the solution was allowed to stir overnight during which time a thick precipitate formed. The mixture was diluted with water (400 mL) and the solids isolated by filtration, washing with water. The solids were dissolved into ethyl acetate, and the solution was washed with saturated NaHC03, brine, dried over MgS04 and concentrated to provide methyl 4-bromo-5-methoxy-2-nitrobenzoate (10.55 g) as an off-white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ 8.27 (s, 1 H), 7.07 (s, 1 H), 4.03 (s, 3H), 3.95 (s, 3H).
Step 4: Methyl 2-amino-4-bromo-5-methoxybenzoate
Zinc powder (6.24 g, 95.4 mmol) was added portion-wise over 30 min (Caution! exotherm possible) to a solution of methyl 4-bromo-5-methoxy-2-nitrobenzoate (3.076 g, 10.60 mmol) in 3:1 DCM/AcOH (55 mL). The mixture was stirred at room temperature for three days. The mixture was filtered through Celite®, washing the filter pad with EtOAc, and concentrated. The crude product was redissolved into EtOAc, washed with saturated NaHC03, brine, dried over MgS04 and concentrated to a yellow oil. Concentration of the oil from toluene (2x) provided methyl 2-amino-4- bromo-5-methoxybenzoate as a yellow solid (2.737 g). MS (M+1 ) = 262.1. 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.37 (s, 1 H), 7.03 (s, 1 H), 3.90 (s, 3H), 3.85 (s, 3H).
Step 5: (2-Amino-4-bromo-5-methoxyphenyl)methanol
To a solution of lithium aluminum hydride (1.0 M in THF, 1 1.5 mL, 1 1.5 mmol) in THF (52 mL) cooled to -78 °C was added 2-amino-4-bromo-5-methoxybenzoate (2.724 g, 10.47 mmol) as a solution in THF (30 mL) dropwise via syringe over 10 min. The cooling bath was allowed to warm to room temperature over 2 h. The reaction was quenched by slow addition of ethyl acetate. The solution was further diluted with ethyl acetate (100 mL) and stirred rapidly for two days with a saturated solution of Rochelle salt (potassium sodium tartrate). The layers were separated and the aqueous phase extracted with ethyl acetate (3x), and ether. The combined organics were dried over MgSO4 and concentrated to provide (2-amino-4-bromo-5-methoxyphenyl)methanol,
Intermediate 10 (2.358 g), as an orange solid which was used without further purification. MS (M+1 ) = 232.1. 1H NMR (400 MHz, METHANOL-^) δ 6.97 (s, 1 H), 6.88 (s, 1 H), 4.53 (s, 2H), 3.78 (s, 3H).
PREPARATION 11 Intermediate 11 : Synthesis of 4-chloro-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000045_0001
Step 1: (Z)-Ethyl 3-((4-bromo-3-methoxyphenyl)amino)but-2-enoate
To a mixture of 4-bromo-3-methoxyaniline (5.00 g, 24.8 mmol) and ethyl acetoacetate (3.1 mL, 25 mmol) was added 3 drops (~ 0.1 mL) of 1 M HCI. The mixture was stirred at room temperature overnight, then diluted with DCM, neutralized with aqueous NaHC03 and extracted with DCM. The extracts were dried over Na2S04 and concentrated. The crude product was purified by silica gel column chromatography (0-20% EtOAc in heptane) to provide (Z)-ethyl 3-((4-bromo-3- methoxyphenyl)amino)but-2-enoate as a white solid (4.05 g). MS (M+1 ) = 315.9. 1H NMR
(CHLOROFORM-d) δ: 10.41 (br s, 1 H), 7.46 (d, J=8.6 Hz, 1 H), 6.61 (s, 1 H), 6.56-6.60 (m, 1 H), 4.73 (br s, 1 H), 4.16 (q, J=7.1 Hz, 2H), 3.88 (s, 3H), 2.03 (s, 3H), 1.30 (t, J=7.1 Hz, 3H).
Step 2: 6-Bromo-7-methoxy-2-methylquinolin-4(1H)-one
A mixture of (Z)-ethyl 3-((4-bromo-3-methoxyphenyl)amino)but-2-enoate (4.0 g, 12 mmol) in diphenyl ether (12 mL) was heated at 190 °C for 1 hour. The brown reaction mixture was cooled to room temperature and diluted with diethyl ether. The solid was filtered, washed with ether and dried in a vaccum oven at 50 °C overnight to provide 6-bromo-7-methoxy-2-methylquinolin-4(1 H)-one as a light brown solid (2.64 g). MS (M+1 ) = 270.1 . 1H NMR (DMSO-d6) δ: 1 1 .54 (br s, 1 H), 8.1 1 (s, 1 H), 6.99 (s, 1 H), 5.86 (s, 1 H), 3.93 (s, 3H), 2.30 (s, 3H).
Step 3: (7-Methoxy-2-methyl-4-oxo-1,4-dihydroquinolin-6-yl)boronic acid and 7-Methoxy-2- methyl-6-(4, 4, 5, 5-tetramethyl-1 ,3, 2-dioxaborolan-2-yl)quinolin-4(1H)-one
To a mixture of 6-bromo-7-methoxy-2-methylquinolin-4(1 H)-one (1 .00 g, 3.73 mmol),
bis(pinacolato) diboron (2.84 g, 1 1.2 mmol), dppf (207 mg, 0.37 mmol), PdCl2(dppf).CH2Cl2 (304 mg, 0.37 mmol) and KOAc (2.20 g, 22.4 mmol) was added 1 ,4-dioxane (15 mL) and DMF (3.0 mL). The reaction mixture was evacuated and backfilled with nitrogen (4x), then heated at 90 °C overnight. The reaction mixture was cooled to room temperature, then filtered through Celite® washing with EtOAc, then 10% MeOH in DCM, and concentrated in vacuo to provide the crude product. A mixture of (7-methoxy-2-methyl-4-oxo-1 ,4-dihydroquinolin-6-yl)boronic acid (262 mg) and 7-methoxy-2-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinolin-4(1 H)-one (269 mg) was obtained as light brown solid after silica gel chromatography (0-15% MeOH in DCM). (7- Methoxy-2-methyl-4-oxo-1 ,4-dihydroquinolin-6-yl)boronic acid: MS (M+1 ) = 234.1. 1H NMR (DMSO- cfe) δ 1 1.39 (br s, 1 H), 8.28 (s, 1 H), 7.72 (s, 2H), 6.76-6.88 (m, 1 H), 5.76 (s, 1 H), 3.86 (s, 3H), 2.29 (s, 3H). 7-Methoxy-2-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinolin-4(1 H)-one: MS (M+1 ) = 316.0. 1H NMR (CHLOROFORM-d) δ 12.03 (br s, 1 H), 8.68 (s, 1 H), 6.98 (s, 1 H), 6.10 (s, 1 H), 3.73 (s, 3H), 2.42 (s, 3H), 1.28 (s, 12H).
Step 4: 7-Methoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amin
3-yl)quinolin-4(1H)-one
(7-Methoxy-2-methyl-4-oxo-1 ,4-dihydroquinolin-6-yl)boronic acid (396 mg, 1.69 mmol), 6- chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 1 , 400 mg, 1.41 mmol), Pd(PPh3)4 (123 mg, 0.1 1 mmol) and Na2C03 (375 mg, 3.54 mmol) were added to a microwave vial followed by addition of 4:1 1 ,4-dioxane/water (7.5 ml_). The reaction mixture was sealed, then evacuated and back-filled with nitrogen (4x), and heated via microwave irradiation at 1 10 °C for 1 hour. The mixture was filtered through Celite® washing with EtOAc. The resulting filtrate was concentrated and acidified to pH 3 by 4 M HCI in 1 ,4-dioxane. SCX purification
(GENERAL METHOD 3-1 ) followed by silica gel chromatography (0-30% MeOH in DCM) provided 7-methoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)-pyridazin-3-yl)quinolin- 4(1 H)-one (475 mg) as a dark purple solid. MS (M+1 ) = 436.1. 1H NMR (METHANOL-^) δ 8.37 (s, 1 H), 7.70 (d, J=9.6 Hz, 1 H), 7.12 (d, J=9.6 Hz, 1 H), 7.02 (s, 1 H), 6.13 (s, 1 H), 5.21 (t, J=12.1 Hz, 1 H), 3.94 (s, 3H), 3.00 (s, 3H), 2.45 (s, 3H), 1.65-1 .73 (m, 2H), 1 .51 -1.61 (m, 2H), 1 .37 (s, 6H), 1 .22 (s, 6H).
Step 5: 6-(4-Chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2,2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
A mixture of 7-methoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-4(1 H)-one (495 mg, 1.14 mmol) and phosphorous oxychloride (2.5 ml_, 27 mmol) was sealed and heated via microwave irradiation at 1 10 °C for 1 hour. The reaction mixture was poured into 50 ml. crushed ice then basified by slow addition of solid K2C03. The mixture was extracted with DCM (3x). The combined organic extracts were dried over Na2S04 and concentrated to afford the crude product. SCX purification (GENERAL METHOD 3-1 , 5 g SWlaBond Propylsulfonic Acid® cartridge) provided 6-(4-chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N- (2,2,6, 6-tetramethylpiperidin-4-yl)pyridazin-3-amine as a yellow solid (350 mg). MS (M+1 ) = 454.0. 1H NMR (METHANOL-^) δ 8.60 (s, 1 H), 8.12 (d, J=10.1 Hz, 1 H), 7.78 (s, 1 H), 7.63 (s, 1 H), 7.53 (d, J=9.6 Hz, 1 H), 5.36 (t, J=12.1 Hz, 1 H), 4.12 (s, 3H), 3.12(s, 3H), 2.85 (s, 3H), 1 .95-2.14 (m, 4H), 1.66 (s, 6H), 1 .57 (s, 6H).
Step 6: 4-Chloro-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino
yl)quinolin-7-ol
6-(4-Chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (195 mg, 0.43 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using boron tribromide (1 M solution in DCM, 2.49 mL, 2.49 mmol). SCX purification (GENERAL METHOD 3-1 , 2 g SiliaSond Propylsulfonic Acid® cartridge) provided 4-chloro-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol, Intermediate 11 , as a light orange solid (172 mg, 93% pure) which was used in subsequent reactions without further purification. LC/MS Rt = 0.50 min. MS (M+1 ) = 440.2. 1 H NMR (METHANOL-^) δ 8.54 (s, 1 H), 8.26 (d, J=9.6 Hz, 1 H), 7.39 (s, 1 H), 7.38 (s, 1 H), 7.34 (d, J=9.6 Hz, 1 H), 5.23 (t, J=12.0 Hz, 1 H), 3.04 (s, 3H), 2.65 (s, 3H), 1.71 -1 .80 (m, 2H), 1 .59-1.71 (m, 2H), 1 .44 (s, 6H), 1 .29 (s, 6H).
PREPARATION 12
Intermediate 12: Synthesis of 4-chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000047_0001
Step 1: 5-( ((4-Bromo-3-methoxyphenyl)amino)methylene)-2, 2-dimethyl-1 , 3-dioxane-4, 6- dione
To a mixture of 4-bromo-3-methoxyaniline (5.10 g, 25.2 mmol) and 2,2-dimethyl-1 ,3- dioxane-4,6-dione (Meldrum's acid, 4.37 g, 30.3 mmol) in EtOH (30 mL) was added triethyl orthoformate (5.0 mL, 30 mmol). The reaction mixture was refluxed overnight. The mixture was cooled to room temperature and the solid was filtered, washed with EtOAc, and dried to provide 5- (((4-bromo-3-methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxane-4,6-dione as a yellow solid (2.93 g). MS (M-1 ) = 355.8. 1H NMR (CHLOROFORM-d) δ 1 1 .25 (d, J=14.1 Hz, 1 H), 8.62 (d, J=14.1 Hz, 1 H), 7.59 (d, J=8.1 Hz, 1 H), 6.72-6.78 (m, 2H), 3.96 (s, 3H), 1 .77 (s, 6H).
Step 2: 6-Bromo-7-methoxyquinolin-4(1H)-one
A mixture of 5-(((4-bromo-3-methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxane- 4,6-dione (2.60 g, 7.30 mmol) in diphenyl ether (15 mL) was heated at 185 °C for 40 minutes. The brown mixture was cooled to room temperature and diluted with diethyl ether. The solid was filtered, washed with diethyl ether and dried in a vaccum oven at 50 °C overnight to provide 6- bromo-7-methoxyquinolin-4(1 H)-one as a light brown solid (1.56 g). MS (M+1 ) = 256.1 . 1H NMR (DMSO-d6) 5 1 1.67 (br s, 1 H), 8.17 (s, 1 H), 7.86 (dd, J=7.6, 5.6 Hz, 1 H), 7.05 (s, 1 H), 5.99 (d, J=7.6 Hz, 1 H), 3.94 (s, 3H).
Step 3: 7-Met oxy-6-(4!4!5!5-tetramet yl-1!3!2-dioxaborolan-2-yl)quinolin-4(1H)-one and, (7-methoxy-4-oxo- 1, 4-dihydroquinolin-6-yl)boronic acid To a mixture of 6-bromo-7-methoxyquinolin-4(1 H)-one (0.92 g, 2.72 mmol), dppf (151 mg, 0.272 mmol), PdCl2(dppf).CH2Cl2 (222 mg, 0.272 mmol) and KOAc (1 .33 g, 13.6 mmol) was added 1 ,4-dioxane (7 ml.) and DMF (0.7 ml_). The reaction mixture was evacuated and backfilled with nitrogen (4x), then heated at 80 °C for 0.5 hours. Bis(pinacolato) diboron (1 .72 g, 6.79 mmol) was then added, and the mixture was heated at 90 °C overnight. The reaction mixture was cooled to room temperature, filtered through Celite®, washed with 9:1 MeOH/DCM, then 1 :1 MeOH/DCM. The filtrates were concentrated in vacuo. 7-Methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)quinolin-4(1 H)-one (380 mg) and (7-methoxy-4-oxo-1 ,4-dihydroquinolin-6-yl)boronic acid (270 mg, -50% pure) were obtained after silica gel chromatography (0-30% MeOH in DCM). MS (M+1 ) = 302.0 and 220.2, respectively .
Step 4: 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-4( 1H)-one
7-Methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinolin-4(1 H)-one (380 mg, 1.69 mmol), 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 1 , 250 mg, 0.88 mmol), Pd(PPh3)4 (51 mg, 0.044 mmol), aqueous Na2C03 (2.5 M.1.1 ml_, 2.2 mmol) and 1 ,4-dioxane (4 ml.) were combined in a microwave vessel. The vessel was evacuated and back-filled with nitrogen (4x), and heated via microwave irradiation at 1 10 °C for 1 .5 hour. The mixture was cooled to room temperature, diluted with MeOH (3 ml_), filtered through Celite®, and washed with 9:1 MeOH/DCM and then with 1 :1 MeOH/DCM. The resulting filtrate was concentrated and acidified to pH 3 with 1 M HCI in diethyl ether. SCX purification (GENERAL METHOD 3-1 ) followed by silica gel chromatography (0-20% 2 M NH3/MeOH in DCM) provided 7-methoxy-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-4(1 H)-one (370 mg) as a light brown solid. MS (M+1 ) = 422.1 . 1H NMR (METHANOL-^) δ 8.45 (s, 1 H), 7.91 (d, J=7.6 Hz, 1 H), 7.76 (d, J=9.6 Hz, 1 H), 7.16 (d, J=9.6 Hz, 1 H), 7.10 (s, 1 H), 6.28 (d, J=l A Hz, 1 H), 5.39 (m, 1 H), 3.96 (s, 3H), 3.01 (s, 3H), 1.78-1.90 (m, 4H), 1 .54 (s, 6H), 1 .41 (s, 6H).
Step 5: 6-(4-Chloro-7-methoxyquinolin-6-yl)-N-methyl-N-(2!2,6fi-tetramethy^
yl)pyridazin-3-amine
A mixture of 7-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-4(1 H)-one (59 mg, 0.14 mmol), phosphorous oxychloride (0.65 ml_, 7.0 mmol) and acetonitrile (0.60 mL) was sealed and heated via microwave irradiation at 100 °C for 1 hour. The reaction mixture was poured into crushed ice and was basified by slow addition of solid K2C03. The mixture was extracted with DCM (3x). The combined organic extracts were dried over Na2S04 and concentrated to afford the crude product. SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge) followed by silica gel chromatography (0-20% MeOH in DCM) provided 6-(4-chloro-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine as a brown solid (70 mg). MS (M+1 ) = 440.1
Step 6: 4-chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino
7-ol
6-(4-Chloro-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (62 mg, 0.14 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using boron tribromide (1 M solution in DCM , 0.70 mL, 0.70 mmol). SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge) followed by silica gel chromatography (0-20% 1.5 M NH3/MeOH in DCM) provided 4-chloro-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol, Intermediate 12, as a white solid (21 mg). LC/MS Rt = 0.50 min. MS (M+1 ) = 426.2. 1H NMR (METHANOL-^) δ 8.63 (d, J=5.1 Hz, 1 H), 8.60 (s, 1 H), 8.28 (d, J=9.6 Hz, 1 H), 7.46 (s, 1 H), 7.46 (d, J=4.0 Hz, 1 H), 7.34 (d, J=9.6 Hz, 1 H), 5.22 (t, J=1 1.9 Hz, 1 H), 3.06 (s, 3H), 1 .69-1.79 (m, 2H), 1.57-1 .68 (m, 2H), 1 .43 (s, 6H), 1.27 (s, 6H).
PREPARATION 13
Intermediate 13: Synthesis of 4-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
Figure imgf000049_0001
Step 1: 5-(((3-Chloro-4-methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxane-4,6- dione
Triethoxymethane (1 1 mL, 64 mmol) and 2,2-dimethyl-1 ,3-dioxane-4,6-dione (Meldrum's acid, 10.7 g, 74.2 mmol) were added to 3-chloro-4-methoxyaniline (10.0 g, 63.5 mmol) in ethanol (60 mL). The mixture was stirred at reflux overnight. An additional portion of triethoxymethane (2.1 mL, 13 mmol) and 2,2-dimethyl-1 ,3-dioxane-4,6-dione (2.74 g, 19.0 mmol) were added to the reaction and the mixture was stirred at reflux for 3 h. The mixture was cooled to room temperature and the precipitate was collected by filtration and washed with ethanol to provide 5-(((3-chloro-4- methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxane-4,6-dione as a beige solid (18.5 g). MS (M-1 ) = 310.2. 1H NMR (400 MHz, DMSO-d6) δ 1 1 .20 (d, J=14.4 Hz, 1 H), 8.47 (d, J=14.5 Hz, 1 H), 7.79 (d, J=2.8 Hz, 1 H), 7.54 (dd, J=8.9, 2.8 Hz, 1 H), 7.19 (d, J=9.0 Hz, 1 H), 3.87 (s, 3H), 1.67 (s, 6H).
Step 2: 7-Chloro-6-methoxyquinolin-4(1H)-one and 5-chloro-6-methoxyquinolin-4(1H)-one 5-(((3-chloro-4-methoxyphenyl)amino)methylene)-2,2-dimethyl-1 ,3-dioxan (12.9 g, 41.4 mmol) in diphenyl ether (100 mL) was heated at 190 °C for 2 hour. The reaction mixture was cooled to room temperature and diluted with diethyl ether (100 mL). The solid was filtered and washed with diethyl ether to provide a mixture of 7-chloro-6-methoxyquinolin-4(1 H)-one and 5- chloro-6-methoxyquinolin-4(1 H)-one (7.54 g) in a ratio of 4:1 (determined by 1H NMR). 7-Chloro-6- methoxyquinolin-4(1 H)-one: MS (M+1 ) = 210.1 . 1H NMR (400 MHz, METHANOL-^) δ 7.95 (d, J=7.2 Hz, 1 H), 7.77 (s, 1 H), 7.69 (s, 1 H), 6.33 (d, J=7.3 Hz, 1 H), 4.01 (s, 3H). 5-Chloro-6- methoxyquinolin-4(1 H)-one: MS (M+1 ) = 210.1 . 1H NMR (400 MHz, METHANOL-^) δ 7.81 (d, J=7.2 Hz, 1 H), 7.57 (d, J=9.2 Hz, 1 H), 7.52 (d, J=9.2 Hz, 1 H), 6.22 (d, J= 7.2 Hz, 1 H), 3.97 (s, 3H).
Step 3: 6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
yl)quinolin-4( 1H)-one
A mixture of 7-chloro-6-methoxyquinolin-4(1 H)-one and 5-chloro-6-methoxyquinolin-4(1 H)- one (1.42 g, 6.67 mmol), tetrahydroxydiboron (1 .82 g, 20.3 mmol), 2nd Generation XPhos
Precatalyst (0.27 g, 0.34 mmol), XPhos (0.32 g, 0.68 mmol) and potassium acetate (1.99 g, 20.3 mmol) were stirred in ethanol (70 mL) at 80 °C for 2 h. An aqueous solution of K2C03 (1 .8 M, 15 mL, 27 mmol) was added to the reaction at room temperature followed by Intermediate 1 (1.92 g, 6.77 mmol). The mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature, filtered and concentrated in vacuo. The residue was acidified to pH 1 using 1 M HCI (60 mL) and filtered. The filtrate was washed with DCM (2x), then basified to pH 1 1 with a saturated solution of K2C03. The aqueous phase was extracted with 9:1 DCM/MeOH (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (2-15% 7 M ammonia in MeOH gradient, in DCM) to afford 6-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-4(1 H)-one (0.45 g) as a brown solid. MS (M+1 ) = 422.4. 1 H NMR (400 MHz, METHANOL-^) δ 7.99 (d, J=7.2 Hz, 1 H), 7.95 (s, 1 H), 7.93 (d, J=9.7 Hz, 1 H), 7.83 (s, 1 H), 7.16 (d, J=9.7 Hz, 1 H), 6.38 (d, J=7.2 Hz, 1 H), 5.21 -5.33 (m, 1 H), 4.00 (s, 3H), 3.03 (s, 3H), 1.72 (dd, J=12.7, 3.5 Hz, 2H), 1.60 (t, J=12.4 Hz, 2H), 1 .40 (s, 6H), 1 .25 (s, 6H).
Step 4: 6-(4-Chloro-6-methoxyquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin- 4(1 H)-one (0.22 g, 0.52 mmol) in phosphorous oxychloride (1 .5 mL) was heated via microwave at 100 °C for 2 h. The reaction mixture was poured into crushed ice and was basified by slow addition of a saturated solution of K2C03. The mixture was extracted with DCM (3x). The combined organic extracts were dried over Na2S04 and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (2-15% 7 M ammonia in MeOH gradient, in DCM) to afford 6-(4-chloro-6-methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-y 3-amine (0.20 g) as a brown solid. MS (M+1 ) = 440.5. 1H NMR (400 MHz, METHANOL-^) δ 8.66 (d, J=4.8 Hz, 1 H), 8.33 (s, 1 H), 7.87 (d, J=9.6 Hz, 1 H), 7.68 (d, J=4.9 Hz, 1 H), 7.66 (s, 1 H), 7.16 (d, J=9.7 Hz, 1 H), 5.26 (t, J=12.3 Hz, 1 H), 4.06 (s, 3H), 3.03 (s, 3H), 1 .72 (dd, J=12.6, 3.6 Hz, 2H), 1 .59 (t, J=12.4 Hz, 2H), 1 .39 (s, 6H), 1 .24 (s, 6H).
Step 5: 4-Chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-6-ol
A solution of BBr3 in DCM (1 M, 0.7 ml_, 0.7 mmol) was added to 6-(4-chloro-6- methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.10 g, 0.24 mmol) in DCM (2.2 ml.) and the reaction was stirred at room temperature overnight. MeOH was added to the reaction and the solvent was concentrated under reduced pressure. The crude material was solubilized with 2 M ammonia in MeOH and DMSO and purified via preparative reverse-phase HPLC (15 to 45% acetonitrile in water, 5 mM ammonium hydroxide modifier) to afford 4-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol (9 mg). LC/MS Rt = 0.56 min. MS (M+1 ) = 426.3. 1H NMR (400 MHz, METHANOL-^) δ 8.60 (d, J=4.8 Hz, 1 H), 8.47 (s, 1 H), 8.33 (d, J=9.8 Hz, 1 H), 7.63 (s, 1 H), 7.61 (d, J=4.8 Hz, 1 H), 7.39 (d, J=9.9 Hz, 1 H), 5.17-5.47 (m, 1 H), 1.78 (dd, J=12.9, 3.7 Hz, 2H), 1.67 (t, J=12.4 Hz, 2H), 1.46 (s, 6H), 1 .30 (s, 6H). GENERAL METHODS
GENERAL METHOD 1 -1
Representative procedure for Suzuki Coupling
A mixture of a chloropyridazine intermediate, such as Intermediate 1 (1.0 equivalents), potassium triphosphate (5.0 equivalents), and 2nd generation XPhos precatalyst (0.07 equivalents), in DMF (0.3 M) is degassed via vacuum/N2 purge (3x), and heated at 50 °C. Next, a 50 or 100 mg/mL solution of a boronic acid, such as Intermediate 7 or Intermediate 8, in DMF is added in 1 ml. aliquots every 2-3 h until Intermediate 1 is consumed. The reaction is cooled to room temperature, filtered through Celite® washing with EtOAc, and partially concentrated. The crude product is purified by SCX (GENERAL METHOD 3-1 ) and silica gel chromatography.
GENERAL METHOD 1 -2
Representative Procedure for Suzuki Coupling
A mixture of a chloropyridazine intermediate, such as Intermediate 1 (1.0 equivalent), boronic acid reagent (1.5-2 equivalents), and Na2C03 (3 equivalents) in 3:1 DME/water (0.12 M) in a microwave vial is degassed with a stream of dry nitrogen. Pd(PPh3)4 (0.1 equivalents) is added to the reaction mixture and the vial is sealed and heated in a Biotage® Initiator microwave reactor at 140 °C for 30 min. The mixture is diluted with water and extracted with DCM (4x). The organic extracts are dried over MgS04, filtered, and concentrated to afford the crude product. The crude product is purified via silica chromatography as described in GENERAL METHOD 4-1.
GENERAL METHOD 2-1
Representative Procedure for Methoxy Deprotection using Pyridine HCI
The methoxy substrate (1 equivalent) and pyridine hydrochloride (20-30 equivalents) are heated at 160-190 °C for 15-120 min in a Biotage® Initiator microwave reactor. The reaction mixture is solubilized with MeOH/DMSO and purified via preparative reverse-phase HPLC (15 to 45% acetonitrile in water, 5 mM ammonium hydroxide modifier).
GENERAL METHOD 2-2
Representative Procedure for Methoxy Deprotection using Pyridine HCI
The methoxy substrate (1 equivalent) and pyridine hydrochloride (20-25 equivalents) are heated at 160-190 °C for 15-120 min in a Biotage® Initiator microwave reactor. The reaction mixture is solubilized with MeOH and the solution loaded onto a mixture of solid sodium bicarbonate (30 equivalents) and silica gel (~6 g/mmol methoxy substrate), and concentrated to dryness. Column chromatography as described in GENERAL METHOD 4-1 provided the product.
GENERAL METHOD 2-3
Representative Procedure for Methoxy Deprotection using Boron Tribromide (BBr3) The methoxy substrate (1.0 equivalent) is dissolved in DCM (0.03 M) and the solution cooled to 0 °C. Boron tribromide (1 M solution in DCM, 3-10 equivalents) is added dropwise. The crude reaction mixture is stirred at room temperature overnight. If the reaction has not gone to completion, it can be heated at reflux for 4-24 h. The reaction mixture is diluted with DCM and water. The organic layer is diluted with EtOAc and washed with saturated NaHC03 (2x), water, brine and dried over Na2S04 or MgS04. The crude product is purified via preparative reverse-phase HPLC, silica gel chromatography (GENERAL METHODS 4-1 ), or by recrystallization.
GENERAL METHOD 2-4
Representative Procedure for Methoxy Deprotection using Thiophenol
Thiophenol (5.0 equivalents) and potassium carbonate (1 .0 equivalent) are added to the methoxy substrate (1.0 equivalent) in NMP (0.2 M) and the reaction is heated at 190 °C for 15-30 min in a Biotage® Initiator microwave reactor. The reaction mixture is purified by catch and release as described in GENERAL METHOD 3-1 , and the product is purified by reverse-phase preparative HPLC or silica gel chromatography.
GENERAL METHOD 3-1
Representative Procedure for Catch-and-Release (SCX) Purification / Free Basing The crude product is dissolved in MeOH and loaded onto a SiliaSond Propylsulfonic Acid® cartridge (Silicycle, Inc.) or a Bond Elut SCX cartridge (Agilent Technologies, Inc.) preconditioned with MeOH or acetonitrile. The cartridge is flushed with MeOH or acetonitrile, and the product released by elution with 2-7 N ammonia in MeOH. Concentration of the eluent provides the purified product. GENERAL METHOD 4-1
Representative Procedure for Silica Gel Chromatography Purification
The crude product (either neat, dissolved into DCM, or loaded onto a solid support such as silica gel or Celite®) is subjected to normal phase flash column chromatography using RediSep®Rf pre-packed silica gel cartridges (Teledyne Isco, Inc.) with an elution gradient of 1-20% ammonia in MeOH (1.5 N, or 3.5 N concentration), in DCM.
GENERAL METHOD 5-1
Representative Procedure for Iridium Catalyzed Borylation
To a degassed mixture of the substrate (1 .0 equivalent), bis(pinacolato)diboron (1 .50 equivalents) and 4,4'-di-ferf-butyl bipyridine (0.1 equivalents) in 1 ,4-dioxane (0.3 M) is added
[lr(COD)(OMe)]2 (0.05 equivalents). The mixture was heated at 85 °C overnight. The resulting boronic acid solution can be used directly in the subsequent reaction, or optionally the boronic acid is isolated and purified. The pinacol boronate ester is typically not observed as a reaction product. GENERAL METHOD 6-1
Representative Procedure for Metal Scavenging
Residual palladium, and certain other metal contaminants, can be scavenged from reaction products by dissolution in an appropriate solvent (e.g., THF, DCM, or acetonitrile) and stirring with SiliaMetS® Dimercaptotriazine (DMT) resin (Silicycle, Inc.) for 4-24 h using 3-5 equivalents relative to the amount of the palladium catalyst (or other metal) used. Filtration, and washing of the resin, followed by concentration provides the purified product.
GENERAL METHOD 7-1
Representative procedure for boronate ester formation.
Bis(pinacolato) diboron (2.0 equivalents), KOAc (6.0 equivalents), and PdCI2(dppf).CH2Cl2 (0.1 equivalents) were added to flask containing an aryl bromide (1 .0 equivalent). DMSO (0.2 M) is added, and the reaction vessel is evacuated then filled with N2 (2x), and heated at 100 °C overnight. The reaction mixture is cooled to room temperature, then filtered through Celite® (prepacked filter funnel) using EtOAc, and concentrated in vacuo to provide the crude product, which is purified by silica gel chromatography (EtOAc/heptane).
EXAMPLES
The following compounds were prepared from chloropyridazine Intermediate 1 , Intermediate 4, or Intermediate 5, and 7-methoxyquinolin-6-yl)boronic acid (Intermediate 6), according to GENERAL METHOD 1 -2, followed by methoxy deprotection and purification as outlined in GENERAL METHOD 2-2. The hydrochloride salt was formed by dissolution of the free base in 1 M aqueous
HCI (3 equivalents) followed by lyophilization.
Figure imgf000054_0001
Example 2-1 : Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000055_0001
To a solution of Intermediate 11 (22 mg, 0.050 mmol) in MeOH (2 ml.) under a nitrogen atmosphere was added palladium on carbon (10 wt %, 5.9 mg, 0.005 mmol) and a drop of concentrated HCI. The mixture was evacuated and back-filled with hydrogen (4x), and stirred rapidly overnight. The mixture was filtered through Celite®, rinsed with MeOH and concentrated. SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge) and preparative reverse-phase HPLC purification provided 2-methyl-6-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol as a light yellow solid (1 1.5 mg). LC/MS Rt = 0.39 min. MS (M+1 ) = 406.0. 1 H NMR (400 MHz, METHANOL-^) δ 8.20 (s, 1 H), 8.16 (d, J=9.6 Hz, 1 H), 8.12 (d, J=8.6 Hz, 1 H), 7.32 (s, 1 H), 7.26 (d, J=9.6 Hz, 1 H), 7.17 (d, J=8.6 Hz, 1 H), 5.09 (t, J=12.1 Hz, 1 H), 2.98 (s, 3H), 2.64 (s, 3H), 1 .62-1.72 (m, 2H), 1 .50-1.60 (m, 2H), 1.38 (s, 6H), 1 .22 (s, 6H).
Example 3-1 : Synthesis of 7-(6-(methyl(1 , 2,2,6, 6-pentamethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000055_0002
Step 1. 6-( 6-Methoxyisoquinolin-7-yl)-N-methyl-N-( 1, 2, 2, 6, 6-pentamethylpiperidin-4- yl)pyridazin-3-amine
The title compound (63 mg, 0.15 mmol) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction of Intermediate 4 (75 mg, 0.25 mmol) and Intermediate 7 (77 mg, 0.38 mmol). MS (M+1 ) = 420.1
Step 2. 7-( 6-(Methyl( 1, 2, 2, 6, 6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6- ol
The title compound (26 mg) was prepared from 6-(6-methoxyisoquinolin-7-yl)-N-methyl-N- (1 ,2,2,6,6-pentamethylpiperidin-4-yl)pyridazin-3-amine (63 mg, 0.150 mmol) following GENERAL METHOD 2-4 for demethylation using thiophenol. The crude residue was purified by preparative reverse-phase HPLC (25 to 50% acetonitrile in water, 5 mM ammonium hydroxide modifier). LC/MS Rt = 0.38 min. MS (M+1 ) = 406.2. 1H NMR (400 MHz, METHANOL-^) δ 9.57 (s, 1 H), 8.98 (s, 1 H), 8.48 (d, J=10.0 Hz, 1 H), 8.40 (d, J=6.8 Hz, 1 H), 8.21 (d, J=6.8 Hz, 1 H), 7.70 (d, J=9.8 Hz, 1 H), 7.64 (s, 1 H), 5.35 (br s, 1 H), 3.18 (s, 3H), 2.94 (s, 3H), 2.24-2.36 (m, 2H), 2.10-2.19 (m, 2H), 1 .66 (s, 6H), 1.61 (s, 6H).
Example 3-2: Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000056_0001
Step 1. 6-( 6-Methoxyisoquinolin-7-yl)-N-methyl-N-( 2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
A 30 mL vial with a septum cap was charged with 6-chloro-N-methyl-N-(2, 2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 1 (200 mg, 0.71 mmol), potassium triphosphate (751 mg, 3.54 mmol), and 2nd generation XPhos precatalyst (39 mg, 0.05 mmol), and DMF (2.5 mL). The mixture was degassed via vacuum/purge (3x), and heated at 50 °C. A solution of Intermediate 7 in DMF (100 mg/mL) was added in 1 mL aliquots every 2-3 h until Intermediate 1 was consumed. The reaction was cooled to room temperature, filtered through a Celite® column washing with EtOAc, and partially concentrated. The mixture was diluted with MeOH and acidified to pH <4 with 1 M HCI. SCX purification (GENERAL METHOD 3-1 ) followed by elution through a silica gel plug (10-50% MeOH/DCM) provided the title compound (130 mg) which was taken on without further purification. MS (M+1 ) = 406.7
Step 2. 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol
6-(6-Methoxyisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine was deprotected following GENERAL METHOD 2-4 for demethylation using thiophenol. SCX purification (GENERAL METHOD 3-1 ) followed by preparative reverse phase HPLC (25 to 50% acetonitrile in water, 5 mM ammonium hydroxide modifier) provided the title compound (63 mg). LC/MS Rt = 0.39 min. MS (M+1 ) = 392.2. 1H NMR (400 MHz, METHANOL-^) δ 9.13 (s, 1 H), 8.59 (s, 1 H), 8.37 (d, J=10.0 Hz, 1 H), 8.28 (d, J=5.8 Hz, 1 H), 7.64 (d, J=6.0 Hz, 1 H), 7.40 (d, J=10.0 Hz, 1 H), 7.30 (s, 1 H), 5.27 (br. s., 1 H), 3.07 (s, 3H), 1 .76-1.85 (m, 2H), 1.65-1 .75 (m, 2H), 1.48 (s, 6H), 1 .33 (s, 6H).
