WO2008063114A9 - Amino- imidazolones and their use as medicament for treating cognitive impairment alzheimer disease, neurodegeneration and dementia - Google Patents

Amino- imidazolones and their use as medicament for treating cognitive impairment alzheimer disease, neurodegeneration and dementia

Info

Publication number
WO2008063114A9
WO2008063114A9 PCT/SE2007/001018 SE2007001018W WO2008063114A9 WO 2008063114 A9 WO2008063114 A9 WO 2008063114A9 SE 2007001018 W SE2007001018 W SE 2007001018W WO 2008063114 A9 WO2008063114 A9 WO 2008063114A9
Authority
WO
WIPO (PCT)
Prior art keywords
dementia
compound
6alkylc
alkylc
compound according
Prior art date
Application number
PCT/SE2007/001018
Other languages
French (fr)
Other versions
WO2008063114A1 (en
Inventor
Stefan Berg
Original Assignee
Astrazeneca Ab
Astex Therapeutics Ltd
Stefan Berg
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Astrazeneca Ab, Astex Therapeutics Ltd, Stefan Berg filed Critical Astrazeneca Ab
Publication of WO2008063114A1 publication Critical patent/WO2008063114A1/en
Publication of WO2008063114A9 publication Critical patent/WO2008063114A9/en

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia

Definitions

  • Amino-imidazolones and their use as medicament for treating cognitive impairment Alzheimer disease, neurodegeneration and dementia.
  • the present invention relates to novel compounds, their pharmaceutical compositions.
  • the present invention relates to therapeutic methods for the treatment and/or prevention of A ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease
  • ⁇ -secretase activity Hussain et al., 1999; Lin et. al, 2000; Yan et. al, 1999; Sinlia et. al., 1999 and Vassar et. al., 1999).
  • ⁇ -secretase is also known in the literature as Asp2 (Yan et. al, 1999), Beta site APP Cleaving Enzyme (BACE) (Vassar et. al., 1999) or memapsin-2 (Lin et al., 2000).
  • BACE was identified using a number of experimental approaches such as EST database analysis (Hussain et al.
  • BACE was found to be a pepsin-like aspartic proteinase, the mature enzyme consisting of the N-terminal catalytic domain, a transmembrane domain, and a small cytoplasmic domain.
  • BACE has an optimum activity at pH 4.0-5.0 (Vassar et al, 1999) and is inhibited weakly by standard pepsin inhibitors such as pepstatin. It has been shown that the catalytic domain minus the transmembrane and cytoplasmic domain has activity against substrate peptides (Lin et al, 2000).
  • BACE is a membrane bound type 1 protein that is synthesized as a partially active proenzyme, and is abundantly expressed in brain tissue.
  • a ⁇ amyloid- ⁇ -protein
  • a ⁇ or amyloid- ⁇ -protein is the major constituent of the brain plaques which are characteristic of Alzheimer's disease (De Strooper et al, 1999).
  • a ⁇ is a 39-42 residue peptide formed by the specific cleavage of a class I transmembrane protein called APP, or amyloid precursor protein.
  • a ⁇ -secretase activity cleaves this protein between residues Met671 and Asp672 (numbering of 770aa isoform of APP) to form the N-terminus of A ⁇ .
  • a second cleavage of the peptide is associated with ⁇ -secretase to form the C-terminus of the A ⁇ peptide.
  • Alzheimer's disease is estimated to afflict more than 20 million people worldwide and is believed to be the most common form of dementia.
  • Alzheimer's disease is a progressive dementia in which massive deposits of aggregated protein breakdown products - amyloid plaques and neurofibrillary tangles accumulate in the brain. The amyloid plaques are thought to be responsible for the mental decline seen in Alzheimer's patients.
  • Alzheimer's disease increases with age, and as the aging population of the developed world increases, this disease becomes a greater and greater problem.
  • this disease becomes a greater and greater problem.
  • any individuals possessing the double mutation of APP known as the Swedish mutation (in which the mutated APP forms a considerably improved substrate for BACE) have a much greater chance of developing AD, and also of developing it at an early age (see also US 6,245,964 and US 5,877,399 pertaining to transgenic rodents comprising APP-Swedish). Consequently, there is also a strong need for developing a compound that can be used in a prophylactic fashion for these individuals.
  • APP The gene encoding APP is found on chromosome 21 , which is also the chromosome found as an extra copy in Down's syndrome.
  • Down's syndrome patients tend to acquire Alzheimer's disease at an early age, with almost all those over 40 years of age showing Alzheimer's-type pathology (Oyama et al., 1994). This is thought to be due to the extra copy of the APP gene found in these patients, which leads to overexpression of APP and therefore to increased levels of APP ⁇ causing the high prevalence of Alzheimer's disease seen in this population.
  • inhibitors of BACE could be useful in reducing Alzheimer's-type pathology in Down's syndrome patients.
  • Drugs that reduce or block BACE activity should therefore reduce A ⁇ levels and levels of fragments of A ⁇ in the brain, or elsewhere where A ⁇ or fragments thereof deposit, and thus slow the formation of amyloid plaques and the progression of AD or other maladies involving deposition of A ⁇ or fragments thereof (Yankner, 1996; De Strooper and Konig, 1999).
  • BACE is therefore an important candidate for the development of drugs as a treatment and/or prophylaxis of A ⁇ -related pathologies such as Downs syndrome and ⁇ - amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ - amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms
  • the compounds of the present invention show beneficial properties compared to the potential inhibitors known in the art, e.g. improved hERG selectivity.
  • A is independently selected from a 5, 6 or 7 membered heterocyclic ring optionally substituted with one or more R 1 ;
  • B is independently selected from a 4, 5, 6 or 7 membered heterocyclic ring, Q ⁇ alkyl, C 2 .6alkenyl, C ⁇ alkynyl, C 0 . 6 alkylaryl, Co- ⁇ alkylheteroaryl, Co-ealkylCs-gcycloalkyl, Co- ⁇ alkylCs- ⁇ cycloalkenyl, Co-6alkylC 3 -6cycloalkynyl, Co- 6 alkylC 3 - 6 heterocyclyl optionally substituted with one or more R 2 ;
  • C is independently selected from phenyl or a 5 or 6 membered heteroaromatic ring optionally substituted with one or more R 3 ;
  • R 1 is independently selected from halogen, cyano, nitro, OR 6 , d- ⁇ alkyl, C 2 - 6 a.ken.yl, C 2 .6alkynyl, Co- ⁇ alkylaryl, Co- ⁇ alkylheteroaryl, Co- ⁇ alkylCs- ⁇ cycloalkyl, Co-6alkylC 3 _ ecycloalkenyl, C0.salkylC3.6cycloalky1.yl, Co-6alkylC 3 -6heterocyclyl, NR 6 R 7 , CONR 6 R 7 , NR 6 (CO)R 7 , 0(CO)R 6 , CO 2 R 6 , COR 6 , (SO 2 )NR 6 R 7 , NR 6 (SO 2 )R 7 , SOR 6 , SO 2 R 6 , OSO 2 R 6 and SO 3 R 6 wherein said Q ⁇ alkyl, C 2- 6alkenyl, C 2 -6alkynyl
  • R 2 , R 3 and R 4 are independently selected from halogen, cyano, nitro, OR 6 , C ⁇ alkyl, C 2 . ⁇ alkenyl, Ca- ⁇ alkynyl, Co- ⁇ alkylaryl, Co- ⁇ alkylheteroaryl, Co- 6 alkylC 3 .
  • R 5 is independently selected from hydrogen, cyano, OR 6 , C ⁇ alkyl, C 2-6 alkenyl, C 2 -6alkynyl, Co-ealkylaryl, Co- ⁇ alkylheteroaryl, Co- ⁇ alkylCs- ⁇ cycloalkyl, Co-6alkylC3. ⁇ cycloalkenyl, Co.6alkylC 3 . 6 cycloalkynyl, CONR 6 R 7 , CO 2 R 6 , COR 6 , SO 2 R 6 and SO 3 R 6 wherein said Ci- 6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 0 .
  • D is independently selected from halogen, nitro, CN, OR 0 , C ⁇ alkyl, C 2 _ 6 atkenyl, C 2- 6alkynyl, Co- ⁇ alkylaryl, Co-ealkylheteroaryl, Co-ealkylCs.
  • R b and R 7 are independently selected from hydrogen, C h alky!, fluoromethyl, difluoromethyl, trifluoromethyl, C 2 . 6 alkenyl, Ca- ⁇ alkynyl, Co- ⁇ alkylaryl, Co- 6 alkylheteroaryl, Co- ⁇ alkylC 3 .(;cycloalkyl, Co-6alkylC 3 .6cycloalkenyl, Co-salkylC 3 . ⁇ cycloalkynyl, Co-ealkylheterocyclyl; or R 6 and R 7 may together form a 5 or 6 membered heterocyclic ring containing one or more heteroatoms selected from N, O or S;
  • n 0, 1, 2 or 3
  • p 0, 1, 2 or 3
  • q 0, 1, 2 or 3
  • the present invention further provides pharmaceutical compositions comprising as active ingredient a therapeutically effective amount of a compound of formula I in association with pharmaceutically acceptable excipients, carriers or diluents.
  • the present invention farther provides methods of modulating activity of BACE comprising contacting the BACE enzyme with a compound of formula I.
  • the present invention further provides methods of treating or preventing an A ⁇ -related pathology in a patient, comprising administering to the patient a therapeutically effective amount of a compound of formula I.
  • the present invention further provides a compound described herein for use as a medicament.
  • A is a 6 membered heterocyclic ring
  • B is Co-6alkylC3-6cycloalkyl
  • C is phenyl substituted with one or more R 3 ;
  • R 3 is halogen
  • Some compounds of formula I may have stereogenic centres and/or geometric isomeric centres (E- and Z- isomers), and it is to be understood that the invention encompasses all such optical isomers, enantiomers, diastereoisomers, atropisomers and geometric isomers.
  • the present invention relates to the use of compounds of formula I as hereinbefore defined as well as to the salts thereof.
  • Salts for use in pharmaceutical compositions will be pharmaceutically acceptable salts, but other salts may be useful in the production of the compounds of formula I.
  • the present invention provides compounds of formula I, or pharmaceutically acceptable salts, tautomers or in vzvo-hydrolysable precursors thereof, for use as medicaments.
  • the present invention provides compounds described here in for use as medicaments for treating or preventing an A ⁇ -related pathology.
  • the A ⁇ -related pathology is Downs syndrome, a ⁇ -amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, a disorder associated with cognitive impairment, MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with Alzheimer disease, dementia of mixed vascular origin, dementia of degenerative origin, pre-senile dementia, senile dementia, dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
  • MCI mimild cognitive impairment
  • the present invention provides use of compounds of formula I or pharmaceutically acceptable salts, tautomers or in vzvo-hydrolysable precursors thereof, in the manufacture of a medicament for the treatment or prophylaxis of A ⁇ -related pathologies.
  • the A ⁇ -related pathologies include such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
  • MCI mimild cognitive impairment
  • the present invention provides a method of inhibiting activity of BACE comprising contacting the BACE with a compound of the present invention.
  • BACE is thought to represent the major ⁇ -secretase activity, and is considered to be the rate- limiting step in the production of amyloid- ⁇ -protein (A ⁇ ).
  • a ⁇ amyloid- ⁇ -protein
  • BACE is an important candidate for the development of drugs as a treatment and/or prophylaxis of A ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated
  • the present invention provides a method for the treatment of A ⁇ - related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment' " ), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, presenile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration, comprising administering to a mammal (including human) a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, tautomer or in vz ' r ⁇ -hydrolysable precursor thereof.
  • a ⁇ - related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid
  • the present invention provides a method for the prophylaxis of A ⁇ - related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, presenile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration comprising administering to a mammal (including human) a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, tautomer or in vzvo-hydrolysable precursors.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy,
  • the present invention provides a method of treating or preventing A ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre- senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration by administering to a mammal (including human) a compound of formula I or a pharmaceutically acceptable salt, tautomer or in vzvo-hydrolysable precursors and a cognitive and/or memory enhancing agent.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid ang
  • the present invention provides a method of treating or preventing A ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, presenile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration by administering to a mammal (including human) a compound of formula I or a pharmaceutically acceptable salt, tautomer or in vzvo-hydrolysable precursors thereof wherein constituent members are provided herein, and a choline esterase inhibitor or anti-inflammatory agent.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as
  • the present invention provides a method of treating or preventing A ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration, or any other disease, disorder, or condition described herein, by administering to a mammal (including human) a compound of the present inventionand an atypical antipsychotic agent.
  • a ⁇ -related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders
  • Atypical antipsychotic agents includes, but not limited to, Olanzapine (marketed as Zyprexa), Aripiprazole (marketed as Abilify), Risperidone (marketed as Risperdal), Quetiapine (marketed as Seroquel), Clozapine (marketed as Clozaril), Ziprasidone (marketed as Geodon) and Olanzapine/Fluoxetine (marketed as Symbyax).
  • the mammal or human being treated with a compound of the invention has been diagnosed with a particular disease or disorder, such as those described herein. In these cases, the mammal or human being treated is in need of such treatment. Diagnosis, however, need not be previously performed.
  • the present invention also includes pharmaceutical compositions which contain, as the active ingredient, one or more of the compounds of the invention herein together with at least one pharmaceutically acceptable carrier, diluent or excipent.
  • a variety of compounds in the present invention may exist in particular geometric or stereoisomeric forms.
  • the present invention takes into account all such compounds, including cis- and trans isomers, R- and S- enantiomers, diastereomers, (D)-isomers, (L)- isomers, the racemic mixtures thereof, and other mixtures thereof, as being covered within the scope of this invention.
  • Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
  • the compounds herein described may have asymmetric centers. Compounds of the present invention containing an asymmetrically substituted atom may be isolated in optically active or racemic forms.
  • optically active forms such as by resolution of racemic forms, by synthesis from optically active starting materials, or synthesis using optically active reagents.
  • separation of the racemic material can be achieved by methods known in the art.
  • Cis and trans geometric isomers of the compounds of the present invention are described and may be isolated as a mixture of isomers or as separated isomeric forms. All chiral, diastereomeric, racemic forms and all geometric isomeric forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated.