Example 3-3: Synthesis of 7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)isoquinolin-6-ol
Figure imgf000056_0002
Step 1. 6-Methoxy-7-(6-((2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinolin The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction from Intermediate 3 (200 mg, 0.741 mmol) and Intermediate 7 (256 mg, 1 .26 mmol). MS (M+1 ) = 393.0
Step 2. 7-(6-((2,2^^-Tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinolin-6-ol
The title compound (66 mg) was prepared from 6-methoxy-7-(6-((2,2,6,6-tetramethylpiperidin-4- yl)oxy)pyridazin-3-yl)isoquinoline following GENERAL METHOD 2-4 for demethylation using thiophenol. LC/MS Rt = 0.38 min. MS (M+1 ) = 379.2. 1H NMR (400 MHz, METHANOL-^) δ 9.50 (s, 1 H), 8.98 (s, 1 H), 8.49 (d, J=9.5 Hz, 1 H), 8.28-8.33 (m, 1 H), 8.12 (d, J=6.8 Hz, 1 H), 7.57 (s, 1 H), 7.41 (d, J=9.3 Hz, 1 H), 5.80 (tt, J=4.1 , 10.7 Hz, 1 H), 2.43 (dd, J=4.0, 13.8 Hz, 2H), 1 .80 (dd, J=1 1.2, 13.4 Hz, 2H), 1 .56 (s, 6H), 1 .48 (s, 6H).
Example 3-4: Synthesis of 7-(6-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol- 2(1 H)-yl)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000057_0001
Step 1. 6-Methoxy-7-(6-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)- yl)pyridazin-3-yl)isoquinoline
The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction from Intermediate 5 (75 mg, 0.314 mmol) and Intermediate 7 (108 mg, 0.534 mmol). MS (M+1 ) = 362.5
Step 2. 7-(6-((3aR,6aS)-5-Methylhexahydropyrrdo[3,4-c]pyrrol-2(1H)-yl)py
yl)isoquinolin-6-ol
The title compound (21 mg) was prepared from 6-methoxy-7-(6-((3aR,6aS)-5- methylhexahydropyrrolo[3,4-c]pyrrol-2(1 H)-yl)pyridazin-3-yl)isoquinoline following GENERAL METHOD 2-4 for demethylation using thiophenol. LC/MS Rt = 0.33 min. MS (M+1 ) = 348.2. 1 H NMR (400 MHz, METHANOL-^) δ 9.03 (s, 1 H), 8.51 (s, 1 H), 8.28 (d, J=9.8 Hz, 1 H), 8.18 (d, J=6.0 Hz, 1 H), 7.53 (d, J=6.0 Hz, 1 H), 7.20 (t, J=4.9 Hz, 2H), 3.63-3.73 (m, 2H), 3.53-3.62 (m, 2H), 3.09- 3.17 (m, 2H), 2.99-3.08 (m, 2H), 2.78 (dd, J=2.9, 10.2 Hz, 2H), 2.49 (s, 3H).
Example 3-5: Synthesis of 1 -cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000058_0001
Step 1. 3-(Benzyloxy)-4-iodobenzoic acid
Benzyl bromide (7 mL, 57.8 mmol) was added to a mixture of 3-hydroxy-4-iodobenzoic acid (5.13 g, 18.9 mmol) and K2C03 (8.0 g, 58 mmol) in acetone (200 mL). The mixture was then heated at reflux overnight. The mixture was cooled to room temperature then filtered through a small Celite® pad. The filtrate was concentrated in vacuo. MeOH (30 mL) was added to the residue followed by 1 M NaOH (94 mL, 94 mmol). The resulting mixture was heated in a 70 °C oil bath for 1 h. Next, the mixture was cooled to room temperature then concentrated in vacuo to approximately one half the starting volume to remove the MeOH, . The resulting solution was diluted with water (20 mL) then extracted with Et20 (2 x 100 mL). The organic layer was washed with 1 M NaOH (50 mL), and the combined aqueous layers were slowly acidified to pH 2 using concentrated HCI. Ethyl acetate (150 mL) was added to the resulting precipitate. After separation, the aqueous layer was extracted with EtOAc (3 x 150 mL). The combined organic layers were dried over MgS04, filtered, and concentrated in vacuo to afford the title compound as a white solid (6.6 g). MS (M+1 ) = 353.5. 1H NMR (400 MHz, DMSO-d6) δ 12.93 (br s, 1 H), 7.93 (d, J=8.1 Hz, 1 H), 7.48-7.57 (m, 3H), 7.41 (t, J=7.3 Hz, 2H), 7.28-7.35 (m, 2H), 5.26 (s, 2H).
Step 2. (3-(Benzyloxy)-4-iodophenyl)methanol
Borane tetrahydrofuran complex solution (1.0 M in THF, 7.83 ml, 7.83 mmol) was slowly added to a cooled solution (ice bath) of 3-(benzyloxy)-4-iodobenzoic acid (1.98 g, 5.59 mmol) in THF (50 mL). After complete addition, the reaction mixture was allowed to warm to room
temperature overnight. The reaction mixture was cooled in an ice bath for 5 min then slowly quenched with 1 M HCI (10 mL). The reaction was then diluted with water (20 mL), and extracted with Et20 (3 x 50 mL). The organic layer was washed successively with saturated aqueous
NaHC03, and brine. The organics were dried over MgS04, filtered, and concentrated in vacuo affording the title compound as a thick colorless oil (1.9 g). MS (M-OH) = 323.0
Step 3. 2-(3-(Benzyloxy)-4-iodobenzyloxy)tetrahydro-2H-pyran
p-Toluenesulfonic acid monohydrate (0.215 g, 1.129 mmol) was added to a solution of (3- (benzyloxy)-4-iodophenyl)methanol (1.92 g, 5.64 mmol) and 3,4-dihydro-2H-pyran (0.64 mL, 6.8 mmol) in DCM (50 mL) at room temperature. After 2 h, the solution was diluted with DCM (30 mL), then washed with saturated aqueous NaHC03 solution (2 x 20 mL). The organic layer was washed with brine, dried over MgS04, filtered, and concentrated in vacuo. The resulting oil was stored under high vacuum overnight. The residue was purified by silica gel chromatography (80 g silica gel, 2-30% EtOAc/heptane) to afford the desired product as a thick colorless oil. (2.0 g, 4.7 mmol). MS (M+H20) = 442.2. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.76 (d, J=7.6 Hz, 1 H), 7.51 (d, J=7.6 Hz, 2H), 7.39 (t, J=7.3 Hz, 2H), 7.29-7.35 (m, 1 H), 6.92 (d, J=2.0 Hz, 1 H), 6.74 (dd, J=8.1 , 1 .52 Hz, 1 H), 5.18 (s, 2H), 4.71 (d, J=12.6 Hz, 1 H), 4.66 (t, J=3.5 Hz, 1 H), 4.47 (d, J=12.1 Hz, 1 H), 3.82-3.93 (m, 1 H), 3.46-3.58 (m, 1 H), 1.80-1 .94 (m, 1 H), 1.68-1 .79 (m, 1 H), 1 .50-1.68 (m, 4H).
Step 4. 2-(2-(Benzyloxy)-4-((tetra ydro-2H-pyran-2-yloxy)met yl)p enyl)-4,4, 5, 5- tetramethyl-1 , 3, 2-dioxaborolane
Following GENERAL METHOD 7-1 for boronate ester formation with 2-(3-(benzyloxy)-4- iodobenzyloxy)tetrahydro-2H-pyran (1 .96 g, 4.62 mmol) afforded the title compound (1.96 g). MS (M+H20) = 442.3. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.68 (d, J=8.1 Hz, 1 H), 7.61 (d, J=7.6 Hz, 2H), 7.36 (t, J=7.3 Hz, 2H), 7.30 (d, J=7.1 Hz, 1 H), 6.94-7.00 (m, 2H), 5.14 (s, 2H), 4.77 (d, J=12.6 Hz, 1 H), 4.68 (t, J=3.5 Hz, 1 H), 4.54 (d, J=12.6 Hz, 1 H), 3.85-3.95 (m, 1 H), 3.48-3.57 (m, 1 H), 1.82-1 .94 (m, 1 H), 1.50-1.78 (m, 5H), 1 .37 (s, 12H).
Step 5. (3-(Benzyloxy)-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi yl)phenyl)methanol
6-Chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 1 , 503 mg, 1.78 mmol), was added to a round bottom flask containing 2-(2-(benzyloxy)-4-((tetrahydro- 2H-pyran-2-yloxy)methyl)phenyl)-4, 4, 5, 5-tetramethyl-1 , 3, 2-dioxaborolane (1.96 g, 3.23 mmol). Na2C03 (566 mg, 5.34 mmol) and Pd(PPh3)4 (230 mg, 0.20 mmol) were added followed by addition of 4:1 DME/water (15 ml_). The reaction flask was equipped with a reflux condenser, and the flask evacuated and filled with N2 (2x) then heated in a 100 °C oil bath overnight (18 h). The mixture was cooled to room temperature then filtered through Celite® with a MeOH wash. The filtrate was acidified to pH 2 using 1 M HCI then heated at 70 °C for 1 h. The mixture was cooled to room temperature then filtered through Celite® washing with MeOH. SCX purification (GENERAL
METHOD 3-1 , 20 g cartridge) afforded the title compound (750 mg) as a yellow amorphous solid after storing under high vacuum for several hours. MS (M+1 ) = 461 .4. 1H NMR (400 MHz, DMSO- d6) δ ppm 7.74 (d, J=9.6 Hz, 1 H), 7.68 (d, J=8.1 Hz, 1 H), 7.41 (d, J=7.6 Hz, 1 H), 7.36 (t, J=7.3 Hz, 2H), 7.27-7.33 (m, 1 H), 7.19 (s, 1 H), 6.94-7.06 (m, 2H), 5.16 (s, 2H), 5.09 (t, J=5.8 Hz, 2H), 4.54 (d, J=5.6 Hz, 2H), 2.90 (s, 3H), 1 .38-1 .63 (m, 5H), 1.28 (br s, 6H), 1.13 (br s, 6H).
Step 6. 3-(Benzyloxy)-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)benzaldehyde
To a 50 ml. round bottom flask containing (3-(benzyloxy)-4-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)phenyl)methanol (218 mg, 0.47 mmol) and DCM (2.4 ml.) was added manganese dioxide (259 mg, 2.98 mmol). The reaction was stirred at room temperature for 18 h and additional manganese dioxide was added (123 mg, 1 .42 mmol). The reaction was stirred at room temperature for 24 h then filtered through Celite® and concentrated in vacuo to afford 3-(benzyloxy)-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)benzaldehyde (217 mg). MS (M+1 ) = 459.5
Step 7. 1-(3-(Benzyloxy)-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)am
yl)phenyl)-2-nitroethanol
To a 50 ml. round bottom flask containing 3-(benzyloxy)-4-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)benzaldehyde (200 mg, 0.44 mmol) and MeOH (2.2 ml.) was added nitromethane (0.47 ml_, 0.87 mmol) and 1 M aqueous NaOH (1.3 ml_, 1.31 mmol). The reaction was stirred at room temperature for 18 h then the pH was adjusted to 6-7 with 1 M aqueous HCI. SCX purification (GENERAL METHOD 3-1 ) and concentration of the product- containing fractions afforded 1-(3-(benzyloxy)-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)phenyl)-2-nitroethanol (176 mg). MS (M+1 ) = 520.5
Step 8. 2-Amino-1-(3-methoxy-4-(6-(methyl(2,2,6^-tetramethylpiperM^
yl)amino)pyridazin-3-yl)phenyl)ethanol
To a 50 ml. round bottom flask containing 1 -(3-(benzyloxy)-4-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)phenyl)-2-nitroethanol (176 mg, 0.40 mmol) in MeOH (5.0 ml.) was added 10% palladium on carbon (21 1 mg, 0.20 mmol). The resulting suspension was stirred under hydrogen at atmospheric pressure and room temperature for 18 h. The suspension was filtered through Celite®and the filter pad was washed with MeOH. SCX purification (GENERAL METHOD 3-1 ) and concentration of the product-containing fractions afforded 2-amino-1 -(3- methoxy-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)phenyl)ethanol (164 mg). MS (M+1 ) = 414.5
Step 9. N-(2-Hydroxy-2-(3-methoxy-4-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)phenyl)ethyl)cyclopropanecarboxamide
In a 50 ml. round bottom flask, 2-amino-1-(3-methoxy-4-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)phenyl)ethanol (250 mg, 0.61 mmol),
cyclopropanecarboxylic acid (104 mg, 1 .21 mmol), DIPEA (0.42 ml_, 2.42 mmol), and HATU (230 mg, 0.61 mmol) in DCM (1 .5 ml.) and DMF (1 .5 ml.) were combined. The mixture was stirred at room temperature for 45 min, acidified with excess acetic acid and concentrated in vacuo. SCX purification (GENERAL METHOD 3-1 ) and concentration of the product-containing fractions afforded N-(2-hydroxy-2-(3-methoxy-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)phenyl)ethyl)cyclopropanecarboxamide (1 12 mg). MS (M+1 ) = 482.6
Step 10. 6-( 1 -Cyclopropyl-6-methoxyisoquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
To a 50 ml. round bottom flask containing N-(2-hydroxy-2-(3-methoxy-4-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)phenyl)ethyl)cyclopropanecarboxamide (68 mg, 0.14 mmol) in acetonitrile (1.4 mL) was added phosphorus oxychloride (POCI3, 0.66 mL, 0.71 mmol). The mixture was heated at 60 °C for 1 h then additional POCI3 (0.66 mL, 0.71 mmol) was added and the reaction mixture was heated at 80 °C for 18 h. Additional POCI3 (0.66 mL, 0.71 mmol) and toluene (1.4 mL) were added and the reaction heated at 90 °C for 1 h. The mixture was then cooled to room temperature, quenched with a minimal amount of water and concentrated in vacuo. SCX purification (GENERAL METHOD 3-1 ) and concentration of the product-containing fractions afforded 6-(1 -cyclopropyl-6-methoxyisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (63 mg). MS (M+1 ) = 446.1
Step 11. 1-Cyclopropyl-7-(6-(methyl(2,2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinolin-6-ol
Following standard GENERAL METHOD 2-3 for methoxy deprotection using boron tribromide, the title compound was afforded. LC/MS Rt = 0.43 min. MS (M+1 ) = 430.4 1H NMR (400 MHz, METHANOL-^) δ 8.74 (s, 1 H), 8.18 (d, J=10.0 Hz, 1 H), 8.01 (d, J=5.8 Hz, 1 H), 7.26 (d, J=6.0 Hz, 1 H), 7.13-7.20 (m, 2H), 4.99-5.16 (m, 1 H), 2.97 (s, 3H), 2.69-2.78 (m, 1 H), 1 .66-1.74 (m, 2H), 1.45-1 .57 (m, 2H), 1.35 (s, 6H), 1 .20 (s, 7H), 1 .06-1.14 (m, 4H).
Example 3-6: Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 ,6-diol
Figure imgf000061_0001
Step 1. 7-Bromo-1-ethoxy-6-methoxyisoquinoline
In a 30 mL vial, 7-bromo-1-chloro-6-methoxyisoquinoline (200 mg, 0.734 mmol) was partially dissolved in sodium ethoxide in ethanol (3 M, 2 mL, 6.00 mmol) and the mixture was heated for 18 h at 80 °C. The mixture was cooled, and the solvent was removed under vacuum. The crude residue was purified by silica gel chromatography (5-30% EtOAc/heptane) to provide the title compound (157 mg). MS (M+1 ) = 284.0. 1H NMR (400 MHz, CHLOROFORM-d) δ 8.46 (s, 1 H), 7.95 (d, J=6.0 Hz, 1 H), 7.09 (d, J=6.0 Hz, 1 H), 7.03 (s, 1 H), 4.53 (q, J=7.2 Hz, 2H), 4.02 (s, 3H), 1.50 (t, J=7.2 Hz, 3H).
Step 2: 1-Ethoxy-6-methoxy- 7-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)isoquinoline Following GENERAL METHOD 7-1 for boronate ester formation, 7-bromo-1-ethoxy-6- methoxyisoquinoline (53 mg, 0.188 mmol) afforded the title compound (60 mg). MS (M+1 ) = 330.0 Step 3. 6-( 1-Ethoxy-6-methoxyisoquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction from Intermediate 1 (35 mg, 0.12 mmol) and 1-ethoxy-6-methoxy-7-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline (60 mg). MS (M+1 ) = 450.2
Step 4. 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline- 1, 6- diol
The title compound (1 1 mg, 0.024 mmol) was prepared from 6-(1 -ethoxy-6- methoxyisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine following GENERAL METHOD 2-4 for deprotection using thiophenol. LC/MS Rt = 0.50 min. MS (M+1 ) = 408.3. 1H NMR (400 MHz, METHANOL-^) δ 8.57 (s, 1 H), 8.48 (d, J=9.8 Hz, 1 H), 8.08 (d, J=10.0 Hz, 1 H), 7.25 (d, J=7.3 Hz, 1 H), 7.17 (s, 1 H), 6.60 (d, J=7.0 Hz, 1 H), 4.8-5.1 (the methine signal is obscured by a broad H20 peak), 3.21 (s, 3H), 2.12 (d, J=12.5 Hz, 2H), 1 .99-2.07 (m, 2H), 1.66 (s, 6H), 1.59 (s, 6H)).
Example 3-7: Synthesis of 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 -carbonitrile
Figure imgf000062_0001
Step 1. 7-Bromo-6-methoxyisoquinoline 2-oxide
To a 50 mL round bottom flask containing 7-bromo-6-methoxyisoquinoline (prepared as described in WO2007000240) (200 mg, 0.84 mmol) in DCM (4.2 mL) was added m- chloroperoxybenzoic acid (mCPBA, 269 mg, 1 .09 mmol). The mixture was stirred at room temperature for 18 h, diluted with DCM and washed with saturated aqueous sodium bicarbonate solution, then saturated aqueous sodium thiosulfate, and brine. The resulting organic layer was dried with sodium sulfate and concentrated in vacuo to afford 7-bromo-6-methoxyisoquinoline 2- oxide (213 mg). MS (M+1 ) = 256.3
Step 2. 7-Bromo-6-methoxyisoquinoline-1-carbonitrile
To a 50 mL round bottom flask containing 7-bromo-6-methoxyisoquinoline 2-oxide (181 mg,
0.71 mmol) in acetonitrile (3.6 mL) was added TEA (0.20 mL, 1 .43 mmol) and
trimethylsilanecarbonitrile (0.29 mL, 2.14 mmol). The mixture was refluxed at 82 °C for 2 h, cooled, and concentrated in vacuo. The residue was basified with saturated aqueous Na2C03, extracted with DCM, dried with MgS04, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (0-20% EtOAc/heptane) to afford 7-bromo-6-methoxyisoquinoline-1 - carbonitrile (184 mg). MS (M+1 ) = 265.0
Step 3. 6-Methoxy-7-(4, 4, 5, 5-tetramethyl-1 , 3, 2-dioxaborolan-2-yl)isoquinoline-1-carbonitrile Following GENERAL METHOD 7-1 for boronate ester formation, 7-bromo-6- methoxyisoquinoline-1-carbonitrile (184 mg, 0.70 mmol) afforded 6-methoxy-7-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)isoquinoline-1 -carbonitrile (217 mg). MS (M+1 ) = 31 1.5
Step 4. 6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
yl)isoquinoline- 1 -carbonitrile
Following GENERAL METHOD 1 -1 for Suzuki coupling, 6-methoxy-7-(4,4,5,5-tetramethyl- 1 , 3, 2-dioxaborolan-2-yl)isoquinoline-1 -carbonitrile (100 mg, 0.32 mmol)
and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 1 , 61 mg, 0.22 mmol), affords 6-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)isoquinoline-1-carbonitrile (15 mg). MS (M+1 ) = 431.6
Step 5. 6-Hydroxy- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline- 1 -carbonitrile
Following standard GENERAL METHOD 2-1 for methoxy deprotection, the title compound was afforded. LC/MS Rt = 0.51. MS (M+1 ) = 417.2. 1H NMR (400 MHz, METHANOL-^) δ 8.63 (s, 1 H), 8.55 (d, J=5.8 Hz, 1 H), 8.50 (d, J=9.8 Hz, 1 H), 7.89-8.04 (m, 2H), 7.49 (s, 1 H), 5.05-5.24 (m, 1 H), 3.23 (s, 3H), 2.01 -2.21 (m, 4H), 1.69 (s, 6H), 1 .59 (s, 6H).
Example 4-1 : Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-7-ol
Figure imgf000063_0001
Step 1. 6-(7-Methoxyisoquinolin-6-yl)-N-methyl-N-(2,2^,6-tetramethylpi
yl)pyridazin-3-amine
The title compound (300 mg) was prepared from Intermediate 1 (240 mg, 0.849 mmol) and Intermediate 7 (350 mg, 1.724 mmol) following GENERAL METHOD 1 -1 for Suzuki coupling. MS (M+1 ) = 406.5
Step 2. 6-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-7-ol
The title compound (72 mg) was prepared from 6-(7-methoxyisoquinolin-6-yl)-N-methyl-N- (2,2,6, 6-tetramethylpiperidin-4-yl)pyridazin-3-amine (300 mg, 0.740 mmol) following GENERAL METHOD 2-4 for demethylation using thiophenol. LC/MS Rt = 0.39 min. MS (M+1 ) = 392.3. 1H NMR (400 MHz, METHANOL-^) δ 8.97 (s, 1 H), 8.33 (s, 1 H), 8.26 (d, J=9.8 Hz, 1 H), 8.16 (d, J=5.8 Hz, 1 H), 7.71 (d, J=5.8 Hz, 1 H), 7.40 (s, 1 H), 7.29 (d, J=10.0 Hz, 1 H), 5.23 (br s, 1 H), 2.97 (s, 3H), 1.59-1.79 (m, 4H), 1 .41 (s, 6H), 1 .26 (s, 6H). Example 5-1 : Synthesis of 8-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000064_0001
Step 1: 7-(Benzyloxy)-8-bromoquinoline
To a mixture of 8-bromoquinolin-7-ol (Chen, X., et al, WO 2010054006; 1257 mg, 5.61 mmol) and K2C03 (2.32 g, 16.8 mmol) in DMF (12 ml.) was added benzyl bromide (730 uL, 6.2 mmol), and the reaction mixture stirred overnight. The mixture was heated to 50 °C for 3 h to drive the reaction to completion. After cooling to room temperature, the mixture was diluted with 1 :1 EtOAc/diethyl ether, washed with water (6x), brine, dried over MgS04, and concentrated to a brown liquid. Column chromatography (DCM elution, followed by 0-10% EtOAc in DCM) provided 7- (benzyloxy)-8-bromoquinoline (801 mg) as an off-white solid. MS (M+1 ) = 314.1 . 1H NMR (400 MHz, CHLOROFORM-d) δ 9.05 (dd, J=1.8, 4.29 Hz, 1 H), 8.13 (dd, J=1.8, 8.34 Hz, 1 H), 7.77 (d, J=8.6 Hz, 1 H), 7.56 (d, J=l A Hz, 2H), 7.31-7.47 (m, 5H), 5.39 (s, 2H).
Step 2: (7-(Benzyloxy)quinolin-8-yl)boronic acid
Butyllithium (2.5 M in heptane, 0.42 ml_, 1.1 mmol) was added dropwise to a solution of 7-
(benzyloxy)-8-bromoquinoline (300 mg, 0.95 mmol) cooled to -78 °C. The solution was stirred for 0.5 h after which time trimethyl borate (0.270 ml_, 2.4 mmol) was added and the solution slowly warmed to room temperature and stirred overnight. LC/MS analysis shows complete conversion of the bromide to the boronic acid. The solution was rotovapped to dryness and concentrated from heptane (2x) to provide the crude (7-(benzyloxy)quinolin-8-yl)boronic acid (276 mg) as an off-white solid. MS (M+1 ) = 280.2. Based on LC/MS analysis, the yield of boronic acid was estimated to be near quantitative and this material was used directly in the subsequent Suzuki coupling.
Step 3: 6-(7-(Benzyloxy)quinolin-8-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
A mixture of 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine,
(Intermediate 1 , 200 mg, 0.707 mmol), the crude (7-(benzyloxy)quinolin-8-yl)boronic acid
(estimated 0.95 mmol) and sodium carbonate (300 mg, 2.83 mmol) in 3:1 DME/water (8 mL) was degassed with a stream of dry nitrogen for 5 min. Tetrakis(triphenylphosphine)palladium(0) was added (123 mg, 0.106 mmol) and the mixture heated at 140 °C under microwave irradiation for 30 min. The reaction was diluted with DCM/water, and extracted with DCM (4x). The extracts were acidified with HCI (4 equivalents) in MeOH and concentrated. The crude material was subjected to catch and release purification as described in GENERAL METHOD 3-1 , followed by silica gel chromatography as described in GENERAL METHOD 4-1 to provide 6-(7-(benzyloxy)quinolin-8-yl)- N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine as a light brown foam (214 mg). MS (M+1 ) = 482.5
Step 4: 8-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol To a solution of 6-(7-(benzyloxy)quinolin-8-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (176 mg, 0.365 mmol) in 1 :1 EtOAc/MeOH (3.6 ml.) under a nitrogen atmosphere was added palladium on carbon (10 wt %, 39 mg, 0.037 mmol). The atmosphere was replaced with hydrogen (balloon) and the mixture stirred rapidly overnight. The flask was flushed with nitrogen, diluted with DCM, filtered through Celite®, and concentrated. Silica gel
chromatography as described in GENERAL METHOD 4-1 provided 8-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol as a yellow foam (100 mg). LC/MS Rt = 0.41 min. MS (M+1 ) = 392.3. 1 H NMR (400 MHz, METHANOL-^) δ 8.83 (d, J=10.1 Hz, 1 H), 8.75 (dd, J=4.3, 1.8 Hz, 1 H), 8.22 (dd, J=8.1 , 1.5 Hz, 1 H), 7.81 (d, J=9.1 Hz, 1 H), 7.24-7.38 (m, 3H), 5.12 (t, J=12.4 Hz, 1 H), 3.04 (s, 3H), 1 .68-1.77 (m, 2H), 1.52-1.65 (m, 2H), 1.39 (s, 6H), 1 .24 (s, 6H).
Example 6-1 : Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
Figure imgf000065_0001
Step 1: 7-Bromo-6-methoxyquinoline
According to the general procedure of Martinez, R., et al, J. Org. Chem. 2008, 73, 9778-9780, in a 100 ml. round bottom flask, a mixture of (2-amino-4-bromo-5-methoxyphenyl)methanol
(Intermediate 10, 2.36 g, 9.14 mmol), benzophenone (3.33 g, 18.3 mmol), KOtBu (2.05 g, 18.3 mmol), and ethanol (0.53 ml, 9.1 mmol) in 1 ,4-dioxane (30 ml.) was heated at 80 °C for 2 h. The reaction was diluted with EtOAc and extracted with 1 M HCI (3x). The acidic extracts were washed with ether, basified to pH 10 and extracted with EtOAc (5x). The extracts were washed with brine, dried over MgS04 and concentrated to an orange oil. Silica gel chromatography (2-60% EtOAc in DCM) provided 7-bromo-6-methoxyquinoline (603 mg) as a yellow solid. MS (M+1 ) = 238.1 . 1H NMR (400 MHz, CHLOROFORM-d) δ 8.80 (dd, J=4.5, 1 .5 Hz, 1 H), 8.44 (s, 1 H), 8.12 (d, J=8.1 Hz, 1 H), 7.43 (dd, J=8.3, 4.3 Hz, 1 H), 7.12 (s, 1 H), 4.04 (s, 3H).
Step 2: (6-Methoxyquinolin-7-yl)boronic acid
Butyllithium (1.6 M in heptane, 1.90 ml_, 3.04 mmol) was added drop-wise over the course of one hour to a solution of 7-bromo-6-methoxyquinoline (0.603 g, 2.53 mmol) and triisopropyl borate (0.823 ml, 3.55 mmol) in THF (12.6 ml.) cooled to -78 °C. After the addition was complete, the cooling bath was removed and the slurry allowed to warm to room temperature. LC/MS analysis shows near quantitative conversion to the boronic acid. The mixture was concentrated, then the residue was concentrated sequentially from toluene and from heptane, to provide (6- methoxyquinolin-7-yl)boronic acid as an orange foam which was used immediately without purification. MS (M+1 ) = 204.2
Step 3. 6-( 6-Methoxyquinolin- 7-yl)-N-methyl-N-( 2, 2, 6, 6-tetramethylpiperidin-4-yl)pyridazin-3- amine
Suzuki coupling between 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine, (Intermediate 1 , 100 mg, 0.354 mmol) and the crude (6-methoxyquinolin-7-yl)boronic acid (0.530 mmol) using GENERAL METHOD 1 -2, SCX purification (GENERAL METHOD 3-1 , 2 g SWlaBond Propylsulfonic Acid® cartridge), silica gel chromatography (GENERAL METHOD 4-1), and palladium scavenging (GENERAL METHOD 6-1 , SiliaMetS® DMT (232 mg, 0.141 mmol)) provided 6-(6-methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine as a light yellow solid (105 mg). MS (M+1 ) = 406.3. 1 H NMR (400 MHz, METHANOL-^) δ 8.73 (dd, J=4.5, 1.5 Hz, 1 H), 8.34 (d, J=7.6 Hz, 1 H), 8.25 (s, 1 H), 7.83 (d, J=9.6 Hz, 1 H), 7.49-7.55 (m, 1 H), 7.48 (s, 1 H), 7.15 (d, J=9.6 Hz, 1 H), 5.25 (t, J=12.4 Hz, 1 H), 4.01 (s, 3H), 3.02 (s, 3H), 1 .66-1.76 (m, 2H), 1 .51 -1.64 (m, 2H), 1.38 (s, 6H), 1.23 (s, 6H).
Step 4. 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol 6-(6-Methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1. The crude material was subjected to silica gel chromatography (GENERAL METHOD 4-1 ) two times, to provide 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol as a light yellow solid (35 mg). LC/MS Rt = 0.47 min. MS (M+1 ) = 392.3. 1H NMR (400 MHz, METHANOL-^) 5 8.67 (dd, J=4.0, 1 .5 Hz, 1 H), 8.40 (s, 1 H), 8.30 (d, J=10.1 Hz, 1 H), 8.18 (d, J=8.1 Hz, 1 H), 7.45 (dd, J=8.3, 4.3 Hz, 1 H), 7.37 (d, J=10.1 Hz, 1 H), 7.33 (s, 1 H), 5.09-5.24 (m, 1 H), 3.05 (s, 3H), 1 .67- 1.76 (m, 2H), 1.53-1 .65 (m, 2H), 1 .40 (s, 6H), 1 .24 (s, 6H).
Example 6-2: Synthesis of 2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
Figure imgf000066_0001
Step 1. 7-Bromo-6-methoxy-2-methylquinoline According to the general procedure of Matsugi, M, et al, Tetrahedron Lett, 2000, 41, 8523, to a solution of 3-bromo-4-methoxyaniline (1 .00 g, 4.95 mmol) in 6 M HCI (25 ml.) and toluene (6 ml.) heated to 100 °C was added dropwise crotonaldehyde (0.407 ml_, 4.95 mmol). Stirring was continued for 2 h at 100 °C and the mixture then allowed to cool to room temperature. The toluene layer was removed and the aqueous layer basified to pH 7 by slow addition of 2 M NaOH. The aqueous phase was extracted with DCM (6x) and concentrated to a green residue. Following the general procedure for quinoline purification described by Leir, C. M. J. Org. Chem., 1977, 42, 91 1 , the residue was dissolved into 1 M HCI (75 ml.) and zinc chloride (1 .349 g, 9.90 mmol) was added resulting in immediate formation of a dark gummy solid. Sonication and stirring of the mixture transformed the gummy solid to a light brown precipitate. The solids were isolated by filtration, washing with 1 M HCI, 2-propanol, then water. The solids were partitioned between 1 :1
EtOAc/diethyl ether and 1 :1 water/concentrated ammonium hydroxide. The organic phase was washed with water, brine, dried over MgS04 and concentrated to a dark brown residue consisting of a mixture of the two possible cyclization regioisomers. Silica gel chromatography (3-40% gradient of EtOAc in DCM) provided the more mobile title compound, 7-bromo-6-methoxy-2- methylquinoline (365 mg), TLC (4:1 DCM/EtOAc) Rf 0.6, MS (M+1 ) = 254.0, 1 H NMR (400 MHz, METHANOL-^) δ 8.10-8.18 (m, 1 H), 7.38 (d, J=8.6 Hz, 1 H), 7.32 (s, 1 H), 3.99 (s, 3H), 2.66 (s, 3H), and the less mobile 5-bromo-6-methoxy-2-methylquinoline (70 mg), TLC (4:1 DCM/EtOAc) Rf 0.4, MS (M+1 ) = 254.0, 1H NMR (400 MHz, METHANOL-^) δ 8.49 (d, J=8.6 Hz, 1 H), 7.98 (d, J=9.1 Hz, 1 H), 7.67 (d, J=9.1 Hz, 1 H), 7.48 (d, J=8.6 Hz, 1 H), 4.03 (s, 3H), 2.71 (s, 3H).
Step 2. (6-Methoxy-2-methylquinolin-7-yl)boronic acid
Butyllithium (1.6 M in heptane, 0.41 mL, 0.65 mmol) was added dropwise over the course of 1 h to a solution of 7-bromo-6-methoxy-2-methylquinoline (150 mg, 0.60 mmol) and triisopropyl borate (0.180 mL, 0.77 mmol) cooled to -78 °C. After addition was complete, the reaction was stirred at -78 °C for one hour and then allowed to warm to room temperature and stir overnight. LC/MS analysis shows a 9:1 ratio of the title boronic acid to the debrominated side product (6- methoxy-2-methylquinoline). Solvent was removed via rotary evaporation, and the resulting solid concentrated from heptane (2x) to provide the crude (6-methoxy-2-methylquinolin-7-yl)boronic acid (MS (M+1 ) = 218.1 ) which was used directly in the subsequent Suzuki coupling estimating 90% yield based on LC/MS analysis.
Step 3: 6-( 6-Methoxy-2-methylquinolin- 7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
6-Chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, Intermediate 1 (107 mg, 0.38 mmol) and the crude (6-methoxy-2-methylquinolin-7-yl)boronic acid were subjected to Suzuki coupling using GENERAL METHOD 1 -2. The reaction was partitioned between DCM/water, extracted with DCM (3x), dried over MgS04 and concentrated. The crude material was subjected to silica gel chromatography (GENERAL METHOD 4-1 ) to provide 6-(6-methoxy-2- methylquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (149 mg) as an off-white solid. MS (M+1 ) = 420.1. 1H NMR (400 MHz, METHANOL-^) δ 8.21 (d, J=8.6 Hz, 1 H), 8.17 (s, 1 H), 7.81 (d, J=9.6 Hz, 1 H), 7.38-7.46 (m, 2H), 7.15 (d, J=9.6 Hz, 1 H), 5.25 (t, J=12.4 Hz, 1 H), 3.98 (s, 3H), 3.02 (s, 3H), 2.69 (s, 3H), 1.67-1 .76 (m, 2H), 1 .54-1.64 (m, 2H), 1 .39 (s, 6H), 1 .24 (s, 6H).
Step 4. 2-Methyl- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-6-ol
6-(6-Methoxy-2-methylquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (140 mg, 0.317 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-2. The crude material was subjected to silica gel chromatography
(GENERAL METHOD 4-1 ) to provide 2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol (97 mg) as a light yellow solid. LC/MS Rt = 0.47 min. MS (M+1 ) = 406.1. 1H NMR (400 MHz, METHANOL-^) δ 8.31 (s, 1 H), 8.28 (d, J=10.1 Hz, 1 H), 8.07 (d, J=8.6 Hz, 1 H), 7.38 (d, J=9.6 Hz, 1 H), 7.35 (d, J=8.6 Hz, 1 H), 7.29 (s, 1 H), 5.09-5.25 (m, 1 H), 3.05 (s, 3H), 2.68 (s, 3H), 1 .67-1.77 (m, 2H), 1.54-1 .65 (m, 2H), 1.40 (s, 6H), 1.24 (s, 6H).
Example 7-1 : Synthesis of 3-chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol hydrochloride salt
Figure imgf000068_0001
Step 1 : 6-(3-Chloro- 7-methoxyquinolin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
6-(7-Methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 9, 50 mg, 0.123 mmol), was borylated according to GENERAL METHOD 5-1 , with the exception that the reaction was heated at 65 °C overnight. To the crude boronic acid solution was added water (400 uL) and CuCI2 (49.7 mg, 0.370 mmol). The mixture was heated at 65 °C for 1 h, then allowed to stir at room temperature overnight. The mixture was partitioned between
DCM/water, and was washed with DCM (6x). The aqueous phase was acidified to pH 1 and concentrated to dryness. SCX purification of the resulting solids (GENERAL METHOD 3-1 ) followed by silica gel chromatography (GENERAL METHOD 4-1 ) provided 6-(3-chloro-7- methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (18 mg) as a light yellow solid. MS (M+1 ) = 440.3 Step 2: 3-Chloro-6-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol
6-(3-Chloro-7-methoxyquinolin-6-yl)-N-m
3-amine (18 mg) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-2. The crude material was subjected to silica gel chromatography (GENERAL
METHOD 4-1 ) to provide the title compound. The solid was dissolved into aqueous HCI (1.0 M, 0.21 ml_, 0.205 mmol)/water (300 uL) and the solution was lyophilized overnight to provide 3- chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
hydrochloride salt (14 mg) as a yellow/orange solid. LC/MS Rt = 0.53 min. MS (M+1 ) = 426.3. Free base: 1H NMR (400 MHz, METHANOL-^) δ 8.68 (d, J=2.5 Hz, 1 H), 8.33-8.39 (m, 2H), 8.27 (d, J=10.1 Hz, 1 H), 7.44 (s, 1 H), 7.35 (d, J=10.1 Hz, 1 H), 5.14-5.25 (m, 1 H), 3.05 (s, 3H), 1.68-1 .77 (m, 2H), 1.54-1 .66 (m, 2H), 1 .41 (s, 6H), 1 .25 (s, 6H).