  • substitution means that substitution is optional and therefore it is possible for the designated atom or moiety to be unsubstituted.
  • substitution means that any number of hydrogens on the designated atom or moiety is replaced with a selection from the indicated group, provided that the normal valency of the designated atom or moiety is not exceeded, and that the substitution results in a stable compound.
  • a substituent is methyl (i.e., CH 3 )
  • 3 hydrogens on the carbon atom can be replaced.
  • alkyl used alone or as a suffix or prefix, is intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having from 1 to 12 carbon atoms or if a specified number of carbon atoms is provided then that specific number would be intended.
  • “Co- 6 alkyl” denotes alkyl having 0, 1, 2, 3, 4, 5 or 6 carbon atoms.
  • alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, pentyl, and hexyl.
  • a subscript is the integer 0 (zero) the group to which the subscript refers to indicates that the group may be absent, i.e. there is a direct bond between the groups.
  • alkenyl used alone or as a suffix or prefix is intended to include both branched and straight-chain alkene or olefin containing aliphatic hydrocarbon groups having from 2 to 12 carbon atoms or if a specified number of carbon atoms is provided then that specific number would be intended.
  • “Ca- ⁇ alkenyl” denotes alkenyl having 2, 3, 4, 5 or 6 carbon atoms.
  • alkenyl examples include, but are not limited to, vinyl, allyl, 1-pro ⁇ enyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylbut-2-enyl, 3-methylbut- 1-enyl, 1-pentenyl, 3-pentenyl and 4-hexenyl.
  • alkynyl used alone or as a suffix or prefix is intended to include both branched and straight-chain alkyne containing aliphatic hydrocarbon groups having from 2 to 12 carbon atoms or if a specified number of carbon atoms is provided then that specific number would be intended.
  • C ⁇ alkynyl denotes alkynyl having 2, 3, 4, 5 or 6 carbon atoms.
  • alkynyl include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 3-butynyl, -pentynyl, hexynyl and l-methylpent-2-ynyl.
  • aromatic refers to hydrocarbonyl groups having one or more unsaturated carbon ring(s) having aromatic characters, (e.g. 4n + 2 delocalized electrons) and comprising up to about 14 carbon atoms.
  • hetero aromatic refers to groups having one or more unsaturated rings containing carbon and one or more heteroatoms such as nitrogen, oxygen or sulphur having aromatic character (e.g. 4n + 2 delocalized electrons).
  • aryl refers to an aromatic ring structure made up of from 5 to 14 carbon atoms. Ring structures containing 5, 6, 7 and 8 carbon atoms would be single-ring aromatic groups, for example, phenyl. Ring structures containing 8, 9, 10, 11, 12, 13, or 14 would be poly cyclic, for example naphthyl. The aromatic ring can be substituted at one or more ring positions with such substituents as described above.
  • aryl also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "fused rings") wherein at least one of the rings is aromatic, for example, the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls.
  • ortho, meta and para apply to 1,2-, 1,3- and 1 ,4-disubstituted benzenes, respectively.
  • the names 1 ,2-dimethylbenzene and ortho-dimethylbenzene are synonymous.
  • cycloalkyl is intended to include saturated ring groups, having the specified number of carbon atoms. These may include fused or bridged polycyclic systems. Preferred cycloalkyls have from 3 to 10 carbon atoms in their ring structure, and more preferably have 3, 4, 5, and 6 carbons in the ring structure. For example, “C 3 - 6 cycloalkyl” denotes such groups as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
  • cycloalkenyl refers to ring-containing hydrocarbyl groups having at least one carbon-carbon double bond in the ring, and having from 4 to 12 carbons atoms.
  • cycloalkynyl refers to ring-containing hydrocarbyl groups having at least one carbon-carbon triple bond in the ring, and having from 7 to 12 carbons atoms.
  • halo or halogen refers to fluoro, chloro, bromo, and iodo.
  • Counterion is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, sulfate, tosylate, benezensulfonate, and the like.
  • heterocyclyl or “heterocyclic” or “heterocycle” refers to a saturated, unsaturated or partially saturated, monocyclic, bicyclic or tricyclic ring (unless otherwise stated) containing 3 to 20 atoms of which 1, 2, 3, 4 or 5 ring atoms are chosen from nitrogen, sulphur or oxygen, which may, unless otherwise specified, be carbon or nitrogen linked, wherein a -CH 2 - group is optionally be replaced by a -C(O)-; and where unless stated to the contrary a ring nitrogen or sulphur atom is optionally oxidised to form the N-oxide or S-oxide(s) or a ring nitrogen is optionally quarternized; wherein a ring -NH is optionally substituted by acetyl, formyl, methyl or mesyl; and a ring is optionally substituted by one or more halo.
  • heterocyclyl group when the total number of S and O atoms in the heterocyclyl exceeds 1 , then these heteroatoms are not adjacent to one another. If the said heterocyclyl group is bi- or tricyclic then at least one of the rings may optionally be a heteroaromatic or aromatic ring provided that at least one of the rings is non-heteroaromatic. If the said heterocyclyl group is monocyclic then it must not be aromatic.
  • heterocyclyls include, but are not limited to, piperidinyl, N- acetylpiperidinyl, N-methylpiperidinyl, JV-forrnylpiperazinyl, iV-mesylpiperazinyl, homopiperazinyl, piperazinyl, azetidinyl, oxetanyl, morpholinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, indolinyl, tetrahydropyranyl, dihydro-2H-pyranyl, tetrahydrofuranyl and 2,5-dioxoimidazolidinyl.
  • heteroaryl refers to an aromatic heterocycle having at least one heteroatom ring member such as sulfur, oxygen, or nitrogen.
  • Heteroaryl groups include monocyclic and polycyclic (e.g., having 2, 3 or 4 fused rings) systems. Examples of heteroaryl groups include without limitation, pyridyl (i.e., pyridinyl), pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl (i.e.
  • furanyl quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl, pyrryl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, fluorenonyl, benzimidazolyl, indolinyl, and the like.
  • the heteroaryl group has from 1 to about 20 carbon atoms, and in further embodiments from about 3 to about 20 carbon atoms. In some embodiments, the heteroaryl group contains 3 to about 14, 4 to about 14, 3 to about 7, or 5 to 6 ring-forming atoms. In some embodiments, the heteroaryl or heteroaromatic group has 1 to about 4, 1 to about 3, or 1 to 2 heteroatoms. In some embodiments, the heteroaryl or heteroaromatic group has 1 heteroatom.
  • protecting group means temporary substituents which protect a potentially reactive functional group from undesired chemical transformations.
  • protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones respectively.
  • the field of protecting group chemistry has been reviewed (Greene, T.W.; Wuts, P.G.M. Protective Groups in Organic Synthesis, 3 rd ed.; Wiley: New York, 1999).
  • pharmaceutically acceptable is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salts refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof.
  • pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
  • the pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
  • such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid.
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods.
  • such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used.
  • tautomer means other structural isomers that exist in equilibrium resulting from the migration of a hydrogen atom. For example, keto-enol tautomerism where the resulting compound has the porperties of both a ketone and an unsturated alchol.
  • stable compound and “stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
  • Compounds of the invention further include hydrates and solvates.
  • the present invention further includes isotopically-labeled compounds of the invention.
  • An “isotopically” or “radio-labeled” compound is a compound of the invention where one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occu ⁇ ing).
  • Suitable radionuclides that may be incorporated in compounds of the present invention include but are not limited to 2 H (also written as D for deuterium), 3 H (also written as T for tritium), 11 C, 13 C 5 14 C, 13 N, 15 N, 15 0, 17 O, 18 0, 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I and 131 I.
  • the radionuclide that is incorporated in the instant radio- labeled compounds will depend on the specific application of that radio-labeled compound. For example, for in vitro receptor labeling and competition assays, compounds that incorporate 3 H, 14 C, 82 Br, 125 1 , 131 1, 35 S or will generally be most useful. For radio- imaging applications 11 C, 18 F, 125 I 5 123 1, 124 I 3 131 I 5 75 Br, 76 Br or 77 Br will generally be most useful.
  • a "radio-labeled compound” is a compound that has incorporated at least one radionuclide.
  • the radionuclide is selected from the group consisting of 3 H, 14 C, 125 1 , 35 S and 82 Br.
  • the anti-dementia treatment defined herein may be applied as a sole therapy or may involve, in addition to the compound of the invention, conventional chemotherapy.
  • chemotherapy may include one or more of the following categories of agents: acetyl cholinesterase inhibitors, anti-inflammatory agents, cognitive and/or memory enhancing agents or atypical antipsychotic agents.
  • Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment.
  • Such combination products employ the compounds of this invention.
  • Compounds of the present invention may be administered orally, parenteral, buccal, vaginal, rectal, inhalation, insufflation, sublingually, intramuscularly, subcutaneously, topically, intranasally, intraperitoneally, intrathoracially, intravenously, epidurally, ⁇ intrathecally, intracerebroventricularly and by injection into the joints.
  • the dosage will depend on the route of administration, the severity of the disease, age and weight of the patient and other factors normally considered by the attending physician, when determining the individual regimen and dosage level as the most appropriate for a particular patient.
  • An effective amount of a compound of the present invention for use in therapy of dementia is an amount sufficient to symptomatically relieve in a warm-blooded animal, particularly a human the symptoms of dementia, to slow the progression of dementia, or to reduce in patients with symptoms of dementia the risk of getting worse.
  • inert, pharmaceutically acceptable carriers can be either solid or liquid.
  • Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories.
  • a solid carrier can be one or more substances, which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material.
  • the carrier is a finely divided solid, which is in a mixture with the finely divided active component.
  • the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
  • a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient sized molds and allowed to cool and solidify.
  • Suitable carriers include magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.
  • the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for the therapeutic treatment (including prophylactic treatment) of mammals including humans, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.
  • the pharmaceutical composition of this invention may also contain, or be co-administered (simultaneously or sequentially) with, one or more pharmacological agents of value in treating one or more disease conditions referred to herein.
  • composition is intended to include the formulation of the active component or a pharmaceutically acceptable salt with a pharmaceutically acceptable carrier.
  • this invention may be formulated by means known in the art into the form of, for example, tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely divided powders or aerosols or nebulisers for inhalation, and for parenteral use (including intravenous, intramuscular or infusion) sterile aqueous or oily solutions or suspensions or sterile emulsions.
  • Liquid form compositions include solutions, suspensions, and emulsions.
  • Sterile water or water-propylene glycol solutions of the active compounds may be mentioned as an example of liquid preparations suitable for parenteral administration.
  • Liquid compositions can also be formulated in solution in aqueous polyethylene glycol solution.
  • Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired.
  • Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.
  • the pharmaceutical compositions can be in unit dosage form.
  • the composition is divided into unit doses containing appropriate quantities of the active component.
  • the unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparations, for example, packeted tablets, capsules, and powders in vials or ampoules.
  • the unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms.
  • compositions may be formulated for any suitable route and means of administration.
  • Pharmaceutically acceptable carriers or diluents include those used in formulations suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) administration.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.
  • conventional non-toxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, cellulose, cellulose derivatives, starch, magnesium stearate, sodium saccharin, talcum, glucose, sucrose, magnesium carbonate, and the like may be used.
  • Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc, an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline aqueous dextrose, glycerol, ethanol, and the like, to thereby form a solution or suspension.
  • the pharmaceutical composition to be administered may also contain minor amounts of non- toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanol amine sodium acetate, sorbitan monolaurate, triethanolamine oleate, etc.
  • auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanol amine sodium acetate, sorbitan monolaurate, triethanolamine oleate, etc.
  • the compounds of the invention may be derivatised in various ways.
  • “derivatives" of the compounds includes salts (e.g. pharmaceutically acceptable salts), any complexes (e.g.
  • prodrugs is meant for example any compound that is converted in vivo into a biologically active compound.
  • Salts of the compounds of the invention are preferably physiologically well tolerated and non toxic. Many examples of salts are known to those skilled in the art. All such salts are within the scope of this invention, and references to compounds include the salt forms of the compounds.
  • the compounds may contain an amine function, these may form quaternary ammonium salts, for example by reaction with an alkylating agent according to methods well known to the skilled person. Such quaternary ammonium compounds are within the scope of the invention.
  • Compounds containing an amine function may also form TV-oxides.
  • a reference herein to a compound that contains an amine function also includes the JV-oxide.
  • one or more than one nitrogen atom may be oxidised to form an JV-oxide.
  • TV-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle.
  • TV-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4 th Edition, Wiley Interscience, pages. More particularly, TV-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compound is reacted with /w-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as dichloromethane. Where the compounds contain chiral centres, all individual optical forms such as enantiomers, epimers and diastereoisomers, as well as racemic mixtures of the compounds are within the scope of the invention.
  • an oxidizing agent such as hydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid)
  • MCPBA /w-chloroper
  • the quantity of the compound to be administered will vary for the patient being treated and will vary from about 100 ng/kg of body weight to 100 mg/kg of body weight per day and preferably will be from 10 pg/kg to 10 mg/kg per day.
  • dosages can be readily ascertained by those skilled in the art from this disclosure and the knowledge in the art.
  • the skilled artisan can readily determine the amount of compound and optional additives, vehicles, and/or carrier in compositions and to be administered in methods of the invention.
  • Beta secretase including BACE
  • Inhibitors of beta secretase have been shown to be useful in blocking formation or aggregation of A ⁇ peptide and therefore have beneficial effects in treatment of Alzheimer's Disease and other neurodegenerative diseases associated with elevated levels and/or deposition of A ⁇ peptide. Therefore, it is believed that the compounds of the present invention may be used for the treatment of Alzheimer disease and disease associated with dementia
  • compounds of the present invention and their salts are expected to be active against age-related diseases such as Alzheimer, as well as other A ⁇ related pathologies such as Downs syndrome and ⁇ -amyloid angiopathy. It is expected that the compounds of the present invention would most likely be used as single agents but could also be used in combination with a broad range of cognition deficit enhancement agents.
  • the present invention also relates to processes for preparing the compound of formula (I) as a free base or a pharmaceutically acceptable salt thereof.
  • suitable protecting groups will be added to, and subsequently removed from the various reactants and intermediates in a manner that will be readily understood by one skilled in the art of organic synthesis.
  • Conventional procedures for using such protecting groups as well as examples of suitable protecting groups are for example described in Protective Groups in Organic Synthesis by T.W. Greene, P.G.M Wutz, 3 rd Edition, Wiley-Interscience, New York, 1999. It is understood that microwaves can be used for the heating of reaction mixtures.
  • the reaction may be carried out by treating an appropriate amine with nitrous acid followed by treating the formed diazonium salt with an appropriate cuprous halide such as copper(I) bromide or copper(I) chloride, or with copper and hydrobromic acid or hydrochloric acid.
  • the reactions may be preformed in a suitable solvent such as water at a temperature between -20 0 C and reflux.
  • halo represents halogen such as bromine or chlorine
  • R 8 represents hydrogen, alkyl, aryl or two R 8 may form a cyclic boronic ester.
  • the reaction may be carried out by: a) an alkyllithium such as butyllithium, or magnesium, and a suitable boron compound such as trimethyl borate or triisopropyl borate.
  • the reaction may be performed in a suitable solvent such as tetrahydrofuran, hexane or dichloromethane in a temperature range between -78 0 C and +20 0 C; or, b) a suitable boron species such as 4,4,4',4 l ,5 J 5,5 r ,5'-octamethyl-2,2'-bi-l ; 3,2- dioxaborolane, biscatecholatodiboron, or pinacolborane in the presence of a suitable palladium catalyst such as tris(dibenzylideneaceton)dipalladium(0), [1,1'- bis(diphenylphos ⁇ hino)ferrocene]palladium(II) chloride, palladium(O)
  • the reaction may be performed in a solvent such as dioxane, toluene, acetonitrile, water, ethanol or 1,2- dimethoxyethane, or mixtures thereof, at temperatures between +20 0 C and +160 0 C.
  • a solvent such as dioxane, toluene, acetonitrile, water, ethanol or 1,2- dimethoxyethane, or mixtures thereof, at temperatures between +20 0 C and +160 0 C.
  • the reaction may be carried out by treating the compound of formula V with an alkyllithium, such as butyllithium, or magnesium followed by addition of a compound of formula VI.
  • the reaction may be preformed in a suitable solvent such as diethyl ether or tetrahydrofuran at a temperature between -78 0 C and reflux.
  • the reaction may be carried out by reduction using an appropriate reducing agent such as sodium borohydride, cyanoborohydride or lithium aluminium hydride.
  • the reaction may be preformed in a suitable solvent such as methanol, ethanol, diethyl ether or tetrahydrofuran at a temperature between -78 0 C and reflux.
  • the reaction may be carried out by treating the compound of formula VIII with a suitable thiocarbonyl transfer reagent such as O,O-dipyridine-2-yl thiocarbonate or thiophosgene and with or without a suitable base such as sodium bicarbonate.
  • a suitable thiocarbonyl transfer reagent such as O,O-dipyridine-2-yl thiocarbonate or thiophosgene and with or without a suitable base such as sodium bicarbonate.
  • the reaction may be preformed in a suitable solvent such as dichloromethane or chloroform at a temperature between -78 0 C and reflux.
  • the reaction may be carried out by treating the appropriate isothiocyanate and carbon disulfide with a suitable base such as potassium ter/-butoxide in a suitable solvent such as tetrahydrofuran or diethyl ether at a temperature between -78 0 C and reflux.
  • a suitable base such as potassium ter/-butoxide
  • a suitable solvent such as tetrahydrofuran or diethyl ether
  • the reaction may be carried out by treating a compound of formula X with an appropriate diamine such as 1,3-diaminopropane or ethyl enediamine.
  • the reaction may be preformed in a suitable solvent such as ethanol or methanol at a temperature between 0 0 C and reflux.
  • the reaction may be carried out by treating the appropriate thione such as a compound of formula XI with an appropriate oxidazing agent such as tert-butyl hydroperoxide and aqueous ammonia.
  • an appropriate oxidazing agent such as tert-butyl hydroperoxide and aqueous ammonia.
  • the reaction may be performed in a suitable solvent such as methanol at a temperature between 0 0 C and reflux.
  • the reaction may be carried out by treating the methyl ether with a suitable Lewis acid such as boron tribromide in a suitable solvent such as dichloromethane at a temperature between -78 0 C and reflux.
  • a suitable Lewis acid such as boron tribromide
  • a suitable solvent such as dichloromethane
  • the reaction may be carried out by treating the appropriate alcohol with a suitable sulfonyl chloride or anhydride such as methanesulfonyl chloride, 1-propanesulfonyl chloride, cyclopropanesulfonyl chloride or methanesulfonic anhydride in the presence of a suitable base such as triethylamine.
  • a suitable sulfonyl chloride or anhydride such as methanesulfonyl chloride, 1-propanesulfonyl chloride, cyclopropanesulfonyl chloride or methanesulfonic anhydride in the presence of a suitable base such as triethylamine.
  • the reaction ma ⁇ ' be carried out in a suitable solvent such as dichloromethane at a temperature between 0 0 C and reflux.
  • Another object of the invention is the process for the preparation of compounds of general Formula (I), wherein A, B, C, D, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 unless otherwise specified, are defined as hereinbefore, and salts thereof.
  • the free base may be treated with an acid such as a hydrogen halide such as hydrogen chloride in a suitable solvent such as tetrahydrofuran, diethyl ether, methanol, ethanol, chloroform or dichloromethane or mixtures thereof and the reaction may occur between -30 0 C to +50 0 C.
  • reaction of process (a) may be carried out by coupling of a suitable compound such as a compound of formula XVI with an appropriate aryl boronic acid or ester of formula IV wherein R 8 represents hydrogen, alkyl, aryl or two R 8 may form a cyclic boronic ester.
  • the reaction may be carried out using a suitable palladium catalyst such as, [1,1'- bis(diphenylphosphino)ferrocene]palladium(II) chloride, tetrakis(triphenyl ⁇ hosphine)palIadium(0), palladium diphenylphosphineferrocene dichloride, ⁇ alladium(II) acetate or bis(dibenzylideneacetone) palladium (0), together with, or without, a suitable ligand such as triphenylphosphine, tri-tert-butylphosphine or 2- (dicyclohexylphosphino)biphenyl, or using a nickel catalyst such as nickel on charcoal or l,2-bis(diphenylphosphino)ethanenickel dichloride together with zinc and sodium triphenylphosphinetrimetasulfonate.
  • a suitable palladium catalyst such as, [1,1'- bis(diphen
  • a suitable base such as cesium fluoride, an alkyl amine such as triethyl amine, or an alkali metal or alkaline earth metal carbonate or hydroxide such as potassium carbonate, sodium carbonate, cesium carbonate, or sodium hydroxide may be used in the reaction, which may be performed in a temperature range between +20 0 C and +160 0 C, in a suitable solvent such as toluene, tetrahydrofuran, dioxane, dimethoxyethane, water, ethanol or ⁇ N-dimethylformamide, or mixtures thereof.
  • a suitable solvent such as toluene, tetrahydrofuran, dioxane, dimethoxyethane, water, ethanol or ⁇ N-dimethylformamide, or mixtures thereof.
  • LC-MS analyses were performed on an LC-MS system consisting of a Waters Alliance 2795 HPLC, a Waters PDA 2996 diode array detector, a Sedex 75 ELS detector and a ZMD single quadrupole mass spectrometer.
  • the mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode.
  • the capillary voltage was set to 3.2 kV and the cone voltage to 30 V, respectively.
  • the mass spectrometer was scanned between m/z 100-600 by a scan time of 0.7s.
  • the diode array detector was scanned from 200-400 nm.
  • the temperature of the ELS detector was adjusted to 40 0 C and the pressure was set to 1.9 bar.
  • the mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode.
  • ES electrospray ion source
  • the capillary voltage was set to 3.2 kV and the cone voltage to 30 V 5 respectively.
  • the mass spectrometer was scanned between mfz 100-700 with a scan time of 0.3s.
  • the diode array detector scanned from 200-400 nm.
  • the temperature of the ELS detector was adjusted to 40 0 C and the pressure was set to 1.9 bar.
  • the mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode.
  • the capillary voltage was set to 3.2 kV and the cone voltage to 30 V, respectively.
  • the mass spectrometer scanned between m/z 100-700 with a scan time of 0.3s.
  • the diode array detector scanned from 200-400 nm.
  • the temperature of the ELS detector was adjusted to 40 0 C and the pressure was set to 1.9 bar. Separation was performed on an X-Terra MS C8, 3.0 mm x 50 mm, 3.5 ⁇ m (Waters) run at a flow rate of 1 mL/min.
  • a linear gradient was applied starting at 100% A (A: 1OmM ammonium acetate in 5% acetonitrile, or S mM formic acid in 5% acetonitrile) ending at 100% B (B: acetonitrile).
  • B acetonitrile
  • the column oven temperature was set to 40 0 C, or
  • LC-MS analyses were preformed on a Water Acquity system with PDA (Waters 2996) and Waters ZQ mass spectrometer. Column; Acquity UPLCTM BEH C 8 1.7 ⁇ m 2.1 x 50mm. The column temperature was set to 65°C. A linear 2 min gradient from 100% A (A: 95% 0.01M ammonium acetate in MiIIiQ water and 5% acetonitrile) to 100% B (5% 0.01 M ammonium acetate in MiIIiQ water and 95% acetonitrile) was applied for LC-separation at flow rate 1.2 mL/min. The PDA was scanned from 210-350nm and 254nm was extracted for purity determination.
  • the ZQ mass spectrometer was run with ES in pos/neg switching mode.
  • the Capillary Voltage was 3kV and the Cone Voltage was 30V or LC-MS analyses were performed on a LC-MS consisting of a Waters sample manager 2111 C, a Waters 1525 ⁇ binary pump, a Waters 1500 column oven, a Waters ZQ single quadrupole mass spectrometer, a Waters PDA2996 diode array detector and a Sedex 85 ELS detector.
  • the mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode.
  • the mass spectrometer scanned between m/z 100-700 with a scan time of 0.3s.
  • the capillary voltage was set to 3.4 kV and the cone voltage was set to 30 V, respectively.
  • the diode array detector scanned from 200-400 nm. The temperature of the ELS detector was adjusted to 40 0 C and the pressure was set to 1.9 bar. For separation a linear gradient was applied starting at 100 % A (A: 10 mM NH4OAc in 5 % CH3CN or 8 mM HCOOH in 5% CH3CN) and ending at 100 % B (B: CH3CN).
  • the column used was a Gemini CIS, 3.0 mm x 50 mm, 3 ⁇ m, (Phenomenex) which was run at a flow rate of 1 ml/min.
  • the column oven temperature was set to 40 0 C or
  • LC-MS analyses were performed on a LC-MS consisting of a Waters sample manager 2777 1 C, a Waters 1525 ⁇ binary pump, a Waters 1500 column oven, a Waters ZQ single quadrupole mass spectrometer, a Waters PDA2996 diode array detector and a Sedex 85 ELS detector.
  • the mass spectrometer was configured with an atmospheric pressure chemical ionisation (APCI) ion source which was further equipped with atmospheric pressure photo ionisation (APPI) device.
  • APCI atmospheric pressure chemical ionisation
  • APPI atmospheric pressure photo ionisation
  • the mass spectrometer scanned in the positive mode, switching between APCI and APPI mode.
  • the mass range was set to m/z 120-800 using a scan time of 0.3 s.
  • the APPI repeller and the APCI corona were set to 0.86 kV and 0.80 ⁇ A, respectively.
  • the desolvation temperature (300 0 C), desolvation gas (400 L/Hr) and cone gas (5 L/Hr) were constant for both APCI and APPI mode. Separation was performed using a Gemini column CIS, 3.0 mm x 50 mm, 3 ⁇ m, (Phenomenex) and run at a flow rate of 1 ml/min. A linear gradient was used starting at 100 % A (A: 10 mM NH4OAc in 5% MeOH) and ending at 100% B (MeOH). The column oven temperature was set to 40 0 C.
  • GC-MS Compound identification was performed on a GC-MS system (GC 6890, 5973N MSD) supplied by Agilent Technologies. The column used was a VF-5 MS 3 ID 0.25 mm x 15m, 0.25 ⁇ m (Varian Inc.). A linear temperature gradient was applied starting at 40 0 C (hold 1 min) and ending at 300 °C (hold 1 min), 25 °C/minute.
  • the mass spectrometer was equipped with a chemial ionisation (CI) ion source and the reactant gas was methane.
  • the mass spectrometer was equipped with an electron impact (EI) ion source and the electron voltage was set to 70 eV.
  • CI chemial ionisation
  • EI electron impact
  • the mass spectrometer scanned between m/z 50-500 and the scan speed was set to 3.25 scan/s, or Compound identification was performed on a GC-MS system (GC 6890, 5973N MSD) supplied by Agilent Technologies.
  • the mass spectrometer was equipped with a Direct Inlet Probe (DIP) interface manufactured by SIM GmbH.
  • the mass spectrometer was configured with a chemical ionisation (CI) ion source and the reactant gas was methane.
  • the mass spectrometer was equipped with an electron impact (EI) ion source and the electron voltage was set to 70 eV.
  • the mass spectrometer scanned between m/z 50-500 and the scan speed was set to 3.25 scan/s.
  • a linear temperature gradient was applied starting at 40 0 C (hold 1 min) and ending at 300 0 C (hold 1 min), 25 °C/minute.
  • the column used was a VF-5 MS, ID 0.25 mm x 30m, 0.25 ⁇ m (Varian Inc.).
  • Preparative-HPLC Preparative chromatography was run on Waters auto purification HPLC with a diode array detector. Column: XTerra MS C8, 19 x 300 mm, 10 ⁇ m.
  • Preparative-HPLC was run on a Waters FractionLynx system with a Autosampler combined Automated Fraction Collector (Waters 2767), Gradient Pump (Waters 2525),
  • Microwave heating was performed in a Creator or Initiaror or Smith Synthesizer Single- mode microwave cavity producing continuous irradiation at 2450 MHz.
  • TLC Thin layer chromatography
  • 3-bromobenzonitrile (2.73 g, 15 mmol) was dissolved in dry tetrahydroforan (15 mL) and added drop wise to a cyclopropanemagnesiumbromide (36 mL, 18 mmol, 0.5 M in tetrahydroftiran) under an atmosphere of argon.