Example 7-2: Synthesis of 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000069_0001
Step 1: 6-(3-Bromo-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
6-(7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 9, 100 mg, 0.247 mmol), was borylated according to GENERAL METHOD 5-1. To the crude boronic acid solution was added water (800 uL) and CuBr2 (165 mg, 0.740 mmol). The resulting mixture was stirred at room temperature overnight. The mixture was partitioned between DCM/water. The aqueous phase was extracted with EtOAc (5x), DCM (2x), and the combined organic extracts were concentrated. Silica gel chromatography (GENERAL METHOD 4-1 ) provided 6-(3-bromo-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine (73 mg) as a light yellow solid. MS (M+1 ) = 484.3. 1H NMR (400 MHz, METHANOL-^) δ 8.84 (d, J=2.5 Hz, 1 H), 8.57 (d, J=2.0 Hz, 1 H), 8.1 1 (s, 1 H), 7.84 (d, J=9.6 Hz, 1 H), 7.53 (s, 1 H), 7.15 (d, J=9.6 Hz, 1 H), 5.27 (t, J=12.1 Hz, 1 H), 4.03 (s, 3H), 3.02 (s, 3H), 1.66-1 .75 (m, 2H), 1.52- 1 .63 (m, 2H), 1.38 (s, 6H), 1.23 (s, 6H).
Step 2. 3-Bromo-6-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol
6-(3-Bromo-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazi 3-amine (73 mg) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-2. The crude material was subjected to silica gel chromatography (GENERAL
METHOD 4-1 ) to provide 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol (36 mg) as a bright yellow solid. LC/MS Rt = 0.53 min. MS (M+1 ) = 472.2. 1 H NMR (400 MHz, METHANOL-^) δ ppm 8.74 (d, J=2.0 Hz, 1 H), 8.51 (s, 1 H), 8.32 (s, 1 H), 8.26 (d, J=10.1 Hz, 1 H), 7.41 (s, 1 H), 7.34 (d, J=10.1 Hz, 1 H), 5.13-5.27 (m, 1 H), 3.04 (s, 3H), 1 .69-1.78 (m, 2H), 1.55-1.67 (m, 2H), 1 .41 (s, 6H), 1 .26 (s, 6H).
Example 7-3: Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-3-carbonitrile
Figure imgf000070_0001
To a vial containing 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-
3-yl)quinolin-7-ol (Example 7-2, 52 mg, 0.1 1 mmol), Pd(PPh3)4 (13 mg, 0.01 mmol), and zinc cyanide (26 mg, 0.22 mmol) was added DMF (1 ml_). The mixture was sealed and heated in a Biotage® Initiator microwave reactor at 120 °C for 1 h. Copper iodide (4 mg, 0.02 mmol) was added and the reaction mixture was heated under microwave irradiation at 120 °C for 1 h. The crude reaction mixture was cooled to room temperature, filtered through Celite55 and purified via reverse phase preparative HPLC (15 to 45% acetonitrile in water, 5 mM ammonium hydroxide modifier). The product-containing fractions were concentrated in vacuo to afford 7-hydroxy-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3-carbonitrile as a brown solid (5 mg). LC/MS Rt = 0.51 min. MS (M+1 ) = 417.2. 1H NMR (400 MHz, DMSO-d6) δ 9.01 (d, J=2.0 Hz, 1 H), 8.87 (d, J=2.0 Hz, 1 H), 8.64 (s, 1 H), 8.20-8.36 (m, 2H), 7.38-7.46 (m, 2H), 5.10 (br s, 1 H), 2.99 (s, 3H), 1 .44-1.74 (m, 4H), 1.32 (s, 6H), 1 .17 (s, 6H).
Example 7-4: Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(1 -methyl-1 H-imidazol-4-yl)quinolin-7-ol
Figure imgf000070_0002
Step 1. (7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amin
yl)quinolin-3-yl)boronic acid
The title compound was synthesized from 6-(7-methoxyquinolin-6-yl)-N-methyl-N-(2, 2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 9), following GENERAL METHOD 5-1 for borylation, and purified following GENERAL METHOD 3-1 to afford (7-methoxy-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-3-yl)boronic acid. MS (M+1 ) = 450.5
Step 2. 6-(7-Methoxy-3-(1 -methyl- 1H-imidazol-4-yl)quinolin-6-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-3- yl)boronic acid (0.24 g, 0.53 mmol), 4-bromo-1 -methyl-1 H-imidazole (0.26 g, 1.6 mmol), and Na2C03 (0.20 g, 1 .9 mmol) were added to a microwave vial. Pd(PPh3)4 (0.05 g, 0.04 mmol) was then added followed by addition of 1 ,4-dioxane (2.2 ml.) and water (0.6 ml_). The reaction mixture was sealed and heated in a Biotage® Initiator microwave reactor at 130 °C for 1 h. A solution of NaHC03was added to the reaction and the aqueous phase was extracted with DCM (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (1-10% 7 M ammonia in MeOH gradient, in DCM) to give 6-(7-methoxy-3-(1 -methyl-1 H-imidazol-4-yl)quinolin-6- yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine as a yellow solid (25 mg). MS (M+1 ) = 486.6
Step 3. 6-(6-(Methyl(2,2^^-tetramethylpiperidin-4-yl)amino)pyridaz^
imidazol-4-yl)quinolin- 7-ol
Following GENERAL METHOD 2-1 , methoxy deprotection of 6-(7-methoxy-3-(1 -methyl-1 H- imidazol-4-yl)quinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.05 g, 0.10 mmol) afforded 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(1 - methyl-1 H-imidazol-4-yl)quinolin-7-ol as a brown solid (5 mg). LC/MS Rt = 0.44 min. MS (M+1 ) = 472.3. 1H NMR (400 MHz, DMSO-d6) δ 13.52 (br s, 1 H), 9.19 (d, J=2.0 Hz, 1 H), 8.55 (s, 1 H), 8.54 (d, J=2.0 Hz, 2H), 8.34 (d, J=10.0 Hz, 1 H), 7.80 (d, J=1 .0 Hz, 1 H), 7.73 (d, J=1 .0 Hz, 1 H), 7.43 (d, J=10.0 Hz, 1 H), 7.35 (s, 1 H), 5.02 (br s, 1 H), 3.74 (s, 3H), 2.99 (s, 3H), 1.55 (dd, J=12.0, 3.5 Hz, 2H), 1.45 (t, J=12.0 Hz, 2H), 1 .27 (s, 6H), 1 .10 (s, 6H).
Example 7-5: Synthesis of 3-(1 H-imidazol-1 -yl)-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol hydrochloride salt
Figure imgf000071_0001
Step 1. 6-(3-(1H-lmidazol-1-yl)-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
yl)boronic acid (Example 7-4 Step 1 , 0.06 g, 0.14 mmol), imidazole (0.06 g, 0.8 mmol) and copper(ll) nitrate hemi(pentahydrate) (0.04 g, 0.2 mmol) were added to tetramethylethylenediamine (0.03 ml_, 0.20 mmol) in methanol (2 ml_). The reaction mixture was heated at 40 °C overnight. A 28% ammonium hydroxide solution was added to the reaction and the aqueous phase was extracted with DCM (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (1 -10% 7 M ammonia in MeOH gradient, in DCM) to provide 6-(3-(1 H-imidazol-1- yl)-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine. MS (M+1 ) = 472.5. 1H NMR (400 MHz, METHANOL-^) δ 9.13 (d, J=2.6 Hz, 1 H), 8.60 (d, J=2.6 Hz, 1 H), 8.36 (s, 1 H), 8.28 (s, 1 H), 7.89 (d, J=9.7 Hz, 1 H), 7.79 (s, 1 H), 7.65 (s, 1 H), 7.27 (s, 1 H), 7.19 (d, J=9.6 Hz, 1 H), 5.31 (br s, 1 H), 4.08 (s, 3H), 3.05 (s, 3H), 1.75 (dd, J=12.3, 2.9 Hz, 2H), 1 .56- 1 .70 (m, 2H), 1.42 (s, 6H), 1.27 (s, 6H).
Step 2. 3-(1H-lmidazol-1-yl)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin
3-yl) quinolin- 7-ol hydrochloride salt
and 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qu
3-ol
Following GENERAL METHOD 2-1 , methoxy deprotection of 6-(7-methoxy-3-(1 -methyl-1 H- imidazol-4-yl)quinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.04 g, 0.07 mmol) afforded a mixture of 7-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-3-ol (25 mg, MS (M+1 ) = 422.5), and 6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(1 -methyl-1 H-imidazol-4-yl)quinolin-7-ol which were separated by preparative reverse-phase HPLC. 6-(6-(Methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(1-methyl-1 H-imidazol-4-yl)quinolin-7-ol was suspended in
acetonitrile/H20 (3/1 ml_). 1 M Aqueous HCI (3 equivalents) was then added and solvent was concentrated in vacuo to afford 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)- 3-(1-methyl-1 H-imidazol-4-yl)quinolin-7-ol hydrochloride salt as a yellow solid (9.1 mg). LC/MS Rt = 0.42 min. MS (M+1 ) = 458.4. 1H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1 H), 9.19 (d, J=2.5 Hz, 1 H), 9.10-9.19 (m, 1 H), 8.74 (d, J=2.5 Hz, 1 H), 8.62 (s, 1 H), 8.36 (s, 1 H), 8.33 (d, J=10.0 Hz, 1 H), 8.17- 8.26 (m, 1 H), 7.96 (s, 1 H), 7.52-7.65 (m, 2H), 5.08-5.24 (m, 1 H), 3.04 (s, 3H), 2.04 (t, J=12.5 Hz, 2H), 1.80 (d, J=12.5 Hz, 2H), 1 .55 (s, 6H), 1 .49 (s, 6H).
Example 7-6: Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-3,7-diol formate salt.
Figure imgf000073_0001
A 1 M solution of BBr3 in DCM (1.2 ml_, 1.2 mmol) was added to 7-methoxy-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-3-ol (Example 7-5 Step 2, 50 mg, 0.12 mmol) in DCM (0.6 ml.) and the reaction was stirred at room temperature for 3 h.
MeOH was added to the reaction and the solvent was concentrated under reduced pressure. The crude material was purified via preparative reverse-phase HPLC (5 to 20% acetonitrile in water, 0.1 % formic acid modifier). The solvent was concentrated in vacuo to afford the formate salt of 6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3,7-diol (3 mg). LC/MS Rt = 0.42 min. MS (M+1 ) = 408.3. 1H NMR (400 MHz, DMSO-d6) δ 13.07 (br s, 1 H), 8.49 (d, J=2.5 Hz, 1 H), 8.37 (s, 1 H), 8.32 (d, J=10.0 Hz, 1 H), 8.29 (s, 2H), 7.50 (d, J=2.5 Hz, 1 H), 7.41 (d, J=10.0 Hz, 1 H), 7.29 (s, 1 H), 4.91 -5.18 (m, 1 H), 2.99 (s, 3H), 1 .62 (d, J=7.5 Hz, 4H), 1.35 (s, 6H), 1 .20 (s, 6H).
Example 8-1 : Synthesis of 3-ethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000073_0002
Step 1: 6-Bromo-3-ethyl-7-methoxyquinoline
Following the general procedure of Mierde, V. and Verpoort, et al, Tetrahedron Lett., 2009, 50, 201 , a solution of (2-amino-5-bromo-4-methoxyphenyl)methanol (Ramurthy, S., et al, WO 2008079988; 0.886 g, 3.82 mmol) and butyraldehyde (0.344 ml, 3.82 mmol) was heated at 80 °C in 1 ,4-dioxane (9 ml_) for 2 h. Benzophenone (0.765 g, 4.20 mmol) and potassium ferf-butoxide (0.514 g, 4.58 mmol) were added and the mixture heated at 120 °C for 0.5 h. The mixture was cooled to room temperature, diluted with EtOAc, washed with saturated aqueous sodium bicarbonate and brine. The organic phase was extracted with 1 M HCI (3x). The acidic extracts were basified to pH 12 by addition of 2 M NaOH and extracted with EtOAc (3x) and diethyl ether (2x). The organic extracts were concentrated to a yellow residue which was subjected to silica gel chromatography (2-30% EtOAc in DCM) providing 6-bromo-3-ethyl-7-methoxyquinoline (433 mg) as a yellow solid. MS (M+1 ) = 267.9. 1H NMR (400 MHz, METHANOL-^) δ ppm 8.69 (d, J=2.0 Hz, 1 H), 8.14 (s, 1 H), 8.04 (d, J=1.0 Hz, 1 H), 7.40 (s, 1 H), 4.03 (s, 3H), 2.84 (q, J=7.6 Hz, 2H), 1 .34 (t, J=7.6 Hz, 3H).
Step 2: (3-Ethyl-7-methoxyquinolin-6-yl)boronic acid From 6-bromo-3-ethyl-7-methoxyquinoline (150 mg, 0.564 mmol), 3-ethyl-7- methoxyquinolin-6-yl)boronic acid (0.564 mmol, estimated quantitative conversion based on LC/MS analysis. MS (M+1 ) = 232.1 ), was produced in a manner similar to that outlined in Example 5-1 , Step 2. This material was used directly in the subsequent Suzuki coupling.
Step 3: 6-(3^thyl-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetra^
yl)pyridazin-3-amine
6-Chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 1 , 1 15 mg, 0.407 mmol), and the crude 3-ethyl-7-methoxyquinolin-6-yl)boronic acid (132 mg, 0.564 mmol) were subjected to Suzuki coupling using GENERAL METHOD 1 -2. The reaction was partitioned between DCM/water, extracted with DCM (3x), dried over MgS04 and concentrated to a light yellow solid. The crude material was subjected to silica gel chromatography (GENERAL METHOD 4-1 ) to provide 6-(3-ethyl-7-methoxyquinolin-6-yl)-N-methyl-N-(2, 2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine (100 mg). MS (M+1 ) = 434.2. 1H NMR (400 MHz, METHANOL-^) δ 8.70 (d, J=2.0 Hz, 1 H), 8.17 (d, J=1.5 Hz, 1 H), 8.10 (s, 1 H), 7.83 (d, J=9.6 Hz, 1 H), 7.49 (s, 1 H), 7.15 (d, J=9.6 Hz, 1 H), 5.26 (t, J=12.4 Hz, 1 H), 4.01 (s, 3H), 3.01 (s, 3H), 2.81 - 2.92 (m, 2H), 1.66-1 .75 (m, 2H), 1 .51 -1.62 (m, 2H), 1 .32-1.42 (m, 9H), 1.23 (s, 6H).
Step 4. 3^thyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qu
7-ol
6-(3-Ethyl-7-methoxyquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine (100 mg, 0.231 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-2. The crude material was subjected to silica gel chromatography
(GENERAL METHOD 4-1 ) to provide 3-ethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol as a yellow solid (81 mg). LC/MS Rt = 0.44 min. MS (M+1 ) = 420.2. 1H NMR (400 MHz, METHANOL-^) δ 8.63 (d, J=2.0 Hz, 1 H), 8.33 (s, 1 H), 8.27 (d, J=9.6 Hz, 1 H), 8.13 (s, 1 H), 7.41 (s, 1 H), 7.35 (d, J=9.6 Hz, 1 H), 5.10-5.23 (m, 1 H), 3.04 (s, 3H), 2.84 (q, J=7.6 Hz, 2H), 1 .67-1.75 (m, 2H), 1.54-1 .65 (m, 2H), 1.40 (s, 6H), 1 .36 (t, J=7.6 Hz, 3H), 1.24 (s, 6H).
Example 8-2: Synthesis of 3-isopropyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000074_0001
Using procedures described in Example 8-1 , Steps 1 -4, beginning with 3-methyl butanal and (2-amino-5-bromo-4-methoxyphenyl)methanol, 3-isopropyl-6-(6-(methyl(2,2,6,6-tetramethyl- piperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol was produced as a light yellow solid. LC/MS Rt = 0.47 min. MS (M+1 ) = 434.5. 1H NMR (400 MHz, METHANOL-^) δ 8.66 (d, J=2.0 Hz, 1 H), 8.34 (s, 1 H), 8.27 (d, J=10.1 Hz, 1 H), 8.15 (d, J=2.0 Hz, 1 H), 7.40 (s, 1 H), 7.34 (d, J=10.1 Hz, 1 H), 5.15 (t, J=12.4 Hz, 1 H), 3.13 (m, 1 H), 3.04 (s, 3H), 1.67-1 .75 (m, 2H), 1 .53-1.64 (m, 2H), 1.36-1.42 (m, 12H), 1 .23 (s, 6H).
Example 9-1 : Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one
Figure imgf000075_0001
Step 1: 6-Bromo-7-methoxyquinoline 1 -oxide
To a solution of 6-bromo-7-methoxyquinoline (PREPARATION 6 Step 1 , 2 g, 8.40 mmol) in
DCM (42 mL) was added methyltrioxorhenium(VII) (0.209 g, 0.840 mmol). The solution was stirred for 5 min after which time hydrogen peroxide (30 wt % in water, 1 .03 mL, 10.1 mmol) was added. The solution was allowed to stir overnight during which time a thick precipitate formed. The mixture was diluted with heptane and the solids were isolated by filtration, washing with heptane. Drying of the solids in vacuo provided 6-bromo-7-methoxyquinoline 1-oxide (1.76 g). MS (M+1 ) = 255.9. 1H NMR (400 MHz, METHANOL-^) δ 8.57 (dd, J=6.1 , 1 .0 Hz, 1 H), 8.28 (s, 1 H), 7.95 (d, J=8.6 Hz, 1 H), 7.93 (s, 1 H), 7.34 (dd, J=8.3, 6.3 Hz, 1 H), 4.01 (s, 3H).
Step 2: 6-Bromo-7-methoxyquinolin-2(1H)-one
Trifluoroacetic anhydride (1 .4 mL, 9.9 mmol) was added dropwise to 6-bromo-7- methoxyquinoline 1-oxide (0.26 g, 1.03 mmol) in DMF (7 mL) and the mixture was stirred at room temperature overnight. Water (20 mL) was added to the reaction and the resulting solid was collected by filtration. The solid was triturated with hot acetonitrile and filtered to afford 6-bromo-7- methoxyquinolin-2(1 H)-one (0.17 g). MS (M+1 ) = 256.0. 1H NMR (400 MHz, DMSO-d6) δ 8.60 (d, J=6.1 Hz, 1 H), 8.49 (s, 1 H), 7.98 (s, 1 H), 7.86 (d, J=8.4 Hz, 1 H), 7.38 (dd, J=8.4, 6.1 Hz, 1 H), 4.05 (s, 3H).
Step 3: (7-Methoxy-2-oxo-1 ,2-dihydroquinolin-6-yl)boronic acid and 7-methoxy-6-(4,4,5,5- tetramethyl-1 , 3, 2-dioxaborolan-2-yl)quinolin-2( 1H)-one
Bis(pinacolato)diboron (0.33 g, 1.31 mmol), KOAc (0.19 g, 2.0 mmol), PdCI2(dppf).CH2CI2 (0.03 g, 0.03 mmol) and 6-bromo-7-methoxyquinolin-2(1 H)-one (0.17 g, 0.65 mmol) in DMSO (3.3 mL) were heated to 80 °C for 3 h. Additional portions of bis(pinacolato)diboron (0.17 g, 0.66 mmol) and KOAc (0.19 g, 2.0 mmol) were added and the mixture was heated at 80 °C overnight. The mixture was cooled to room temperature and the precipitate was collected by filtration to afford 7- methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinolin-2(1 H)-one. The filtrate was diluted with water and the aqueous phase was extracted with 3:1 chloroform/propan-2-ol (2x). To the combined organic phases was added to the previously collected 7-methoxy-6-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)quinolin-2(1 H)-one, and the solvent was concentrated in vacuo to afford a mixture (0.18 g) of 7-methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinolin-2(1 H)-one (MS (M+1 ) = 302.2) and (7-methoxy-2-oxo-1 ,2-dihydroquinolin-6-yl)boronic acid (MS (M+1 ) = 220.1 ).
Step 4:7-Methoxy-6-(6-(methyl(2,2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-2(1H)-one
The mixture of (7-methoxy-2-oxo-1 ,2-dihydroquinolin-6-yl)boronic acid and 7-methoxy-6- (4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinolin-2(1 H)-one (0.18 g), (Intermediate 1 , 0.17 g, 0.60 mmol), Na2C03 (0.19 g, 1.8 mmol) and Pd(PPh3)4 (0.07 g, 0.06 mmol) in 6:1 1 ,4- dioxane/water (3 mL) were heated in a Biotage® Initiator microwave reactor at 130 °C for 1 h.
Dichloromethane and 1 M HCI were added. The aqueous phase was washed with DCM (2x), then basified to pH 9 with a saturated solution of K2C03. The aqueous phase was extracted with DCM (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (2-10% 7 M ammonia in MeOH gradient, in DCM) to afford 7-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin- 4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one (16 mg). MS (M+1 ) = 422.3
Step 5: 7-Hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-2( 1H)-one
Following GENERAL METHOD 2-1 , methoxy deprotection of 7-methoxy-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one (16 mg, 0.038 mmol) afforded 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-2(1 H)-one (3.6 mg). LC/MS Rt = 0.44 min. MS (M+1 ) = 408.2. 1H NMR (400 MHz, METHANOL-^) δ 8.21 (d, J=10.0 Hz, 1 H), 8.14 (s, 1 H), 7.95 (d, J=9.5 Hz, 1 H), 7.36 (d, J=10.0 Hz, 1 H), 6.87 (s, 1 H), 6.44 (d, J=9.5 Hz, 1 H), 5.09-5.23 (m, 1 H), 3.04 (s, 3H), 1 .76 (dd, J=12.5, 3.5 Hz, 2H), 1.65 (t, J=12.5 Hz, 2H), 1 .44 (s, 6H), 1 .29 (s, 6H).
Example 9-2: Synthesis of 7-hydroxy-1 -methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one hydrochloride salt
Figure imgf000076_0001
Step 1. 6-Chloro-7-methoxyquinoline
The title compound was synthesized from 4-chloro-3-methoxyaniline in a manner analogous to PREPARATION 6 Step 1. MS (M+1 ) = 194.0. 1H NMR (400 MHz, METHANOL-^) δ 8.80 (dd, J=4.5, 1 .5 Hz, 1 H), 8.26 (dd, J=8.5, 1 .5 Hz, 1 H), 8.02 (s, 1 H), 7.49 (s, 1 H), 7.45 (dd, J=8.5, 4.5 Hz, 1 H), 4.07 (s, 3H).
Step 2. 6-Chloro-7-methoxyquinoline 1 -oxide
Methyltrioxorhenium (0.03 g, 0.12 mmol) was added to 6-chloro-7-methoxyquinoline (1 .69 g, 8.73 mmol) in DCM (40 mL). A solution of hydrogen peroxide (50 wt % in water, 1.0 mL, 17 mmol) was then added at 5 °C and the mixture was stirred at room temperature overnight. An additional portion of methyltrioxorhenium (0.03 g, 0.12 mmol) was added followed by hydrogen peroxide (50 wt % in water, 1.0 mL, 17 mmol) and the reaction was stirred for 4 h. The precipitate was collected by filtration to afford 6-chloro-7-methoxyquinoline 1 -oxide as a beige solid (1 .8 g). MS (M+1 ) = 210.0. 1H NMR (400 MHz, DMSO-d6) δ 8.59 (d, J=6.0 Hz, 1 H), 8.32 (s, 1 H), 8.03 (s, 1 H), 7.85 (d, J=8.5 Hz, 1 H), 7.39 (dd, J=8.5, 6.0 Hz, 1 H), 4.05 (s, 3H).
Step 3. 6-Chloro-7-methoxyquinolin-2(1H)-one
Trifluoroacetic anhydride (6.0 mL, 42 mmol) was added dropwise to 6-chloro-7- methoxyquinoline 1-oxide (1.18 g, 5.63 mmol) in DMF (8.0 mL) at 5 °C. The reaction mixture was heated in a Biotage® Initiator microwave reactor at 100 °C for 30 min. Water (20 mL) was added to the reaction and the resulting solid was collected by filtration. The solid was triturated with hot acetonitrile and filtered to afford 6-chloro-7-methoxyquinolin-2(1 H)-one (0.39 g). MS (M+1 ) = 210.1. 1H NMR (400 MHz, DMSO-d6) δ 1 1 .70 (br s, 1 H), 7.79 (d, J=9.5 Hz, 1 H), 7.79 (s, 1 H), 6.95 (s, 1 H), 6.37 (dd, J=9.5, 2.0 Hz, 1 H), 3.89 (s, 3H).
Step 4. 6-Chloro-7-methoxy-1 -methylquinolin-2(1H)-one
Potassium bis(trimethylsilyl)amide (0.5 M in toluene, 2.7 mL, 1.4 mmol) was added to 6- chloro-7-methoxyquinolin-2(1 H)-one (0.20 g, 0.95 mmol) in toluene (5 mL) and the mixture was stirred for 10 min at room temperature, lodomethane (0.07 mL, 1 mmol) was added to the reaction and the mixture was stirred 2 h at 50 °C. An additional portion of potassium bis(trimethylsilyl)amide (0.5 M in toluene, 2.7 mL, 1.4 mmol) was added and the mixture was stirred for 10 min at room temperature, lodomethane (0.08 mL, 1 mmol) was added to the reaction and the mixture was stirred at 50 °C overnight. Water was added to the reaction and the aqueous phase was extracted with DCM (3x). The combined organic phases were concentrated under reduced pressure. The crude material was purified by silica gel chromatography (gradient of 50-100% 3:1 EtOAc/EtOH, in heptane) to give 6-chloro-7-methoxy-1-methylquinolin-2(1 H)-one (0.14 g). MS (M+1 ) = 224.1 . 1H NMR (400 MHz, METHANOL-^) δ 7.82 (d, J=9.3 Hz, 1 H), 7.75 (s, 1 H), 7.1 1 (s, 1 H), 6.58 (d, J=9.4 Hz, 1 H), 4.08 (s, 3H), 3.78 (s, 3H). Step 5. 7-Methoxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinolin-2(1H)-one.
6-Chloro-7-methoxy-1-methylquinolin-2(1 H)-one (0.14 g, 0.62 mmol), tetrahydroxydiboron (0.22 g, 2.5 mmol), 2nd Generation XPhos Precatalyst (0.01 g, 0.02 mmol), XPhos (0.02 g, 0.04 mmol) and potassium acetate (0.24 g, 2.47 mmol) were stirred in ethanol (6.5 ml.) at 80 °C for 30 min. An aqueous solution of K2C03 (1 .8 M, 1 .1 ml_, 2.0 mmol) was added to the reaction at room temperature followed by Intermediate 1 (0.23 g, 0.80 mmol). The mixture was stirred at 80 °C for 2 h, then a 4% solution of Na2C03was added to the reaction and the aqueous phase was extracted with DCM (3x). The combined organic phases were extracted with 2 M HCI (3x). The combined acidic aqueous phases were basified to pH 1 1 with 6 M NaOH then extracted with DCM (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (2-15% 7 N ammonia in MeOH gradient, in DCM) to give an inseparable mixture of 7-methoxy-1 -methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one and Intermediate 1 (0.26 g). MS (M+1 ) = 436.5
Step 6. 7-Hydroxy- 1 -methyl-6-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinolin-2(1H)-one hydrochloride salt.
A 1 M solution of BBr3 in DCM (0.2 ml_, 3 mmol) was added to the mixture of 7-methoxy-1- methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one and Intermediate 1 (0.03 g) in DCM (0.6 ml.) at 5 °C and the reaction was stirred at 5 °C for 2 h. The solvent was concentrated under reduced pressure. The crude material was purified via preparative reverse-phase HPLC (5 to 20% acetonitrile in water, 7.5% formic acid modifier). The product- containing fractions were free based following GENERAL METHOD 3-1. The resulting solid was suspended in acetonitrile/H20 (3/1 ml_). 1 M aqueous HCI (3 equivalents) was added and solvent was concentrated in vacuo to afford 7-hydroxy-1 -methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one hydrochloride as a yellow solid (5 mg). LC/MS Rt = 0.48 min. MS (M+1 ) = 422.2. 1H NMR (400 MHz, DMSO-d6) δ 8.95-9.08 (m, 1 H), 8.21 -8.41 (m, 2H), 7.99-8.16 (m, 1 H), 7.84 (d, J=9.5 Hz, 1 H), 7.49-7.74 (m, 1 H), 6.99 (s, 1 H), 6.45 (d, J=9.5 Hz, 1 H), 5.00-5.20 (m, 1 H), 3.57 (s, 3H), 3.01 (s, 3H), 1 .99 (t, J=13.0 Hz, 2H), 1.80 (d, J=13.0 Hz, 2H), 1.53 (s, 6H), 1.46 (s, 6H).
Example 10-1 : Synthesis of 4-methoxy-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000079_0001
To a mixture of Intermediate 11 (150 mg, 0.32 mmol) in MeOH (1 .5 ml.) was added NaOMe (25 wt % in methanol, 0.36 ml_, 1 .6 mmol). The mixture was heated at 120 °C under microwave irradiation for 1 h. An additional portion of NaOMe (25 wt % in methanol, 0.2 ml.) was added and the mixture was then heated at 130 °C for 2 h. The mixture was diluted with aqueous NaHC03 solution and extracted with DCM. The combined organic extracts were dried over Na2S04, and concentrated in vacuo to give a light brown solid. The crude product was purified by silica gel chromatography (0-10% 2 M NH3 in MeOH /DCM), then by preparative reverse-phase HPLC to obtain the title compound as a white solid (82 mg). LC/MS Rt = 0.41 min. MS (M+1 ) = 436.3. 1H NMR (METHANOL-^) δ 8.39 (s, 1 H), 8.1 1 (d, J=10.1 Hz, 1 H), 7.27 (d, J=10.1 Hz, 1 H), 7.24 (s, 1 H), 6.66 (s, 1 H), 5.09 (t, J=12.1 Hz, 1 H), 4.06 (s, 3H), 3.00 (s, 3H), 2.59 (s, 3H), 1 .64-1.74 (m, 2H), 1.51 -1 .62 (m, 2H), 1.39 (s, 6H), 1.23 (s, 6H).
Example 10-2: Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(pyrrolidin-1 -yl)quinolin-7-ol
Figure imgf000079_0002
To a mixture of Intermediate 11 (22 mg, 0.046 mmol) in NMP (0.8 ml.) was added pyrrolidine (26.3 mg, 0.370 mmol). The mixture was heated at 130 °C under microwave irradiation for 2 h. The mixture was cooled to room temperature, acidified by addition of 1 M HCI in ether and subjected to SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge). The crude product was further purified by preparative reverse-phase HPLC to obtain 2- methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(pyrrolidin-1 - yl)quinolin-7-ol as a light yellow solid (7.5 mg). LC/MS Rt = 0.44 min. MS (M+1 ) = 475.2. 1 H NMR (METHANOL-^) δ 8.59 (s, 1 H), 8.14 (d, J=9.6 Hz, 1 H), 7.14 (d, J=9.6 Hz, 1 H), 6.77 (s, 1 H), 6.23 (s, 1 H), 5.20 (t, J=12.4 Hz, 1 H), 3.91 (br s, 4H), 2.99 (s, 3H), 2.48 (s, 3H), 2.03-2.17 (m, 4H), 1 .71 (d, J=3.5 Hz, 2H), 1.51 -1 .64 (m, 2H), 1 .39 (s, 6H), 1 .24 (s, 6H).
The following example compounds were prepared from the Intermediate 11 and the appropriate sodium alkoxide or amine according to the preparations of Example 10-1 or Example 10-2.
Figure imgf000080_0001
Example 11 -1 : Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(1 -methyl-1 H-pyrazol-4-yl)quinolin-7-ol
Figure imgf000080_0002
Step 1 : 6-(7-Methoxy-2-methyl-4-( 1 -methyl- 1 H-pyrazol-4-yl)quinolin-6-yl)-N-methyl-N-
(2, 2, 6, 6-tetramethylpiperidin-4-yl)pyridazin-3-amine
A mixture of 6-(4-chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2, 2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine (35 mg, 0.077 mmol) from PREPARATION 11 Step 5, 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1-methyl-pyrazole (48.1 mg, 0.231 mmol), XPhos Precatalyst (CAS No. 1028206-56-5, 5.70 mg, 7.71 umol) and cesium carbonate (100 mg, 0.308 mmol) in 1 ,4-dioxane (1 mL) and water (0.25 mL) was evacuated, filled with N2 (4x) and heated under microwave irradiation at 120 °C for 1 h. The reaction mixture was concentrated and acidified to pH 3 by addition of 4 M HCI in 1 ,4-dioxane. SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge) provided the crude product as a -2:1 mixture (23 mg) of 6- (7-methoxy-2-methyl-4-(1-methyl-1 H-pyrazol-4-yl)quinolin-6-yl)-N-methyl-N-(2,2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine (MS (M+1 ) = 500.2) and 7-methoxy-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)-pyridazin-3-yl)quinolin-4(1 H)-one.
Step 2: 2-Methyl-6-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-( methyl- 1H-pyrazol-4-yl)quinolin-7-ol
The mixture of 6-(7-methoxy-2-methyl-4-(1 -methyl-1 H-pyrazol-4-yl)quinolin-6-yl)-N-methyl- N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine and 7-methoxy-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)-pyridazin-3-yl)quinolin-4(1 H)-one (21 mg) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 using pyridine HCI. Preparative reverse-phase HPLC purification provided 2-methyl-6-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(1 -methyl-1 H-pyrazol-4-yl)quinolin-7-ol as a yellow solid (4.6 mg). LC/MS Rt = 0.42 min. MS (M+1 ) = 486.3. 1H NMR (METHANOL-^) δ 8.43 (s, 1 H), 8.14 (s, 1 H), 8.02 (d, J=9.6 Hz, 1 H), 7.92 (s, 1 H), 7.36 (s, 1 H), 7.24 (d, J=9.6 Hz, 1 H), 7.20 (s, 1 H), 5.14 (t, J=12.1 Hz, 1 H), 4.04 (s, 3H), 2.99 (s, 3H), 2.64 (s, 3H), 1.66-1.79 (m, 2H), 1 .53-1.66 (m, 2H), 1.41 (s, 6H), 1.26 (s, 6H).
Example 12-1 : Synthesis of 4-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol formate salt
Figure imgf000081_0001
To 4-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol (Intermediate 13, 0.05 g, 0.12 mmol) in MeOH (0.2 mL) was added NaOMe (25 wt % in methanol, 0.3 mL, 10 mmol). The mixture was heated at 165 °C under microwave irradiation for 1 h. The reaction mixture was diluted with DMSO (5 mL), acidified with formic acid (0.5 mL) and purified via preparative reverse-phase HPLC (5 to 20% acetonitrile in water, 7.5% formic acid as modifier) to afford 4-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol formate salt (6 mg) as a yellow solid. LC/MS Rt = 0.42 min. MS (M+1 ) = 422.5. 1H NMR
(METHANOL-^) δ 8.57 (d, J=5.5 Hz, 1 H), 8.40 (br s, 2H), 8.35 (s, 1 H), 8.33 (d, J=10.0 Hz, 1 H), 7.61 (s, 1 H), 7.42 (d, J=10.0 Hz, 1 H), 6.96 (d, J=5.5 Hz, 1 H), 5.48 (m, 1 H), 4.12 (s, 3H), 3.08 (s, 3H), 2.05-1 .94 (m, 4H), 1.68 (s, 6H), 1.53 (s, 6H).