  • the mixture was heated to 50 0 C for 4h and then cooled to 0 0 C.
  • Dry methanol (50 mL) was added, the mixture was stirred for 30 min and sodium borohydride (1.14 g, 30 mmol) was added. After 2.5h was saturated aqueous ammonium chloride added and the mixture was extracted with dichloromethane.
  • Example 3 4-(3-Bromophenyl)-4-cyclopropyl-1.3-thiazolidine-2,5-dithione l-Bromo-3-[cyclopropyl(isothiocyanato)methyl]benzene (0.35 g, 1.32 mmol) and carbon disulfide (0.16 mL, 2.63 mmol) in dry tetrahydrofuran (8 mL) was added drop wise to a stirred mixture of potassium tert-butoxide (0.27 g, 2.38 mmol) in dry tetrahydroruran (8 mL) at -78 0 C. After the addition, the mixture was allowed to reach room temperature overnight.
  • 1,3-dibromobenzene (2.42 mL, 20.0 mmol) was dissolved in dry diethyl ether (30 mL) and cooled to -78 0 C.
  • n-Butyllithium (8.0 mL, 20.0 mmol, 2.5 M in hexane) was added drop wise and the mixture was stirred for 70 min.
  • Cyclobutanecarbonitrile (1.62 g, 20.0 mmol) in dry diethyl ether (10 mL) was added and the mixture was stirred at -78 0 C for 15 min and at 0 0 C for 40 min.
  • the enzyme used in the IGEN Cleavage-, Fluorescent-, TR-FRET- and BiaCore assays is described as follows: The soluble part of the human ⁇ -Secretase (AA 1 — AA 460) was cloned into the ASP2- Fc 10-1 -IRES-GFP -neoK mammalian expression vector. The gene was fused to the Fc domain of IgGl (affinity tag) and stably cloned into HEK 293 cells. Purified sBACE-Fc is stored in Tris buffer, pH 9.2 and has a purity of 95%.
  • the enzyme was diluted to 43 ⁇ g/ml in 40 mM MES pH 5.0.
  • the IGEN substrate was diluted to 12 ⁇ M in 40 mM MES pH 5.0.
  • Compounds were diluted to the desired concentration in dimethyl sulfoxide (final dimethyl sulfoxide concentration in assay is 5%).
  • the assay was performed in a 96 well PCR plate from Greiner (#650201). Compound in dimethyl sulfoxide (3 ⁇ L) and enzyme (27 ⁇ L) were added to the plate, and pre- incubated for 10 min. The reaction was started with substrate (30 ⁇ L). The final dilution of enzyme was 20 ⁇ g/ml and the final concentration of substrate was 6 ⁇ M.
  • reaction was stopped by removing 10 ⁇ L of the reaction mix and diluting it 1 :25 in 0.2 M Trizma-HCl, pH 8.0.
  • the product was quantified by adding 50 ⁇ L of a 1:5000 dilution of the neoepitope antibody to 50 ⁇ L of the 1:25 dilution of the reaction mix (all antibodies and the strep tavidin coated beads were diluted in PBS containing 0.5% BSA and 0.5% Tween20).
  • the enzyme was diluted to 52 ⁇ g/ml in 40 mM MES pH 5.0.
  • the substrate (Dabcyl-Edans) was diluted to 30 ⁇ M in 40 mM MES pH 5.0.
  • Compounds were diluted to the desired concentration in dimethyl sulfoxide (final dimethyl sulfoxide concentration in assay is 5%).
  • the assay is done in a Corning 384 well round bottom, low volume, non-binding surface plate (Corning #3676).
  • Enzyme (9 ⁇ L) together with 1 ⁇ L of compound in dimethyl sulfoxide were added to the plate and pre-incubated for 10 min.
  • Substrate (10 ⁇ L) was added and the reaction proceeded in the dark at RT for 25 min.
  • the final dilution of enzyme was 23 ⁇ g/ml, and the final concentration of substrate was 15 ⁇ M (Km of 25 ⁇ M).
  • the fluorescence of the product was measured on a Victor II plate reader with an excitation wavelength of 360 nm and an emission wavelength of 485 nm using a protocol for labelled Edans peptide.
  • the dimethyl sulfoxide control defined 100% activity level and 0% activity was defined by exclusion of the enzyme (using 40 mM MES pH 5.0 buffer instead).
  • Enzyme was diluted to 6 ⁇ g/mL and the substrate (Europium)CEVNLDAEFK(Qsy7) to 200 nM in reaction buffer (NaAcetate, chaps, triton x-100, EDTA pH 4.5). Compounds were diluted to the desired concentration in dimethyl sulfoxide (final dimethyl sulfoxide concentration in assay is 5%). The assay was done in a Costar 384 well round bottom, low volume, non-binding surface plate (Corning #3676). Enzyme (9 ⁇ L) and 1 ⁇ L of compound in dimethyl sulfoxide was added to the plate, mixed and pre-incubated for 10 min.
  • Substrate (10 ⁇ L) was added and the reaction proceeded in the dark for 15 min at RT.
  • the reaction was stopped with the addition of 7 ⁇ L NaAcetate, pH 9.
  • the fluorescence of the product was measured on a Victor II plate reader with an excitation wavelength of 340 5 nm and an emission wavelength of 615 nm.
  • the final concentration of the enzyme was 2.7 ⁇ g/ml and the final concentration of the substrate was 100 nM (Km of 290 nM).
  • the dimethyl sulfoxide control defined the 100% activity level and 0% activity was defined by exclusion of the enzyme (using reaction buffer instead).
  • BACE was assayed on a Biacore3000 instrument by attaching either a peptidic transition state isostere (TSI) or a scrambled version of the peptidic TSI to the surface of a Biacore CM5 sensor chip.
  • TSI transition state isostere
  • the surface of a CM5 sensor chip has 4 distinct channels that can be used to couple the peptides.
  • the scrambled peptide KPES-statine-ETIAEVENV was is coupled to channel 1 and the TSI inhibitor KTEEISEVN-statine-VAEF was coupled to channel 2 of the same chip.
  • the two peptides were dissolved at 0.2 mg/mL in 20 mM sodium acetate pH 4.5, and then the solutions were centrifuged at 14K rpm to remove any particulates.
  • Carboxyl groups on the dextran layer were activated by injecting a one to one mixture of 0.5 M N-ethyl-N' (3-dimethylaminopropyl)-carbodiimide and 0.5 M N-
  • the BACE Biacore assay was done by diluting BACE to 0.5 ⁇ M in sodium acetate buffer at pH 4.5 (running buffer minus dimethyl sulfoxide). The diluted BACE was mixed with dimethyl sulfoxide or compound diluted in dimethyl sulfoxide at a final concentration of 5% dimethyl sulfoxide. The BACE/inhibitor mixture was incubated for 30 minutes at RT before being injected over channel 1 and 2 of the CM5 Biacore chip at a rate of 20 ⁇ L/min.
  • the pcDNA3.1 plasmid encoding the cDNA of human full-length APP695 was stably transfected into HEK-293 cells using the Lipofectamine transfection reagent according to manufacture's protocol (Invitrogen). Colonies were selected with 0.1-0.5 mg/mL of zeocin. Limited dilution cloning was performed to generate homogeneous cell lines. Clones were characterized by levels of APP expression and A ⁇ secreted in the conditioned media using an ELISA assay developed in-house.
  • HEK293 cells stably expressing human wild-type APP were grown at 37 0 C, 5% CO 2 in DMEM containing 4500 g/L glucose, GlutaMAX and sodium pyruvate supplemented with 10% FBS, 1% non-essential amino acids and 0.1 mg/mL of the selection antibiotic zeocin.
  • HEK293-APP695 cells were harvested at 80-90% confluence and seeded at a concentration of 0.2x10 6 cells/mL, 100 mL cell suspension/well, onto a black clear bottom 96-well poly-D-lysine coated plate. After over night incubation at 37 0 C, 5% CO 2 , the cell medium was replaced with cell culture medium with penicillin and streptomycin (100 LVmL, 100 ⁇ g/mL, respectively) containing test compounds in a final dimethyl sulfoxide concentration of 1%. Cells were exposed to the test compounds for 24 h at 37 0 C, 5% CO 2 .
  • test plate 100 ⁇ L cell medium was transferred to a round bottom polypropylene 96-well plate (assay plate). The cell plate was saved for the ATP assay, as described below.
  • 50 ⁇ L of primary detection solution containing 0.5 ⁇ g/mL of the rabbit anti-A ⁇ 40 antibody and 0.5 ⁇ g/mL of the biotinylated monoclonal mouse 6E10 antibody in DPBS with 0.5 %BSA and 0.5% Tween-20 was added per well and incubated over night at 4 0 C.
  • SH-SY5Y cells were grown 37 0 C with 5% CO 2 in DMEM/F-12 1:1 containing GlutaMAX supplemented with 1 mM HEPES 3 10% FBS and 1% non-essential amino acids.
  • SH-SY5Y cells were harvested at 80-90% confluence and seeded at a concentration of 1.5xlO 6 cells/mL, 100 mL cell suspension/well, onto a black clear flat bottom 96-well tissue culture plate. After 7 hours of incubation at 37 0 C, 5% CO 2 , the cell medium was replaced with 90 ⁇ l cell culture medium with penicillin and streptomycin (100 XJ /mL, 100 ⁇ g/mL, respectively) containing test compounds in a final dimethyl sulfoxide concentration of 1%. Cells were exposed to the test compounds for IS h at 37 0 C, 5% CO 2 .
  • sAPP ⁇ microplates from Meso Scale Discovery were used and the assay was performed according to the manufacture's protocol. Briefly, 25 ⁇ L cell medium was transferred to a previously blocked MSD sAPP ⁇ microplate. The cell plate was saved for the ATP assay, as described below. The sAPP ⁇ was captured during shaking at RT for 1 hour, by antibodies spotted in the wells of the microplate. After multiple washes, SULFO-TAG labeled detection antibody was added (25 ⁇ L/well, final concentration InM) to the assay plate and the plate was incubated with shaking at RT for 1 hour. Following multiple washes, 150 ⁇ l/well of Read Buffer T was added to the plate. After 10 minutes at RT the plate was read in the SECTORTM Imager for electro-chemiluminescence.
  • MSD Meso Scale Discovery
  • ATP assay As indicated above, after transferring medium for analysis of A ⁇ 40 or sAPP ⁇ from the cell plate, the plate was used to analyze cytotoxicity using the ViaLightTM Plus cell proliferation/cytotoxicity kit from Cambrex BioScience that measures total cellular ATP. The assay was performed according to the manufacture's protocol. Briefly, 50 ⁇ L cell lysis reagent was added per well. The plates were incubated at RT for 10 min. Two min after addition of 100 ⁇ L reconstituted ViaLightTM Plus ATP reagent, the luminescence was measured in a Wallac Victor 2 1420 multilabel counter.
  • the hERG-expressing Chinese hamster ovary Kl (CHO) cells described by (Persson, Carlsson, Duker, & Jacobson, 2005) were grown to semi-confluence at 37 0 C in a humidified environment (5% CO 2 ) in F- 12 Ham medium containing L-glutamine, 10% foetal calf serum (FCS) and 0.6 mg/ml hygromycin (all Sigma-Aldrich). Prior to use, the monolayer was washed using a pre-warmed (37°C) 3 ml aliquot of Versene 1:5,000 (Invitrogen).
  • CHO-KvI.5 cells which were used to adjust the voltage offset on Ion WorksTM HT, were maintained and prepared for use in the same way.
  • PatchPlate in which a recording is attempted in each well by using suction to position and hold a cell on a small hole separating two isolated fluid chambers. Once sealing has taken place, the solution on the underside of the PatchPlate is changed to one containing amphotericin B. This permeablises the patch of cell membrane covering the hole in each well and, in effect, allows a perforated, whole-cell patch clamp recording to be made.
  • a ⁇ -test IonWorksTM HT from Essen Instrument was used. There is no capability to warm solutions in this device hence it was operated at room temperature ( ⁇ 21°C), as follows.
  • the reservoir in the "Buffer” position was loaded with 4 ml of PBS and that in the "Cells” position with the CHO-hERG cell suspension described above.
  • Each compound plate was laid-out in 12 columns to enable ten, 8- point concentration-effect curves to be constructed; the remaining two columns on the plate were taken up with vehicle (final concentration 0.33% DMSO), to define the assay baseline, and a supra-maximal blocking concentration of cisapride (final concentration 10 mM) to define the 100% inhibition level.
  • the fiuidics-head (F-Head) of IonWorksTM HT then added 3.5 ⁇ l of PBS to each well of the PatchPlateTM and its underside was perfused with "internal" solution that had the following composition (in mM): K-Gluconate 100, KCl 40, MgCl 2 3.2, EGTA 3 and HEPES 5 (all Sigma-Aldrich; pH 7.25-7.30 using 10 M KOH).
  • the electronics-head (E-head) then moved round the PatchPlateTM performing a hole test (i.e. applying a voltage pulse to determine whether the hole in each well was open).
  • the F-head then dispensed 3.5 ⁇ l of the cell suspension described above into each well of the PatchPlateTM and the cells were given 200 seconds to reach and seal to the hole in each well. Following this, the E-head moved round the PatchPlateTM to determine the seal resistance obtained in each well.
  • the solution on the underside of the PatchPlateTM was changed to "access" solution that had the following composition (in mM): KCl 140, EGTA 1, MgCl 2 1 and HEPES 20 (pH 7.25-7.30 using 10 M KOH) plus 100 ⁇ g/ml of amphotericin B (Sigma-Aldrich).
  • the E-head moved round the PatchPlateTM 48 wells at a time to obtain pre-compound hERG current measurements.
  • the F-head then added 3.5 Dl of solution from each well of the compound plate to 4 wells on the PatchPlateTM (the final DMSO concentration was 0.33% in every well). This was achieved by moving from the most dilute to the most concentrated well of the compound plate to minimise the impact of any compound carry-over.
  • the E-head then moved around all 384-wells of the PatchPlateTM to obtain post-compound hERG current measurements. In this way, non-cumulative concentration-effect curves could be produced where, providing the acceptance criteria were achieved in a sufficient percentage of wells (see below), the effect of each concentration of test compound was based on recording from between 1 and 4 cells.
  • the pre- and post-compound hERG current was evoked by a single voltage pulse consisting of a 20 s period holding at -70 mV, a 160 ms step to -60 mV (to obtain an estimate of leak), a 100 ms step back to -70 mV, a 1 s step to + 40 mV, a 2 s step to -30 mV and finally a 500 ms step to -7OmV.
  • a single voltage pulse consisting of a 20 s period holding at -70 mV, a 160 ms step to -60 mV (to obtain an estimate of leak), a 100 ms step back to -70 mV, a 1 s step to + 40 mV, a 2 s step to -30 mV and finally a 500 ms step to -7OmV.
  • Currents were leak-subtracted based on the estimate of current evoked during the +1OmV step at the start of the
  • any voltage offsets in Ion WorksTM HT were adjusted in one of two ways.
  • a depolarising voltage ramp was applied to CHO- KvI.5 cells and the voltage noted at which there was an inflection point in the current trace (i.e. the point at which channel activation was seen with a ramp protocol).
  • the voltage at which this occurred had previously been determined using the same voltage command in conventional electrophysiology and found to be -15 mV (data not shown); thus an offset potential could be entered into the Ion WorksTM HT software using this value as a reference point.
  • any offset was adjusted by determining the hERG tail current reversal potential in IonWorksTM HT, comparing it with that found in conventional electrophysiology (-82 mV) and then making the necessary offset adjustment in the IonWorksTM HT software.
  • the current signal was sampled at 2.5 kHz.
  • Pre- and post-scan hERG current magnitude was measured automatically from the leak subtracted traces by the IonWorksTM HT software by taking a 40 ms average of the current during the initial holding period at -70 mV (baseline current) and subtracting this from the peak of the tail current response.
  • the acceptance criteria for the currents evoked in each well were: pre-scan seal resistance >60 M ⁇ , pre-scan hERG tail current amplitude >150 pA; post-scan seal resistance >60 M ⁇ .
  • the degree of inhibition of the hERG current was assessed by dividing the post-scan hERG current by the respective pre-scan hERG current for each well.
  • Typical IC50 values for the compounds of the present invention are in the range of about 1 to about 10,000 nM.
  • Biological data on exemplified final compounds is given below in Table 1.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Psychiatry (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hospice & Palliative Care (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

This invention relates to novel compounds having the structural formula (I) below: and to their pharmaceutically acceptable salt, compositions and methods of use. These novel compounds provide a treatment or prophylaxis of cognitive impairment, Alzheimer Disease, neurodegeneration and dementia.

Description

Amino-imidazolones and their use as medicament for treating cognitive impairment Alzheimer disease, neurodegeneration and dementia.
The present invention relates to novel compounds, their pharmaceutical compositions. In addition, the present invention relates to therapeutic methods for the treatment and/or prevention of Aβ-related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
Background of the invention
Several groups have identified and isolated aspartate proteinases that have β-secretase activity (Hussain et al., 1999; Lin et. al, 2000; Yan et. al, 1999; Sinlia et. al., 1999 and Vassar et. al., 1999). β-secretase is also known in the literature as Asp2 (Yan et. al, 1999), Beta site APP Cleaving Enzyme (BACE) (Vassar et. al., 1999) or memapsin-2 (Lin et al., 2000). BACE was identified using a number of experimental approaches such as EST database analysis (Hussain et al. 1999); expression cloning (Vassar et al. 1999); identification of human homologs from public databases of predicted C. elegans proteins (Yan et al. 1999) and finally utilizing an inhibitor to purify the protein from human brain (Sinha et al. 1999). Thus, five groups employing three different experimental approaches led to the identification of the same enzyme, making a strong case that BACE is a β- secretase. Mention is also made of the patent literature: WO96/40885, EP871720, U.S. Patents Nos. 5,942,400 and 5,744,346, EP855444, US 6,319,689, WO99/64587, WO99/31236, EP1037977, WO00/17369, WO01/23533, WO0047618, WO00/58479, WO00/69262, WO01/00663, WO01/00665, US 6,313,268.
BACE was found to be a pepsin-like aspartic proteinase, the mature enzyme consisting of the N-terminal catalytic domain, a transmembrane domain, and a small cytoplasmic domain. BACE has an optimum activity at pH 4.0-5.0 (Vassar et al, 1999) and is inhibited weakly by standard pepsin inhibitors such as pepstatin. It has been shown that the catalytic domain minus the transmembrane and cytoplasmic domain has activity against substrate peptides (Lin et al, 2000). BACE is a membrane bound type 1 protein that is synthesized as a partially active proenzyme, and is abundantly expressed in brain tissue. It is thought to represent the major β-secretase activity, and is considered to be the rate-limiting step in the production of amyloid-β-protein (Aβ). It is thus of special interest in the pathology of Alzheimer's disease, and in the development of drugs as a treatment for Alzheimer's disease.
Aβ or amyloid-β-protein is the major constituent of the brain plaques which are characteristic of Alzheimer's disease (De Strooper et al, 1999). Aβ is a 39-42 residue peptide formed by the specific cleavage of a class I transmembrane protein called APP, or amyloid precursor protein. Aβ-secretase activity cleaves this protein between residues Met671 and Asp672 (numbering of 770aa isoform of APP) to form the N-terminus of Aβ. A second cleavage of the peptide is associated with γ-secretase to form the C-terminus of the Aβ peptide.
Alzheimer's disease (AD) is estimated to afflict more than 20 million people worldwide and is believed to be the most common form of dementia. Alzheimer's disease is a progressive dementia in which massive deposits of aggregated protein breakdown products - amyloid plaques and neurofibrillary tangles accumulate in the brain. The amyloid plaques are thought to be responsible for the mental decline seen in Alzheimer's patients.
The likelihood of developing Alzheimer's disease increases with age, and as the aging population of the developed world increases, this disease becomes a greater and greater problem. In addition to this, there is a familial link to Alzheimer's disease and consequently any individuals possessing the double mutation of APP known as the Swedish mutation (in which the mutated APP forms a considerably improved substrate for BACE) have a much greater chance of developing AD, and also of developing it at an early age (see also US 6,245,964 and US 5,877,399 pertaining to transgenic rodents comprising APP-Swedish). Consequently, there is also a strong need for developing a compound that can be used in a prophylactic fashion for these individuals.
The gene encoding APP is found on chromosome 21 , which is also the chromosome found as an extra copy in Down's syndrome. Down's syndrome patients tend to acquire Alzheimer's disease at an early age, with almost all those over 40 years of age showing Alzheimer's-type pathology (Oyama et al., 1994). This is thought to be due to the extra copy of the APP gene found in these patients, which leads to overexpression of APP and therefore to increased levels of APPβ causing the high prevalence of Alzheimer's disease seen in this population. Thus, inhibitors of BACE could be useful in reducing Alzheimer's-type pathology in Down's syndrome patients.
Drugs that reduce or block BACE activity should therefore reduce Aβ levels and levels of fragments of Aβ in the brain, or elsewhere where Aβ or fragments thereof deposit, and thus slow the formation of amyloid plaques and the progression of AD or other maladies involving deposition of Aβ or fragments thereof (Yankner, 1996; De Strooper and Konig, 1999). BACE is therefore an important candidate for the development of drugs as a treatment and/or prophylaxis of Aβ-related pathologies such as Downs syndrome and β- amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
It would therefore be useful to inhibit the deposition of Aβ and portions thereof by inhibiting BACE through inhibitors such as the compounds provided herein.
The therapeutic potential of inhibiting the deposition of Aβ has motivated many groups to isolate and characterize secretase enzymes and to identify their potential inhibitors {see, e.g., WO01/23533 A2, EP0855444, WO00/17369, WO00/58479, WO00/47618, WO00/77030, WO01/00665, WO01/00663, WO01/29563, WO02/25276, US5,942,400, US6,245,884, US6,221,667, US6,211,235, WO02/02505, WO02/02506, WO02/02512, WO02/02518, WO02/02520, WO02/14264, WO05/05S311, WO05/097767,
WO06/041404, WO06/041405, WO06/0065204, WO06/0065277, US2006287294, WO06/138265, US20050282826, US20050282825, US20060281729, WO06/138217, WO06/138230, WO06/138264, WO06/138265, WO06/138266, WO06/099379, WO06/076284, US20070004786, US20070004730, WO07/011833, WO07/011810, US20070099875, US20070099898, WO07/058601, WO07/058581, WO07/058580, WO07/058583, WO07/058582, WO07/058602, WO07/073284, WO07/049532, WO07/038271, WO07/016012, WO07/005366, WO07/005404, WO06/0009653).
The compounds of the present invention show beneficial properties compared to the potential inhibitors known in the art, e.g. improved hERG selectivity.
Disclosure of the invention
Provided herein are novel BACE inhibitors. Thus in a first aspect of the invention there is provided compounds of structural formula I:
Figure imgf000005_0001
wherein
A is independently selected from a 5, 6 or 7 membered heterocyclic ring optionally substituted with one or more R1;
B is independently selected from a 4, 5, 6 or 7 membered heterocyclic ring, Q^alkyl, C2.6alkenyl, C^alkynyl, C0.6alkylaryl, Co-βalkylheteroaryl, Co-ealkylCs-gcycloalkyl, Co- βalkylCs-βcycloalkenyl, Co-6alkylC3-6cycloalkynyl, Co-6alkylC3-6heterocyclyl optionally substituted with one or more R2;
C is independently selected from phenyl or a 5 or 6 membered heteroaromatic ring optionally substituted with one or more R3;
R1 is independently selected from halogen, cyano, nitro, OR6, d-βalkyl, C2-6a.ken.yl, C2.6alkynyl, Co-βalkylaryl, Co-βalkylheteroaryl, Co-βalkylCs-βcycloalkyl, Co-6alkylC3_ ecycloalkenyl, C0.salkylC3.6cycloalky1.yl, Co-6alkylC3-6heterocyclyl, NR6R7, CONR6R7, NR6(CO)R7, 0(CO)R6, CO2R6, COR6, (SO2)NR6R7, NR6(SO2)R7, SOR6, SO2R6, OSO2R6 and SO3R6 wherein said Q^alkyl, C2-6alkenyl, C2-6alkynyl, Co-ealkylaryl, Co- βalkylheteroaryl, C0.6alkylC3.6cycloa.kyl, Co-βalkylCa-ecycloalkenyl, Co-6alkylC3- βcycloalkynyl and Co-6alkylC3-6heterocyclyI may be optionally substituted with one or more D; or two R1 substituents may together with the atom to which they are attached form a cyclic or heterocyclic ring optionally substituted with one or more D;
R2, R3 and R4 are independently selected from halogen, cyano, nitro, OR6, C^alkyl, C2.όalkenyl, Ca-βalkynyl, Co-βalkylaryl, Co-βalkylheteroaryl, Co-6alkylC3.όcycloa]lcyl, Co- βalkylCs-βcycloalkenyl, C0-6alkylC3_6cycloalkynyl, Co-βalkylCs-βheterocyclyl, NR6R7, CONR6R7, NR6(CO)R7, 0(CO)R6, CO2R6, COR6, (SO2)NR6R7, NR6(SO2)R7, SO2R6, SOR6, OSO2R6 and SO3R6 wherein said C1-όalkyl, C2-6alkenyl, C2-6alkynyl, Co^alkylaryl, Co-ealkylheteroaryl, Co-ealkylCs-βcycloalkyl, Co-βalkylCs-βcycloalkenyl, Co-OaIlCyIC3- δcycloalkynyl and Co-όalkylCs-δheterocyclyl is optionally substituted with one or more D; or two R2, R3 or R4 substituents may together with the atoms to which they are attached foπn a cyclic or heterocyclic ring optionally substituted with one or more D;
R5 is independently selected from hydrogen, cyano, OR6, C^alkyl, C2-6alkenyl, C2-6alkynyl, Co-ealkylaryl, Co-βalkylheteroaryl, Co-βalkylCs-βcycloalkyl, Co-6alkylC3. βcycloalkenyl, Co.6alkylC3.6cycloalkynyl,
Figure imgf000006_0001
CONR6R7, CO2R6, COR6, SO2R6 and SO3R6 wherein said Ci-6alkyl, C2-6alkenyl, C2-6alkynyl, C0.6alkylaryl, C0, 6alkylheteroaryl, Co.ealkylCs.gcycloalkyl, Co-6alkyIC3.6cycloalkenyl, Co-ealkylCs. 6cycloalkynyl, Co-6alkylC3-6heterocyclyl may be optionally substituted with one or more D;
D is independently selected from halogen, nitro, CN, OR0, C^alkyl, C2_6atkenyl, C2-6alkynyl, Co-βalkylaryl, Co-ealkylheteroaryl,
Figure imgf000007_0001
Co-ealkylCs. δcycloalkenyl, Co-ealkylQ-όcycloalkynyl, Co-βalkylheterocyclyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoromethoxy, difluoromethoxy, trifluoromethoxy, NR6R7, CONR6R7, NR6(CO)R7, 0(CO)R6, CO2R6, COR6, (SO2)NR6R7, NR6SO2R7, SO2R6, SOR6, OSO2R6 and SO3R6, wherein said Ci-6alkyl, C2.6alkenyl, Cz-βalkynyl, C0- βalkylaryl, Co-βalkylheteroaryl, Co-6alkylC3-6cycloalkyl, Co-βalky IC3-6Cy cloalkenyl, C0. δalkylCs.όcycloalkynyl or Co-ealkylheterocyclyl may be optionally substituted with one or more substituents independently selected from halo, nitro, cyano, OR6, Ci-βalkyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoromethoxy, difluoromethoxy and trifluoromethoxy;
Rb and R7 are independently selected from hydrogen, Chalky!, fluoromethyl, difluoromethyl, trifluoromethyl, C2.6alkenyl, Ca-βalkynyl, Co-βalkylaryl, Co- 6alkylheteroaryl, Co-δalkylC3.(;cycloalkyl, Co-6alkylC3.6cycloalkenyl, Co-salkylC3. βcycloalkynyl, Co-ealkylheterocyclyl; or R6 and R7 may together form a 5 or 6 membered heterocyclic ring containing one or more heteroatoms selected from N, O or S;
m = 0, 1, 2 or 3; n = 0, 1, 2 or 3; p = 0, 1, 2 or 3; q = 0, 1, 2 or 3;
as a free base or a pharmaceutically acceptable salt, solvate or solvate of a salt thereof.