Example 13-1 : Synthesis of 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoxalin-6-ol hydrochloride salt
Figure imgf000082_0001
Step 1: 4-Bromo-5-ethoxybenzene-1 ,2-diamine
Acetic acid (2 mL) was added drop-wise to 4-bromo-5-ethoxy-2-nitroaniline (5.00 g, 19.2 mmol) and zinc (6.47 g, 99.0 mmol) in CH2CI2 (100 mL) at 4 °C (Caution! Reaction is exothermic). After the exotherm, an additional portion of AcOH (2 mL) was added at 4 °C and the reaction was stirred at room temperature for three days. The reaction mixture was filtered and the filtrate was concentrated in vacuo to provide 4-bromo-5-ethoxybenzene-1 ,2-diamine (8.41 g). MS (M+1 ) = 231 .2
Step 2: 6-Bromo-7-ethoxyquinoxaline
Glyoxal (8.8 M in water, 1.7 mL, 15.0 mmol) was added to 4-bromo-5-ethoxybenzene-1 ,2- diamine (1.16 g, 5.00 mmol) in THF (50 mL) and the reaction mixture was stirred for 3 h. Silica gel (6 g) was added to the reaction and solvent was concentrated in vacuo. The material was purified by silica gel chromatography (gradient of 10-50% 3:1 EtOAc/EtOH, in heptane) to give 6-bromo-7- ethoxyquinoxaline (0.94 g). MS (M+1 ) = 255.2, 1H NMR (400 MHz, DMSO-d6) δ 8.91 (d, J=2.0 Hz, 1 H), 8.81 (d, J=2.0 Hz, 1 H), 8.38 (s, 1 H), 7.57 (s, 1 H), 4.32 (q, J=7.0 Hz, 2H), 1.46 (t, J=7.0 Hz, 3H).
Step 3: 6-(7-Ethoxyquinoxalin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4-yl)pyridazin- 3-amine
Following the general procedure of Bernhardt, S.; Manolikakes, G.; Kunz, T.; Knochel, P.
Angew. Chem. Int. Ed. 2011 , 50, 9205 -9209, Zn(OPiv)2-2 LiCI (1 .64 g, 4.64 mmol) was placed in a Schlenk-flask equipped with a magnetic stir bar and a septum, and dried for 5 min under vaccum with a heat gun. The zinc salt was dissolved in dry THF (10 mL). 6-Bromo-7-ethoxyquinoxaline
(0.94 g, 3.71 mmol) was added and the mixture was stirred for 5 min at room temperature.
Magnesium turnings (0.23 g, 9.28 mmol) were added. The reaction mixture was stirred at 50 °C for
4 h. The solution was transferred, via canula, to another reaction flask. Intermediate 1 (0.88 g, 3.09 mmol) and [1 ,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(ll) dichloride (0.21 g, 0.31 mmol) were added and the reaction mixture was heated at 50 °C for 5 days.
The reaction mixture was cooled to room temperature and the volatiles were removed under vacuum. 2 M aqueous HCI was added to the crude material and the aqueous phase was washed with DCM and basified to pH 1 1 with 6 M aqueous NaOH. The precipitate was collected by filtration and the filtrate was extracted with 9:1 DCM/MeOH (3x). The organic extracts and the collected precipitate were mixed and volatiles were removed under vacuum. The crude material was purified by silica gel chromatography (2-15% 7 M ammonia in MeOH gradient, in DCM) followed by a second purification via reverse-phase preparative HPLC (25-50% acetonitrile in water, 5 mM NH4OH as modifier). 6-(7-Ethoxyquinoxalin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (0.05 g) was provided as a brown solid. MS (M+1 ) = 421 .2. 1H NMR (400 MHz, METHANOL-^) δ 8.84 (d, J=2.0 Hz, 1 H), 8.78 (d, J=2.0 Hz, 1 H), 8.34 (s, 1 H), 7.95 (d, J=9.5 Hz, 1 H), 7.58 (s, 1 H), 7.20 (d, J=9.5 Hz, 1 H), 5.24-5.49 (m, 1 H), 4.35 (q, J=7.0 Hz, 2H), 3.05 (s, 3H), 1.76 (dd, J=12.5, 3.5 Hz, 2H), 1.64 (t, J=12.5 Hz, 2H), 1.51 (t, J=7.0 Hz, 3H), 1.43 (s, 6H), 1.28 (s, 6H).
Step 4: 7-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol hydrochloride salt.
A solution of BBr3 in DCM (1 M, 1 .0 mL, 1 mmol) was rapidly added to 6-(7- ethoxyquinoxalin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.04 mg, 0.10 mmol) in DCM (1 mL). The reaction mixture was stirred at 40 °C for 4 days. MeOH was added to the reaction at 0 °C and the solvent was concentrated under reduced pressure. The crude material was dissolved with 2:1 DMSO/7 N ammonia in MeOH (6 mL) and was purified via reverse- phase preparative HPLC (15-40% acetonitrile in water, 5 mM ammonium hydroxide as modifier). The product-containing fractions were concentrated in vacuo and the resulting solid was dissolved in 4:1 acetonitrile/H20 (5 mL). 1 M aqueous HCI (3 equivalents) was added and solvent was concentrated in vacuo to afford 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol hydrochloride as a brown solid (26 mg). LC/MS Rt = 0.48 min. MS (M+1 ) = 393.2. 1H NMR (400 MHz, METHANOL-^) δ 8.89 (d, J=2.0 Hz, 1 H), 8.83 (d, J=2.0 Hz, 1 H), 8.54 (d, J=10.0 Hz, 1 H), 8.44 (s, 1 H), 8.1 1 (d, J=10.0 Hz, 1 H), 7.58 (s, 1 H), 4.97 (br s, 1 H), 3.25 (s, 3H), 1 .95-2.29 (m, 4H), 1 .67 (s, 6H), 1 .59 (s, 6H).
PREPARATION 14
Intermediate 14: Synthesis of 7-bromo-6-methoxyisoquinolin-1 -ol
Figure imgf000083_0001
In a 10 mL microwave vial, 7-bromo-1-chloro-6-methoxyisoquinoline (240 mg, 0.881 mmol) was dissolved in acetic acid (5041 μΙ, 88 mmol). The mixture was heated to 150 °C for 180 min in microwave. Water was then added to the reaction vessel. The resulting precipitate was collected, washed with water, and then allowed to dry under vacuum to provide 7-bromo-6- methoxyisoquinolin-1 -ol, Intermediate 14 (171 mg, 0.660 mmol, 74.9 % yield). MS (M+1 ) = 256.3. 1H NMR (400 MHz, DMSO-d6) δ 1 1.27 (d, J=3.26 Hz, 1 H), 8.26 (s, 1 H), 7.30 (s, 1 H), 7.20 (dd, J=5.77, 7.03 Hz, 1 H), 6.52 (d, J=7.03 Hz, 1 H), 3.96 (s, 3H).
PREPARATION 15
Intermediate 15: Synthesis of 2-hydroxy-4-methoxy-5-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)benzaldehyde
Figure imgf000084_0001
Step 1: 5-Bromo-2-hydroxy-4-methoxybenzaldehyde
In a 250 mL flask, 2-hydroxy-4-methoxybenzaldehyde (5 g, 32.9 mmol) was dissolved in acetic acid (65.7 mL), and cooled to 0 °C, then Br2 (1 .862 ml, 36.1 mmol) in 10 mL acetic acid was added slowly. Upon complete addition, the reaction mixture was allowed to warm to room temperature, and stirred for 2 h. 100 mL of water was added to reaction flask, and the resulting white precipitate was filtered and washed with water. Drying under vacuum afforded the title compound (6.8 g, 29.4 mmol, 90 % yield). MS (M+1 ) = 233.3. 1H NMR (400 MHz,
CHLOROFORM-d) δ 1 1.45 (s, 1 H), 9.70 (s, 1 H), 7.69 (s, 1 H), 6.49 (s, 1 H), 3.96 (s, 3H).
Step 2: 2-Hydroxy-4-methoxy-5-(4, 4, 5, 5-tetramethyl- 1, 3, 2-dioxaborolan-2-yl)benzaldehyde
The title compound (4.9 g, 17.6 mmol, 82 % yield) was prepared following GENERAL METHOD 7-1 for boronate ester formation from 5-bromo-2-hydroxy-4-methoxybenzaldehyde (5.0 g, 21.64 mmol) and purified by silica gel chromatography (0-5% MeOH/DCM). MS (M+1 ) = 279.3. PREPARATION 16
Intermediate 16: Synthesis of 2-formyl-5-methoxy-4-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)phenyl trifluoromethanesulfonate
Figure imgf000084_0002
Step 1: 2-Hydroxy-4-methoxy-5-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)benzaldehyde The title compound (3.5 g, 8.78 mmol, 53%) was prepared following GENERAL METHOD 9-1 for Suzuki reaction of Intermediate 1 (4.7 g, 16.62 mmol) and Intermediate 15 (7.86 g, 28.3 mmol). MS (M+1 ) = 399.2.
Step 2: 2-Formyl-5-methoxy-4-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)ami
3-yl)phenyl trifluoromethanesulfonate
In a 100 mL flask, 2-hydroxy-4-methoxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)benzaldehyde (2.07 g, 5.19 mmol) and N-phenyltrifluoromethane sulfonimide (3.71 g, 10.39 mmol) were dissolved in DCM (26.0 mL) under N2. TEA (2.90 ml, 20.78 mmol) was added, and the heterogenous mixture was stirred for several hours at RT until complete dissolution was observed. The reaction was stirred overnight, then taken up in minimum amount of DCM and purified by silica gel chromatography (1-20% MeOH/DCM, elutes at 5% MeOH, column pretreated with TEA). The collected fractions were concentrated, and placed under high vacuum for several days to provide the title compound, (2.3 g, 4.03 mmol, 78 % yield). MS (M+1 ) = 399.2. PREPARATION 17
Intermediate 17: Synthesis of 2-formyl-5-methoxy-4-(6-((2,2,6,6-tetramethylpiperidin-
4-yl)oxy)pyridazin-3-yl)phenyl trifluoromethanesulfonate
Figure imgf000085_0001
Step 1: 2-Hydroxy-4-methoxy-5-( 6-((2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)benzaldehyde
The title compound (3.5 g, 8.78 mmol, 53%) was prepared following GENERAL METHOD
9-1 for Suzuki reaction of Intermediate 15 (2.63 g, 9.45 mmol) and Intermediate 3 (1 .5 g, 5.56 mmol). MS (M + 1 ) = 386.5. 1 H NMR (400 MHz, CHLOROFORM-d) δ 9.82 (s, 1 H), 8.20 (s, 1 H), 7.85 (d, J = 9.35 Hz, 1 H), 6.92 (d, J = 9.09 Hz, 1 H), 6.56 (s, 1 H), 5.76-5.90 (m, 1 H), 3.93 (s, 3H), 2.24 (dd, J = 4.04, 12.38 Hz, 2H), 1.35 (m, 8H), 1 .14-1.27 (s, 6H).
Step 2. 2-Formyl-5-methoxy-4-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)phenyl trifluoromethanesulfonate
2-Hydroxy-4-methoxy-5-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)benzaldehyde (500 mg, 1 .297 mmol) and N-phenyltrifluoromethane sulfonimide (927 mg, 2.59 mmol) were dissolved in DCM (6486 μΙ) under N2. Triethylamine (723 μΙ, 5.19 mmol) was added, and the heterogenous mixture was stirred for several hour at room temperature until complete dissolution was observed. After stirring overnight, the reaction was taken up in a minimum amount of dichloromethane, and purified by silica gel chromatography (1 -20% MeOH/DCM, elutes at 5% MeOH, column pretreated with TEA) to provide the title compound (650 mg, 1 .256 mmol, 97 % yield). MS (M + 1 ) = 518.1.
GENERAL METHOD 8-1 :
Representative Procedure for MP-Carbonate Free Basing
The trifluoroacetic acid salt of the product from preparatory reverse phase HPLC purification is dissolved in MeOH and loaded onto PL-HC03 MP® cartridge (Agilent Technologies)
preconditionsed with MeOH. The cartridge is then flushed with excess MeOH to provided the product as a free base. MP-carbonate, a resin-bound base, is also available from Biotage (part number 800267 for 10 grams). The chemical name for MP-carbonate is macroporous
triethylammonium methyl polystyrene carbonate.
GENERAL METHOD 9-1 for Suzuki reaction
Representative Procedure for Suzuki Coupling with Pd2(dba)3 and SPhos
A mixture of a chloropyridazine intermediate, such as Intermediate 1 (1.0 equivalent), a boronic acid reagent (1.5-2 equivalents), SPhos (0.2 equivalents), Pd2(dba)3 (0.05 equivalents), and K3P04 (3 equivalents) in 4:1 dioxane/water (0.2 M) was heated in an oil bath at 100 °C for 18 h. The mixture was concentrated in vacuo and the crude residue was purified via silica gel
chromatography (1 -30% MeOH/DCM, column pretreated with TEA).
GENERAL METHOD 10-1 for Sonogashira reaction
Representative Procedure for Sonogashira reaction
Intermediate 16 or Intermediate 17 (1 equivalent) was dissolved in dry acetonitrile (5A MS) and TEA (0.1 M, Ratio: 3:1 ). Acetylene (4 equivalents), Cul (0.6 equivalent), and Pd(PPh3)2CI2 (0.3 equivalents) were added. The vessel was purged with N2, and stirred at room temperature for 6 h. The mixture was concentrated in vacuo. The crude residue was taken up in dichloromethane, and purified by silica gel chromatography (1-20% MeOH/DCM, elutes at 10% MeOH, column pretreated with TEA), and used in next step without further purification.
GENERAL METHOD 11 -1 for Isoquinoline formation
Representative Procedure for aldehyde cyclization
In a microwave vial, the product from Sonogashira reaction (1 equivalent) and NH3 (2M in MeOH) (2-5 ml.) were combined and heated in the microwave for 30 min at 150 °C. The crude product was concentrated, diluted with a minimum amount of MeOH, and purified by prep-HPLC (0.1 % TFA/MeCN/H20) or taken on without further purification.
Example 14-1 : Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(tetrahydro-2H-pyran-4-yl)quinolin-7-ol
Figure imgf000087_0001
Using procedures described in Example 8-1 , Steps 1 -4, beginning with 2-(tetrahydro-2H- pyran-4-yl)acetaldehyde and (2-amino-5-bromo-4-methoxyphenyl)methanol, 6-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(tetrahydro-2H-pyran-4-yl)quinolin-7-ol was produced as a light yellow solid. LC/MS Rt = 0.48 min. MS (M+1 ) = 476.5. 1 H NMR (400 MHz, METHANOL-^) δ ppm 8.71 (d, J=2.0 Hz, 1 H), 8.40 (s, 1 H), 8.31 (d, J=9.6 Hz, 1 H), 8.20 (d, J=2.0 Hz, 1 H), 7.43 (s, 1 H), 7.38 (d, J=9.6 Hz, 1 H), 5.15-5.29 (m, 1 H), 4.12 (d, J=1 1.1 Hz, 2H), 3.57-3.73 (m, 2H), 3.07 (m, 4H), 1 .87-2.00 (m, 4H), 1.72-1 .81 (m, 2H), 1.60-1 .71 (m, 2H), 1 .45 (s, 6H), 1 .29 (s, 6H).
Example 15-1 : Synthesis of 3-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
Figure imgf000087_0002
Using procedures described in Example 7-1 , Steps 1 -2, beginning with 6-(6- methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Example 6- 1 Step 3, 50 mg, 0.123 mmol), 3-chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol (6 mg) was obtained as a light yellow solid. LC/MS Rt = 0.57 min. MS (M+1 ) = 426.2.
Example 15-2: Synthesis of 3-bromo-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
Using procedures described in Example 7-2, Steps 1 -2, beginning with 6-(6- methoxyquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Example 6- 1 Step 3, 200 mg, 0.493 mmol), 3-bromo-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol (48 mg) was obtained as a light yellow solid. LC/MS Rt = 0.60 min. MS (M+1 ) = 472.1. 1H NMR (400 MHz, METHANOL-^) δ 8.71 (d, J=2.3 Hz, 1 H), 8.44 (s, 1 H), 8.42 (d, J=2.2 Hz, 1 H), 8.34 (d, J=9.9 Hz, 1 H), 7.40 (d, J=9.9 Hz, 1 H), 7.30 (s, 1 H), 5.37-5.17 (m, 1 H), 3.08 (s, 3H), 1.75 (dd, J=36.1 , 12.5 Hz, 4H), 1 .47 (s, 6H), 1 .32 (s, 6H).
Example 15-3: Synthesis of 3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol
Figure imgf000088_0001
Step 1. 7-Bromo-6-methoxy-3-methylquinoline
According to the general procedure of Matsugi, M, et al, Tetrahedron Lett, 2000, 41, 8523, to a solution of 3-bromo-4-methoxyaniline (1 .00 g, 4.95 mmol) in 6 M HCI (25 mL) and toluene (6 mL) heated to 100 °C was added dropwise methacrolein (0.408 mL, 4.95 mmol). Heating was continued for two hours. The reaction was cooled to room temperature and the phases separated. The aqueous phase was basified to pH 12 by addition of 2 M NaOH, extracted with DCM (5x), and ethyl acetate (2x). The combined organics were dried over MgS04 and concentrated to provide a green residue. Silica gel chromatography (3-40% gradient of EtOAc in DCM) provided the more mobile title compound, 7-bromo-6-methoxy-3-methylquinoline (373 mg) as an off-white solid, TLC (9:1 DCM/EtOAc) Rf 0.6, MS (M+1 ) = 254.2, 1H NMR (400 MHz, METHANOL-^) δ ppm 8.57 (d, J=2.0 Hz, 1 H), 8.18 (s, 1 H), 8.07 (s, 1 H), 7.32 (s, 1 H), 4.01 (s, 3 H), 2.50 (s, 3 H), and the less mobile isomer, 5-bromo-6-methoxy-3-methylquinoline, (245 mg) as a brown solid, TLC (9:1 DCM/EtOAc) Rf 0.6, MS (M+1 ) = 254.2, 1H NMR (400 MHz, METHANOL-^) δ ppm 8.63 (d, J=2.0 Hz, 1 H), 8.35-8.43 (m, 1 H), 8.03 (d, J=8.6 Hz, 1 H), 7.66 (d, J=9.1 Hz, 1 H), 4.06 (s, 3H), 2.59 (s, 3H).
Step 2. (6-Methoxy-3-methylquinolin-7-yl)boronic acid
Using procedures described in Example 6-2, Step 2, starting with 7-bromo-6-methoxy-3- methylquinoline (373 mg), (6-methoxy-3-methylquinolin-7-yl)boronic acid was obtained as an off- white solid (176 mg) after purification of the crude product by silica gel chromatography (12 g silica gel, 2-30% MeOH in DCM over 30 column volumes). MS (M+1 ) = 218.3. 1 H NMR (400 MHz, METHANOL-^) δ ppm 8.54 (d, J=2.0 Hz, 1 H), 8.04 (s, 1 H), 7.83 (s, 1 H), 7.21 (s, 1 H), 3.95 (s, 3H), 2.52 (s, 3H).
Step 3. 6-( 6-Methoxy-3-methylquinolin- 7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
Using procedures described in Example 6-2, Step 3, starting with (6-methoxy-3- methylquinolin-7-yl)boronic acid (172 mg, 0.79 mmol), 6-(6-methoxy-3-methylquinolin-7-yl)-N- methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (144 mg) was obtained as a light yellow solid. MS (M+1 ) = 420.6. 1 H NMR (400 MHz, METHANOL-^) δ ppm 8.59 (d, J=2.0 Hz, 1 H), 8.19 (s, 1 H), 8.10 (s, 1 H), 7.81 (d, J=9.6 Hz, 1 H), 7.38 (s, 1 H), 7.14 (d, J=9.6 Hz, 1 H), 5.23 (t, J=12.4 Hz, 1 H), 3.99 (s, 3H), 3.01 (s, 3H), 2.54 (s, 3H), 1.66-1.74 (m, 2H), 1 .51-1.62 (m, 2H), 1.38 (s, 6H), 1.23 (s, 6H).
Step 4. 3-Methyl- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-6-ol
6-(6-Methoxy-3-methylquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (144 mg, 0.34 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-2. The crude material was subjected to silica gel chromatography
(GENERAL METHOD 4-1 ) to provide 3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-6-ol as a yellow solid (107 mg). LC/MS Rt = 0.49 min MS (M+1 ) = 406.6. 1H NMR (400 MHz, METHANOL-^) δ ppm 8.54 (d, J=2.0 Hz, 1 H), 8.35 (s, 1 H), 8.28 (d, J=10.1 Hz, 1 H), 7.95 (br. s., 1 H), 7.37 (d, J=9.6 Hz, 1 H), 7.24 (s, 1 H), 5.08-5.24 (m, 1 H), 3.04 (s, 3H), 2.51 (s, 3H), 1.68-1 .77 (m, 2H), 1.54-1.66 (m, 2H), 1 .40 (s, 6H), 1 .24 (s, 6H).
Example 16-1 : Synthesis of 5-bromo-3-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol
Figure imgf000089_0001
To a solution of 3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-6-ol (Example 15-3, 36 mg, 0.089 mmol) in DCM (1 ml.) was added N- bromosuccinimide (19 mg, 0.1 1 mmol). After stirring for one hour, the mixture was diluted with ethyl acetate, washed with saturated sodium bisulfite, brine, dried over magnesium sulfate and concentrated under reduced pressure. The crude material was purified by silica gel chromatography according to GENERAL METHOD 4-1 to provide 5-bromo-3-methyl-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol (20 mg) as a yellow solid. LC/MS Rt = 0.58 min. MS (M+1 ) = 486.4. 1H NMR (400 MHz, METHANOL-^) δ ppm 8.58 (d, J=2.0 Hz, 1 H), 8.38 (s, 1 H), 8.34 (d, J=10.1 Hz, 1 H), 8.32 (s, 1 H), 7.40 (d, J=10.1 Hz,1 H), 5.22 (m, 1 H), 3.07 (s, 3H), 2.59 (s, 3H), 1 .73-1 .82 (m, 2H), 1.60-1.72 (m, 2H), 1 .46 (s, 6H), 1 .30 (s, 6H).
Example 17-1 : Synthesis of 6-hydroxy-1 -methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-4(1H)-one
Figure imgf000090_0001
Step 1: 6-methoxy-1-methyl-7-(6-(methyl(2,2, 6, 6-tetramethylpiperidin-4-yl)amino)pyrM 3-yl)quinolin-4(1H)-one
6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
4(1 H)-one (PREPARATION 13, Step 3) (0.07 g, 0.17 mmol), iodomethane (0.05 g, 0.32 mmol) and KH (50% by weight in paraffin, 0.02 g, 0.03 mmol) were stirred in dry TH F (2.0 mL) and DMF (0.6 mL) at room temperature under dry nitrogen for 1 h. A solution of sodium bicarbonate was added to the reaction mixture. The aqueous phase was extracted with 9: 1 DCM/MeOH (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (2-15% 7 M ammonia in MeOH gradient, in DCM) to afford 6-methoxy-1 -methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-4(1 H)-one as a brown solid (90 mg). MS (M+1 ) = 436.5. 1 H NMR (400 MHz, METHANOL-^) δ 8.10 (s, 1 H), 8.02 (d, J=7.4 Hz, 1 H), 7.98 (d, J=9.8 Hz, 1 H), 7.92 (s, 1 H), 7.18 (d, J=9.7 Hz, 1 H), 6.33 (d, J=7.4 Hz, 1 H), 5.47-5.30 (m, 1 H), 4.01 (s, 3H), 4.01 (s, 3H), 3.03 (s, 3H), 1 .79 (d, J=13.1 Hz, 2H), 1 .70 (t, J=12.9 Hz, 2H), 1 .47 (s, 6H), 1 .32 (s, 6H).
Step 2: 6-Hydroxy- 1 -methyl- 7-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinolin-4(1H)-one
A solution of BBr3 (1 .0 M in DCM, 1 .0 mL, 1 .0 mmol) was added to 6-methoxy-1 -methyl-7- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-4(1 H)-one (0.09 g, 0.20 mmol) in DCM (2.0 mL) and the reaction was stirred at reflux for 5 h. The reaction mixture was then added dropwise to MeOH (10 mL) and the solvent was concentrated under reduced pressure. The crude material was purified via preparative reverse-phase HPLC (15 to 40% acetonitrile in water, 5 mM ammonium hydroxide modifier) and the product-containing fractions were concentrated in vacuo. The resulting solid was suspended in acetonitrile/water (3/1 mL) and 1 M aqueous HCI (3 equivalents) was then added. The solvent was concentrated in vacuo to afford 6-hydroxy-1 -methyl- 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-4(1 /-/)-one hydrochloride salt as a yellow solid (28 mg). LC/MS Rt = 0.43 min. MS (M+1 ) = 422.2. H NMR (400 MHz, METHANOL-^) δ 8.80 (d, J=7.0 Hz, 1 H), 8.58 (d, J=10.2 Hz, 1 H), 8.57 (s, 1 H), 8.06 (d, J=10.0 Hz, 1 H), 7.94 (s, 1 H), 7.13 (d, J=6.7 Hz, 1 H), 5.02 (br.s, 1 H), 4.43 (s, 3H), 3.26 (s, 3H), 2.27-2.02 (m, 4H), 1 .68 (s, 6H), 1 .59 (s, 6H). Example 18-1 : Synthesis of 2,3-dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoxalin-6-ol
Figure imgf000091_0001
Step 1: 6-Bromo-7-ethoxy-2,3-dimethylquinoxaline
Biacetyl (0.51 ml_, 5.85 mmol) was added to 4-bromo-5-ethoxybenzene-1 ,2-diamine
(Example 13-1 , Step 1) (1 .04 g, 4.50 mmol) and ammonium chloride (0.24 g, 4.50 mmol) in MeOH (45 ml.) and the reaction was stirred at reflux for 3 h. Silica gel (6 g) was added and the solvent was removed in vacuo. Silica gel chromatography (gradient of 5-40% 3: 1 EtOAc/EtOH, in heptane) provided 6-bromo-7-ethoxy-2,3-dimethylquinoxaline as an orange solid (1 .04 g). MS = 281 .2. 1 H NMR (400 MHz, METHANOL-^) δ 8.15 (s, 1 H), 7.34 (s, 1 H), 4.28 (q, J=7.0 Hz, 2H), 2.70 (s, 3H), 2.70 (s, 3H), 1 .55 (t, J=7.0 Hz, 3H).
Step 2: (7-Ethoxy-2,3-dimethylquinoxalin-6-yl)boronic acid
Bis(pinacolato) diboron (0.50 g, 2.0 mmol), KOAc (0.29 g, 3.0 mmol), and PdCI2(dppf).CH2Cl2 (0.06 g, 0.07 mmol) were added to 6-bromo-7-ethoxy-2,3-dimethylquinoxaline (0.28 g, 0.99 mmol) in dioxane (5 ml_). The mixture heated at 80 °C overnight under an atmosphere of nitrogen. The reaction mixture was cooled to RT, filtered through Celite® (pre-packed filter funnel) washing with EtOAc, and concentrated in vacuo to afford (7-ethoxy-2,3-dimethylquinoxalin- 6-yl)boronic acid as a black solid. MS (M+1 ) = 247.1 .
Step 3: 6-(7-Ethoxy-2, 3-dimethylquinoxalin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
(7-Ethoxy-2,3-dimethylquinoxalin-6-yl)boronic acid (0.49 g, 1 .98 mmol), 6-chloro-N-methyl- N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.85 g, 3.00 mmol), and Na2C03 (0.81 g, 3.9 mmol) were added to a microwave vial. Pd(PPh3)4 (0.23 g, 0.20 mmol) was then added followed by addition of 1 ,4-dioxane (8.5 ml.) and water (1 .4 ml_). The reaction mixture was sealed and heated in a Biotage® Initiator microwave reactor at 130 °C for 1 h. A solution of NaHC03 was added to the reaction and the aqueous phase was extracted with DCM (3x). The combined organic phases were dried over anhydrous Na2S04 filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (1 -15% 7 M ammonia in MeOH gradient, in DCM) followed by a second purification via reverse-phase preparative HPLC (5-20% acetonitrile in water, 7.5% formic acid modifier). The product-containing fractions were free-based by catch and release using SiliaBond Propylsulphonic Acid® (4 eq, acetonitrile as eluent and a 2 N ammonia solution in MeOH to release the material). The solvent was concentrated in vacuo to give 6-(7- ethoxy-2,3-dimethylquinoxalin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidi
as a yellow solid (0.35 g). MS (M+1 ) = 449.5. 1 H NMR (400 MHz, METHANOL-^) δ 8.23 (s, 1 H), 7.93 (d, J=9.6 Hz, 1 H), 7.47 (s, 1 H), 7.20 (d, J=9.7 Hz, 1 H), 5.31 (t, J=12.6 Hz, 1 H), 4.32 (q, J=7.0 Hz, 2H), 3.06 (s, 3H), 2.77 (s, 3H), 2.75 (s, 3H), 1 .77 (dd, J=12.6, 3.5 Hz, 2H), 1 .64 (t, J=12.4 Hz, 2H), 1 .50 (t, J=7.0 Hz, 3H), 1 .44 (s, 6H), 1 .29 (s, 6H).
Step 4: 2, 3-Dimethyl-7-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol
A solution of BBr3 (1 .0 M in DCM, 7.7 mL, 7.7 mmol) was added to 6-(7-ethoxy-2,3- dimethylquinoxalin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.35 g, 0.77 mmol) in DCM (7 mL). The reaction mixture was stirred at 40 °C overnight. The reaction mixture was added to MeOH (50 mL) at 0 °C and the solvent was concentrated under reduced pressure. The crude material was purified by silica gel chromatography (1 -15% 7 M ammonia in MeOH gradient, in DCM). The product-containing fractions were concentrated in vacuo and the resulting solid was dissolved in 4: 1 acetonitrile/water (15 mL). 1 M aqueous HCI (3 equivalents) was added and solvent was concentrated in vacuo to afford the hydrochloride salt of 2,3-dimethyl- 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol as a yellow solid (0.26 g). LC/MS Rt = 0.53 min. MS (M+1 ) = 421 .3. 1 H NMR (400 MHz, METHANOL-^) δ 8.52 (d, J=8.2 Hz, 1 H), 8.43 (s, 1 H), 8.1 1 (br.s, 1 H), 7.56 (s, 1 H), 4.92 (partially obscured by residual water peak, 1 H), 3.25 (s, 3H), 2.86 (s, 3H), 2.81 (s, 3H), 2.36-1 .95 (m, 4H), 1 .66 (s, 6H), 1 .58 (s, 6H).
Examples 18-2 and 18-3: Synthesis of 2-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol and 3-methyl-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin-6-ol
Figure imgf000092_0001
Step 1: 6-Bromo-7-ethoxy-2-methylquinoxaline and 7-bromo-6-ethoxy-2-methylquinoxaline 2-Oxopropanal (8.26 g, 45.8 mmol) was added dropwise to 4-bromo-5-ethoxybenzene-1 ,2- diamine (Example 13-1 Step 1) (8.41 g, 36.4 mmol) in THF (360 mL) and the reaction was stirred overnight. The solvent was removed in vacuo and the crude material purified by silica gel chromatography (gradient of 25-50% 3:1 EtOAc/EtOH, in heptane) to give a mixture of 6-bromo-7- ethoxy-2-methylquinoxaline and 7-bromo-6-ethoxy-2-methylquinoxaline (2.04 g). MS (M+1 ) = 269.1 . 1 H NMR (400 MHz, CHLOROFORM-d) as a 1 .7:1 .0 mixture of regioisomers δ 8.67 (s) and 8.60 (s) (1 H), 8.30 (s) and 8.26 (s) (1 H), 7.36 (s) and 7.35 (s) (1 H), 4.26 (q, J=7.0 Hz) and 4.26 (q, J=7.0 Hz) (2H), 2.71 (s, 3H), 1 .57 (t, J=6.9 Hz) and 1 .54 (t, 7.0 Hz) (3H). Step 2: 6-Ethoxy-2-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinoxaline, (7- ethoxy-3-methylquinoxalin-6-yl)boronic acid, and 7-ethoxy-2-methyl-6-(4, 4, 5, 5-tetramethyl- 1, 3, 2- dioxaborolan-2-yl)quinoxaline and (7-ethoxy-2-methylquinoxalin-6-yl)boronic acid
Bis(pinacolato)diboron (4.85 g, 19.1 mmol), KOAc (2.25 g, 22.3 mmol), PdCI2(dppf).CH2CI2 (0.47 g, 0.08 mmol), 6-bromo-7-ethoxy-2-methylquinoxaline and 7-bromo-6-ethoxy-2- methylquinoxaline (2.04 g, 7.64 mmol) in dioxane (75 ml.) were heated at 80 °C overnight. The reaction mixture was cooled to RT, filtered through Celite® (pre-packed filter funnel) washing with EtOAc, and concentrated in vacuo. Silica gel chromatography (gradient of 5-30% MeOH in DCM) afforded a 2.1 :1.0 mixture of (7-ethoxy-3-methylquinoxalin-6-yl)boronic acid, (7-ethoxy-2- methylquinoxalin-6-yl)boronic acid and 6-ethoxy-2-methyl-7-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)quinoxaline, 7-ethoxy-2-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)quinoxaline as a dark oil. (M+1 ) = 233.3 and 315.3.
Step 3: 6-(7-Ethoxy-3-methylquinoxalin-6-yl)-N-methyl-N-(2,2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine, and 6-(7-ethoxy-2-methylquinoxalin-6-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
A mixture of 6-ethoxy-2-methyl-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinoxaline, (7-ethoxy-3-methylquinoxalin-6-yl)boronic acid, 7-ethoxy-2-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)quinoxaline and (7-ethoxy-2-methylquinoxalin-6-yl)boronic acid, 6-chloro-N- methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 1 , 2.52 g, 8.90 mmol), aqueous tripotassium phosphate (0.5 M, 36 ml_, 18 mmol), 2nd generation XPhos precatalyst (0.21 g, 0.27 mmol) and XPhos (0.12 g, 0.27 mmol) in 1 :1 THF/water (64 ml.) was stirred at 50 °C overnight. The reaction mixture was concentrated in vacuo and diluted with water. The aqueous phase was extracted with 9:1 DCM/MeOH (3x). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. Silica gel chromatography (1-15% 7 M ammonia in MeOH gradient, in DCM) provided 6-(7-ethoxy-3-methylquinoxalin-6-yl)-N- methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine and 6-(7-ethoxy-2-methylquinoxalin- 6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (2.60 g) as a 2:1 mixture of regioisomers). MS (M+1 ) = 435.5. 1H NMR (400 MHz, METHANOL-^) δ 8.75 (s) and 8.68 (s) (1 H), 8.27 (s) and 8.23 (s) (1 H), 7.91 (d, J=9.7 Hz) and 7.90 (d, J=9.7 Hz) (1 H), 7.52 (s) and 7.46 (s) (1 H), 7.20-7.10 (m, 1 H), 5.27 (t, J=12.6 Hz, 1 H), 4.31 (q, J=7.0 Hz) and 4.30 (q, J=6.9 Hz) (2H), 3.02 (s) and 2.93 (s) (3H), 2.75 (s) and 2.74 (s) (3H), 1 .72 (dd, J=12.5, 3.5 Hz) and 1.68-1 .50 (m) (4H), 1.48 (t, J=7.0 Hz) and 1 .48 (t, J=7.0 Hz) (3H), 1.39 (s) and 1 .33 (s) (6H), 1.24 (s) and 1 .20 (s) (6H). Step 4: 2-Methyl-7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol, and 3-methyl- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoxalin-6-ol
A solution of BBr3 (1.0 M in DCM, 16 mL, 16 mmol) was added to 6-(7-ethoxy-2- methylquinoxalin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine and 6-(7- ethoxy-3-methylquinoxalin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (0.70 g, 1.61 mmol) in DCM (16 mL) and the reaction was stirred at reflux overnight. The mixture was then added dropwise to MeOH (50 mL) at 0 °C and the solvent concentrated under reduced pressure. The crude material was purified via preparative reverse-phase HPLC (15 to 40% acetonitrile in water, 5 mM ammonium hydroxide modifier) and the regioisomers were separated via preparative SFC (AS-H 21x250mm column, 15% MeOH 10mM NH4OH in C02). Residual palladium was scavenged from each using GENERAL METHOD 6-1. The two solids were separately suspended in 3:1 acetonitrile/water (8 mL). 1 M aqueous HCI (3 equivalents) was added and the solvent concentrated in vacuo to afford two yellow solids, the major regioisomer (56 mg) and the minor regioisomer (19 mg). NMR structure determination attempts were not able to conclusively distinguish the two regioisomers.
Major regioisomer, 18-3 LC/MS Rt = 0.49 min. MS (M+1 ) = 407.2. 1H NMR (400 MHz, METHANOL-^) δ 8.80 (s, 1 H), 8.53 (d, J=9.9 Hz, 1 H), 8.43 (s, 1 H), 8.08 (d, J=10.0 Hz, 1 H), 7.50 (s, 1 H), 5.00 (partially obscured by residual water peak, 1 H) 3.23 (s, 3H), 2.80 (s, 3H), 2.19-1.99 (m, 4H), 1.66 (s, 6H), 1.58 (s, 6H).
Minor regioisomer, 18-2 LC/MS Rt = 0.50 min; MS (M+1 ) = 407.3. 1 H NMR (400 MHz, METHANOL-^) δ 8.86 (s, 1 H), 8.54 (d, J=9.9 Hz, 1 H), 8.34 (s, 1 H), 8.10 (d, J=9.9 Hz, 1 H), 7.56 (s, 1 H), 4.97 (partially obscured by residual water peak, 1 H), 3.24 (s, 3H), 2.77 (s, 3H), 2.23-1.98 (m, 4H), 1.66 (s, 6H), 1.58 (s, 6H).
Example 19-1 : Synthesis of 4-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000094_0001
To a mixture of Intermediate 12 (19.5 mg, 0.046 mmol) in MeOH (0.8 mL) was added NaOMe (25 wt % in methanol, 0.084 mL, 0.37 mmol). The mixture was heated at 120 °C under microwave irradiation for 1 h. An additional portion of NaOMe (25 wt % in methanol, 0.08 mL) was added and the mixture was then heated at 130 °C for 2 h. The mixture was cooled to room temperature, acidified by addition of 1 M HCI in ether and subjected to SCX purification (GENERAL METHOD 3-1 , 1 g S\\\aBond Propylsulfonic Acid® cartridge). The crude product was further purified by preparative reverse-phase HPLC to obtain the title compound as a white solid (8 mg). LC/MS Rt = 0.39 min. MS (M+1 ) = 422.3. 1H NMR (METHANOL-^) δ 8.50 (s, 1 H), 8.47 (d, J=5.6 Hz, 1 H), 8.14 (d, J=10.1 Hz, 1 H), 7.25 (s, 1 H), 7.24 (d, J=10.1 Hz, 1 H), 6.74 (d, J=5.6 Hz, 1 H), 5.07 (t, J=12.1 Hz, 1 H), 4.02 (s, 3H), 2.94 (s, 3H), 1.58-1 .70 (m, 2H), 1 .46-1.58 (m, 2H), 1 .32 (s, 6H), 1.16 (s, 6H).
Example 20-1 : Synthesis of 4-(azetidin-1 -yl)-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000095_0001
To a mixture of Intermediate 11 (31 mg, 0.070 mmol) in NMP (0.8 mL) was added azetidine
(0.047 mL, 0.70 mmol). The mixture was heated at 130 °C under microwave irradiation for 2 h. The mixture was cooled to room temperature, acidified by addition of 1 M HCI in ether and subjected to SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge). The crude product was further purified by preparative reverse-phase HPLC to obtain 4-(azetidin-1 -yl)-2- methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol as a light yellow solid (16 mg). LC/MS Rt = 0.48 min. MS (M+1 ) = 461 .3. 1H NMR (METHANOL-^) δ 8.17 (s, 1 H), 8.1 1 (d, J=9.6 Hz, 1 H), 7.14 (d, J=9.6 Hz, 1 H), 6.74 (s, 1 H), 5.81 (s, 1 H), 5.22 (t, J=12.4 Hz, 1 H), 4.62 (t, J=7.6 Hz, 4H), 2.99 (s, 3H), 2.56 (dt, J=15.5, 7.6 Hz, 2H), 2.44 (s, 3H), 1 .66-1.75 (m, 2H), 1.54-1 .66 (m, 2H), 1.40 (s, 6H), 1.25 (s, 6H).
Example 20-2: Synthesis of 7-hydroxy-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-4-carbonitrile
Figure imgf000095_0002
A mixture of Intermediate 11 (40 mg, 0.091 mmol), zinc cyanide (21 mg, 0.18 mmol) and tetrakis(triphenylphosphine)palladium(0) (1 1 mg, 9.1 μηηοΙ) in NMP (1 mL) was evacuated, filled with N2 (4x) and heated under microwave irradiation at 150 °C for 1.5 h. The reaction mixture was filtered through celite, washed with MeOH, concentrated, and subjected to SCX purification (GENERAL METHOD 3-1 , 1 g S\\\aBond Propylsulfonic Acid® cartridge). The crude product was purified by preparative reverse-phase HPLC to obtain 7-hydroxy-2-methyl-6-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyrida as a yellow solid (22 mg).
LC/MS Rt = 0.54 min. MS (M+1 ) = 431 .3. 1 H NMR (METHANOL-^) δ 8.37 (s, 1 H), 8.24 (d, J=9.6 Hz, 1 H), 7.60 (s, 1 H), 7.42 (s, 1 H), 7.33 (d, J=9.6 Hz, 1 H), 5.23 (t, J=1 1.9 Hz, 1 H), 3.05 (s, 3H), 2.71 (s, 3H), 1 .70-1.81 (m, 2H), 1 .58-1.69 (m, 2H), 1.43 (s, 6H), 1.27 (s, 6H).
Example 20-3: Synthesis of 4-cyclopropyl-2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000096_0001
Step 1: 6-(4-Cyclopropyl-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2, 2, 6, 6- tetramethylpiperidin-4-yl)pyridazin-3-amine
A flask containing a mixture of 6-(4-chloro-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-
(2,2,6, 6-tetramethylpiperidin-4-yl)pyridazin-3-amine (PREPARATION 11 Step 5, 48 mg, 0.1 1 mmol), cyclopropyltrifluoroborate (24 mg, 0.16 mmol), n-butyl di-1 -adamantylphosphine (5 mg, 0.013 mmol) and cesium carbonate (103 mg, 0.32 mmol) in toluene (1 mL) and water (0.1 mL) was evacuated, filled with N2 (4x) and heated at 100 °C for 7 h. The reaction mixture was concentrated and acidified by addition of 1 M HCI in ether. SCX purification (GENERAL METHOD 3-1 , 2 g
SiliaSond Propylsulfonic Acid® cartridge) provided the crude product as a light brown solid (50 mg, 93% pure).
Step 2: 4-Cyclopropyl-2-met yl-6-(6-(met yl(2,2, 6, 6-tetramet ylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol
6-(4-Cyclopropyl-7-methoxy-2-methylquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (50 mg, 0.10 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using BBr3. Preparative reverse-phase HPLC purification provided 4-cyclopropyl-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol as a beige solid (28 mg). LC/MS Rt = 0.46 min. MS (M+1 ) = 446.3. 1H NMR
(METHANOL-^) δ 8.70 (s, 1 H), 8.29 (d, J=9.6 Hz, 1 H), 7.36 (s, 1 H), 7.32 (d, J=9.6 Hz, 1 H), 6.86 (s, 1 H), 5.17 (t, J=12.4 Hz, 1 H), 3.04 (s, 3H), 2.60 (s, 3H), 2.53-2.65 (m, 1 H), 1.67-1 .80 (m, 2H), 1 .54-1.67 (m, 2H), 1.41 (s, 6H), 1 .26 (s, 6H), 1.21-1 .24 (m, 2H), 0.88-0.95 (m, 2H).
Example 20-4: Synthesis of 4-(3,6-dihydro-2H-pyran-4-yl)-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol
Figure imgf000097_0001
A mixture of Intermediate 11 (27 mg, 0.061 mmol), 3,6 dihydro-2H-pyran-4-boronic acid pinacol ester (38.7 mg, 0.18 mmol), tetrakis(triphenylphosphine)palladium(0) (7 mg, 6 μηηοΙ) and NaHC03 (1 M aqueous solution, 0.18 ml, 0.18 mmol) in 1 ,4-dioxane (0.8 mL) was evacuated, filled with N2 (4x) and heated under microwave irradiation at 120 °C for 1 h. The reaction mixture was filtered through Celite®, washed with MeOH, concentrated, and subjected to SCX purification
(GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge). The crude product was further purified by Silica Gel Chromatography (GENERAL METHOD 4-1 ) to obtain 4-(3,6-dihydro- 2H-pyran-4-yl)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol as a yellow solid (27 mg). LC/MS Rt = 0.43 min. MS (M+1 ) = 488.3. 1 H NMR
(METHANOL-^) δ 8.36 (s, 1 H), 8.12 (d, J=10.1 Hz, 1 H), 7.38 (s, 1 H), 7.30 (d, J=10.1 Hz, 1 H), 7.08 (s, 1 H), 6.01 (t, J=1 .5 Hz, 1 H), 5.14 (t, J=12.1 Hz, 1 H), 4.41 (q, J=2.5 Hz, 2H), 4.04 (t, J=5.3 Hz, 2H), 3.03 (s, 3H), 2.65 (s, 3H), 2.56 (dd, J=4.5, 2.5 Hz, 2H), 1 .67-1 .74 (m, 2H), 1 .52-1 .64 (m, 2H), 1 .39 (s, 6H), 1 .20-1 .27 (m, 6H).
Example 20-5: Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)quinolin-7-ol formate salt
Figure imgf000097_0002
A mixture of Example 20-4 (14 mg, 0.029 mmol), Pd-C (3 mg, 10 wt % on carbon, 3 μηιοΙ), Pd(OH)2 (2.0 mg, 20 wt % on carbon, 3 μιηοΙ) and one drop of concentrated HCI aqueous solution in methanol (10 mL) was evacuated, filled with H2 (4x) and shaken under H2 (50 psi) on a Parr shaker hydrogenator at room temperature overnight. The reaction mixture was filtered through Celite®, washed with MeOH, concentrated, and subjected to SCX purification (GENERAL
METHOD 3-1 , 1 g SWlaBond Propylsulfonic Acid® cartridge). The crude product was further purified by preparative reverse-phase HPLC to obtain 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(tetrahydro-2H-pyran-4-yl)quinolin-7-ol formate salt as a yellow solid (6 mg). LC/MS Rt = 0.42 min. MS (M+1 ) = 490.4. 1 H NMR (METHANOL-^) δ 8.42 (s, 1 H), 8.40 (s, 1 H), 8.23 (d, J=9.8 Hz, 1 H), 7.30 (s, 1 H), 7.25 (d, J=9.8 Hz, 1 H), 7.1 1 (s, 1 H), 5.40 (p, J=8.0 Hz, 1 H), 4.07-3.97 (m, 2H), 3.68 (m, 3H), 2.97 (s, 3H), 2.57 (s, 3H), 1 .94-1 .75 (m, 8H), 1 .57 (s, 6H), 1.42 (s, 6H).
Example 20-6: Synthesis of 2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(oxetan-3-yl)quinolin-7-ol
Figure imgf000098_0001
To a mixture of Intermediate 11 (46 mg, 0.1 1 mmol), 3-iodooxetane (27 mg, 0.15 mmol), FeS04.7H20 (10 mg, 0.034 mmol) and H2S04 (0.024 mL, 0.45 mmol, ~ 1 drop) in DMSO (0.5 mL) was added H202 (30% aqueous solution, 0.035 mL, 0.34 mmol) at room temperature. After 30 min, another portion of FeS04.7H20 (9.5 mg, 0.034 mmol) was added and the mixture was stirred at room temperature for 40 min. Further FeS04.7H20 (9.5 mg, 0.034 mmol) and H202 (30% aqueous solution, 0.035 mL, 0.34 mmol) was added and the mixture was stirred at room temperature for 60 min, treated with Na2S203 (0.5 mL, 20% aqueous solution), and subjected to SCX purification
(GENERAL METHOD 3-1 , 2 g SiliaSond Propylsulfonic Acid® cartridge). The crude product was further purified by preparative reverse-phase HPLC to obtain 2-methyl-6-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(oxetan-3-yl)quinolin-7-ol (5 mg) as a yellow solid. MS (M+1 ) = 462.3. 1H NMR (METHANOL-^) δ 8.22 (d, J=10.1 Hz, 1 H), 8.05 (s, 1 H), 7.39 (s, 1 H), 7.26-7.35 (m, 2H), 5.29 (dd, J=8.1 , 5.6 Hz, 2H), 5.08 (q, J=7.9, 1 H), 4.97 (dd, J = 7.1 , 5.7 Hz, 2H), 3.03 (s, 3H), 2.69 (s, 3H), 1 .71 (dd, J = 12.6, 3.6 Hz, 2H), 1.59 (t, J = 12.4 Hz, 2H), 1 .40 (s, 6H), 1.24 (s, 6H).
Example 21 -1 : Synthesis of 4-(dimethylamino)-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol di-formate salt
Figure imgf000098_0002
To a mixture of Intermediate 12 (45 mg, 0.094 mmol) in NMP (1 mL) was added
dimethylamine (2 M in THF, 0.25 mL, 0.50 mmol). The mixture was heated at 140 °C under microwave irradiation for 1 .5 h. The mixture was cooled to room temperature, acidified by addition of 1 M HCI in ether and subjected to SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge). The crude product was further purified by preparative reverse- phase HPLC to obtain 4-(dimethylamino)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol di-formate as a light yellow solid (25 mg). LC/MS Rt = 0.43 min. MS (M+1 ) = 435.3. 1H NMR (METHANOL-^) 8.71 (s, 1 H), 8.46 (s, 2H), 8.28 (d, J = 9.9 Hz, 1 H), 8.21 (d, J = 7.1 Hz, 1 H), 7.39 (d, J = 9.8 Hz, 1 H), 7.23 (s, 1 H), 6.82 (d, J = 7.1 Hz, 1 H), 5.49 (tt, J = 1 1 .0, 5.0 Hz, 1 H), 3.54 (s, 6H), 3.08 (s, 3H), 2.05 - 1 .93 (m, 5H), 1.66 (s, 6H), 1.52 (s, 6H) Example 22-1 : Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin-4(1 H)-one
Figure imgf000099_0001
Step 1: 2-Amino-5-bromo-4-methoxybenzoic acid
To a mixture of 4-methoxyanthranilic acid (0.67 g, 4.0 mmol) in DCM (10 mL) and DMF (3 mL) was added NBS (0.71 g, 4.0 mmol) at 0 °C. The mixture was stirred at room temperature for 4 h, then concentrated and subjected to silica gel chromatography (5-100% EtOAc/Heptane) to provide 2-amino-5-bromo-4-methoxybenzoic acid as a beige solid (1 .30 g, containing 1 equivalent succinamide). MS (M-1 ) = 244.3/246.3. 1H NMR (DMSO-d6) δ 7.76 (s, 1 H), 6.42 (s, 1 H), 3.79 (s, 3H). This material was used without further purification.
Step 2: 6-Bromo- 7-methoxyquinazolin-4( 1 H)-one
A mixture of 2-amino-5-bromo-4-methoxybenzoic acid (1 .25 g, 3.6 mmol) and
formimidamide acetate (0.75 g, 7.2 mmol) in EtOH (15 mL) was refluxed for 24 h. The mixture was cooled to room temperature and the solid was filtered, washed with water, and dried to provide 6- bromo-7-methoxyquinazolin-4(1 H)-one as a white solid (0.858 g). MS (M+1 ) = 255.0/257.0. 1H NMR (DMSO-d6) δ 12.30 (br. s, 1 H), 8.21 (s, 1 H), 8.12 (s, 1 H), 7.25 (s, 1 H), 4.00 (s, 3H).
Step 3: 7-Methoxy-6-(4, 4, 5, 5-tetramethyl-1 ,3, 2-dioxaborolan-2-yl)quinazolin-4( 1H)-one A mixture of 6-bromo-7-methoxyquinazolin-4(1 H)-one (385 mg, 1.51 mmol), dppf (84 mg, 0.15 mmol), PdCI2(dppf).CH2CI2 (123 mg, 0.15 mmol), and KOAc (899 mg, 9.1 mmol) in 1 ,4- dioxane (6 mL) and DMF (1 mL) was evacuated and backfilled with nitrogen (4x), then heated at 90 °C for 45 min. Bis(pinacolato) diboron (1.15 g, 4.5 mmol) was then added, and the mixture was heated at 100 °C for 21 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. After silica gel chromatography (0-20% MeOH in DCM) 7-methoxy-6-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)quinazolin-4(1 H)-one (305 mg, -61 % pure) was obtained as a brown solid. MS (M+1 ) = 303.1 .
Step 4: 7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazolin-4( 1H)-one 7-Methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazolin-4(1 H)-one (305 mg, -61 % pure, 0.62 mmol ), 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine, (Intermediate 1 , 145 mg, 0.51 mmol), Pd(PPh3)4 (47 mg, 0.041 mmol), aqueous Na2C03 (2.0 M, 0.64 ml_, 1.3 mmol) and 1 ,4-dioxane (2.4 ml.) were combined in a microwave vessel. The vessel was evacuated and back-filled with nitrogen (4x), and heated via microwave irradiation at 1 10 °C for 1 h. The mixture was cooled to room temperature and acidified to pH 2 with 4 M HCI in 1 ,4- dioxane. SCX purification (GENERAL METHOD 3-1 ) followed by silica gel chromatography (0-20% MeOH in DCM) provided 7-methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)quinazolin-4(1 H)-one (137 mg) as a light brown solid. MS (M+1 ) = 423.1. 1H NMR
(METHANOL-^) δ 8.47 (s, 1 H), 8.14 (s, 1 H), 7.81 (d, J=9.6 Hz, 1 H), 7.29 (s, 1 H), 7.15 (d, J=9.6 Hz, 1 H), 5.25 (t, J=12.4 Hz, 1 H), 4.02 (s, 3H), 3.03 (s, 3H), 1.72 (m, 2H), 1 .60 (m, 2H), 1 .41 (s, 6H), 1 .26 (s, 6H).
Step 5: 7-Hydroxy-6-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazolin-4( 1H)-one
7-Methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin- 4(1 H)-one (32 mg, 0.076 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using boron tribromide (1 .0 M solution in DCM , 0.38 ml_, 0.38 mmol). SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge) followed by preparative reverse-phase HPLC provided 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin- 4-yl)amino)pyridazin-3-yl)quinazolin-4(1 H)-one as a white solid (10 mg). LC/MS Rt = 0.45 min. MS (M+1 ) = 409.2. 1H NMR (METHANOL-^) δ 8.63 (s, 1 H), 8.22 (d, J=9.6 Hz, 1 H), 8.05 (s, 1 H), 7.37 (d, J=9.6 Hz, 1 H), 7.12 (s, 1 H), 5.19 (t, J=12.0 Hz, 1 H), 3.03 (s, 3H), 1 .71 -1.81 (m, 2H), 1 .59-1 .71 (m, 2H), 1.44 (s, 6H), 1.29 (s, 6H).
Example 23-1 : Synthesis of 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinazolin-7-ol
Figure imgf000100_0001
Step 1: 2-Amino-4-met oxybenzalde yde
To a suspension of 2-amino-4-methoxybenzyl alcohol (2 g, 13.1 mmol) in DCM (30 ml.) and was added Mn02 (6.81 g, 78 mmol) at room temperature. After stirring overnight, the mixture was filtered through Celite®, rinsed with DCM, concentrated and purified by silica gel chromatography (5-50% EtOAc/Heptane) to provide 2-amino-4-methoxybenzaldehyde (0.99 g) as a yellow solid. MS (M+1 ) = 152.2. 1H NMR (Chloroform-d) δ9.74 (s, 1 H), 7.40 (d, J=8.6 Hz, 1 H), 6.35 (dd, J=8.7, 2.4 Hz, 1 H), 6.23 (s, 2H), 6.09 (d, J=2.3 Hz, 1 H), 3.84 (d, J = 4.1 Hz, 3H).
Step 2: 2-Amino-5-bromo-4-methoxybenzaldehyde
To a solution of 2-amino-4-methoxybenzaldehyde (0.99 g, 6.6 mmol) in THF (20 mL) was added NBS (1.17 g, 6.6 mmol) at 0 °C. The mixture was stirred at room temperature for 3 h, then quenched with Na2S203 (5 mL, 20% aqueous solution). After stirring for 5 min, the mixture was diluted with an aqueous NaHC03 solution and extracted with EtOAc. The combined organic extracts were washed with brine, dried over Na2S04, concentrated in vacuo, and subjected to silica gel chromatography (5-50% EtOAc/Heptane) to provide 2-amino-5-bromo-4-methoxybenzaldehyde (0.79 g) as a yellow solid. MS (M+1 ) = 230.1/232.1 . 1H NMR (Chloroform-d) δ 9.59 (s, 1 H), 7.52 (s, 1 H), 6.21 (s, 2H), 6.03 (s, 1 H), 3.83 (s, 3H).
Step 3: 6-Bromo-7-methoxyquinazoline
A mixture of 2-amino-5-bromo-4-methoxybenzaldehyde (350 mg, 1.52 mmol) and formimidamide acetate (238 mg, 2.28 mmol) in EtOH (10 mL) was refluxed for 16 h. The mixture was cooled to room temperature and concentrated. The residue was basified with an aqueous
NaHC03 solution and extracted with DCM. The combined organic extracts were dried over Na2S04, concentrated in vacuo, and subjected to silica gel chromatography (0-30% EtOAc/Heptane) to provide 6-bromo-7-methoxyquinazoline (253 mg) as a yellow solid. MS (M+1 ) = 239.0/241.0. 1 H NMR (Chloroform-d) δ 9.28 (s, 1 H), 9.23 (s, 1 H), 8.18 (s, 1 H), 7.39 (s, 1 H), 4.1 1 (s, 3H).
Step 4: 7-Methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinazoline
To a mixture of 6-bromo-7-methoxyquinazoline (253 mg, 0.95 mmol), dppf (53 mg, 0.095 mmol), PdCI2(dppf).CH2CI2 (78 mg, 0.095 mmol), and KOAc (561 mg, 5.7 mmol) was added 1 ,4- dioxane (5 mL). The reaction mixture was evacuated and backfilled with nitrogen (4x), then heated at 90 °C for 30 min. Bis(pinacolato) diboron (726 mg, 2.9 mmol) was then added, and the mixture was heated at 90 °C for 16 hr. Further bis(pinacolato) diboron (242 mg, 0.95 mmol), dppf (26.4 mg, 0.048 mmol) and PdCI2(dppf).CH2CI2 (39 mg, 0.048 mmol) were added and the mixture was heated at 95 °C for 23 h. The mixture was cooled to room temperature and filtered through Celite® washing with 9:1 MeOH/DCM. The filtrates were concentrated in vacuo. After silica gel chromatography (0- 20% MeOH in DCM), 7-methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazoline (180 mg, -67% pure) was obtained as a brown solid. MS (M+1 ) = 287.2.
Step 5: 6-(7-Methoxyquinazolin-6-yl)-N-methyl-N-(2,2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
7-Methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazoline (180 mg, -67% pure, 0.42 mmol) was coupled with 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine (Intermediate 1 , 100 mg, 0.35 mmol), using the same method as Example Example 20-6 Step 4 for Suzuki coupling. The crude product was purified by SCX purification (GENERAL
METHOD 3-1 ) followed by silica gel chromatography as described in GENERAL METHOD 4-1 to provide 6-(7-methoxyquinazolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine (28.9 mg) as a light brown solid. MS (M+1 ) = 407.3. 1H NMR (METHANOL-^) δ 9.41 (s, 1 H), 9.14 (s, 1 H), 8.29 (s, 1 H), 7.82 (d, J=9.6 Hz, 1 H), 7.47 (s, 1 H), 7.12 (d, J=9.6 Hz, 1 H), 5.24 (t, J=12.1 Hz, 1 H), 4.07 (s, 3H), 3.00 (s, 3H), 1 .62-1.73 (m, 2H), 1 .48-1.60 (m, 2H), 1.36 (s, 6H), 1 .17- 1.26 (m, 6H).
Step 6: 6-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin-7-ol 6-(7-Methoxyquinazolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3- amine (28 mg, 0.069 mmol) was subjected to methoxy deprotection conditions as described in
GENERAL METHOD 2-3 using boron tribromide (1 .0 M solution in DCM , 0.41 ml_, 0.41 mmol). SCX purification (GENERAL METHOD 3-1 , 1 g SiliaSond Propylsulfonic Acid® cartridge) followed by preparative reverse-phase HPLC provided 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinazolin-7-ol as a yellow solid (16 mg). LC/MS Rt = 0.46 min. MS (M+1 ) = 393.2. 1H NMR (METHANOL-^) δ 9.20 (s, 1 H), 8.91 (s, 1 H), 8.40 (s, 1 H), 8.25 (d, J=9.6 Hz, 1 H), 7.29 (d, J=9.6 Hz, 1 H), 7.20 (s, 1 H), 5.18-5.36 (m, 1 H), 3.03 (s, 3H), 1 .75-1.83 (m, 2H), 1.64-1 .74 (m, 2H), 1.46 (s, 6H), 1.31 (s, 6H).
Example 24-1 : Synthesis of 7-hydroxy-1 -methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3,4-dihydroquinolin-2(1 H)-one
Figure imgf000102_0001
Step 1: 6-Bromo-7-hydroxy-3,4-dihydroquinolin-2(1H)-one
To a solution of 7-hydroxy-3,4-dihydroquinolin-2(1 H)-one (590 mg, 3.5 mmol) in DMF (3 mL) was added NBS (655 mg, 3.7 mmol) at 0 °C in three portions over 30 min. The mixture was stirred at room temperature overnight, then quenched with Na2S203 (5 mL, 20% aqueous solution). After stirring for 5 min, the mixture was diluted with aqueous NaHC03 and extracted with EtOAc. The combined organic extracts were washed with brine, dried over Na2S04, concentrated in vacuo, and purified by silica gel chromatography (5-50% EtOAc/Heptane) to provide 6-bromo-7-hydroxy-3,4- dihydroquinolin-2(1 H)-one (0.82 g) as a yellow solid. MS (M+1 ) = 241 .9/243.9. 1H NMR (Methanol- d4) δ 7.24 (s, 1 H), 6.47 (s, 1 H), 2.84 (t, J=7.6 Hz, 2H), 2.47-2.56 (m, 2H).
Step 2: 6-Bromo-7-methoxy-1 -methyl-3,4-dihydroquinolin-2(1H)-one To a solution of 6-bromo-7-hydroxy-3,4-dihydroquinolin-2(1 H)-one (650 mg, 2.7 mmol) in DMF (3 ml.) was added K2C03 (557 mg, 4.0 mmol) at room temperature. After 5 min, Mel (0.185 ml, 2.9 mmol) was added and the mixture was stirred overnight. The mixture was quenched with water and extracted with EtOAc (3x). The combined organic extracts were washed with brine, dried over Na2S04 and concentrated to afford the crude product. 6-Bromo-7-methoxy-1-methyl-3,4- dihydroquinolin-2(1 H)-one (174 mg) and 6-bromo-7-methoxy-3,4-dihydroquinolin-2(1 H)-one (396 mg) were obtained after silica gel chromatography purification (5-50% EtOAc/Heptane). MS (M+1 ) = 269.9/271 .9 and 255.9/257.9, respectively .
Step 3: 7-Methoxy- 1-methyl-6-(4, 4, 5, 5-tetramethyl- 1, 3, 2-dioxaborolan-2-yl)-3, 4- dihydroquinolin-2( 1H)-one
To a mixture of 6-bromo-7-methoxy-1-methyl-3,4-dihydroquinolin-2(1 H)-one (170 mg, 0.63 mmol), dppf (34.9 mg, 0.063 mmol), PdCI2(dppf).CH2CI2 (51 .4 mg, 0.063 mmol), and KOAc (371 mg, 3.78 mmol) was added 1 ,4-dioxane (2 ml_). The reaction mixture was evacuated and backfilled with nitrogen (4x), then heated at 90 °C for 20 min. Bis(pinacolato) diboron (479 mg, 1.89 mmol) was then added, and the mixture was heated at 98 °C for 16 h. The reaction mixture was cooled to room temperature and filtered through Celite® washing with 9:1 MeOH/DCM. The filtrates were concentrated in vacuo. After silica gel chromatography (0-10% MeOH in DCM) 7- methoxy-1-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-3,4-dihydroquinolin-2(1 H)-one (500 mg, -38% pure) was obtained as a brown solid. MS (M+1 ) = 318.4.
Step 4: 7-Methoxy- 1-methyl-6-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-
3- yl)-3, 4-dihydroquinolin-2( 1H)-one
7-Methoxy-1-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-3,4-dihydroquinolin- 2(1 H)-one (470 mg, -38% pure, 0.56 mmol) was coupled with 6-chloro-N-methyl-N-(2,2,6,6- tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 1 , 1 10 mg, 0.39 mmol), using the same method as Example 20-6 Step 4 for Suzuki coupling. The crude product was purified by SCX purification (GENERAL METHOD 3-1 ) followed by silica gel chromatography as described in
GENERAL METHOD 4-1 to provide 7-methoxy-1 -methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-
4- yl)amino)pyridazin-3-yl)-3,4-dihydroquinolin-2(1 H)-one (92 mg) as a white solid. MS (M+1 ) = 438.2. 1H NMR (METHANOL-^) δ 7.84 (d, J=9.6 Hz, 1 H), 7.49 (s, 1 H), 7.16 (d, J=9.6 Hz, 1 H), 6.85 (s, 1 H), 5.42-5.55 (m, 1 H), 3.91 (s, 3H), 3.43 (s, 3H), 3.01 (s, 3H), 2.92 (t, J=7.3 Hz, 2H), 2.60-2.70 (m, 2H), 1.90-2.02 (m, 4H), 1 .64 (s, 6H), 1 .52 (s, 6H).
Step 5: 7-Hydroxy- 1-methyl-6-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)-3, 4-dihydroquinolin-2( 1H)-one
7-Methoxy-1 -methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3,4- dihydroquinolin-2(1 H)-one (91 mg, 0.21 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 using boron tribromide (1.0 M solution in DCM, 1.04 mL, 1.04 mmol). SCX purification (GENERAL METHOD 3-1 , 2 g SiliaSond Propylsulfonic Acid® cartridge) followed by preparative reverse-phase HPLC provided 7-hydroxy-1-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3,4-dihydroquinolin-2(1 H)-one as a yellow solid (69 mg). LC/MS Rt = 0.51 min. MS (M+1 ) = 424.4. 1H NMR (METHANOL-^) δ 8.04 (d, J=9.6 Hz, 1 H), 7.57 (s, 1 H), 7.28 (d, J=9.6 Hz, 1 H), 6.69 (s, 1 H), 5.03 (t, J=12.4 Hz, 1 H), 3.34 (s, 3H), 2.99 (s, 3H), 2.90 (t, J=l A Hz, 2H), 2.63 (t, J=l A Hz, 2H), 1 .63-1.72 (m, 2H), 1.50-1.61 (m, 2H), 1.38 (s, 6H), 1.22 (s, 6H).
Example 25-1 : Synthesis of 7-hydroxy-1 -methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3,4-dihydroquinolin-2(1 H)-one
Figure imgf000104_0001
Step 1: N-(4-Bromo-2-formyl-5-methoxyphenyl)acetamide
To a mixture of 2-amino-5-bromo-4-methoxybenzaldehyde (1.1 g, 4.2 mmol) in DCM (30 mL) and was added pyridine (1 .68 mL, 20.8 mmol) at room temperature. The mixture was cooled to 0 °C, and acetyl chloride (1 .48 mL, 20.8 mmol) was added. The mixture was stirred at room temperature overnight, quenched with water, and extracted with DCM (3x). The combined organic extracts were dried over Na2S04 and concentrated to afford the crude product. Silica gel chromatography (5-50% EtOAc/Heptane) provided N-(4-bromo-2-formyl-5- methoxyphenyl)acetamide (498 mg). MS (M+1 ) = 271.9/273.9. 1H NMR (Chloroform-d) δ 1 1.37 (s, 1 H), 9.73 (s, 1 H), 8.52 (s, 1 H), 7.79 (s, 1 H), 4.02 (s, 3H), 2.28 (s, 3H).
Step 2: 6-Bromo-7-methoxy-2-methylquinazoline
The mixture of N-(4-bromo-2-formyl-5-methoxyphenyl)acetamide (138 mg, 0.41 mmol) and ammonia (2.0 M methanol solution, 4.1 mL, 8.1 mmol) was heated at 75 °C for 4 h in a sealed vial vented with a 22 gauge needle. The mixture was cooled to room temperature, concentrated, and subjected to silica gel chromatography (5-60% EtOAc/Heptane) to provide 6-bromo-7-methoxy-2- methylquinazoline (103 mg) as a yellow solid. MS (M+1 ) = 252.9/254.9. 1H NMR (Chloroform-d) δ 9.04 (s, 1 H), 8.02 (d, J=3A Hz, 1 H), 7.21 (s, 1 H), 3.99 (d, J=3.2 Hz, 3H), 2.78 (s, 3H).
Step 3: 7-Methoxy-2-methyl-6-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)quinazoline To a mixture of 6-bromo-7-methoxy-2-methylquinazoline (103 mg, 0.41 mmol), dppf (23 mg, 0.041 mmol), PdCI2(dppf).CH2CI2 (33 mg, 0.041 mmol), and KOAc (240 mg, 2.44 mmol) was added 1 ,4-dioxane (2.5 mL). The reaction mixture was evacuated and backfilled with nitrogen (4x), then heated at 90 °C for 20 min. Bis(pinacolato) diboron (310 mg, 1 .22 mmol) was then added, and the mixture was heated at 98 °C for 16 h. Further bis(pinacolato) diboron (207 mg, 0.81 mmol), dppf (23 mg, 0.041 mmol) and PdCI2(dppf).CH2CI2 (33 mg, 0.041 mmol) were added and the mixture was heated at 98 °C for 16 h. The reaction mixture was cooled to room temperature and filtered through Celite®, washing with 9:1 MeOH/DCM. The filtrates were concentrated in vacuo. After silica gel chromatography (0-10% MeOH in DCM) 7-methoxy-2-methyl-6-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)quinazoline (157 mg, -72% pure) was obtained as a brown solid. MS (M+1 ) = 301.0.
Step 4: 6-(7-Methoxy-2-methylquinazolin-6-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
7-Methoxy-2-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazoline (180 mg, -72% pure, 0.37 mmol) was coupled with 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (Intermediate 1 , 80 mg, 0.283 mmol), using the same method as Example 20-6 Step 4 for Suzuki coupling. The crude product was purified by SCX purification (GENERAL METHOD 3-1 ) followed by silica gel chromatography as described in GENERAL METHOD 4-1 to provide 6-(7-methoxy-2-methylquinazolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (90 mg) as a yellow solid. MS (M+1 ) = 421.1 . 1H NMR (METHANOL-^) δ 9.34 (s, 1 H), 8.26 (s, 1 H), 7.82 (d, J=9.6 Hz, 1 H), 7.40 (s, 1 H), 7.15 (d, J=9.6 Hz, 1 H), 5.25 (t, J=12.9 Hz, 1 H), 4.06 (s, 3H), 3.01 (s, 3H), 2.82 (s, 3H), 1 .70 (dd, J=12.5, 3.6 Hz, 2H), 1 .58 (t, J=12.4 Hz, 2H), 1.38 (s, 6H), 1.23 (s, 6H).
Step 5: 6-( 6-(Methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin-7-ol 6-(7-Methoxy-2-methylquinazolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (92 mg, 0.21 mmol) was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-4 using thiophenol (0.028 ml_, 0.27 mmol). SCX purification (GENERAL METHOD 3-1 , 2 g S\\\aBond Propylsulfonic Acid® cartridge) followed by preparative reverse-phase HPLC provided 6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazolin-7-ol as a yellow solid (27 mg). LC/MS Rt = 0.47 min. MS (M+1 ) = 407.2. 1H NMR (METHANOL-^) δ 9.17 (s, 1 H), 8.41 (s, 1 H), 8.25 (d, J=9.7 Hz, 1 H), 7.32 (d, J=9.8 Hz, 1 H), 7.17 (s, 1 H), 5.25 (t, J=13.1 Hz, 1 H), 3.03 (s, 3H), 2.74 (s, 3H), 1 .79 (dd, J=12.9, 3.7 Hz, 2H), 1 .69 (t, J=12.5 Hz, 2H), 1.47 (s, 6H), 1.32 (s, 6H).
Example 26-1 : Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 -carbonitrile
Figure imgf000106_0001
Step 1: 6-Bromo-7-methoxyisoquinoline 2-oxide
To a 50 mL round bottom flask containing 6-bromo-7-methoxyisoquinoline (prepared as described in in Example 3-7 (100 mg, 0.42 mmol) in DCM (2.1 mL) was added
methyltrioxorhenium (MTO, 4.2 mg, 0.02 mmol). The mixture cooled to 0°C and 50% aqueous hydrogen peroxide (57 mg, 0.84 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 18 h. Then 5-10 mg managanese dioxide was added and the reaction stirred for 2 h then filtered through celite. The filtrate was concentrated in vacuo to afford 6-bromo-7- methoxyisoquinoline 2-oxide (213 mg). MS (M+1 ) = 256.3
Step 2: 7^ydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline- 1 -carbonitrile
7-Hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline- 1 -carbonitrile was prepared from 6-bromo-7-methoxyisoquinoline 2-oxide according to the synthesis of 6-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline-1 -carbonitrile (as described in Example 3-7; LC/MS Rt = 0.54 MS (M+1 ) = 417.2 1H NMR (400 MHz, Methanol-d4) δ 8.44 (d, J= 5.4 Hz, 2H), 8.25 (d, J= 9.9 Hz, 1 H), 8.00 (d, J= 5.5 Hz, 1 H), 7.70 (s, 1 H), 7.25 (d, J= 9.9 Hz, 1 H), 5.34-5.04 (m, 1 H), 3.06 (s, 3H), 1.78 (dd, J= 12.7, 3.5 Hz, 2H), 1.58 (t, J= 12.5 Hz, 2H), 1 .43 (s, 6H), 1.27 (s, 6H).