The present invention further provides pharmaceutical compositions comprising as active ingredient a therapeutically effective amount of a compound of formula I in association with pharmaceutically acceptable excipients, carriers or diluents. The present invention farther provides methods of modulating activity of BACE comprising contacting the BACE enzyme with a compound of formula I.
The present invention further provides methods of treating or preventing an Aβ-related pathology in a patient, comprising administering to the patient a therapeutically effective amount of a compound of formula I.
The present invention further provides a compound described herein for use as a medicament.
In one aspect of the present invention, there is provided a compound according to formula I, wherein A is a 6 membered heterocyclic ring.
In another aspect of the present invention, there is provided a compound according to formula I, wherein B is a Co-galkylCj-scycloalkyl.
In another aspect of the present invention, there is provided a compound according to formula I, wherein B is represented by Cs-βcycloalkyl.
In yet another aspect of the present invention, there is provided a compound according to formula I, wherein B is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
In yet another aspect of the present invention, there is provided a compound according to formula I, wherein R5 is hydrogen.
In yet another aspect of the present invention, there is provided a compound according to formula I5 wherein m is 0.
In yet another aspect of the present invention, there is provided a compound according to formula I, wherein n is 0. In yet another aspect of the present invention, there is provided a compound according to formula I, wherein q is 0.
In yet another aspect of the present invention, there is provided a compound according to formula I, wherein C is phenyl.
In yet another aspect of the present invention, there is provided a compound according to foπnula I, wherein C is phenyl substituted with one or more R3.
In yet another aspect of the present invention, there is provided a compound according to formula I, wherein R3 is halogen.
In yet another aspect of the present invention, there is provided a compound according to formula I, wherein said halogen is chlorine.
In yet another aspect of the present invention, there is provided a compound according to foπnula I, wherein
A is a 6 membered heterocyclic ring;
B is Co-6alkylC3-6cycloalkyl; C is phenyl substituted with one or more R3;
R3 is halogen;
R5 is hydrogen; m = 0; n = 0; p = 2; q = 0.
In yet another aspect of the present invention, there is provided a compound selected from: S-Cyclopropyl-8-(3',5'-dichlorobiphenyl-3-yl)-2,334,8-tetrahydroimidazo[l,5-α]pyrimidin- 6-amine acetate;
8-Cyclohexyl-8-(3',5'-dichlorobiphenyl-3-yl)-2,3,4,8-tetrahydroimidazo[l,5-α]pyrimidin-6- amine acetate; 8-Cyclobutyl-8-(3',5'-dichlorobiphenyl-3-yl)-2,3,4,8-tetrahydroimidazo[l,5-α]pyrimidin-6- amine acetate and
8-Cyclopentyl-8-(3',5l-dichlorobiphenyl-3-yl)-2,3,4>8-tetrahydroimidazo[l,5-α]pyrimidin- 6-amine acetate;
as a free base or a pharmaceutically acceptable salt, solvate or solvate of a salt thereof.
Some compounds of formula I may have stereogenic centres and/or geometric isomeric centres (E- and Z- isomers), and it is to be understood that the invention encompasses all such optical isomers, enantiomers, diastereoisomers, atropisomers and geometric isomers.
The present invention relates to the use of compounds of formula I as hereinbefore defined as well as to the salts thereof. Salts for use in pharmaceutical compositions will be pharmaceutically acceptable salts, but other salts may be useful in the production of the compounds of formula I.
It is to be understood that the present invention relates to any and all tautomeric forms of the compounds of formula I.
Compounds of the invention can be used as medicaments. In some embodiments, the present invention provides compounds of formula I, or pharmaceutically acceptable salts, tautomers or in vzvo-hydrolysable precursors thereof, for use as medicaments. In some embodiments, the present invention provides compounds described here in for use as medicaments for treating or preventing an Aβ-related pathology. In some further embodiments, the Aβ-related pathology is Downs syndrome, a β-amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, a disorder associated with cognitive impairment, MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with Alzheimer disease, dementia of mixed vascular origin, dementia of degenerative origin, pre-senile dementia, senile dementia, dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration. In some embodiments, the present invention provides use of compounds of formula I or pharmaceutically acceptable salts, tautomers or in vzvo-hydrolysable precursors thereof, in the manufacture of a medicament for the treatment or prophylaxis of Aβ-related pathologies. In some further embodiments, the Aβ-related pathologies include such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
In some embodiments, the present invention provides a method of inhibiting activity of BACE comprising contacting the BACE with a compound of the present invention. BACE is thought to represent the major β-secretase activity, and is considered to be the rate- limiting step in the production of amyloid-β-protein (Aβ). Thus, inhibiting BACE through inhibitors such as the compounds provided herein would be useful to inhibit the deposition of Aβ and portions thereof. Because the deposition of Aβ and portions thereof is linked to diseases such Alzheimer Disease, BACE is an important candidate for the development of drugs as a treatment and/or prophylaxis of Aβ-related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration. In some embodiments, the present invention provides a method for the treatment of Aβ- related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment'"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, presenile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration, comprising administering to a mammal (including human) a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt, tautomer or in vz'rø-hydrolysable precursor thereof.
In some embodiments, the present invention provides a method for the prophylaxis of Aβ- related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, presenile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration comprising administering to a mammal (including human) a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, tautomer or in vzvo-hydrolysable precursors.
In some embodiments, the present invention provides a method of treating or preventing Aβ-related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre- senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration by administering to a mammal (including human) a compound of formula I or a pharmaceutically acceptable salt, tautomer or in vzvo-hydrolysable precursors and a cognitive and/or memory enhancing agent.
In some embodiments, the present invention provides a method of treating or preventing Aβ-related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, presenile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration by administering to a mammal (including human) a compound of formula I or a pharmaceutically acceptable salt, tautomer or in vzvo-hydrolysable precursors thereof wherein constituent members are provided herein, and a choline esterase inhibitor or anti-inflammatory agent.
In some embodiments, the present invention provides a method of treating or preventing Aβ-related pathologies such as Downs syndrome and β-amyloid angiopathy, such as but not limited to cerebral amyloid angiopathy, hereditary cerebral hemorrhage, disorders associated with cognitive impairment, such as but not limited to MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with diseases such as Alzheimer disease or dementia including dementia of mixed vascular and degenerative origin, pre-senile dementia, senile dementia and dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration, or any other disease, disorder, or condition described herein, by administering to a mammal (including human) a compound of the present inventionand an atypical antipsychotic agent. Atypical antipsychotic agents includes, but not limited to, Olanzapine (marketed as Zyprexa), Aripiprazole (marketed as Abilify), Risperidone (marketed as Risperdal), Quetiapine (marketed as Seroquel), Clozapine (marketed as Clozaril), Ziprasidone (marketed as Geodon) and Olanzapine/Fluoxetine (marketed as Symbyax).
In some embodiments, the mammal or human being treated with a compound of the invention has been diagnosed with a particular disease or disorder, such as those described herein. In these cases, the mammal or human being treated is in need of such treatment. Diagnosis, however, need not be previously performed.
The present invention also includes pharmaceutical compositions which contain, as the active ingredient, one or more of the compounds of the invention herein together with at least one pharmaceutically acceptable carrier, diluent or excipent.
The definitions set forth in this application are intended to clarify terms used throughout this application. The term "herein" means the entire application.
A variety of compounds in the present invention may exist in particular geometric or stereoisomeric forms. The present invention takes into account all such compounds, including cis- and trans isomers, R- and S- enantiomers, diastereomers, (D)-isomers, (L)- isomers, the racemic mixtures thereof, and other mixtures thereof, as being covered within the scope of this invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention. The compounds herein described may have asymmetric centers. Compounds of the present invention containing an asymmetrically substituted atom may be isolated in optically active or racemic forms. It is well known in the art how to prepare optically active forms, such as by resolution of racemic forms, by synthesis from optically active starting materials, or synthesis using optically active reagents. When required, separation of the racemic material can be achieved by methods known in the art. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and may be isolated as a mixture of isomers or as separated isomeric forms. All chiral, diastereomeric, racemic forms and all geometric isomeric forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated.
When a bond to a substituent is shown to cross a bond connecting two atoms in a ring, then such substituent may be bonded to any atom on the ring. When a substituent is listed without indicating the atom via which such substituent is bonded to the rest of the compound of a given formula, then such substituent may be bonded via any atom in such substituent. Combinations of substituents, positions of substituents and/or variables are permissible only if such combinations result in stable compounds.
As used in this application, the term "optionally substituted," means that substitution is optional and therefore it is possible for the designated atom or moiety to be unsubstituted. In the event a substitution is desired then such substitution means that any number of hydrogens on the designated atom or moiety is replaced with a selection from the indicated group, provided that the normal valency of the designated atom or moiety is not exceeded, and that the substitution results in a stable compound. For example when a substituent is methyl (i.e., CH3), then 3 hydrogens on the carbon atom can be replaced. Examples of such substituents include, but are not limited to: halogen, CN, NH2, OH, SO, SO2, COOH, Od- βalkyl, CH2OH, SO2H, d.6alkyl, OC^alkyl, C(=O)C1-6alkyl, C(=O)OC1-6alkyl, C(=0)NH2, C(=O)NHCi.6alkyl, C(=O)N(C]-6alkyl)2, SO2C1-6alkyl, SO2NHC1-όalkyl.,
SO2N(C1-6alkyl)2, NH(C1-5alkyl), N(Ci-6alkyl)2, NHC(=O)Cwalkyl, NC(=O)(Ci.6alkyl)2, Cs-βaryl, OC5.6aryl, C(=O)C5.6aryl, C(=O)OC5.6aryl, C(=O)NHC5-6aryl, C(=0)N(C5- 6aryl)2, SO2C5.6aryl, SO2NHC5-6aryl, SO2N(C5-5aryl)2, NH(C5-6aryl), N(C5-6aryl)2, NC(=O)C5-6aryl, NC(=O)(C5.6aryl)2, C5-6heterocyclyl, OCs-όheterocyclyl, C(=O)C5- eheterocyclyl, C(=0)OC5-6heterocyclyl5 C(=O)NHC5-6heterocyclyl, C(O)N(C5-
6heterocyclyl)2, SO2Cs.6heterocyclyl,
Figure imgf000015_0001
SO2N(C5.όheterocyclyl)2, NH(C5.6heterocyclyI), N(C5.6heterocyclyl)2, NC(=O)C5.6heterocyclyl, NC(O)(C5. 6heterocyclyl)2.
As used herein, "alkyl", used alone or as a suffix or prefix, is intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having from 1 to 12 carbon atoms or if a specified number of carbon atoms is provided then that specific number would be intended. For example "Co-6 alkyl" denotes alkyl having 0, 1, 2, 3, 4, 5 or 6 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, pentyl, and hexyl. In the case where a subscript is the integer 0 (zero) the group to which the subscript refers to indicates that the group may be absent, i.e. there is a direct bond between the groups.
As used herein, "alkenyl" used alone or as a suffix or prefix is intended to include both branched and straight-chain alkene or olefin containing aliphatic hydrocarbon groups having from 2 to 12 carbon atoms or if a specified number of carbon atoms is provided then that specific number would be intended. For example "Ca-βalkenyl" denotes alkenyl having 2, 3, 4, 5 or 6 carbon atoms. Examples of alkenyl include, but are not limited to, vinyl, allyl, 1-proρenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylbut-2-enyl, 3-methylbut- 1-enyl, 1-pentenyl, 3-pentenyl and 4-hexenyl.
As used herein, "alkynyl" used alone or as a suffix or prefix is intended to include both branched and straight-chain alkyne containing aliphatic hydrocarbon groups having from 2 to 12 carbon atoms or if a specified number of carbon atoms is provided then that specific number would be intended. For example "C^alkynyl" denotes alkynyl having 2, 3, 4, 5 or 6 carbon atoms. Examples of alkynyl include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 3-butynyl, -pentynyl, hexynyl and l-methylpent-2-ynyl.
As used herein, "aromatic" refers to hydrocarbonyl groups having one or more unsaturated carbon ring(s) having aromatic characters, (e.g. 4n + 2 delocalized electrons) and comprising up to about 14 carbon atoms. In addition "hetero aromatic" refers to groups having one or more unsaturated rings containing carbon and one or more heteroatoms such as nitrogen, oxygen or sulphur having aromatic character (e.g. 4n + 2 delocalized electrons).
As used herein, the term "aryl" refers to an aromatic ring structure made up of from 5 to 14 carbon atoms. Ring structures containing 5, 6, 7 and 8 carbon atoms would be single-ring aromatic groups, for example, phenyl. Ring structures containing 8, 9, 10, 11, 12, 13, or 14 would be poly cyclic, for example naphthyl. The aromatic ring can be substituted at one or more ring positions with such substituents as described above. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "fused rings") wherein at least one of the rings is aromatic, for example, the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls. The terms ortho, meta and para apply to 1,2-, 1,3- and 1 ,4-disubstituted benzenes, respectively. For example, the names 1 ,2-dimethylbenzene and ortho-dimethylbenzene are synonymous.
As used herein, the term "cycloalkyl" is intended to include saturated ring groups, having the specified number of carbon atoms. These may include fused or bridged polycyclic systems. Preferred cycloalkyls have from 3 to 10 carbon atoms in their ring structure, and more preferably have 3, 4, 5, and 6 carbons in the ring structure. For example, "C3-6 cycloalkyl" denotes such groups as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
As used herein, "cycloalkenyl" refers to ring-containing hydrocarbyl groups having at least one carbon-carbon double bond in the ring, and having from 4 to 12 carbons atoms.
As used herein, "cycloalkynyl" refers to ring-containing hydrocarbyl groups having at least one carbon-carbon triple bond in the ring, and having from 7 to 12 carbons atoms.
As used herein, "halo" or "halogen" refers to fluoro, chloro, bromo, and iodo. "Counterion" is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, sulfate, tosylate, benezensulfonate, and the like.