Example 26-2: Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2-carbonitrile
Figure imgf000106_0002
7-Hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2- carbonitrile was prepared from 6-chloro-7-methoxyquinoline according to the synthesis of Example 26-1. LC/MS Rt =0.51. MS (M+1 ) = 417.2 1H NMR (400 MHz, DMSO-d6) δ 8.70 (s, 1 H), 8.54 (dd, J= 8.5, 0.8 Hz, 1 H), 8.33 (d, J= 9.9 Hz, 1 H), 7.82 (d, J= 8.3 Hz, 1 H), 7.51 -7.37 (m, 2H), 5.21-4.94 (m, 1 H), 2.99 (s, 3H), 1.68-1 .42 (m, 4H), 1 .30 (s, 6H), 1 .14 (s, 6H).
Example 26-3: Synthesis of 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2-carbonitrile
Figure imgf000107_0001
6-Hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin
carbonitrile was prepared from 7-Bromo-6-methoxyquinoline according to the synthesis of Example 26-1. LC/MS Rt = 0.54. MS (M+1 ) = 417.2 1H NMR (400 MHz, DMSO-d6) δ 8.71 (s, 1 H), 8.50 (d, J= 9.9 Hz, 1 H), 8.41 (d, J= 8.5 Hz, 1 H), 7.90 (d, J= 8.5 Hz, 1 H), 7.48-7.39 (m, 2H), 5.26-4.78 (m, 1 H), 3.00 (s, 3H), 1.66-1 .41 (m, 5H), 1.29 (s, 6H), 1.12 (s, 6H).
Example 26-4: Synthesis of 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 -carboxamide
Figure imgf000107_0002
Step 1: 6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline-1-carboxamide
A mixture of 6-methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline-1 -carbonitrile (87 mg, 0.202 mmol), potassium hydroxide (10% aqueous, 0.15 mL,
0.263 mmol) and hydrogen peroxide (33% aqueous, 0.08 mL, 0.909 mmol) was heated at 40°C for 1 h. SCX purification (GENERAL METHOD 3-1 ) afforded 6-methoxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline-1 -carboxamide (91 mg). MS (M+1 ) =
449.5
Step 2: 6-Hydroxy- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline- 1 -carboxamide
6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline- 1 -carboxamide was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 followed by free basing via GENERAL METHOD 8-1 to afford the title compound (10.9 mg). LC/MS Rt = 0.43. MS (M+1 ) = 435.2 1 H NMR (400 MHz, METHANOL-d4) δ ppm 8.90 (br. s., 1 H), 8.04-8.20 (m, 2H), 7.48 (d, J=6.02 Hz, 1 H), 7.29-7.44 (m, 1 H), 7.18 (d, J=9.54 Hz, 1 H), 7.00 (s, 1 H), 5.06-5.35 (m, 1 H), 3.02 (s, 3H), 1 .65-1.76 (m, 2H), 1.49-1 .62 (m, 2H), 1 .39 (s, 6H), 1.23 (s, 6H).
Example 26-5: Synthesis of 7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2-carboxamide
Figure imgf000108_0001
7-Hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui carboxamide was isolated as a by-product from the methoxy deprotection of 7-methoxy-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)py (Example 26- 2) using GENERAL METHOD 2-1. LC/MS Rt = 0.48. MS (M+1 ) = 435.3 1 H NMR (400 MHz,
DMSO-d6) δ 8.64 (s, 1 H), 8.45 (d, J= 8.4 Hz, 1 H), 8.35 (d, J= 9.9 Hz, 1 H), 8.24 (d, J= 2.9 Hz, 1 H), 7.96 (d, J= 8.4 Hz, 1 H), 7.71 (d, J= 2.8 Hz, 1 H), 7.50-7.38 (m, 2H), 5.31 -4.88 (m, 1 H), 2.99 (s, 3H), 1 .57 (s, 4H), 1 .30 (s, 6H), 1 .14 (s, 6H).
Example 26-6: Synthesis of 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2-carboxamide
Figure imgf000108_0002
6-Hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-^ carboxamide was isolated as a by-product from the methoxy deprotection of 6-methoxy-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2-carbonitrile (Example 26- 3) using GENERAL METHOD 2-1. LC/MS Rt = 0.52. MS (M+1 ) = 435.3 1 H NMR (400 MHz,
Methanol-d4) δ 8.53 (s, 1 H), 8.28 (d, J= 9.9 Hz, 1 H), 8.23 (s, 1 H), 8.14 (d, J= 8.5 Hz, 1 H), 7.38 (s, 1 H), 7.28 (d, J= 9.8 Hz, 1 H), 5.22 (m, 1 H), 3.06 (s, 3H), 1.80 (dd, J= 12.8, 3.4 Hz, 2H), 1.63 (t, J= 12.0 Hz, 2H), 1 .45 (s, 6H), 1.29 (s, 6H).
Example 26-7: Synthesis of methyl 6-hydroxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2-carboxylate
Figure imgf000108_0003
Methyl 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinoline-2-carboxylate was isolated as a by-product in the deprotection of 6-methoxy-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2-carbonitrile (Example 26- 3) by GENERAL METHOD 2-1. LC/MS Rt = 0.57. MS (M+1 ) = 450.3 1H NMR (400 MHz,
Methanol-d4) δ 8.53 (s, 1 H), 8.35-8.19 (m, 2H), 8.06 (dd, J= 8.5, 1 .5 Hz, 1 H), 7.42-7.29 (m, 2H), 5.35-5.08 (m, 1 H), 4.04 (d, J= 1 .5 Hz, 3H), 3.05 (s, 3H), 1 .74 (dd, J= 12.7, 3.6 Hz, 2H), 1 .63 (t, J= 12.5 Hz, 2H), 1 .43 (s, 6H), 1.27 (s, 6H).
Example 27-1 : Synthesis of 6-hydroxy-7-(6-(piperazin-1 -yl)pyridazin-3-yl)quinoline-2- carbonitrile
Figure imgf000109_0001
Step 1: ferf-Butyl 4-(6-(2-cyano-6-methoxyquinolin-7-yl)pyridazin-3-yl)piperazine-1 - carboxylate
ferf-Butyl 4-(6-(2-cyano-6-methoxyquinolin-7-yl)pyridazin-3-yl)piperazine-1 -carboxylate was prepared from (2-cyano-6-methoxyquinolin-7-yl)boronic acid and ferf-butyl 4-(6-chloropyridazin-3- yl)piperazine-1 -carboxylate via Suzuki conditions described in GENERAL METHOD 1 -1 (170 mg). MS (M+1 ) = 447.3.
Step 2: 6-Hydroxy- 7-(6-(piperazin-1-yl)pyridazin-3-yl)quinoline-2-carbonitrile
ferf-Butyl 4-(6-(2-cyano-6-methoxyquinolin-7-yl)pyridazin-3-yl)piperazine-1 -carboxylate was subjected to Boc and methoxy deprotection conditions as described in GENERAL METHOD 2-1. The resulting solid was suspended in acetonitrile:H20 (3:1 ml_). 1 M aqueous HCI (3 equivalents) was added and solvent was concentrated in vacuo to afford the title compound (8.6 mg). LC/MS Rt = 0.48. MS (M+1 ) = 333.1 1H NMR (400 MHz, DMSO-d6) δ 8.72 (s, 1 H), 8.56 (d, J= 9.8 Hz, 1 H), 8.46 (d, J= 8.5 Hz, 1 H), 7.95 (d, J= 8.5 Hz, 1 H), 7.75 (d, J= 9.8 Hz, 1 H), 7.51 (s, 1 H), 4.00 (m, 4H), 3.27 (bs, 4H).
Example 27-2: Synthesis of 7-hydroxy-6-(6-(piperazin-1 -yl)pyridazin-3-yl)quinoline-2- carbonitrile
Figure imgf000109_0002
Step 1: tert-Butyl 4-(6-(2-cyano-7-methoxyquinolin-6-yl)pyridazin-3-yl)piperazine-1- carboxylate
ferf-Butyl 4-(6-(2-cyano-7-methoxyquinolin-6-yl)pyridazin-3-yl)piperazine-1 -carboxylate was prepared from 7-methoxy-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinoline-2-carbonitrile and ferf-butyl 4-(6-chloropyridazin-3-yl)piperazine-1 -carboxylate via Suzuki conditions described in GENERAL METHOD 1 -1 (100 mg). MS (M+1 ) = 447.4
Step 2: 7-Hydroxy-6-(6-(piperazin-1-yl)pyridazin-3-yl)quinoline-2-carbonitrile ferf-butyl 4-(6-(2-cyano-7-methoxyquinolin-6-yl)pyridazin-3-yl)piperazine-1 -carboxylate was subjected to Boc and methoxy deprotection conditions as described in GENERAL METHOD 2-1. Free basing via SCX, as described in GENERAL METHOD 3-1 , afforded the title compound (1 .9 mg). LC/MS Rt = 0.45. MS (M+1 ) = 333.1 1 H NMR (400 MHz, Methanol-d4) δ 8.47-8.36 (m, 2H), 8.27 (d, J= 9.9 Hz, 1 H), 7.62 (d, J= 8.3 Hz, 1 H), 7.55 (s, 1 H), 7.42 (d, J= 9.8 Hz, 1 H), 3.90-3.70 (m, 4H), 3.20-2.97 (m, 4H).
Example 27-3: Synthesis of -(6-(piperazin-1 -yl)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000110_0001
Step 1: tert-Butyl 4-(6-(6-methoxyisoquinolin-7-yl)pyridazin-3-yl)piperaz
ferf-Butyl 4-(6-(6-methoxyisoquinolin-7-yl)pyridazin-3-yl)piperazine-1 -carboxylate was prepared from 6-methoxy-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline-1-carbonitrile and ferf-butyl 4-(6-chloropyridazin-3-yl)piperazine-1 -carboxylate via Suzuki conditions described in GENERAL METHOD 1 -1 (340 mg). MS (M+1 ) = 422.6
Step 2: 7-(6-(Piperazin-1-yl)pyridazin-3-yl)isoquinolin-6-ol
ferf-Butyl 4-(6-(6-methoxyisoquinolin-7-yl)pyridazin-3-yl)piperazine-1 -carboxylate was subjected to Boc and methoxy deprotection conditions as described in GENERAL METHOD 2-1 to afford the title compound (1 1.3 mg). LC/MS Rt = 0.31. MS (M+1 ) = 308.1 1H NMR (400 MHz, DMSO-d6) δ 9.17 (s, 1 H), 8.76 (s, 1 H), 8.41 (d, J= 9.9 Hz, 1 H), 8.35 (d, J= 5.8 Hz, 1 H), 7.68-7.57 (m, 2H), 7.29 (s, 1 H), 3.74-3.55 (m, 4H), 2.94-2.73 (m, 4H).
Example 28-1 : Synthesis of 7-(6-(1 ,2,3,6-tetrahydropyridin-4-yl)pyridazin-3-yl)quinolin-
6-ol
Figure imgf000110_0002
Step 1: tert-Butyl 4-(6-chloropyridazin-3-yl)-5,6-dihydropyridine-1(2H)-carboxylate
A mixture of 3,6-dichloropyridazine (265 mg, 1 .78 mmol), ferf-butyl 4-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1 (2H)-carboxylate (500 mg, 1.62 mmol), and Na2C03 (514 mg, 4.85 mmol) in dioxane (3.1 ml.) and water (0.5 ml.) was degassed with a stream of dry nitrogen for 5 min. PdCI2(dppf)-CH2Cl2 adduct (66 mg, 0.08 mmol) was added and the mixture was degassed for an addition 5 min with dry nitrogen and heated at 80°C for 3 h. The reaction mixture was concentrated in vacuo, diluted with water and extracted with DCM. The organic exact was dried over Na2S04 and concentrated in vacuo. The crude product was purified by silica gel chromatography (0-100% EtOAc/Hep) to afford ferf-butyl 4-(6-chloropyridazin-3-yl)-5,6- dihydropyridine-1 (2H)-carboxylate (192 mg). MS - ferf-butyl (M+1 ) = 240.4
Step 2: tert-Butyl 4-( 6-( 6-methoxyquinolin- 7-yl)pyridazin-3-yl)-5, 6-dihydropyridine- 1 ( 2H)- carboxylate
ferf-Butyl 4-(6-(6-methoxyquinolin-7-yl)pyridazin-3-yl)-5,6-dihydropyridine-1 (2H)-carboxylate was prepared from ferf-butyl 4-(6-chloropyridazin-3-yl)-5,6-dihydropyridine-1 (2H)-carboxylate and 6-methoxy-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinoline (prepared as in Example 6-1 ) via Suzuki conditions described in GENERAL METHOD 1 -1 (175 mg). MS (M+1 ) = 419.1
Step 3: 7-( 6-( 1, 2, 3, 6-Tetrahydropyridin-4-yl)pyridazin-3-yl)quinolin-6-ol
ferf-Butyl 4-(6-(6-methoxyquinolin-7-yl)pyridazin-3-yl)-5,6-dihydropyridine-1 (2H)-carboxylate was subjected to Boc and methoxy deprotection conditions as described in GENERAL METHOD 2- 3 to afford the title compound (2.0 mg). LC/MS Rt = 0.40. MS (M+1 ) = 305.1 1 H NMR (400 MHz, Methanol-d4) δ 8.97 (d, J = 4.7 Hz, 1 H), 8.81 (d, J = 8.4 Hz, 1 H), 8.74 (s, 1 H), 8.59 (d, J = 9.0 Hz, 1 H), 8.23 (d, J = 8.9 Hz, 1 H), 7.89 (dd, J = 8.3, 4.7 Hz, 1 H), 7.66 (s, 1 H), 7.04 - 6.77 (m, 1 H), 4.03 (d, J = 3.5 Hz, 2H), 3.57 (t, J = 6.0 Hz, 2H), 3.14 (d, J = 6.3 Hz, 2H).
Example 29-1 : Synthesis of 1 -methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-7-ol
Figure imgf000111_0001
Step 1: 6-Bromo-7-methoxy-1-methylisoquinoline
To a mixture of 6-bromo-7-methoxyisoquinoline 2-oxide (160 mg, 0.630 mmol), prepared in the same manner as in Example 26-1 , and THF (3.2 mL) cooled to 0 °C was added
Bis(cyclopentadienyl)-μ-chloro(dimethylaluminum)-μ-methylenetitanium (Tebbe Reagent) (0.5 M in toluene, 1 .4 mL, 0.693 mmol). The mixture was stirred at 0°C for 1 h, quenched with 5.0 N aqueous NaOH, and extracted with DCM (3x). The combined organic exacts were dried over Na2S04 and concentrated in vacuo. The crude product was purified by silica gel chromatography (0-5%
MeOH/DCM) to afford 6-bromo-7-methoxy-1 -methylisoquinoline (43 mg). MS (M+1 ) = 254.3
Step 2: 7 -Methoxy- 1-methyl-6-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)isoquinoline 7-Methoxy-1-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline was prepared from 6-bromo-7-methoxy-1 -methylisoquinoline via boronate ester formation as described in GENERAL METHOD 7-1 (132 mg). MS (M+1 ) = 300.5
Step 3: 6-(7-Methoxy-1-methylisoquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine 6-(7-methoxy-1-methylisoquinolin-6-yl)-N-m
yl)pyridazin-3-amine was prepared from 7-methoxy-1-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)isoquinoline and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (Intermediate 1 ) via Suzuki conditions described in GENERAL METHOD 1 -1 (177 mg). MS (M+1 ) = 420.6
Step 4: 1-Methyl-6-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinolin- 7-oi
6-(7-Methoxy-1-methylisoquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 followed by free basing via GENERAL METHOD 8-1 to afford the title compound (8.0 mg). LC/MS Rt = 0.42. MS (M+1 ) = 406.3 1 H NMR (400 MHz, DMSO-d6) δ 8.52 (s, 1 H), 8.38 (d, J= 9.9 Hz, 1 H), 8.15 (d, J= 5.7 Hz, 1 H), 7.58 (d, J= 5.7 Hz, 1 H), 7.49 (s, 1 H), 7.39 (d, J= 9.9 Hz, 1 H), 2.98 (s, 3H), 2.79 (s, 3H), 1.65-1 .36 (m, 4H), 1.26 (s, 6H), 1 .09 (s, 6H).
Example 29-2: Synthesis of 1 -methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoli
Figure imgf000112_0001
Step 1: N-(1-(3-Bromo-4-methoxyphenyl)ethyl)-2, 2-dimethoxyethanamine
A mixture of 2, 2-dimethoxyethanamine (2.5 g, 24.01 mmol) and 1-(3-bromo-4- methoxyphenyl)ethanone (5.0 g, 21 .83 mmol) in toluene in a 250 ml. flask equipped with a dean- stark adaptor and condenser was heated to 140°C for 18 h. The reaction mixture was cooled to ambient temperature and concentrated in vacuo. To the resulting oil was added MeOH (1 12 ml.) followed by NaBH4 (1.6 g, 41.9 mmol). The mixture was stirred at room temperature for 3.5 h, cooled to 0°C and acidified with 1 N aqueous HCI then concentrated in vacuo to remove MeOH.
The resulting aqueous mixture was washed with DCM, basified with 1 N aqueous NaOH and extracted with DCM. The organic extract was dried over Na2S04 and concentrated in vacuo to afford N-(1-(3-bromo-4-methoxyphenyl)ethyl)-2, 2-dimethoxyethanamine (2.1 g). MS (M+1 ) = 319.7.
Step 2: 7-Bromo-6-methoxy-1-methylisoquinoline
To a 2 ml. microwave vial containing N-(1-(3-bromo-4-methoxyphenyl)ethyl)-2,2- dimethoxyethanamine (250 mg, 0.786 mmol) cooled to -78°C was slowly added chlorosulfonic acid (0.53 ml_, 7.86 mmol). The mixture was then allowed to stir at room temperature for 18 h, then added dropwise to ice water. The resulting aqeuous mixture was washed with diethyl ether, basified with saturated aqueous Na2C03, and extracted with DCM (2x). The resulting organic extracts were dried over Na2S04 and concentrated in vacuo to afford 7-bromo-6-methoxy-1- methylisoquinoline (86 mg). MS (M+1 ) = 254.0.
Step 3: 6-Methoxy- 1 -methyl- 7-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)isoquinoline 6-Methoxy-1-methyl-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline was prepared from 7-bromo-6-methoxy-1 -methylisoquinoline via boronate ester formation as decribed in
GENERAL METHOD 7-1. MS (M+1 ) = 300.5.
Step 4: 6-(6-Methoxy-1-methylisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine
6-(6-Methoxy-1-methylisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine was prepared from 6-methoxy-1-methyl-7-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)isoquinoline and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (Intermediate I) via Suzuki conditions described in GENERAL METHOD 1 -1 (156 mg). MS (M+1 ) = 420.6.
Step 5: 1 -Methyl- 7-( 6-(methyl( 2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinolin-6-ol
6-(6-Methoxy-1-methylisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 followed by free basing via GENERAL METHOD 8-1 to afford the title compound (6.0 mg). LC/MS Rt = 0.41 . MS (M+1 ) = 404.1 1 H NMR (400 MHz, Methanol-d4) δ 8.95 (s, 1 H), 8.55 (d, J= 9.9 Hz, 1 H), 8.21 (d, J= 6.8 Hz, 1 H), 8.03 (d, J= 6.8 Hz, 1 H), 7.68 (d, J= 9.9 Hz, 1 H), 7.58 (s, 1 H), 5.55-5.23 (m, 1 H), 3.22 (s, 3H), 3.14 (s, 3H), 2.07-1 .98 (m, 4H), 1 .66 (s, 6H), 1 .54 (s, 6H).
Example 29-3: Synthesis of 1 ,3-dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000113_0001
Step 1: N-(1, 1-Dimethoxypropan-2-ylidene)-1-phenylmethanamine
A mixture of 1 , 1 -dimethoxypropan-2-one (12.7 ml_, 105 mmol), phenylmethanamine (10 ml_, 91 mmol) and magnesium sulfate (1 1 .0 g, 91 mmol) in DCM (91 ml.) was stirred at room
temperature for 18 h. The reaction mixture was filtered through celite and concentrated in vacuo to afford N-(1 ,1 -dimethoxypropan-2-ylidene)-1 -phenylmethanamine. 1H NMR (400 MHz, Chloroform- d) 5 7.34 (d, J= 5.0 Hz, 4H), 7.32-7.18 (m, 1 H), 4.56 (d, J= 1.6 Hz, 2H), 3.44 (s, 6H), 1.95 (s, 3H).
Step 2: N-Benzyl-1, 1-dimethoxypropan-2-amine
To a mixture of N-(1 , 1-dimethoxypropan-2-ylidene)-1-phenylmethanamine (18.9 g, 91 mmol) in MeOH (101 ml) cooled to -78°C was added NaBH4 (4.0 g, 105 mmol) portionwise over 20 min. The resulting suspension was stirred at room temperature for 3 days and concentrated in vacuo. The resulting slurry was taken up in toluene, washed with water, saturated aqueous ammonium chloride, then dried over Na2S04 and concentrated in vacuo to afford N-benzyl-1 ,1 - dimethoxypropan-2-amine (19.0 g). MS (M+1 ) = 210.0.
Step 3: 1, 1-Dimethoxypropan-2-amine
A mixture of N-benzyl-1 , 1 -dimethoxypropan-2-amine (19.0 g, 91 mmol) and 10% palladium on carbon (0.97 g, 0.910 mmol in MeOH (182 ml) was stirred at 50 psi under hydrogen for 18 h. The suspension was then filtered through celite and concentrated in vacuo to afford 1 , 1- dimethoxypropan-2-amine (10.8 g). 1 H NMR (400 MHz, Chloroform-d) δ 3.93 (d, J= 5.9 Hz, 1 H), 3.38 (s, 3H), 3.35 (s, 3H), 2.95 (p, J= 6.4 Hz, 1 H), 1 .04 (d, J= 6.5 Hz, 3H).
Step 4: N-(1-(3-Bromo-4-methoxyphenyl)ethyl)-1, 1-dimethoxypropan-2-amine
A mixture of 1 , 1 -dimethoxypropan-2-amine (0.5 g, 4.20 mmol), 1-(3-bromo-4- methoxyphenyl)ethanone (1.2 g, 5.0 mmol) and titanium(IV) isopropoxide (2.5 ml, 8.39 mmol) in THF (5 ml) was stirred at room temperature for 18 h. The mixture was then cooled to 0°C, NaBH4 (0.48 g, 12.59 mmol) was added followed by EtOH (5 mL) and the mixture was stirred at room temperature 4 h. 1 N aqueous ammonium hydroxide was added and the reaction was filtered through celite. The filtrate was extracted with ether and the organic extract was extracted with 1 N aqueous HCI. The aqueous mixture was basified with 1 N aqueous NaOH and extracted with ether (2x). The organic extracts were dried over Na2S04 and concentrated in vacuo to afford N-(1 -(3- bromo-4-methoxyphenyl)ethyl)-1 ,1 -dimethoxypropan-2-amine (0.94 g). MS (M+1 ) = 334.1.
Step 5: 7-Bromo-6-methoxy-1,3-dimethylisoquinoline
To a 5 mL microwave vial containing N-(1-(3-bromo-4-methoxyphenyl)ethyl)-1 , 1- dimethoxypropan-2-amine (1 g, 1.81 mmol) cooled to -78°C was slowly added chlorosulfonic acid 2.2 mL, 32.5 mmol). The mixture was then allowed to stir at room temperature for 18 h, then added dropwise to ice water. The resulting aqueous material was washed with ether, basified with saturated aqueous Na2C03, and extracted with DCM (2x). The resulting organic extracts were dried over Na2S04 and concentrated in vacuo. The crude product was purified by silica gel chromatography (20-80% EtOAc/Hep) to afford 7-bromo-6-methoxy-1 ,3-dimethylisoquinoline (298 mg). MS (M+1 ) = 268.0.
Step 6: 6-Methoxy- 1, 3-dimethyl-7-(4, 4, 5, 5-tetramethyl-1 , 3, 2-dioxaborolan-2-yl)isoquinoline
6-methoxy-1 ,3-dimethyl-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline was prepared from 7-bromo-6-methoxy-1 ,3-dimethylisoquinoline via boronate ester formation as described in GENERAL METHOD 7-1 (135 mg). MS (M+1 ) = 314.1 .
Step 7: 6-( 6-Methoxy-1, 3-dimethylisoquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin- 4-yl)pyridazin-3-amine 6-(6-Methoxy-1 ,3-dimethylisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethyl^
yl)pyridazin-3-amine was prepared from 6-methoxy-1 ,3-dimethyl-7-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)isoquinoline and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin- 3-amine (Intermediate 1 ) via Suzuki conditions described in GENERAL METHOD 1 -1 (253 mg). MS (M+1 ) = 434.2.
Step 8: 1,3-Dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amin
yl)isoquinolin-6-ol
6-(6-Methoxy-1 ,3-dimethylisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 followed by free basing via GENERAL METHOD 3-1 to afford the title compound (9.7 mg). LC/MS Rt = 0.43. MS (M+1 ) = 420.2 1 H NMR (400 MHz, DMSO-d6) δ 8.58 (s, 1 H), 8.48 (d, J = 9.9 Hz, 1 H), 7.38 (d, J = 10.0 Hz, 1 H), 7.27 (s, 1 H), 7.16 (s, 1 H), 5.21 - 4.85 (m, 1 H), 3.33 (s, 3H), 2.99 (s, 3H), 2.88 (s, 3H), 1.63-1 .37 (m, 4H), 1.28 (s, 6H), 1 .1 1 (s, 6H).
Example 30-1 : Synthesis of 7-hydroxy-3-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline-1 -carbonitrile
Figure imgf000115_0001
Step 1: N-(4-Bromo-3-methoxybenzyl)-1 , 1 -dimethoxypropan-2-amine
To a mixture of 4-bromo-3-methoxybenzaldehyde (500 mg, 2.325 mmol) and 1 , 1 - dimethoxypropan-2-amine (prepared as in Example 29-3) (610 mg, 5.12 mmol) in THF (4.7 mL) was added sodium triacetoxyborohydride (749 mg, 3.53 mmol). The reaction mixture was stirred at room temperature for 18 h, diluted with EtOAc, washed with saturated aqueous sodium
bicarbonate, water, and brine, and dried over MgS04 to afford N-(4-bromo-3-methoxybenzyl)-1 , 1- dimethoxypropan-2-amine (700 mg). MS (M+1 ) = 319.9
Step 2: 6-Bromo-7-methoxy-3-methylisoquinoline
To a 5 mL microwave vial containing N-(4-bromo-3-methoxybenzyl)-1 , 1 -dimethoxypropan-2-amine
(5.8 g, 18.26 mmol) cooled to -78°C was slowly added chlorosulfonic acid (12.2 ml, 183 mmol).
The mixture was then allowed to stir at room temperature for 18 h and added dropwise to ice water.
The resulting aqeuous was washed with diethyl ether, basified with saturated aqueous Na2C03, and extracted with DCM (2x). The resulting organic extracts were dried over Na2S04 and concentrated in vacuo. The crude product was purified by silica gel chromatography (20-80%
EtOAc/Hep) to afford 6-bromo-7-methoxy-3-methylisoquinoline (630 mg). MS (M+1 ) = 254.0 Step 3: 6-Bromo-7-methoxy-3-methylisoquinoline 2-oxideTo mixture of 6-bromo-7-methoxy-
3- methylisoquinoline (630 mg, 2.50 mmol) and MTO (24.9 mg, 0.100 mmol) in DCM (12.5 ml.) cooled to 0°C was added hydrogen peroxide (0.31 ml_, 5.00 mmol). The resulting mixture was stirred at room temperature for 3 days. 5-10 mg of Mn02 was added and the reaction mixture was stirred for 2 h. The suspension was filtered through celite and the filtrate was concentrated in vacuo to afford 6-bromo-7-methoxy-3-methylisoquinoline 2-oxide (610 mg). MS (M+1 ) = 269.9
Step 4: 6-Bromo- 7-methoxy-3-methylisoquinoline- 1 -carbonitrile
To a mixture of 6-bromo-7-methoxy-3-methylisoquinoline 2-oxide (585 mg, 2.18 mmol) and trimethylsilyl cyanide (351 ml_, 2.62 mmol) in DCM (10.9 ml.) was added dimethylcarbamoyl chloride (200 ml_, 2.18 mmol). The reaction mixture was stirred at room temperature for 18 h, quenched with excess saturated aqueous Na2C03 and extracted with DCM (2X). The organic extracts were washed with brine, dried over Na2S04 and concentrated in vacuo to afford 6-bromo- 7-methoxy-3-methylisoquinoline-1-carbonitrile (756 mg). MS (M+1 ) = 279.0
Step 5: 7-Methoxy-3-methyl-6-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)isoquinoline-1- carbonitrile
7-Methoxy-3-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline-1 - carbonitrile was prepared from 6-bromo-7-methoxy-3-methylisoquinoline-1 -carbonitrile via boronate ester formation as decribed in GENERAL METHOD 7-1 (702 mg). MS (M+1 ) = 325.1.
Step 6: 7-Methoxy-3-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)ami
3-yl)isoquinoline-1 -carbonitrile
7-Methoxy-3-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline-1 -carbonitrile was prepared from 7-methoxy-3-methyl-6-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)isoquinoline-1 -carbonitrile and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-
4- yl)pyridazin-3-amine (Intermediate 1 ) via Suzuki conditions described in GENERAL METHOD 1 - 1 (960 mg). MS (M+1 ) = 445.4.
Step 7: 7-Hydroxy-3-methyl-6-( 6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)isoquinoline-1 -carbonitrile
7-Methoxy-3-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinoline-1 -carbonitrile was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 followed by free basing via GENERAL METHOD 3-1 to afford the title compound (1 1.6 mg). LC/MS Rt = 0.46. MS (M+1 ) = 431 .4 1H NMR (400 MHz, Methanol-d4) δ 8.34 (s, 1 H), 8.24 (d, J= 9.8 Hz, 1 H), 7.84 (s, 1 H), 7.64 (s, 1 H), 7.26 (d, J= 9.8 Hz, 1 H), 5.18 (d, J= 12.0 Hz, 1 H), 3.06 (s, 3H), 2.68 (s, 3H), 1.76 (dd, J= 12.7, 3.4 Hz, 2H), 1.57 (t, J= 12.6 Hz, 2H), 1 .41 (s, 6H), 1 .25 (s, 6H). Example 31 -1 : Synthesis of 1 -amino-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoli
Figure imgf000117_0001
Step 1: 2-(7-Bromo-6-methoxyisoquinolin-1-yl)isoindoline-1,3-dione
To a mixture of 7-bromo-6-methoxyisoquinoline 2-oxide (prepared as described in in
Example 3-7 (300 mg, 1 .18 mmol), phthalimide (191 mg, 1.29 mmol) and tributylamine (0.6 ml_, 2.48 mmol) in DCM (5.2 mL) was added benzoyl chloride (0.2 ml_, 1.71 mmol) in DCM (0.74 ml_). The reaction mixture was stirred at room temperature for 0.5 h, quenched with water and extracted with DCM. The organic extracts were washed with brine and concentrated in vacuo. The crude product was purified by silica gel chromatography (0-100% EtOAc/Hep) to afford 2-(7-bromo-6- methoxyisoquinolin-1 -yl)isoindoline-1 ,3-dione (257 mg). MS (M+1 ) = 385.0
Step 2: 2-( 6-Methoxy- 7-(4, 4, 5, 5-tetramethyl-1 ,3, 2-dioxaborolan-2-yl)isoquinolin- 1- yl)isoindoline- 1, 3-dione
2-(6-Methoxy-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinolin-1-yl)isoindoline-1 ,3- dione was prepared from 2-(7-bromo-6-methoxyisoquinolin-1-yl)isoindoline-1 , 3-dione via boronate ester formation as described in GENERAL METHOD 7-1. The resulting solid was stirred in diethyl ether and filtered, washing with diethyl ether to afford the product (1 10 mg). MS (M+1 ) = 430.9
Step 3: 6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinolin- 1 -amine
A mixture of 2-(6-methoxy-7-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinolin-1 - yl)isoindoline-1 , 3-dione (1 10 mg, 0.256 mmol), 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine (108 mg, 0.383 mmol), potassium acetate (271 mg, 1.28 mmol) and Xphos precatalyst (20.1 mg, 0.026 mmol) in DMF (1 .3 mL) was stirred at room temperature for 18 h. The mixture was diluted with excess MeOH, filtered through celite and concentrated in vacuo. To the crude material was added hydrazine (35% aq, 68.8 μΙ_, 0.767 mmol) and MeOH (2 mL). The mixture was heated at 60°C for 48 h and purified via preparative reverse-phase HPLC (acetonitrile in water, 5mM ammonium hydroxide modifier) to afford 6-methoxy-7-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-1-amine (15 mg). MS (M+1 ) = 421.3
Step 4: 1-Amino-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinolin-6-ol
6-Methoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-1 - amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-1 followed by free basing via GENERAL METHOD 3-1 to afford the title compound (5.1 mg). LC/MS Rt = 0.43. MS (M+1 ) = 405.3 1 H NMR (400 MHz, Methanol-d4) δ 8.55 (s, 1 H), 8.32 (d, J = 10.0 Hz, 1 H), 7.61 (d, J = 6.1 Hz, 1 H), 7.31 (d, J = 9.9 Hz, 1 H), 7.07 (s, 1 H), 6.81 (d, J = 6.1 Hz, 1 H), 5.28 - 5.04 (m, 1 H), 3.01 (s, 3H), 1.78 - 1.50 (m, 4H), 1.41 (s, 6H), 1.25 (s, 6H).
Example 32-1 : Synthesis of 7-hydroxy-1 ,3-dimethyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazoline-2,4(1 H,3H)-dione
Figure imgf000118_0001
Step 1: 7-(Benzyloxy)-6-bromoquinazoline-2,4(1H,3H)-dione
A mixture of 2-amino-4-(benzyloxy)-5-bromobenzoic acid (500 mg, 1 .55 mmol) and urea (3.7 g, 62.1 mmol) was heated at 150°C for 48 h. The reaction was then cooled to 100°C, excess water was added and the mixture was stirred for 5 min, then cooled to room temperature. The product precipitated and was filtered to afford 7-(benzyloxy)-6-bromoquinazoline-2,4(1 H,3H)-dione (341 mg). MS (M+1 ) = 348.9.
Step 2: 7-(Benzyloxy)-6-bromo-1,3-dimethylquinazoline-2,4(1H,3H)-dione
A mixture of 7-(benzyloxy)-6-bromoquinazoline-2,4(1 H,3H)-dione (292 mg, 0.841 mmol),
K2C03 (581 mg, 4.21 mmol) and Mel (0.174 ml_, 2.78 mmol) in DMF (4.2 ml.) was stirred at room temperature for 3 h. The mixture was diluted with water and the resulting precipitate was filtered and washed with ether to afford 7-(benzyloxy)-6-bromo-1 ,3-dimethylquinazoline-2,4(1 H,3H)-dione (207 mg). MS (M+1 ) = 377.0.
Step 3: 7-(Benzyloxy)-1, 3-dimethyl-6-(4, 4, 5, 5-tetramethyl- 1, 3, 2-dioxaborolan-2- yl)quinazoline-2,4( 1H, 3H)-dione
7-(Benzyloxy)-1 ,3-dimethyl-6-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazoline- 2,4(1 H,3H)-dione was prepared from 7-(benzyloxy)-6-bromo-1 ,3-dimethylquinazoline-2,4(1 H,3H)- dione via boronate ester formation as decribed in GENERAL METHOD 7-1 (1 15 mg). MS (M+1 ) = 423.2.
Step 4: 7-(Benzyloxy)-1 , 3-dimethyl-6-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinazoline-2,4(1H, 3H)-dione
7-(Benzyloxy)-1 ,3-dimethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazoline-2,4(1 H,3H)-dione was prepared from 7-(benzyloxy)-1 ,3-dimethyl-6-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)quinazoline-2,4(1 H,3H)-dione and 6-chloro-N-methyl-N- (2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 1 ) via Suzuki conditions described in GENERAL METHOD 1 -2 (129 mg). MS (M+1 ) = 543.2. Step 5: 7 -Hydroxy- 1, 3-dimethyl-6-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinazoline-2,4(1H, 3H)-dione
To a 50 mL flask containing 7-(benzyloxy)-1 ,3-dimethyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazoline-2,4(1 H,3H)-dione (93 mg, 0.171 mmol) was added HBr (33% in AcOH, 1 .4 mL, 8.57 mmol). The resulting suspension was stirred at room temperature for 1 h, quenched with excess water and purified by SCX via GENERAL METHOD 3-1 to afford the title compound (16.8 mg). LC/MS Rt = 0.51. MS (M+1 ) = 453.2 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1 H), 8.28 (d, J= 9.9 Hz, 1 H), 7.35 (d, J= 9.8 Hz, 1 H), 6.85 (s, 1 H), 4.99 (m, 1 H), 3.49 (s, 3H), 3.30 (s, 3H), 2.95 (s, 3H), 1.59-1 .37 (m, 4H), 1 .27 (s, 6H), 1 .1 1 (s, 6H).