As used herein, the term "heterocyclyl" or "heterocyclic" or "heterocycle" refers to a saturated, unsaturated or partially saturated, monocyclic, bicyclic or tricyclic ring (unless otherwise stated) containing 3 to 20 atoms of which 1, 2, 3, 4 or 5 ring atoms are chosen from nitrogen, sulphur or oxygen, which may, unless otherwise specified, be carbon or nitrogen linked, wherein a -CH2- group is optionally be replaced by a -C(O)-; and where unless stated to the contrary a ring nitrogen or sulphur atom is optionally oxidised to form the N-oxide or S-oxide(s) or a ring nitrogen is optionally quarternized; wherein a ring -NH is optionally substituted by acetyl, formyl, methyl or mesyl; and a ring is optionally substituted by one or more halo. It is understood that when the total number of S and O atoms in the heterocyclyl exceeds 1 , then these heteroatoms are not adjacent to one another. If the said heterocyclyl group is bi- or tricyclic then at least one of the rings may optionally be a heteroaromatic or aromatic ring provided that at least one of the rings is non-heteroaromatic. If the said heterocyclyl group is monocyclic then it must not be aromatic. Examples of heterocyclyls include, but are not limited to, piperidinyl, N- acetylpiperidinyl, N-methylpiperidinyl, JV-forrnylpiperazinyl, iV-mesylpiperazinyl, homopiperazinyl, piperazinyl, azetidinyl, oxetanyl, morpholinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, indolinyl, tetrahydropyranyl, dihydro-2H-pyranyl, tetrahydrofuranyl and 2,5-dioxoimidazolidinyl.
As used herein, "heteroaryl" or "heteroaromatic" refers to an aromatic heterocycle having at least one heteroatom ring member such as sulfur, oxygen, or nitrogen. Heteroaryl groups include monocyclic and polycyclic (e.g., having 2, 3 or 4 fused rings) systems. Examples of heteroaryl groups include without limitation, pyridyl (i.e., pyridinyl), pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl (i.e. furanyl), quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl, pyrryl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, fluorenonyl, benzimidazolyl, indolinyl, and the like. In some embodiments, the heteroaryl group has from 1 to about 20 carbon atoms, and in further embodiments from about 3 to about 20 carbon atoms. In some embodiments, the heteroaryl group contains 3 to about 14, 4 to about 14, 3 to about 7, or 5 to 6 ring-forming atoms. In some embodiments, the heteroaryl or heteroaromatic group has 1 to about 4, 1 to about 3, or 1 to 2 heteroatoms. In some embodiments, the heteroaryl or heteroaromatic group has 1 heteroatom.
As used herein, the phrase "protecting group" means temporary substituents which protect a potentially reactive functional group from undesired chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones respectively. The field of protecting group chemistry has been reviewed (Greene, T.W.; Wuts, P.G.M. Protective Groups in Organic Synthesis, 3rd ed.; Wiley: New York, 1999).
As used herein, "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid.
The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used.
As used herein, "tautomer" means other structural isomers that exist in equilibrium resulting from the migration of a hydrogen atom. For example, keto-enol tautomerism where the resulting compound has the porperties of both a ketone and an unsturated alchol. As used herein "stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
Compounds of the invention further include hydrates and solvates.
The present invention further includes isotopically-labeled compounds of the invention. An "isotopically" or "radio-labeled" compound is a compound of the invention where one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occuπing). Suitable radionuclides that may be incorporated in compounds of the present invention include but are not limited to 2H (also written as D for deuterium), 3H (also written as T for tritium), 11C, 13C5 14C, 13N, 15N, 150, 17O, 180, 18F, 35S, 36Cl, 82Br, 75Br, 76Br, 77Br, 123I, 124I, 125I and 131I. The radionuclide that is incorporated in the instant radio- labeled compounds will depend on the specific application of that radio-labeled compound. For example, for in vitro receptor labeling and competition assays, compounds that incorporate 3H, 14C, 82Br, 1251 , 1311, 35S or will generally be most useful. For radio- imaging applications 11C, 18F, 125I5 1231, 124I3 131I5 75Br, 76Br or 77Br will generally be most useful.
It is understood that a "radio-labeled compound" is a compound that has incorporated at least one radionuclide. In some embodiments the radionuclide is selected from the group consisting of 3H, 14C, 1251 , 35S and 82Br.
The anti-dementia treatment defined herein may be applied as a sole therapy or may involve, in addition to the compound of the invention, conventional chemotherapy. Such chemotherapy may include one or more of the following categories of agents: acetyl cholinesterase inhibitors, anti-inflammatory agents, cognitive and/or memory enhancing agents or atypical antipsychotic agents.
Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment. Such combination products employ the compounds of this invention.
Compounds of the present invention may be administered orally, parenteral, buccal, vaginal, rectal, inhalation, insufflation, sublingually, intramuscularly, subcutaneously, topically, intranasally, intraperitoneally, intrathoracially, intravenously, epidurally, ■ intrathecally, intracerebroventricularly and by injection into the joints.
The dosage will depend on the route of administration, the severity of the disease, age and weight of the patient and other factors normally considered by the attending physician, when determining the individual regimen and dosage level as the most appropriate for a particular patient.
An effective amount of a compound of the present invention for use in therapy of dementia is an amount sufficient to symptomatically relieve in a warm-blooded animal, particularly a human the symptoms of dementia, to slow the progression of dementia, or to reduce in patients with symptoms of dementia the risk of getting worse.
For preparing pharmaceutical compositions from the compounds of this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories.
A solid carrier can be one or more substances, which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material.
In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
For preparing suppository compositions, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient sized molds and allowed to cool and solidify.
Suitable carriers include magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.
In some embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for the therapeutic treatment (including prophylactic treatment) of mammals including humans, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.
In addition to the compounds of the present invention, the pharmaceutical composition of this invention may also contain, or be co-administered (simultaneously or sequentially) with, one or more pharmacological agents of value in treating one or more disease conditions referred to herein.
The term composition is intended to include the formulation of the active component or a pharmaceutically acceptable salt with a pharmaceutically acceptable carrier. For example this invention may be formulated by means known in the art into the form of, for example, tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely divided powders or aerosols or nebulisers for inhalation, and for parenteral use (including intravenous, intramuscular or infusion) sterile aqueous or oily solutions or suspensions or sterile emulsions.
Liquid form compositions include solutions, suspensions, and emulsions. Sterile water or water-propylene glycol solutions of the active compounds may be mentioned as an example of liquid preparations suitable for parenteral administration. Liquid compositions can also be formulated in solution in aqueous polyethylene glycol solution. Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.
The pharmaceutical compositions can be in unit dosage form. In such form, the composition is divided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparations, for example, packeted tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms.
Compositions may be formulated for any suitable route and means of administration. Pharmaceutically acceptable carriers or diluents include those used in formulations suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.
For solid compositions, conventional non-toxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, cellulose, cellulose derivatives, starch, magnesium stearate, sodium saccharin, talcum, glucose, sucrose, magnesium carbonate, and the like may be used. Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc, an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline aqueous dextrose, glycerol, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of non- toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanol amine sodium acetate, sorbitan monolaurate, triethanolamine oleate, etc. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, 15th Edition, 1975. The compounds of the invention may be derivatised in various ways. As used herein "derivatives" of the compounds includes salts (e.g. pharmaceutically acceptable salts), any complexes (e.g. inclusion complexes or clathrates with compounds such as cyclodextrins, or coordination complexes with metal ions such as Mn2+ and Zn2+), free acids or bases, polymorphic forms of the compounds, solvates (e.g. hydrates), prodrugs or lipids, coupling partners and protecting groups. By "prodrugs" is meant for example any compound that is converted in vivo into a biologically active compound.
Salts of the compounds of the invention are preferably physiologically well tolerated and non toxic. Many examples of salts are known to those skilled in the art. All such salts are within the scope of this invention, and references to compounds include the salt forms of the compounds.
Where the compounds contain an amine function, these may form quaternary ammonium salts, for example by reaction with an alkylating agent according to methods well known to the skilled person. Such quaternary ammonium compounds are within the scope of the invention.
Compounds containing an amine function may also form TV-oxides. A reference herein to a compound that contains an amine function also includes the JV-oxide.
Where a compound contains several amine functions, one or more than one nitrogen atom may be oxidised to form an JV-oxide. Particular examples of TV-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle.
TV-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience, pages. More particularly, TV-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compound is reacted with /w-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as dichloromethane. Where the compounds contain chiral centres, all individual optical forms such as enantiomers, epimers and diastereoisomers, as well as racemic mixtures of the compounds are within the scope of the invention.
Compounds may exist in a number of different geometric isomeric, and tautomeric forms and references to compounds include all such forms. For the avoidance of doubt, where a compound can exist in one of several geometric isomeric or tautomeric forms and only one is specifically described or shown, all others are nevertheless embraced by the scope of this invention.
The quantity of the compound to be administered will vary for the patient being treated and will vary from about 100 ng/kg of body weight to 100 mg/kg of body weight per day and preferably will be from 10 pg/kg to 10 mg/kg per day. For instance, dosages can be readily ascertained by those skilled in the art from this disclosure and the knowledge in the art. Thus, the skilled artisan can readily determine the amount of compound and optional additives, vehicles, and/or carrier in compositions and to be administered in methods of the invention.
Compounds of the present invention have been shown to inhibit beta secretase (including BACE) activity in vitro. Inhibitors of beta secretase have been shown to be useful in blocking formation or aggregation of Aβ peptide and therefore have beneficial effects in treatment of Alzheimer's Disease and other neurodegenerative diseases associated with elevated levels and/or deposition of Aβ peptide. Therefore, it is believed that the compounds of the present invention may be used for the treatment of Alzheimer disease and disease associated with dementia Hence, compounds of the present invention and their salts are expected to be active against age-related diseases such as Alzheimer, as well as other Aβ related pathologies such as Downs syndrome and β-amyloid angiopathy. It is expected that the compounds of the present invention would most likely be used as single agents but could also be used in combination with a broad range of cognition deficit enhancement agents. Methods of Preparation
The present invention also relates to processes for preparing the compound of formula (I) as a free base or a pharmaceutically acceptable salt thereof. Throughout the following description of such processes it is understood that, where appropriate, suitable protecting groups will be added to, and subsequently removed from the various reactants and intermediates in a manner that will be readily understood by one skilled in the art of organic synthesis. Conventional procedures for using such protecting groups as well as examples of suitable protecting groups are for example described in Protective Groups in Organic Synthesis by T.W. Greene, P.G.M Wutz, 3rd Edition, Wiley-Interscience, New York, 1999. It is understood that microwaves can be used for the heating of reaction mixtures.
Preparation of Intermediates
The process, wherein A, B, C, D, R1, R2, R3, R4, R5, R6 and R7, unless otherwise specified, are as hereinbefore defined, comprises,
(i) diazotization of a compound of formula II to obtain a compound of formula III, wherein halo represents bromine or chloride.
C C V-NH2 ( C Vhalo
II III
The reaction may be carried out by treating an appropriate amine with nitrous acid followed by treating the formed diazonium salt with an appropriate cuprous halide such as copper(I) bromide or copper(I) chloride, or with copper and hydrobromic acid or hydrochloric acid. The reactions may be preformed in a suitable solvent such as water at a temperature between -20 0C and reflux. (ii) borylation of a compound of formula III, wherein halo represents halogen such as bromine or chlorine, to obtain a compound of formula IV, wherein R8 represents hydrogen, alkyl, aryl or two R8 may form a cyclic boronic ester.
Figure imgf000027_0001
in iv
The reaction may be carried out by: a) an alkyllithium such as butyllithium, or magnesium, and a suitable boron compound such as trimethyl borate or triisopropyl borate. The reaction may be performed in a suitable solvent such as tetrahydrofuran, hexane or dichloromethane in a temperature range between -78 0C and +20 0C; or, b) a suitable boron species such as 4,4,4',4l,5J5,5r,5'-octamethyl-2,2'-bi-l;3,2- dioxaborolane, biscatecholatodiboron, or pinacolborane in the presence of a suitable palladium catalyst such as tris(dibenzylideneaceton)dipalladium(0), [1,1'- bis(diphenylphosρhino)ferrocene]palladium(II) chloride, palladium(O) tetrakistriphenylphosphine, palladium diphenylphosphineferrocene dichloride or palladium acetate, with or without a suitable ligand such as tricyclohexylphosphine or 2- (dicyclohexylphosphino)biphenyl, and a suitable base, such as a tertiary amine, such as trietylamine or diisopropylethylamine, or potassium acetate may be used. The reaction may be performed in a solvent such as dioxane, toluene, acetonitrile, water, ethanol or 1,2- dimethoxyethane, or mixtures thereof, at temperatures between +20 0C and +160 0C.
(Hi) reaction of a compound of formula V wherein halo represents halogen e.g. bromide, R9 is aryl or heteroaryl, and a compound of formula VI wherein R10 is defined as B under formula I , to obtain a compound of formula VII.
Figure imgf000028_0001
V VI VII
The reaction may be carried out by treating the compound of formula V with an alkyllithium, such as butyllithium, or magnesium followed by addition of a compound of formula VI. The reaction may be preformed in a suitable solvent such as diethyl ether or tetrahydrofuran at a temperature between -78 0C and reflux.
(iv) reaction of a compound of formula VII to obtain a compound of formula VIII.
Figure imgf000028_0002
VIl VIII
The reaction may be carried out by reduction using an appropriate reducing agent such as sodium borohydride, cyanoborohydride or lithium aluminium hydride. The reaction may be preformed in a suitable solvent such as methanol, ethanol, diethyl ether or tetrahydrofuran at a temperature between -78 0C and reflux.
(v) reaction of a compound of formula VIII to obtain a compound of formula IX.
Figure imgf000028_0003
The reaction may be carried out by treating the compound of formula VIII with a suitable thiocarbonyl transfer reagent such as O,O-dipyridine-2-yl thiocarbonate or thiophosgene and with or without a suitable base such as sodium bicarbonate. The reaction may be preformed in a suitable solvent such as dichloromethane or chloroform at a temperature between -78 0C and reflux.
(\1) reaction of a compound of formula IX to obtain a compound of formula X.
Figure imgf000029_0001
IX X
The reaction may be carried out by treating the appropriate isothiocyanate and carbon disulfide with a suitable base such as potassium ter/-butoxide in a suitable solvent such as tetrahydrofuran or diethyl ether at a temperature between -78 0C and reflux.
(vii) reaction of a compound of formula X to obtain a compound of formula XI.