Example 33-1 : Synthesis of 6-hydroxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)benzo[d]oxazol-2(3H)-one
Figure imgf000119_0001
Step 1: 5-Bromo-6-methoxybenzo[d]oxazol-2(3H)-one
A mixture of 6-methoxybenzo[d]oxazol-2(3H)-one (400 mg, 2.422 mmol) and bromine (0.19 mL, 3.63 mmol) in water (1.3 mL, 72.7 mmol) and AcOH (1.4 mL, 24.22 mmol) was stirred at room temperature for 18 h. The reaction mixture was diluted with water and the product was filtered to afford 5-bromo-6-methoxybenzo[d]oxazol-2(3H)-one (535 mg) as a white precipitate. MS (M+1 ) = 243.9.
Step 2: 6-Met oxy-5-(4, 4, 5, 5-tetramet yl-1 ,3, 2-dioxaborolan-2-yl)benzo[d]oxazol-2(3H)-one 6-Methoxy-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)benzo[d]oxazol-2(3H)-one was prepared from 5-bromo-6-methoxybenzo[d]oxazol-2(3H)-one via boronate ester formation as described in GENERAL METHOD 7-1 (400 mg). MS (M+1 ) = 292.1 .
Step 3: 6-Methoxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin yl)benzo[d]oxazol-2(3H)-one
6-Methoxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)benzo[d]oxazol-2(3H)-one was prepared from 6-methoxy-5-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)benzo[d]oxazol-2(3H)-one and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-
4-yl)pyridazin-3-amine (Intermediate 1 ) via Suzuki conditions described in GENERAL METHOD 1 -
2 (77 mg). MS (M+1 ) = 412.2.
Step 4: 6-hydroxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyrM
yl)benzo[d]oxazol-2(3H)-one
6-Methoxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)benzo[d]oxazol-2(3H)-one was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 to afford the title compound (3.3 mg). LC/MS Rt = 0.46. MS (M+1 ) = 398.2 1H NMR (400 MHz, Methanol-d4) δ 7.92 (d, J= 9.9 Hz, 1 H), 7.35 (s, 1 H), 7.18 (d, J= 9.9 Hz, 1 H), 6.86 (s, 1 H), 5.04 (t, J= 12.5 Hz, 1 H), 3.00 (s, 3H), 1 .73 (dd, J= 12.8, 3.4 Hz, 2H), 1 .53 (t, J= 12.4 Hz, 2H), 1 .39 (s, 6H), 1.23 (s, 6H).
Example 34-1 : Synthesis of 2-methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-2H-indazol-6-ol
Figure imgf000120_0001
Step 1: 5-Bromo-6-methoxy-1 -methyl-1 H-indazole and 5-bromo-6-methoxy-2-methyl-2H- indazole
To a suspension of NaH (85 mg, 2.1 1 mmol) in DMF (4 ml.) at 0°C was added 1 H-indazol-6- ol (400 mg, 1 .76 mmol) in DMF (4 ml_). The mixture was stirred at room temperature for 20 min, cooled to 0°C and Mel (0.17 ml_, 2.64 mmol) was added. The reaction mixture was stirred at room temperature for 2 h, quenched with excess saturated aqeuous NH4CI and extracted with EtOAc. The organic exact was dried over Na2S04 and concentrated in vacuo. The crude product was purified by silica gel chromatography (0-100% EtOAc/Hep) to afford 5-bromo-6-methoxy-1-methyl- 1 H-indazole (276 mg). MS (M+1 ) = 243.0 and 5-bromo-6-methoxy-2-methyl-2H-indazole (76 mg). MS (M+1 ) = 243.0.
Step 2: 6-Methoxy-2-methyl-5-(4, 4, 5, 5-tetramethyl- 1, 3, 2-dioxaborolan-2-yl)-2H-indazole 6-Methoxy-2-methyl-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-2H-indazole was prepared from 5-bromo-6-methoxy-2-methyl-2H-indazole via boronate ester formation as described in GENERAL METHOD 7-1 (88 mg). MS (M+1 ) = 289.1 .
Step 3: 6-(6-Methoxy-2-methyl-2H-indazol-5-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
6-(6-Methoxy-2-methyl-2H-indazol-5-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine was prepared 6-methoxy-2-methyl-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan- 2-yl)-2H-indazole and 6-chloro-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (Intermediate 1 ) via Suzuki conditions described in GENERAL METHOD 1 -2 (101 mg). MS (M+1 ) = 409.3.
Step 4: 2-Methyl-5-(6-(methyl(2, 2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-2H- indazol-6-ol
6-(6-Methoxy-2-methyl-2H-indazol-5-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4- yl)pyridazin-3-amine was subjected to methoxy deprotection conditions as described in GENERAL METHOD 2-3 to afford the title compound (12.5 mg). LC/MS Rt = 0.43. MS (M+1 ) = 395.2. 1 H NMR (400 MHz, Methanol-d4) δ 8.19-8.06 (m, 3H), 7.30 (d, J= 9.9 Hz, 1 H), 6.94 (s, 1 H), 5.08 (t, J= 12.5 Hz, 1 H), 4.14 (s, 3H), 3.01 (s, 3H), 1.69 (dd, J= 12.7, 3.6 Hz, 2H), 1.58 (t, J= 12.5 Hz, 2H), 1 .39 (s, 6H), 1 .23 (s, 6H).
Example 34-2: Synthesis of 1 -methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-1 H-indazol-6-ol
Figure imgf000121_0001
1-Methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-^
was prepared in the same manner as Example 34-1 (9.4 mg). LC/MS Rt = 0.48. MS (M+1 ) = 395.3. 1H NMR (400 MHz, Methanol-d4) δ 8.24-8.12 (m, 2H), 7.95 (d, J= 1.0 Hz, 1 H), 7.32 (d, J= 9.9 Hz, 1 H), 6.93 (d, J= 0.9 Hz, 1 H), 5.14-4.99 (m, 1 H), 3.97 (s, 3H), 3.01 (s, 3H), 1 .69 (dd, J= 12.7, 3.6 Hz, 2H), 1.57 (t, J= 12.4 Hz, 2H), 1 .38 (s, 6H), 1.23 (s, 6H).
Example 35-1 : Synthesis of 6-hydroxy-2-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-1(2H)-one hydrochloride salt
Figure imgf000121_0002
Step 1: 7-Bromo-6-methoxy-2-methylisoquinolin-1(2H)-one
In a 100 mL flask, KHMDS (3.31 mL, 1.653 mmol) was added to Intermediate 14 (210 mg, 0.827 mmol) in dioxane (12 mL) at 0 °C, and stirred for 0.5 hr. Methyl iodide (0.078 mL, 1.240 mmol) was added, and the mixture was allowed to stir at room temperature overnight. Water (50 mL) was added to quench the reaction, then the reaction mixture was diluted with EtOAc (100 mL). The layers were separated, and the aqueous layer was extracted with EtOAc (2X). The combined organic layers were washed with brine, dried over MgS04, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (20-60% EtOAc/Hep, 24 g Isco column) to provide 7-bromo-6-methoxy-2-methylisoquinolin-1 (2H)-one as a yellowish solid, (1 12 mg, 0.409 mmol, 49.5 % yield). MS (M+1 ) = 270.1 . 1H NMR (400 MHz, CHLOROFORM-d) δ 8.61 (s, 1 H), 7.08 (d, J=7.53 Hz, 1 H), 6.86 (s, 1 H), 6.40 (d, J=7.28 Hz, 1 H), 3.99 (s, 3H), 3.59 (s, 3H).
Step 2: (6-Methoxy-2-methyl-1-oxo-1,2-dihydroisoquinolin-7-yl)boronic acid The title compound (product not isolated) was prepared following GENERAL METHOD 7-1 for boronate ester formation from 7-bromo-6-methoxy-2-methylisoquinolin-1 (2H)-one. MS (M+1 ) = 234.2 (boronic acid), 316.2 (boronic ester).
Step 3: 6-Methoxy-2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino
3-yl)isoquinolin-1(2H)-one
The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction of Intermediate 1 (70 mg, 0.25 mmol) and (6-methoxy-2-methyl-1-oxo-1 ,2- dihydroisoquinolin-7-yl)boronic acid (100 mg, 0.43 mmol). MS (M+1 ) = 436.6.
Step 4 : 6-Hydroxy-2-methyl-7-(6-(methyl(2,2, 6, 6-tetramethylpiperidin-4-yl)amino)pyridazin- 3-yl)isoquinolin-1(2H)-one
The title compound (48 mg, 0.103 mmol, 49.7 % yield) was prepared from 6-methoxy-2- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-1 (2H)-one (90 mg, 0.207 mmol) following GENERAL METHOD 2-3 for demethylation using boron tribromide. The crude residue was purified by preparative reverse-phase HPLC (10-30% MeCN/Water, 0.01 % TFA modifier). LC/MS Rt = 0.50 min. MS (M+1 ) = 422.2. 1H NMR (400 MHz, METHANOL-^) δ 8.59 (s, 1 H), 8.48 (d, J=10.04 Hz, 1 H), 8.06 (d, J=10.04 Hz, 1 H), 7.45 (d, J=7.28 Hz, 1 H), 7.10-7.19 (m, 1 H), 6.61 (d, J=7.28 Hz, 1 H), 5.02 (br. s., 1 H), 3.60 (s, 3H), 3.20 (s, 3H), 1 .99-2.15 (m, 4H), 1.65 (s, 6H), 1.58 (s, 6H).
Example 35-2: Synthesis of 2-ethyl-6-hydroxy-7-(6-((2,2,6,6-tetramethylpiperidin-4- yl)oxy)pyridazin-3-yl)isoquinolin-1(2H)-one
Figure imgf000122_0001
Step 1: 7-Bromo-2-ethyl-6-methoxyisoquinolin-1(2H)-one
In a 30 ml. vial, LiHMDS (2.4 ml_, 1 .2 mmol) was added to Intermediate 14 (75 mg, 0.295 mmol) in dioxane (1 .5 ml.) at RT, and stirred for 0.5 hr. Then ethyl iodide (70 μΙ, 0.885 mmol) was added, and the mixture was stirred overnight at 80°C, and allowed to stir overnight. The reaction was cooled to room temperature and water (10 ml.) was added to quench reaction, and the reaction mixture was diluted with EtOAc (15 ml_). The layers were separated and the aqueous layers extracted with EtOAc (2x). The combined organic layers were washed with brine, dried over MgS04, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (20-60% EtoAc/Hep, 12 g Isco column, elutes at 40% EtOAc) to provide the title compound as a yellowish solid (62 mg, 0.220 mmol). MS (M + 1 ) = 283.9. 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1 H), 7.51 (d, J=7.33 Hz, 1 H), 7.29 (s, 1 H), 6.60 (d, J=7.33 Hz, 1 H), 3.90-4.01 (m, 5H), 1.23 (t, J=7.07 Hz, 3H).
Step: 2-Et yl-6-met oxy-7-(4,4, 5, 5-tetramet yl-1 ,3, 2-dioxaborolan-2-yl)isoquinolin- 1(2H)- one
The title compound (product not isolated) was prepared following GENERAL METHOD 7-1 for boronate ester formation from 7-bromo-2-ethyl-6-methoxyisoquinolin-1 (2H)-one. The crude product was used without further purification. MS (M+1 ) = 248.1 (boronic acid), 330.1 (boronic ester).
Step 3: 2-Et yl-6-met oxy- 7-(6-((2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)isoquinolin-1(2H)-one
The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction of Intermediate 3 (50 mg, 0.185 mmol) and 2-ethyl-6-methoxy-7-(4,4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinolin-1 (2H)-one (68.7 mg, 0.278 mmol). MS (M+1 ) = 437.2.
Step 4: 2^thyl-6-hydroxy-7-(6-((2!2fi!6-tetramethylpiperidin-4-yl)oxy)pyridazin-3^ yl)isoquinolin- 1 ( 2H)-one
The title compound (9 mg, 0.019 mmol, 13.18 % yield) was prepared from 2-ethyl-6- methoxy-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinolin-1 (2H)-one (63 mg, 0.144 mmol) following GENERAL METHOD 2-3 for demethylation using boron tribromide. The crude residue was purified by preparative reverse-phase HPLC (10-30% ACN/Water, 0.01 % TFA modifier). LC/MS Rt = 0.53 min. MS (M+1 ) = 423.2. 1H NMR (400 MHz, METHANOL-^) δ 8.75 (d, J=9.35 Hz, 1 H), 8.70 (s, 1 H), 7.97-8.05 (m, 3H), 7.51 (d, J=7.33 Hz, 1 H), 7.20 (s, 1 H), 6.66 (d, J=7.33 Hz, 1 H), 5.69-5.84 (m, 1 H), 4.10 (q, J=7.07 Hz, 2H), 2.53 (dd, J=3.79, 13.64 Hz, 2H), 1 .99 (m, 2H), 1.66 (s, 6H), 1.62 (s, 6H), 1.38 (t, J=7.07 Hz, 3H).
Example 35-3: Synthesis of 1 -ethoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000123_0001
Step 1: 7-Bromo-1-et oxy-6-met oxyisoquinoline
In a 30 ml. vial, 7-bromo-1-chloro-6-methoxyisoquinoline (200 mg, 0.734 mmol) was partially dissolved in 2 ml. sodium ethoxide 3 M solution (2 ml_, 6.00 mmol), and stirred at 80 °C overnight. The mixture was concentrated in vacuo. The crude residue was purified by silica gel chromatography (12g Isco column, 5-30% EtOAc/Hep) to provide the title compound (157 mg, 0.556 mmol, 76 % yield). MS (M+1 ) = 284.0. 1 H NMR (400 MHz, Chloroform-d) δ 8.46 (s, 1 H), 7.95 (d, J=5.8 Hz, 1 H), 7.09 (d, J=6.0 Hz, 1 H), 7.03 (s, 1 H), 4.53 (q, J=l A Hz, 2H), 4.02 (s, 3H), 1 .50 (t, J=7.0 Hz, 3H).
Step 2: 1-Ethoxy-6-methoxy- 7-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)isoquinoline The title compound (product not isolated) was prepared following GENERAL METHOD 7-1 for boronate ester formation from 7-bromo-2-ethyl-6-methoxyisoquinolin-1 (2H)-one. MS (M+1 ) = 248.1 (boronic acid), 330.1 (boronic ester).
Step 3: 6-( 1-Ethoxy-6-methoxyisoquinolin-7-yl)-N-methyl-N-(2, 2, 6, 6-tetramethylpiperidin-4- yl)pyridazin-3-amine
The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction of Intermediate 1 (100 mg, 0.354 mmol) and 1 -ethoxy-6-methoxy-7-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline (150 mg, 0.607 mmol). MS (M+1 ) = 450.1 .
Step 4 : 1-Ethoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)isoquinolin-6-ol
The title compound (10 mg, 0.022 mmol, 10 % yield) was prepared from 6-(1-ethoxy-6- methoxyisoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine (100 mg, 0.222 mmol) following GENERAL METHOD 2-3 for demethylation using boron tribromide. The crude residue was purified by preparative reverse-phase HPLC (10-30% MeCN/Water, 0.01 % TFA modifier). LC/MS Rt = 0.56 min. MS (M+1 ) = 436.3. 1H NMR (400 MHz, METHANOL-^) δ 8.64 (s, 1 H), 8.24 (d, J=10.04 Hz, 1 H), 7.81 (d, J=6.02 Hz, 1 H), 7.36 (d, J=10.04 Hz, 1 H), 7.19 (s, 1 H), 7.12 (d, J=6.02 Hz, 1 H), 5.29 (br. s., 1 H), 4.54 (q, J=7.03 Hz, 2H), 3.05 (s, 3H), 1.69-1 .87 (m, 4H), 1 .48- 1 .56 (m, 9H), 1.36 (s, 6H).
Example 35-4: Synthesis of 7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)isoquinoline-1 ,6-diol
Figure imgf000124_0001
Step 1 : 1 -Ethoxy-6-methoxy- 7-(6-((2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)isoquinoline
The title compound (product not isolated) was prepared following GENERAL METHOD 1 -1 for Suzuki reaction of Intermediate 3 (30 mg, 0.1 1 1 mmol) and 1 -ethoxy-6-methoxy-7-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)isoquinoline (50 mg, 0.151 mmol). MS (M+1 ) = 437.6.
Step 2: 5-Bromo-7-( 6-( ( 2, 2, 6, 6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinoline- 1, 6- diol The title compound (product not isolated) was prepared from 1-ethoxy-6-methoxy-7-(6- ((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinoline (45 mg, 0.103 mmol) following GENERAL METHOD 2-3 for demethylation using boron tribromide. The crude residue was purified by preparative reverse-phase HPLC (15-40% ACN/Water, 5 mM NH4OH modifier). MS (M+1 ) = 475.0.
Step 3. 7-( 6-((2, 2, 6, 6-Tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinoline- 1, 6-diol In a 30 mL vial, 5-bromo-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3- yl)isoquinoline-1 , 6-diol (5 mg, 10.56 μηηοΙ), and Pd-C (1 1 .24 mg, 10.56 μηηοΙ) were combined in ethanol (Volume: 1500 μΙ) with a few drops of 1 N HCI. The vial was purged with H2 for 5 min, and stirred overnight under an H2 balloon at room temperature. The crude residue was purified by preparative reverse-phase HPLC (10-30% MeCN/Water, 0.01 % TFA modifier) to provide the title compound (1 mg, 0.002 mmol, 23 % yield). LC/MS Rt = 0.49 min. MS (M+1 ) = 395.1. 1H NMR (400 MHz, METHANOL-^) δ 8.72 (s, 1 H), 8.36 (d, J=9.54 Hz, 1 H), 7.30 (d, J=9.54 Hz, 1 H), 7.08 (d, J=7.28 Hz, 1 H), 7.04 (s, 1 H), 6.49 (d, J=7.28 Hz, 1 H), 5.75 (t, J=4A 4 Hz, 1 H), 2.42 (dd, J=4.02, 13.80 Hz, 2H), 1.68-1 .80 (m, 2H), 1 .54 (s, 6H), 1 .45 (s, 6H).
Example 36-1 to 36-6:
The following compounds were prepared from triflate Intermediate 16 or Intermediate 17 , and an alkyne, according to GENERAL METHOD 10-2 for Sonogashira reaction, followed by amine cyclization according to GENERAL METHOD 10-3, and then methoxy deprotection as outlined in GENERAL METHOD 2-1. The products were purified by preparative reverse-phase HPLC (10- 30% MeCN/Water, 0.01 % TFA modifier).
Table Examples 36-1 to 36-6:
Figure imgf000125_0001
Figure imgf000126_0001
Figure imgf000127_0001
Example 37-1 : Synthesis of 3-methyl-7-(6-(piperazin-1 -yl)pyridazin-3-yl)isoquinolin-6-ol
Figure imgf000127_0002
Step 1: tert-Butyl 4-(6-(5-formyl-4-hydroxy-2-methoxyphenyl)pyridazin-3-yl)p
carboxylate
The title compound (107 mg, 0.258 mmol, 21.43 % yield) was prepared following
GENERAL METHOD 9-1 for Suzuki reaction of ferf-butyl 4-(6-chloropyridazin-3-yl)piperazine-1- carboxylate (4.7 g, 16.62 mmol) and Intermediate 15 (500 mg, 1 .8 mmol). Dioxane was removed in vacuo and EtOAc/DCM and water were added. The layers were separated and the aqueous layer was washed with DCM. The combined organics were dried over MgS04, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (5-80% EtOAc/Hep). MS (M+1 ) = 415.5. 1H NMR (400 MHz, Chloroform-d) δ 1 1 .57-1 1 .51 (m, 1 H), 9.85 - 9.79 (m, 1 H), 8.26-8.20 (m, 1 H), 7.82-7.73 (m, 1 H), 6.97-6.89 (m, 1 H), 6.58 - 6.52 (m, 1 H), 3.96- 3.90 (m, 3H), 3.70 (s, 4H), 3.65-3.56 (m, 4H), 1 .53-1.47 (m, 9H).
Step 2: tert-Butyl 4-(6-(5-formyl-2-methoxy-4-(((trifluoromethyl)sulfonyl)oxy)- phenyl)pyridazin-3-yl)piperazine- 1 -carboxylate
In a 20 ml. vial, ferf-butyl 4-(6-(5-formyl-4-hydroxy-2-methoxyphenyl)pyridazin-3- yl)piperazine-1 -carboxylate (100 mg, 0.241 mmol) and N-phenyltrifluoromethane sulfonimide (172 mg, 0.483 mmol) were dissolved in DCM (1 .21 ml.) under N2. TEA (135 μΙ, 0.965 mmol) was added, and the heterogenous mixture stirred for several hours at room temperature until complete dissolution was seen. The reaction was stirred overnight, then taken up in minimum amount of DCM and purified by silica gel chromatography (10-80% EtOAc/Hep, column pretreated with TEA). The collected fractions were concentrated, and placed under high vacuum for several days to provide the title compound, (125 mg, 0.229 mmol, 95 % yield). MS (M+1 ) = 547.3. 1H NMR (400 MHz, CHLOROFORM-d) δ 10.14 (s, 1 H), 8.57 (s, 1 H), 7.77 (d, J= 9.54 Hz, 1 H), 6.92-6.98 (m, 2H), 3.98 (s, 3H), 3.57-3.82 (m, 8H), 1.50 (s, 9H).
Step 3: tert-Butyl 4-(6-(5-formyl-2-methoxy-4-(prop-1-yn-1-yl)phenyl)pyridazin-3- yl)piperazine-1-carboxylate
The title compound (product not isolated) was prepared following GENERAL METHOD 10-1 for Sonogashira reaction of ferf-butyl 4-(6-(5-formyl-2-methoxy-4-(((trifluoromethyl)sulfonyl)- oxy)phenyl)pyridazin-3-yl)piperazine-1-carboxylate (50 mg, 0.091 mmol) and prop-1-yne (18 mg, 0.457 mmol). MS (M+1 ) = 437.6.
Step 4: tert-Butyl 4-(6-(6-methoxy-3-methylisoquinolin-7-yl)pyridazin-3-yl)piperazine-1- carboxylate
The title compound (product not isolated) was prepared following GENERAL METHOD 11 -1 for aldehyde cyclization from ferf-butyl 4-(6-(5-formyl-2-methoxy-4-(prop-1 -yn-1-yl)phenyl)pyridazin-
3-yl)piperazine-1-carboxylate (39 mg, 0.089 mmol). MS (M+1 ) = 436.5.
Step 5: 3-Methyl-7-(6-(piperazin-1-yl)pyridazin-3-yl)isoquinolin-6-ol
The title compound (5 mg, 0.015 mmol, 16.85 % yield) was prepared from ferf-butyl 4-(6-(6- methoxy-3-methylisoquinolin-7-yl)pyridazin-3-yl)piperazine-1-carboxylate (39mg, 0.090 mmol) following GENERAL METHOD 2-1 for demethylation using pyridine hydrochloride. The crude residue was purified by preparative reverse-phase HPLC (10-30% ACN/Water, 0.01 % TFA modifier). LC/MS Rt = 0.35 min. MS (M+1 ) = 322.01. 1 H NMR (400 MHz, METHANOL-^) δ 8.89
(s, 1 H), 8.39 (s, 1 H), 8.19 (d, J=9.85 Hz, 1 H), 7.38 (d, J=9.85 Hz, 1 H), 7.31 (s, 1 H), 7.06 (s, 1 H),
3.55-3.68 (m, 4H), 2.85-2.94 (m, 4H), 2.50 (s, 3H).
LC/MS CONDITIONS
Column: Waters Acquity UPLC BEH C18 1.7um, 2.1x30mm (Part#: 186002349)
Flow rate: 1 ml_/min
Temperature: 55 °C (column temp)
Mobile phase compositions:
A. 0.05% formic acid in water.
B. 0.04% formic acid in methanol
Gradient:
Figure imgf000128_0001
0.50 1.000 20.0 80.0
0.60 1.000 5.0 95.0
0.80 1.000 5.0 95.0
0.90 1.000 95.0 5.0
1.15 1.000 95.0 5.0
ABBREVIATIONS:
Figure imgf000129_0001
spectrometry m: multiplet M: Molar
M, as in M+1 : Molecular Mass mM: millimolar
mAB: monoclonal antibody Me: methyl
MeOH: methanol mg: milligram
MHz: mega Hertz min: minute(s)
mm: millimeter mmol: millimole
ml_: milliliter MS: mass spectrometry
N: normal nM: nanomolar
NMR: Nuclear Magnetic Resonance
NMP: N-methylpyrrolidone
Spectroscopy
PBST: Phosphate buffered saline with Tween Ph: phenyl
PhSH: thiophenol pM: picomolar
ppm: parts per million
q: quartet
RIPA: radio-immunoprecipitation assay Rt: retention time
RT: room temperature
s: singlet sat: saturated
SCX: Strong Cation Exchange SFC: Supercritical Fluid Chromatography
SPhos: 2-Dicyclohexylphosphino-2',6'- dimethoxybiphenyl
t: triplet TBAF: tetra-butylammonium fluoride
TBSCI: tert-butyldimethylsilyl chloride tBu: tert-butyl
TEA: triethylamine Tf: triflate
THF: tetrahydrofuran TLC: thin layer chromatography
TMSOTf: trimethylsilyl
TMB: tertramethylbenzidine
trifluoromethanesulfonate
uL: microliter umol: micromol
UV: ultraviolet
wt: weight XPhos Precatalyst: Chloro(2-
XPhos: 2-Dicyclohexylphosphino-2',4',6'- dicyclohexylphosphino-2',4',6'-triisopropyl-1 , 1 '- triisopropylbiphenyl
biphenyl)[2-(2-aminoethyl)phenyl)]palladium(ll)
Xphos Precatalyst, 2nd Generation: chloro(2- dicyclohexylphosphino-2',4',6'-triisopropyl-1 , 1 '- biphenyl)[2-(2'-amino-1 , 1 '- biphenyl)]palladium(ll)
Biological Example 1 :
A cellular SMN ELISA was used to measure the effects of low molecular weight compounds on SMN protein elevation. Cells from a myoblast cell line derived from the SMNdelta7 mouse model (kind gift from Steve Burden, NYU) were seeded into a 384-well plate at a density of 3000 cells / well and treated with compounds for 24 hours. ELISA capture plates were prepared by coating 384- well plates (Immulon 4HBX) with 0.5 ug/mL of anti-SMN mAb (BD Science, Catalog number 610647) at 4 °C overnight. The plates were washed 5 times with 1 10 uL of PBS-Tween (0.05% Tween-20, PBST), blocked with 100 uL of 1 % BSA in PBST for 2 hours and washed (5 times) with 100uL of PBST. After 24 hours of compound treatment cells were lysed in a modified RIPA-buffer, on ice for 1 hour. 20 uL of lysate and 20 uL of 1 % BSA were then added to the ELISA capture plates and incubated at 4 °C overnight. Plates were washed (5 times) with PBST and then incubated with 1 :100 dilution of primary rabbit anti-SMN polyclonal antibody (Santa cruz, Catalog number SC-15320) at room temperature for 1 hour and subsequently washed (5 times) with 1 10 uL of PBST. This was followed by addition of 1 :100 Goat anti-Rabbit IgG-HRP linked (Cell
Signaling, Catalog number 7074) secondary antibody for 1 hour. Plates were then washed with PBST and incubated with 40 uL TMB substrate (Cell Signaling, Catalog number 7004L) at room temperature for 1-10 minutes with shaking. The reaction was stopped by addition of 40 uL of stop solution (Cell signaling, Catalog number 7002L) and absorption was measured at 450 nm. Data was reported as fold activation over DMSO control, and EC50.
ELISA assay condition 1 : compound concentration range 20 nM-10 uM; ELISA assay condition 2: compound concentration 100 pM - 10 uM. Data generated in Biological Example 1 using ELISA conditions 1 or 2. Biological Example 2:
SMN2 Full reporter construct:
pSMN2 Splicing Luciferase reporter was constructed according to Zhang et al., (An in vivo reporter system for measuring increased inclusion of exon 7 in SMN2 mRNA: potential therapy of SMA. Gene Ther. 2001 Oct;8(20):1532-8) via overlapping PCR. The final PCR fragment containing Exon6-lntron6-Exon7(with a C insertion before 49T)-lntron7-Exon8-Luciferase was inserted to BamHI and Notl site of pCAG-IRESblast expression vector (Chen et al., Establishment and
Maintenance of Genomic Methylation Patterns in Mouse Embryonic Stem Cells by Dnmt3a and Dnmt3b. Mol Cell Biol. 2003 Aug;23(16):5594-605).
Human SMN2 exon1 -exon6 cDNA were amplified by PCR (Forward primer:
ACGGATCCATGGCGATGAGCAGCGG; reverse primer:
GCCAGTATGATAGCCACTCATGTACC), and Exon6-lntron6 (Forward primer:
ATAATTCCCCCACCACCTCCC; reverse primer: CATTCCCTACAATCAATTTCAAATCAGAG) were fused by overlap PCR to create Exon1 -lntron6 fragment, and was inserted into pSMN2 splicing Luciferase reporter via BamHI and Hind 111 sites to create pSMN2 exon1-6 splicing reporter. SMN2 promoter 5.1 kb fragment was amplified from human genomic DNA by PCR (Forward primer: CAGCTAGCACGCGTAAGCTCTGATTGGTGAGCGATGGTGG; reverse primer:
CACTCGAGAGCAAACCCGCGGGTGCGCAGCG), and inserted between Mlul site and BamHI site (Blunted using DNA blunting kit, Takara, Cat# 6025) of pSMN2 exon1-6 splicing reporter to replace chicken β-actin promoter and the human cytomegalovirus immediate early enhancer, to finalize the pSMN2-Full-reporter.
SMN2 Full reporter stable clone in NSC34 cells:
pSMN2-Full reporter was stably transfected into NSC34 cells using Lipofectamine 2000, which were subsequently selected in blasticidin-containing medium for two weeks. Blasticidin-resistant colonies were screened by Luciferase signal and response to SAHA as well as by RT-PCR for alternative splicing of Exon7.
Reporter Gene Assay:
NSC34 SMN2 Full reporter stable line was cultured in DMEM (Invitrogen,
Cat:1 1965)+10%FBS+7μg/ml Blasticidin. Cells in culture media without Blasticidin were seeded into a 384-well plate at a density of 6000 cells / well and treated with compounds for 24 hours.
Luciferase signal was assessed by BrightGlo assay. Eequal volume of BrightGlo reagent
(Promega, Cat# E2620) was added to the cells and incubated for 10 minutes at room temperature.
Luminescent signal was read in either BioTek or Envision plate reader.
Reporter gene assay conditions: compound concentration range 3 nM-10 uM.
Activity Table: Data generated in Biological Example 3 were generated using methods described in Biological example 1 or Biological example 2.
Figure imgf000132_0001
4-chloro-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4-
Preparation 1 1 280 nM, 2.6 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
4-chloro-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-
Preparation 12 30 nM, 2.6 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
4-chloro-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-
Preparation 13 7 nM, 2.6 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-
1-1 20 nM, 2.9 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
6-(6-(methyl(1 , 2,2,6,6- pentamethylpiperidin-4-
1-2 340 nM, 2.4 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
6-(6-((3aR,6aS)-5- methylhexahydropyrrolo[3,4-c]pyrrol-
1-3 255 nM, 2.5 fold 2(1 H)-yl)pyridazin-3-yl)quinolin-7-ol
2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-
2-1 20 nM, 3.0 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
7-(6-(methyl(1 , 2,2,6,6- pentamethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-
3-1 70 nM, 3.1 fold ol
7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-
3-2 5 nM, 2.75 fold ol
7-(6-((2,2,6,6-tetramethylpiperidin-4-
3-3 83 nM, 2.5 fold yl)oxy)pyridazin-3-yl)isoquinolin-6-ol
7-(6-((3aR,6aS)-5- methylhexahydropyrrolo[3,4-c]pyrrol-
3-4 60 nM, 2.3 fold 2(1 H)-yl)pyridazin-3-yl)isoquinolin-6-ol
1-cyclopropyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-
3-5 14 nM, 2.54 fold ol
7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-
3-6 8 nM, 2.43 fold 1 ,6-diol
6-hydroxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 -
3-7 2 nM, 2.9 fold carbonitrile
6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-7-
4-1 6 nM, 2.6 fold ol 8-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 300 nM, 2.6 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 7 nM, 2.82 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
2-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--2 4 nM, 3.2 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
3-chloro-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 14 nM, 2.8 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
3-bromo-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--2 8 nM, 2.6 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-3--3 8 nM, 3.1 fold carbonitrile
6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(1 -methyl--4 200 nM, 2.2 fold 1 H-imidazol-4-yl)quinolin-7-ol
3-(1 H-imidazol-1 -yl)-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4--5 50 nM, 2.5 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-3,7--6 300 nM, 2.9 fold diol
3-ethyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 86 nM, 2.9 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
3-isopropyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--2 190 nM, 2.50 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-2(1 H)--1 30 nM, 2.25 fold one
7-hydroxy-1 -methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-2(1 H)--2 7 nM, 2.25 fold one
4-methoxy-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4--1 10 nM, 3.3 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(pyrrolidin--2 65 nM, 2.6 fold 1-yl)quinolin-7-ol 2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4--3 1 10 nM, 2.6 fold morpholinoquinolin-7-ol
4-(dimethylamino)-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4--4 12 nM, 2.7 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
4-ethoxy-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4--5 20 nM, 3.3 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(1 -methyl--1 60 nM, 2.6 fold 1 H-pyrazol-4-yl)quinolin-7-ol
4-methoxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 3 nM, 2.4 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 9 nM, 2.7 fold yl)amino)pyridazin-3-yl)quinoxalin-6-ol
6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3-(tetrahydro--1 63 nM, 2.91 fold 2H-pyran-4-yl)quinolin-7-ol
3-chloro-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 16 nM, 2.68 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
3-bromo-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--2 19 nM, 2.52 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
3-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--3 7 nM, 3.28 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
5-bromo-3-methyl-7-(6-
(methyl(2,2,6,6-tetramethylpiperidin-4--1 258 nM, 2.44 fold yl)amino)pyridazin-3-yl)quinolin-6-ol
6-hydroxy-1 -methyl-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-4(1 H)--1 685 nM, 2.56 fold one
2,3-dimethyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 15 nM, 3.24 fold yl)amino)pyridazin-3-yl)quinoxalin-6-ol
2-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--2 9 nM, 3.31 fold yl)amino)pyridazin-3-yl)quinoxalin-6-ol 3-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--3 1 16 nM, 2.72 fold yl)amino)pyridazin-3-yl)quinoxalin-6-ol
4-methoxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 1 10 nM, 2.65 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
4-(azetidin-1 -yl)-2-methyl-6-(6-
(methyl(2,2,6,6-tetramethylpiperidin-4--1 1 84 nM, 2.86 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
7-hydroxy-2-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-4--2 1 48 nM, 2.68 fold carbonitrile
4-cyclopropyl-2-methyl-6-(6-
(methyl(2,2,6,6-tetramethylpiperidin-4--3 1 65 nM, 2.72 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
4-(3,6-dihydro-2H-pyran-4-yl)-2- methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--4 1 166 nM, 2.72 fold yl)amino)pyridazin-3-yl)quinolin-7-ol
2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(tetrahydro- 2H-pyran-4-yl)quinolin-7-ol formate-5 1 286 nM, 2.69 fold salt
2-methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-4-(oxetan-3--6 1 165 nM, 3.18 fold yl)quinolin-7-ol
4-(dimethylamino)-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol-1 1 60 nM, 3.01 fold di-formate salt
7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinazolin--1 1 1 10 nM, 2.58 fold 4(1 H)-one
6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4--1 1 41 nM, 3.6 fold yl)amino)pyridazin-3-yl)quinazolin-7-ol
7-hydroxy-1 -methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3,4--1 1 71 nM, 1 .74 fold dihydroquinolin-2(1 H)-one
7-hydroxy-1 -methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-3,4--1 1 23 nM, 2.72 fold dihydroquinolin-2(1 H)-one 7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 --1 1 16 nM, 2.58 fold carbonitrile
7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2--2 1 7 nM, 3.14 fold carbonitrile
6-hydroxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2--3 1 6 nM, 3.47 fold carbonitrile
6-hydroxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 --4 1 7 nM, 2.57 fold carboxamide
7-hydroxy-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2--5 1 1 nM, 2.16 fold carboxamide
6-hydroxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2--6 1 7 nM, 2.6 fold carboxamide
methyl 6-hydroxy-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinoline-2--7 1 17 nM, 2.64 fold carboxylate
6-hydroxy-7-(6-(piperazin-1- yl)pyridazin-3-yl)quinoline-2--1 1 46 nM, 2.49 fold carbonitrile
7-hydroxy-6-(6-(piperazin-1- yl)pyridazin-3-yl)quinoline-2--2 1 395 nM, 2.42 fold carbonitrile
7-(6-(piperazin-1 -yl)pyridazin-3--3 1 199 nM, 2.74 fold yl)isoquinolin-6-ol
7-(6-(1 ,2,3,6-tetrahydropyridin-4--1 1 155 nM, 1.9 fold yl)pyridazin-3-yl)quinolin-6-ol
1 -methyl-6-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-7--1 1 14 nM, 2.96 fold ol
1 -methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6--2 1 9 nM, 2.53 fold ol 1 ,3-dimethyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6--3 2 40 nM, Amax 2453 ol
7-hydroxy-3-methyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinoline-1 --1 2 30 nM, Amax 2008 carbonitrile
1 -amino-7-(6-(methyl(2, 2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6--1 2 25 nM, Amax 3014 ol
7-hydroxy-1 ,3-dimethyl-6-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinazoline--1 2 133 nM, Amax 3313 2,4(1 H,3H)-dione
6-hydroxy-5-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3--1 2 3 uM, Amax 2396 yl)benzo[d]oxazol-2(3H)-one
2-methyl-5-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-2H-indazol-6--1 2 150 nM, Amax 3448 ol
1 -methyl-5-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)-1 H-indazol-6--2 2 220 nM, Amax 3534 ol
6-hydroxy-2-methyl-7-(6- (methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin--1 1 3 nM, 2.43 fold 1 (2H)-one hydrochloride salt
2-ethyl-6-hydroxy-7-(6-((2,2,6,6- tetramethylpiperidin-4- yl)oxy)pyridazin-3-yl)isoquinolin--2 2 363 nM, Amax 4321 1 (2H)-one
1 -ethoxy-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6--3 1 13 nM, 2.26 fold ol
7-(6-((2,2,6,6-tetramethylpiperidin-4- yl)oxy)pyridazin-3-yl)isoquinoline-1 ,6--4 1 96 nM, 3.26 fold diol
7-(6-(methyl(2,2,6,6-tetramethyl- piperidin-4-yl)amino)-pyridazin-3-yl)-3--1 1 282 nM, 2.32 fold phenylisoquinolin-6-ol
3-methyl-7-(6-(methyl(2,2,6,6- tetramethylpiperidin-4-yl)amino)--2 1 9 nM, 2.64 fold pyridazin-3-yl)isoquinolin-6-ol 3-cyclopropyl-7-(6-(methyl(2,2,6,6- tetramethyl-piperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-
36-3 2 30 nM, Amax 2690 ol
3-isopropyl-7-(6-(methyl(2,2,6,6- tetramethyl-piperidin-4- yl)amino)pyridazin-3-yl)isoquinolin-6-
36-4 2 190 nM, Amax 1956 ol
3-propyl-7-(6-((2,2,6,6- tetramethylpiperidin-4-yl)oxy)-
36-5 2 580 nM, Amax 1603 pyridazin-3-yl)isoquinolin-6-ol
3-isopropyl-7-(6-((2,2,6,6- tetramethylpiperidin-4-yl)oxy)-
36-6 2 640 nM, Amax 1298 pyridazin-3-yl)isoquinolin-6-ol
3-methyl-7-(6-(piperazin-1 -
37-1 1 124 nM, 2.33 fold yl)pyridazin-3-yl)isoquinolin-6-ol
Comparison of the SMN ELISA (as described in Biological Example 1 ) potency and fold activation for unsubstituted, ortho-hydroxy and ortho-alkoxy 6,6-heterocyclic substituted pyridazines:
Figure imgf000139_0001
Figure imgf000139_0002
Figure imgf000140_0001

Claims

What is claimed is
1. A compound or salt thereof according to Formula (I)
Figure imgf000141_0001
(IA)
wherein
A is bicyclic heteroaryl or heterocyle having 9 or 10 ring atoms and 1 or 2 ring N atoms and 0 or 1 O atoms, which bicyclic heteroaryl or heterocycle is substituted with 0, 1 , 2, 3, 4 or 5 substituents independently selected from -C(0)NH2,-C(0)0-Ci-C4alkyl, aryl, oxo, cyano, halogen, hydroxy, Ci-C4alkyl, C2-C4alkenyl, C2-C4alkynyl, d-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl CrC4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, CrC4alkyl heteroaryl, CrC4alkoxy aryl, Ci-C4alkoxy heterocyclyl, Ci-C4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, Ci-C4alkoxy, amino and mono-and di-Ci-C4alkylamino; and
B is a group of the formula:
Figure imgf000141_0002
wherein
m, n and p are independently selected from 0 or 1 ;
R, R-i, R2, R3, and R4 are independently selected from the group consisting of hydrogen, Ci C4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino;
R5 and R6 are independently selected from hydrogen and fluorine; or
R and R3, taken in combination form a fused 5 or 6 member heterocyclic ring having 0 or 1 additional ring heteroatoms selected from N, O or S;
R-i and R3, taken in combination form a C-|-C3alkylene group;
R-i and R5, taken in combination form a C-|-C3alkylene group;
R3 and R4, taken in combination with the carbon atom to which they attach, form a spirocyclicC3-C6cycloalkyl;
X is CRARB, O, N R7 or a bond; R7 is hydrogen, or d-C4alkyl;
RA and RB are independently selected from hydrogen and Ci-C4alkyl, or RA and RB, taken in combination, form a divalent C2-C5alkylene group;
Z is CR8 or N; when Z is N, X is a bond;
R8 is hydrogen or taken in combination with R6 form a double bond; or
B is a group of the formula:
Figure imgf000142_0001
wherein
p and q are independently selected from the group consisting of 0, 1 , and 2;
R9 and Ri3 are independently selected from hydrogen and CrC4alkyl;
R-io and Ri4 are independently selected from hydrogen, amino, mono- and di-Cr
C4akylamino and Ci-C4alkyl, which alkyl is optionally substituted with hydroxy, amino or mono- and di-Ci-C4akylamino;
Rii is hydrogen, Ci-C4alkyl, amino or mono- and di-Ci-C4akylamino;
Ri2 is hydrogen or CrC4alkyl; or
R9 and Rn, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 CrC4alkyl groups; or
R-n and R12, taken in combination form a saturated azacycle having 4 to 7 ring atoms which is optionally substituted with 1 -3 C1-C4alkyl groups.
2. The compound of claim 1 , or salt thereof, wherein A is selected from:
Figure imgf000143_0001
Figure imgf000143_0002
Figure imgf000143_0003
Figure imgf000143_0004
Figure imgf000143_0005
Figure imgf000143_0006
wherein
u and v are each, independently, 0, 1 , 2 or 3; and
each Ra and R are, independently, selected from , cyano, halogen, hydroxy, C1-C4alkyl, C2- C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci-
C4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, and Ci-C4alkoxy substituted with hydroxy, Ci-C4alkoxy, amino and mono-and di-Ci-C4alkylamino.
3. A compound according to any one of claims 1-2, or salt thereof, wherein A is selected from:
Figure imgf000144_0001
4. A compound, according to any one of claims 1-3, or salt thereof, wherein the compound of formula II:
Figure imgf000144_0002
(II)
wherein Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, CrC4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, d-C4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, C1-C4alkoxy substituted with hydroxy, C1-C4alkoxy, amino and mono-and di-C1-C4alkylamino.
5. A compound, or salt thereof, according to any one of claims 1 -3, wherein the compound is of formula III:
Figure imgf000145_0001
(III)
wherein
Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, CrC4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, CrC4alkoxy, amino and mono-and di-Ci-C4alkylamino.
6. A compound, according to any one of claims 1-3, or salt thereof, wherein the compound is of formula IV:
Figure imgf000145_0002
(IV)
wherein
Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, CrC4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, CrC4alkoxy, amino and mono-and di-Ci-C4alkylamino.
7. A compound, according to any one of claims 1-3, or salt thereof, wherein the compound is of formula V:
Figure imgf000146_0001
(V)
wherein
Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, C1-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C1-C4alkyl heteroaryl, C1-C4alkoxy aryl, C1-C4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, C1-C4alkoxy substituted with hydroxy, C1-C4alkoxy, amino and mono-and di-C1-C4alkylamino.
8. A compound, according to any one of claims 1-3, or salt thereof, wherein the compound is of formula VI:
Figure imgf000146_0002
(VI)
wherein
Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, d-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, CrC4alkoxy, amino and mono-and di-Ci-C4alkylamino.
9. A compound, according to any one of claims 1-3, or salt thereof, wherein the compound is of formula VII:
Figure imgf000146_0003
wherein
Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, CrC4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, C1-C4alkyl aryl, C1-C4alkyl heterocyclyl, C1-C4alkyl heteroaryl, C1-C4alkoxy aryl, C1-C4alkoxy heterocyclyl, C1-C4alkoxy heteroaryl, C1-C4alkoxy substituted with hydroxy, C1-C4alkoxy, amino and mono-and di-C1-C4alkylamino.
10. A compound, according to any one of claims 1-3, or salt thereof, wherein the compound is of formula VIII:
Figure imgf000147_0001
(VIII)
wherein
Rc and Rd are each, independently, selected from hydrogen, cyano, halogen, hydroxy, CrC4alkyl, C2-C4alkenyl, C2-C4alkynyl, d-C4alkoxy, C3-C7cycloalkyl, heterocyclyl, heteroaryl, heterocyclyl Ci- C4alkyl, CrC4alkyl aryl, CrC4alkyl heterocyclyl, Ci-C4alkyl heteroaryl, CrC4alkoxy aryl, CrC4alkoxy heterocyclyl, CrC4alkoxy heteroaryl, CrC4alkoxy substituted with hydroxy, CrC4alkoxy, amino and mono-and di-Ci-C4alkylamino.
1 1. The compound of any one of claims 1 -10, wherein B is selected from the group consisting of
Figure imgf000147_0002
Figure imgf000147_0003
Figure imgf000147_0004
, and wherein X is O or N(Me) or NH; and
R is hydrogen or methyl.
12. A compound, or salt thereof, selected from the group consisting of
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-2(1 H)-one;
6- (6-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1 H)-yl)pyridazin-3-yl)quinolin-7-ol;
7- hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin- 2(1 H)-one;
6- (6-(methyl(1 ,2,2,6,6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-morpholinoquinolin-
7- ol;
4-chloro-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol; 3-bromo-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
3-ethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
3-(1 H-imidazol-1-yl)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7- ol;
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(1-methyl-1 H-imidazol-4- yl)quinolin-7-ol;
3- isopropyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3,7-diol;
7- hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-3- carbonitrile;
6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
4- (dimethylamino)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinolin-7-ol;
3- chloro-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
4- methoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7- ol;
6- (3-(benzyloxy)isoquinolin-6-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine; 8-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-7-ol;
7- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline-1 ,6-diol;
7-(6-(methyl(1 ,2,2,6,6-pentamethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
1-cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol; 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
6-(1 -(benzyloxy)isoquinolin-7-yl)-N-methyl-N-(2,2,6,6-tetramethylpiperidin-4-yl)pyridazin-3-amine; 7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6-ol; 2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidi H- pyrazol-4-yl)quinolin-7-ol;
2- methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin
4-ethoxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
4- chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidi
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3-(tetrahydro-2H-pyran yl)quinolin-7-ol;
3- chloro-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-^^
3-bromo-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin-6
3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinolin
5- bromo-3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
6- hydroxy-1-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
4(1 H)-one;
2,3-dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperi
2- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin
3- methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoxalin
4- methoxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoli
4-(azetidin-1-yl)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazi
yl)quinolin-7-ol;
7- hydroxy-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qu carbonitrile;
4-cyclopropyl-2-methyl-6-(6-(methyl(2,2,6,6-tetra
7- ol;
4-(3,6-dihydro-2H-pyran-4-yl)-2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4- yl)amino)pyridazin-3-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(tetrahyd
pyran-4-yl)quinolin-7-ol;
2-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-4-(oxetan-^ yl)quinolin-7-ol;
4-(dimethylamino)-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-^
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin H)-one;
6- (6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinazolin-7-ol;
7- hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-3 dihydroquinolin-2(1 H)-one; 7-hydroxy-1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyrid
dihydroquinolin-2(1 H)-one;
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquino
carbonitrile;
7-hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoli
carbonitrile;
6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)qui
carbonitrile;
6- hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin
carboxamide;
7- hydroxy-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2^ carboxamide;
6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2- carboxamide;
methyl 6-hydroxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)quinoline-2- carboxylate;
6- hydroxy-7-(6-(piperazin-1 -yl)pyridazin-3-yl)quinoline-2-carbonitrile;
7- hydroxy-6-(6-(piperazin-1 -yl)pyridazin-3-yl)quinoline-2-carbonitrile;
7-(6-(piperazin-1-yl)pyridazin-3-yl)isoquinolin-6-ol;
7-(6-(1 ,2,3,6-tetrahydropyridin-4-yl)pyridazin-3-yl)quinolin-6-ol;
1-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-7-ol;
1-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
1 ,3-dimethyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
7-hydroxy-3-methyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinoline- 1 -carbonitrile;
1- amino-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol;
7-hydroxy-1 ,3-dimethyl-6-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3- yl)quinazoline-2,4(1 H,3H)-dione;
6-hydroxy-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)benzo[d]oxazol- 2(3H)-one;
2- methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-2H-indazol-6-ol;
1-methyl-5-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)-1 H-indazol-6-ol;
6-hydroxy-2-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin- 1 (2H)-one;
2-ethyl-6-hydroxy-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinolin-1 (2H)-one; 1-ethoxy-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)isoquinolin-6-ol; 7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)pyridazin-3-yl)isoquinoline-1 ,6-d
7-(6-(methyl(2,2,6,6-tetramethyl-piperidin-4-yl)amino)-pyridazin-3-yl)-3-phenyli
3-methyl-7-(6-(methyl(2,2,6,6-tetramethylpiperidin-4-yl)amino)pyridazin-3-yl)i
3-cyclopropyl-7-(6-(methyl(2,2,6,6-tetramethyl-piperidin-4-yl)amino)pyridazin-3-yl)i
3-isopropyl-7-(6-(methyl(2,2,6,6-tetramethyl-piperidin-4-yl)amino)pyridazin-3-yl)i
3-propyl-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)-pyridazin-3-yl)isoquinolin-6-ol;
3-isopropyl-7-(6-((2,2,6,6-tetramethylpiperidin-4-yl)oxy)-pyri^ and
3-methyl-7-(6-(piperazin-1 -yl)pyridazin-3-yl)isoquinolin-6-ol;
13. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.
14. A combination comprising a therapeutically effective amount of a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof and one or more therapeutically active co-agents.
15. A method to treat, prevent or ameliorate an SMN-deficiency-related condition, comprising administering to a subject in need thereof an effective amount of a compound or salt thereof of any one of claims 1 to 12.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3053577A1 (en) * 2015-02-09 2016-08-10 F. Hoffmann-La Roche AG Compounds for the treatment of cancer
WO2016128343A1 (en) * 2015-02-09 2016-08-18 F. Hoffmann-La Roche Ag Compounds for the treatment of cancer
WO2017100726A1 (en) 2015-12-10 2017-06-15 Ptc Therapeutics, Inc. Methods for treatng huntington's disease
JP2018519273A (en) * 2015-05-20 2018-07-19 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ Compounds for treating spinal muscular atrophy
US10357543B2 (en) 2015-11-16 2019-07-23 Ohio State Innovation Foundation Methods and compositions for treating disorders and diseases using Survival Motor Neuron (SMN) protein
WO2021014428A1 (en) 2019-07-25 2021-01-28 Novartis Ag Regulatable expression systems
WO2021084495A1 (en) 2019-11-01 2021-05-06 Novartis Ag The use of a splicing modulator for a treatment slowing progression of huntington's disease
US11382918B2 (en) 2017-06-28 2022-07-12 Ptc Therapeutics, Inc. Methods for treating Huntington's Disease
US11395822B2 (en) 2017-06-28 2022-07-26 Ptc Therapeutics, Inc. Methods for treating Huntington's disease
US11407753B2 (en) 2017-06-05 2022-08-09 Ptc Therapeutics, Inc. Compounds for treating Huntington's disease
WO2022208170A1 (en) 2021-03-29 2022-10-06 Novartis Ag The use of the splicing modulator brataplam for slowing progression of huntington's disease
US11608501B2 (en) 2017-06-14 2023-03-21 Ptc Therapeutics, Inc. Methods for modifying RNA splicing
US11672799B2 (en) 2013-07-31 2023-06-13 Novartis Ag 1,4-disubstituted pyridazine quinolne analogs there of and methods for treating SMN-deficiency-related conditions
US11685746B2 (en) 2018-06-27 2023-06-27 Ptc Therapeutics, Inc. Heteroaryl compounds for treating Huntington's disease
US11780839B2 (en) 2018-03-27 2023-10-10 Ptc Therapeutics, Inc. Compounds for treating Huntington's disease
US11858941B2 (en) 2018-06-27 2024-01-02 Ptc Therapeutics, Inc. Heterocyclic and heteroaryl compounds for treating Huntington's disease

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MY174339A (en) 2012-08-13 2020-04-09 Novartis Ag 1,4-disubstituted pyridazine analogs and methods for treating smn-deficiency-related conditions
WO2019000144A1 (en) * 2017-06-26 2019-01-03 深圳市博奥康生物科技有限公司 Cho cell expressing aitr gene and use thereof
WO2019000150A1 (en) * 2017-06-26 2019-01-03 深圳市博奥康生物科技有限公司 Method for constructing recombinant cho cell highly expressing tl6
BR122020005073A2 (en) 2017-08-04 2020-10-13 Skyhawk Therapeutics, Inc. COMPOUND, PHARMACEUTICAL COMPOSITION AND USE OF THE COMPOUND
KR20210135241A (en) 2019-02-05 2021-11-12 스카이호크 테라퓨틱스, 인코포레이티드 Methods and compositions for controlling splicing
WO2020163544A1 (en) 2019-02-06 2020-08-13 Skyhawk Therapeutics, Inc. Methods and compositions for modulating splicing
US11129829B2 (en) 2019-06-17 2021-09-28 Skyhawk Therapeutics, Inc. Methods for modulating splicing
CN110256342B (en) * 2019-07-16 2022-06-07 河南省科学院化学研究所有限公司 Synthetic method of 2-cyano quinoline derivative
EP4119151A4 (en) 2021-06-03 2023-10-25 MD Healthcare Inc. Composition for preventing or treating eye diseases comprising extracellular vesicles derived from lactobacillus paracasei

Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2427943A1 (en) * 1974-06-10 1976-01-08 Thomae Gmbh Dr K 5(6)-Pyridazinyl-benzimidazoles - esp 3-oxo-pyridazinyl derivs having hypotensive, anti-rhombic and cardiotonic activity
WO2001058891A2 (en) * 2000-02-11 2001-08-16 Vertex Pharmaceuticals Incorporated Piperazine and piperidine derivatives for treatment or prevention of neuronal damage
WO2001066129A1 (en) * 2000-03-10 2001-09-13 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Substances for the treatment of spinal muscular atrophy
WO2001094351A1 (en) * 2000-06-07 2001-12-13 Vertex Pharmaceuticals Incorporated Caspase inhibitors and uses thereof
US6376508B1 (en) * 2000-12-13 2002-04-23 Academia Sinica Treatments for spinal muscular atrophy
WO2007137030A2 (en) * 2006-05-19 2007-11-29 Abbott Laboratories Cns active fused bicycloheterocycle substituted azabicyclic alkane derivatives
US20080247964A1 (en) * 2006-05-08 2008-10-09 Yuelian Xu Substituted azaspiro derivatives
WO2009108657A2 (en) * 2008-02-28 2009-09-03 Vertex Pharmaceuticals Incorporated Heteroaryl derivatives as cftr modulators
WO2009137503A1 (en) * 2008-05-05 2009-11-12 Envivo Pharmaceuticals, Inc. Hdac inhibitors and uses thereof
WO2009150139A2 (en) * 2008-06-10 2009-12-17 Neurosearch A/S Indolyl-pyridazinyl-diazabicyclononane derivatives in labelled and unlabelled form and their use in diagnostic methods
WO2010028192A1 (en) * 2008-09-03 2010-03-11 Repligen Corporation Compositions including 6-aminohexanoic acid derivatives as hdac inhibitors
WO2011036167A1 (en) * 2009-09-22 2011-03-31 Novartis Ag Use of nicotinic acetylcholine receptor alpha 7 activators
WO2011063159A1 (en) * 2009-11-18 2011-05-26 Plexxikon, Inc. Compounds and methods for kinase modulation, and indications therefor
WO2012022467A2 (en) * 2010-08-16 2012-02-23 Santhera Pharmaceuticals (Schweiz) Ag Novel benzoquinone derivatives and use thereof as modulators of mitochondrial function
WO2012127393A1 (en) * 2011-03-18 2012-09-27 Novartis Ag COMBINATIONS OF ALPHA 7 NICOTINIC ACETYLCHOLINE RECEPTOR ACTIVATORS AND mGluR5 ANTAGONISTS FOR USE IN DOPAMINE INDUCED DYSKINESIA IN PARKINSON'S DISEASE
WO2014028459A1 (en) * 2012-08-13 2014-02-20 Novartis Ag 1,4-disubstituted pyridazine analogs and methods for treating smn-deficiency-related conditions

Family Cites Families (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2516040C2 (en) * 1974-06-10 1984-12-20 Dr. Karl Thomae Gmbh, 7950 Biberach Benzimidazoles, processes for their preparation and pharmaceuticals containing them
BR9408137A (en) 1993-11-24 1997-08-12 Du Pont Merck Pharma Prodrug ester compound pharmaceutical composition treatment method and inhibition method
US6982259B2 (en) 2002-04-30 2006-01-03 Schering Aktiengesellschaft N-heterocyclic derivatives as NOS inhibitors
JP2006502134A (en) 2002-08-09 2006-01-19 アストラゼネカ アクチボラグ Compounds having activity at metabotropic glutamate receptors
KR20050039846A (en) 2002-08-09 2005-04-29 아스트라제네카 에이비이 Oxadiazoles as modulators of metabotrophic glutamate receptor-5
EP1529045A2 (en) 2002-08-09 2005-05-11 Astra Zeneca AB New compounds
EP1562945B1 (en) 2002-11-11 2006-11-15 Neurosearch A/S 1,4-diazabicyclo(3,2,2)nonane derivatives, preparation and therapeutical use thereof
JP2007524596A (en) 2003-02-28 2007-08-30 トランスフォーム・ファーマシューティカルズ・インコーポレイテッド Co-crystal pharmaceutical composition
US20050075375A1 (en) 2003-05-14 2005-04-07 Anadys Pharmaceuticals, Inc. Heterocyclic compounds for treating hepatitis C virus
EP1642898B1 (en) 2003-06-27 2013-03-27 Msd K.K. Heteroaryloxy nitrogenous saturated heterocyclic derivative
US7666862B2 (en) 2003-08-15 2010-02-23 Merck & Co., Inc. Mitotic Kinesin Inhibitors
US7399765B2 (en) * 2003-09-19 2008-07-15 Abbott Laboratories Substituted diazabicycloalkane derivatives
US7655657B2 (en) * 2003-12-22 2010-02-02 Abbott Laboratories Fused bicycloheterocycle substituted quinuclidine derivatives
WO2005077373A2 (en) 2004-02-03 2005-08-25 Astrazeneca Ab Treatment of gastro-esophageal reflux disease (gerd)
WO2005077368A2 (en) 2004-02-03 2005-08-25 Astrazeneca Ab Treatment of gastro-esophageal reflux disease (gerd)
FR2868780B1 (en) 2004-04-13 2008-10-17 Sanofi Synthelabo DERIVATIVES OF 1-AMINO-PHTHALAZINE, THEIR PREPARATION AND THEIR THERAPEUTIC APPLICATION
CA2567851A1 (en) 2004-05-21 2006-01-05 Merck & Co., Inc. Amino cyclopentyl heterocyclic and carbocyclic modulators of chemokine receptor activity
EP1765348B1 (en) 2004-06-18 2016-08-03 3M Innovative Properties Company Substituted imidazoquinolines, imidazopyridines, and imidazonaphthyridines
CA2580845A1 (en) 2004-09-20 2006-03-30 Xenon Pharmaceuticals Inc. Pyridazine derivatives for inhibiting human stearoyl-coa-desaturase
PL1637141T3 (en) 2004-09-21 2012-04-30 Trobio Ab Stabilized protease composition comprising a serine protease, morpholino derivatives and reversible inhibitors of said serine protease
KR20070073791A (en) 2004-10-18 2007-07-10 암젠 인코포레이티드 Thiadiazole compounds and methods of use
AU2006264043B2 (en) 2005-06-28 2012-04-26 Sanofi-Aventis Isoquinoline derivatives as inhibitors of Rho-kinase
CN103110635A (en) 2005-07-04 2013-05-22 海波因特制药有限责任公司 Histamine H3 receptor antagonist
GB0517184D0 (en) * 2005-08-22 2005-09-28 Glaxo Group Ltd Compounds
TW200813051A (en) * 2006-05-08 2008-03-16 Neurogen Corp Substituted azaspiro derivatives
WO2008005368A2 (en) * 2006-06-30 2008-01-10 Abbott Laboratories Piperazines as p2x7 antagonists
WO2008058064A1 (en) 2006-11-07 2008-05-15 Lexicon Pharmaceuticals, Inc. Amine-linked multicyclic compounds as inhibitors of the proline transporter
CN101652352A (en) 2006-12-22 2010-02-17 诺瓦提斯公司 Quinazolines for PDK1 inhibition
GB0704394D0 (en) 2007-03-07 2007-04-11 Senexis Ltd Compounds
GB0705803D0 (en) * 2007-03-28 2007-05-02 Glaxo Group Ltd Novel compounds
EP2014656A3 (en) 2007-06-11 2011-08-24 High Point Pharmaceuticals, LLC New heteocyclic h3 antagonists
CA2705947C (en) 2007-11-16 2016-08-09 Rigel Pharmaceuticals, Inc. Carboxamide, sulfonamide and amine compounds for metabolic disorders
GB0810902D0 (en) 2008-06-13 2008-07-23 Astex Therapeutics Ltd New compounds
MX2010013876A (en) 2008-06-20 2011-03-04 Metabolex Inc Aryl gpr119 agonists and uses thereof.
US20120101091A1 (en) 2008-10-03 2012-04-26 Boehringer Ingelheim International Gmbh Viral polymerase inhibitors
WO2010045306A2 (en) 2008-10-16 2010-04-22 Schering Corporation Azine derivatives and methods of use thereof
KR101701367B1 (en) 2008-11-04 2017-02-02 케모센트릭스, 인크. Modulators of cxcr7
CN102482277B (en) 2009-05-05 2017-09-19 达纳-法伯癌症研究所有限公司 Epidermal growth factor receptor inhibitor and the method for treating obstacle
EP2456438A2 (en) * 2009-07-23 2012-05-30 Novartis AG Use of azabicycloalkyl derivatives or pyrrolidine-2-one derivatives
JP5658756B2 (en) 2009-09-10 2015-01-28 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ JAK inhibitors
WO2011078143A1 (en) 2009-12-22 2011-06-30 塩野義製薬株式会社 Pyrimidine derivatives and pharmaceutical composition containing same
JP6026284B2 (en) 2010-03-03 2016-11-16 プロビオドルグ エージー Inhibitors of glutaminyl cyclase
US20130096160A1 (en) 2010-04-14 2013-04-18 Secretary, Department Of Health And Human Services Arylthiazolyl piperidines and related compounds as modulators of survival motor neuron (smn) protein production
AR081626A1 (en) 2010-04-23 2012-10-10 Cytokinetics Inc AMINO-PYRIDAZINIC COMPOUNDS, PHARMACEUTICAL COMPOSITIONS THAT CONTAIN THEM AND USE OF THE SAME TO TREAT CARDIAC AND SKELETIC MUSCULAR DISORDERS
EA030631B1 (en) 2013-07-31 2018-09-28 Новартис Аг 1,4-disubstituted pyridazine quinoline derivatives and methods for treating smn-deficiency-related conditions

Patent Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2427943A1 (en) * 1974-06-10 1976-01-08 Thomae Gmbh Dr K 5(6)-Pyridazinyl-benzimidazoles - esp 3-oxo-pyridazinyl derivs having hypotensive, anti-rhombic and cardiotonic activity
WO2001058891A2 (en) * 2000-02-11 2001-08-16 Vertex Pharmaceuticals Incorporated Piperazine and piperidine derivatives for treatment or prevention of neuronal damage
WO2001066129A1 (en) * 2000-03-10 2001-09-13 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Substances for the treatment of spinal muscular atrophy
WO2001094351A1 (en) * 2000-06-07 2001-12-13 Vertex Pharmaceuticals Incorporated Caspase inhibitors and uses thereof
US6376508B1 (en) * 2000-12-13 2002-04-23 Academia Sinica Treatments for spinal muscular atrophy
US20080247964A1 (en) * 2006-05-08 2008-10-09 Yuelian Xu Substituted azaspiro derivatives
WO2007137030A2 (en) * 2006-05-19 2007-11-29 Abbott Laboratories Cns active fused bicycloheterocycle substituted azabicyclic alkane derivatives
WO2009108657A2 (en) * 2008-02-28 2009-09-03 Vertex Pharmaceuticals Incorporated Heteroaryl derivatives as cftr modulators
WO2009137503A1 (en) * 2008-05-05 2009-11-12 Envivo Pharmaceuticals, Inc. Hdac inhibitors and uses thereof
WO2009150139A2 (en) * 2008-06-10 2009-12-17 Neurosearch A/S Indolyl-pyridazinyl-diazabicyclononane derivatives in labelled and unlabelled form and their use in diagnostic methods
WO2010028192A1 (en) * 2008-09-03 2010-03-11 Repligen Corporation Compositions including 6-aminohexanoic acid derivatives as hdac inhibitors
WO2011036167A1 (en) * 2009-09-22 2011-03-31 Novartis Ag Use of nicotinic acetylcholine receptor alpha 7 activators
WO2011063159A1 (en) * 2009-11-18 2011-05-26 Plexxikon, Inc. Compounds and methods for kinase modulation, and indications therefor
WO2012022467A2 (en) * 2010-08-16 2012-02-23 Santhera Pharmaceuticals (Schweiz) Ag Novel benzoquinone derivatives and use thereof as modulators of mitochondrial function
WO2012127393A1 (en) * 2011-03-18 2012-09-27 Novartis Ag COMBINATIONS OF ALPHA 7 NICOTINIC ACETYLCHOLINE RECEPTOR ACTIVATORS AND mGluR5 ANTAGONISTS FOR USE IN DOPAMINE INDUCED DYSKINESIA IN PARKINSON'S DISEASE
WO2014028459A1 (en) * 2012-08-13 2014-02-20 Novartis Ag 1,4-disubstituted pyridazine analogs and methods for treating smn-deficiency-related conditions

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
JINHE LI ET AL: "Role of 7 nicotinic acetylcholine receptors in regulating tumor necrosis factor- (TNF- ) as revealed by subtype selective agonists", JOURNAL OF NEUROIMMUNOLOGY, ELSEVIER SCIENCE PUBLISHERS BV, NL, vol. 239, no. 1, 10 August 2011 (2011-08-10), pages 37 - 43, XP028328732, ISSN: 0165-5728, [retrieved on 20110816], DOI: 10.1016/J.JNEUROIM.2011.08.007 *
LI T ET AL: "Syntheses and structure-activity relationship (SAR) studies of 2,5-diazabicyclo[2.2.1]heptanes as novel alpha7 neuronal nicotinic receptor (NNR) ligands", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, PERGAMON, AMSTERDAM, NL, vol. 20, no. 12, 15 June 2010 (2010-06-15), pages 3636 - 3639, XP027351220, ISSN: 0960-894X, [retrieved on 20100428] *
R. S. BITNER ET AL: "In Vivo Pharmacological Characterization of a Novel Selective 7 Neuronal Nicotinic Acetylcholine Receptor Agonist ABT-107: Preclinical Considerations in Alzheimer's Disease", JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, vol. 334, no. 3, 26 May 2010 (2010-05-26), pages 875 - 886, XP055008274, ISSN: 0022-3565, DOI: 10.1124/jpet.110.167213 *
W. H. BUNNELLE ET AL.: "Octahydropyrrolo[3,4-c]pyrrole: A Diamie Scaffold for Construction of either alpha4beta2 or alpha7-Selective Nicotinic Acetylcholine Receptor (nAChR) Ligands. Substitutions that Switch Subtype Selectivity", JOURNAL OF MEDICINAL CHEMISTRY, vol. 52, no. 14, 2009, pages 4126 - 4141, XP002732635 *

Cited By (33)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11672799B2 (en) 2013-07-31 2023-06-13 Novartis Ag 1,4-disubstituted pyridazine quinolne analogs there of and methods for treating SMN-deficiency-related conditions
WO2016128343A1 (en) * 2015-02-09 2016-08-18 F. Hoffmann-La Roche Ag Compounds for the treatment of cancer
US11066400B2 (en) 2015-02-09 2021-07-20 Hoffmann-La Roche Inc. Compounds for the treatment of cancer
CN107428729A (en) * 2015-02-09 2017-12-01 豪夫迈·罗氏有限公司 Compound for treating cancer
CN107428729B (en) * 2015-02-09 2021-07-16 豪夫迈·罗氏有限公司 Compounds for the treatment of cancer
EP3053577A1 (en) * 2015-02-09 2016-08-10 F. Hoffmann-La Roche AG Compounds for the treatment of cancer
JP2018519273A (en) * 2015-05-20 2018-07-19 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ Compounds for treating spinal muscular atrophy
US10357543B2 (en) 2015-11-16 2019-07-23 Ohio State Innovation Foundation Methods and compositions for treating disorders and diseases using Survival Motor Neuron (SMN) protein
EP3386511A4 (en) * 2015-12-10 2019-01-02 PTC Therapeutics, Inc. Methods for treating huntington's disease
CN110946865B (en) * 2015-12-10 2024-01-26 Ptc医疗公司 Methods for treating huntington's disease
CN110946865A (en) * 2015-12-10 2020-04-03 Ptc医疗公司 Methods for treating huntington's disease
AU2020201380B2 (en) * 2015-12-10 2020-10-08 Ptc Therapeutics, Inc. Methods for treating Huntington's disease
US10874672B2 (en) 2015-12-10 2020-12-29 Ptc Therapeutics, Inc. Methods for treating Huntington's disease
US10881658B2 (en) 2015-12-10 2021-01-05 Ptc Therapeutics, Inc. Methods for treating Huntington's disease
US11638706B2 (en) 2015-12-10 2023-05-02 Ptc Therapeutics, Inc. Methods for treating Huntington's disease
KR20180103045A (en) * 2015-12-10 2018-09-18 피티씨 테라퓨틱스, 인크. How to treat Huntington's disease
CN108697709A (en) * 2015-12-10 2018-10-23 Ptc医疗公司 Method for treating Huntington disease
EP3848035A1 (en) 2015-12-10 2021-07-14 PTC Therapeutics, Inc. 1,4-disubstituted pyridazine compounds for treating huntington's disease
AU2016366694C1 (en) * 2015-12-10 2021-03-25 Ptc Therapeutics, Inc. Methods for treating huntington's disease
WO2017100726A1 (en) 2015-12-10 2017-06-15 Ptc Therapeutics, Inc. Methods for treatng huntington's disease
CN113717154A (en) * 2015-12-10 2021-11-30 Ptc医疗公司 Methods for treating huntington's disease
KR102488323B1 (en) * 2015-12-10 2023-01-12 피티씨 테라퓨틱스, 인크. Composition for treating or ameliorating Huntington's disease
AU2016366694B2 (en) * 2015-12-10 2020-02-06 Ptc Therapeutics, Inc. Methods for treating huntington's disease
US11407753B2 (en) 2017-06-05 2022-08-09 Ptc Therapeutics, Inc. Compounds for treating Huntington's disease
US11608501B2 (en) 2017-06-14 2023-03-21 Ptc Therapeutics, Inc. Methods for modifying RNA splicing
US11395822B2 (en) 2017-06-28 2022-07-26 Ptc Therapeutics, Inc. Methods for treating Huntington's disease
US11382918B2 (en) 2017-06-28 2022-07-12 Ptc Therapeutics, Inc. Methods for treating Huntington's Disease
US11780839B2 (en) 2018-03-27 2023-10-10 Ptc Therapeutics, Inc. Compounds for treating Huntington's disease
US11858941B2 (en) 2018-06-27 2024-01-02 Ptc Therapeutics, Inc. Heterocyclic and heteroaryl compounds for treating Huntington's disease
US11685746B2 (en) 2018-06-27 2023-06-27 Ptc Therapeutics, Inc. Heteroaryl compounds for treating Huntington's disease
WO2021014428A1 (en) 2019-07-25 2021-01-28 Novartis Ag Regulatable expression systems
WO2021084495A1 (en) 2019-11-01 2021-05-06 Novartis Ag The use of a splicing modulator for a treatment slowing progression of huntington's disease
WO2022208170A1 (en) 2021-03-29 2022-10-06 Novartis Ag The use of the splicing modulator brataplam for slowing progression of huntington's disease

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