Figure imgf000029_0002
X XI
The reaction may be carried out by treating a compound of formula X with an appropriate diamine such as 1,3-diaminopropane or ethyl enediamine. The reaction may be preformed in a suitable solvent such as ethanol or methanol at a temperature between 0 0C and reflux.
(viii) reaction of a compound of formula XI to obtain a compound of formula XII.
Figure imgf000030_0001
XI XII
The reaction may be carried out by treating the appropriate thione such as a compound of formula XI with an appropriate oxidazing agent such as tert-butyl hydroperoxide and aqueous ammonia. The reaction may be performed in a suitable solvent such as methanol at a temperature between 0 0C and reflux.
(ix) reaction of a compound of formula XIII wherein E is ring B, C or phenyl in formula I, to a compound of formula XIV.
Figure imgf000030_0002
XIII XTV
The reaction may be carried out by treating the methyl ether with a suitable Lewis acid such as boron tribromide in a suitable solvent such as dichloromethane at a temperature between -78 0C and reflux.
(x) reaction of a compound of formula XIV, wherein E is ring B, C or phenyl in formula I, to a compoud of formula XV, wherein R11 is alkyl.
Figure imgf000030_0003
XIV XV
The reaction may be carried out by treating the appropriate alcohol with a suitable sulfonyl chloride or anhydride such as methanesulfonyl chloride, 1-propanesulfonyl chloride, cyclopropanesulfonyl chloride or methanesulfonic anhydride in the presence of a suitable base such as triethylamine. The reaction ma}' be carried out in a suitable solvent such as dichloromethane at a temperature between 0 0C and reflux.
Methods of Preparation of End products
Another object of the invention is the process for the preparation of compounds of general Formula (I), wherein A, B, C, D, R1, R2, R3, R4, R5, R6 and R7 unless otherwise specified, are defined as hereinbefore, and salts thereof. When it is desired to obtain the acid salt, the free base may be treated with an acid such as a hydrogen halide such as hydrogen chloride in a suitable solvent such as tetrahydrofuran, diethyl ether, methanol, ethanol, chloroform or dichloromethane or mixtures thereof and the reaction may occur between -30 0C to +50 0C.
(a) reaction of a compound of formula XIV, wherein halo represents a halogen such as bromine, to obtain a compound of formula I.
Figure imgf000031_0001
XVI IV I
The reaction of process (a) may be carried out by coupling of a suitable compound such as a compound of formula XVI with an appropriate aryl boronic acid or ester of formula IV wherein R8 represents hydrogen, alkyl, aryl or two R8 may form a cyclic boronic ester. The reaction may be carried out using a suitable palladium catalyst such as, [1,1'- bis(diphenylphosphino)ferrocene]palladium(II) chloride, tetrakis(triphenylρhosphine)palIadium(0), palladium diphenylphosphineferrocene dichloride, ρalladium(II) acetate or bis(dibenzylideneacetone) palladium (0), together with, or without, a suitable ligand such as triphenylphosphine, tri-tert-butylphosphine or 2- (dicyclohexylphosphino)biphenyl, or using a nickel catalyst such as nickel on charcoal or l,2-bis(diphenylphosphino)ethanenickel dichloride together with zinc and sodium triphenylphosphinetrimetasulfonate. A suitable base such as cesium fluoride, an alkyl amine such as triethyl amine, or an alkali metal or alkaline earth metal carbonate or hydroxide such as potassium carbonate, sodium carbonate, cesium carbonate, or sodium hydroxide may be used in the reaction, which may be performed in a temperature range between +20 0C and +160 0C, in a suitable solvent such as toluene, tetrahydrofuran, dioxane, dimethoxyethane, water, ethanol or Λζ N-dimethylformamide, or mixtures thereof.
General Methods
Starting materials used were available from commercial sources, or prepared according to literature procedures.
1H NMR spectra were recorded in the indicated deuterated solvent, using a Bruker DPX400 NMR spectrometer operating at 400 MHz for 1H equipped with a 4-nucleus probehead with Z-gradients or a Bruker av400 NMR spectrometer operating at 400 MHz 1H equipped with a 3mm flow injection SEI 1HZD-13C probehead with Z-gradients, using a BEST 215 liquid handler for sample injection. Chemical shifts are given in ppm. Resonance multiplicities are denoted s, d, t, q, m and br for singlet, doublet, triplet, quartet, multiplet, and broad respectively.
LC-MS analyses were performed on an LC-MS system consisting of a Waters Alliance 2795 HPLC, a Waters PDA 2996 diode array detector, a Sedex 75 ELS detector and a ZMD single quadrupole mass spectrometer. The mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode. The capillary voltage was set to 3.2 kV and the cone voltage to 30 V, respectively. The mass spectrometer was scanned between m/z 100-600 by a scan time of 0.7s. The diode array detector was scanned from 200-400 nm. The temperature of the ELS detector was adjusted to 40 0C and the pressure was set to 1.9 bar. For separation a linear gradient was applied starting at 100% A (A: 10 mM ammonium acetate in 5% acetonitrile) and ending at 100% B (B: acetonitrile). The column used was an X-Terra MS CS, 3.0 mm x 50 mm, 3.5 μm (Waters) run at a flow rate of 1.0 mL/min. The column oven temperature was set to 40 0C, or
LC-MS analyses were performed on a LC-MS system consisting of a Waters Alliance
2795 HPLC, a Waters PDA 2996 diode array detector, a Sedex 75 ELS detector and a ZQ single quadrupole mass spectrometer. The mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode. The capillary voltage was set to 3.2 kV and the cone voltage to 30 V5 respectively. The mass spectrometer was scanned between mfz 100-700 with a scan time of 0.3s. The diode array detector scanned from 200-400 nm. The temperature of the ELS detector was adjusted to 40 0C and the pressure was set to 1.9 bar. Separation was performed on an X-Terra MS C8, 3.0 mm x 50 mm, 3.5 μm (Waters) run at a flow rate of 1 mL/min. A linear gradient was applied starting at 100% A (A: 1OmM ammonium acetate in 5% acetonitrile or 8 mM formic acid in 5% acetonitrile) ending at 100% B (B: acetonitrile). The column oven temperature was set to 40 0C, or LC-MS analyses were performed on a LC-MS system consisting of a Waters Alliance 2795 HPLC, a Waters PDA 2996 diode array detector, a Sedex 85 ELS detector and a ZQ single quadrupole mass spectrometer. The mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode. The capillary voltage was set to 3.2 kV and the cone voltage to 30 V, respectively. The mass spectrometer scanned between m/z 100-700 with a scan time of 0.3s. The diode array detector scanned from 200-400 nm. The temperature of the ELS detector was adjusted to 40 0C and the pressure was set to 1.9 bar. Separation was performed on an X-Terra MS C8, 3.0 mm x 50 mm, 3.5 μm (Waters) run at a flow rate of 1 mL/min. A linear gradient was applied starting at 100% A (A: 1OmM ammonium acetate in 5% acetonitrile, or S mM formic acid in 5% acetonitrile) ending at 100% B (B: acetonitrile). The column oven temperature was set to 40 0C, or
LC-MS analyses were preformed on a Water Acquity system with PDA (Waters 2996) and Waters ZQ mass spectrometer. Column; Acquity UPLC™ BEH C8 1.7μm 2.1 x 50mm. The column temperature was set to 65°C. A linear 2 min gradient from 100% A (A: 95% 0.01M ammonium acetate in MiIIiQ water and 5% acetonitrile) to 100% B (5% 0.01 M ammonium acetate in MiIIiQ water and 95% acetonitrile) was applied for LC-separation at flow rate 1.2 mL/min. The PDA was scanned from 210-350nm and 254nm was extracted for purity determination. The ZQ mass spectrometer was run with ES in pos/neg switching mode. The Capillary Voltage was 3kV and the Cone Voltage was 30V or LC-MS analyses were performed on a LC-MS consisting of a Waters sample manager 2111 C, a Waters 1525 μ binary pump, a Waters 1500 column oven, a Waters ZQ single quadrupole mass spectrometer, a Waters PDA2996 diode array detector and a Sedex 85 ELS detector. The mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative ion mode. The mass spectrometer scanned between m/z 100-700 with a scan time of 0.3s. The capillary voltage was set to 3.4 kV and the cone voltage was set to 30 V, respectively. The diode array detector scanned from 200-400 nm. The temperature of the ELS detector was adjusted to 40 0C and the pressure was set to 1.9 bar. For separation a linear gradient was applied starting at 100 % A (A: 10 mM NH4OAc in 5 % CH3CN or 8 mM HCOOH in 5% CH3CN) and ending at 100 % B (B: CH3CN). The column used was a Gemini CIS, 3.0 mm x 50 mm, 3 μm, (Phenomenex) which was run at a flow rate of 1 ml/min. The column oven temperature was set to 40 0C or
LC-MS analyses were performed on a LC-MS consisting of a Waters sample manager 27771C, a Waters 1525 μ binary pump, a Waters 1500 column oven, a Waters ZQ single quadrupole mass spectrometer, a Waters PDA2996 diode array detector and a Sedex 85 ELS detector. The mass spectrometer was configured with an atmospheric pressure chemical ionisation (APCI) ion source which was further equipped with atmospheric pressure photo ionisation (APPI) device. The mass spectrometer scanned in the positive mode, switching between APCI and APPI mode. The mass range was set to m/z 120-800 using a scan time of 0.3 s. The APPI repeller and the APCI corona were set to 0.86 kV and 0.80 μA, respectively. In addition, the desolvation temperature (3000C), desolvation gas (400 L/Hr) and cone gas (5 L/Hr) were constant for both APCI and APPI mode. Separation was performed using a Gemini column CIS, 3.0 mm x 50 mm, 3 μm, (Phenomenex) and run at a flow rate of 1 ml/min. A linear gradient was used starting at 100 % A (A: 10 mM NH4OAc in 5% MeOH) and ending at 100% B (MeOH). The column oven temperature was set to 40 0C. GC-MS: Compound identification was performed on a GC-MS system (GC 6890, 5973N MSD) supplied by Agilent Technologies. The column used was a VF-5 MS3 ID 0.25 mm x 15m, 0.25 μm (Varian Inc.). A linear temperature gradient was applied starting at 40 0C (hold 1 min) and ending at 300 °C (hold 1 min), 25 °C/minute. The mass spectrometer was equipped with a chemial ionisation (CI) ion source and the reactant gas was methane. The mass spectrometer was equipped with an electron impact (EI) ion source and the electron voltage was set to 70 eV. The mass spectrometer scanned between m/z 50-500 and the scan speed was set to 3.25 scan/s, or Compound identification was performed on a GC-MS system (GC 6890, 5973N MSD) supplied by Agilent Technologies. The mass spectrometer was equipped with a Direct Inlet Probe (DIP) interface manufactured by SIM GmbH. The mass spectrometer was configured with a chemical ionisation (CI) ion source and the reactant gas was methane. The mass spectrometer was equipped with an electron impact (EI) ion source and the electron voltage was set to 70 eV. The mass spectrometer scanned between m/z 50-500 and the scan speed was set to 3.25 scan/s. A linear temperature gradient was applied starting at 40 0C (hold 1 min) and ending at 300 0C (hold 1 min), 25 °C/minute. The column used was a VF-5 MS, ID 0.25 mm x 30m, 0.25 μm (Varian Inc.).
Preparative-HPLC: Preparative chromatography was run on Waters auto purification HPLC with a diode array detector. Column: XTerra MS C8, 19 x 300 mm, 10 μm.
Gradient with acetonitrile/0.1 M ammonium acetate in 5 % acetonitrile in MiIIiQ Water.
Flow rate: 20 mL/min. Alternatively, purification was achieved on a semi preparative
Shimadzu LC-8A HPLC with a Shimadzu SPD-IOA UV-vis.-detector equipped with a
Waters Symmetry® column (C18, 5 μm, 100 mm x 19 mm). Gradient with acetonitrile/0.1 % trifluoroacetic acid in MiIIiQ Water. Flow rate: 10 mL/min.
Alternatively, another column was used; Atlantis C18 19 x 100 mm, 5 μm column.
Gradient with acetonitrile/0.1 M ammonium acetate in 5% acetonitrile in MiIIiQ Water.
Flow rate: 15 mL/min, or
Preparative-HPLC was run on a Waters FractionLynx system with a Autosampler combined Automated Fraction Collector (Waters 2767), Gradient Pump (Waters 2525),
Regeneration Pump (Waters 600), Make Up Pump (Waters 515), Waters Active Splitter, Column Switch (Waters CFO), PDA (Waters 2996) and Waters ZQ mass spectrometer. Column; XBridge™ Prep C8 5μm OBD™ 19 x 100mm, with guard column; XTerra ® Prep MS CS lOμm 19 x 10 mm Cartridge. A gradient from 100% A (95% 0.1 M ammonium acetate in MiIIiQ water and 5% acetonitrile) to 100% B (100% acetonitrile) was applied for LC-separation at flow rate 25 mL/min. The PDA was scanned from 210- 350 nm. The ZQ mass spectrometer was run with ES in positive mode. The Capillary Voltage was 3kV and the Cone Voltage was 30V. Mixed triggering, UV and MS signal, determined the fraction collection.
Microwave heating was performed in a Creator or Initiaror or Smith Synthesizer Single- mode microwave cavity producing continuous irradiation at 2450 MHz.
Thin layer chromatography (TLC) was performed on Merch TLC-plates (Silica gel 60 F254) and UV visualized the spots. Column chromatography was performed on a Combi Flash® Companion™ using RediSep™ normal-phase flash columns or using Merck Silica gel 60 (0.040-0.063 mm).
Compounds have been named using ACD/Name, version 9.0, software from Advanced Chemistry Development, Inc. (ACD/Labs), Toronto ON, Canada, www.acdlabs.com, 2005.
EXAMPLES
Below follows a number of non-limiting examples of compounds of the invention.
Example 1
1 -f 3-Bromophenyl)-l -cvclopropylmethanamine
Figure imgf000036_0001
3-bromobenzonitrile (2.73 g, 15 mmol) was dissolved in dry tetrahydroforan (15 mL) and added drop wise to a cyclopropanemagnesiumbromide (36 mL, 18 mmol, 0.5 M in tetrahydroftiran) under an atmosphere of argon. The mixture was heated to 50 0C for 4h and then cooled to 0 0C. Dry methanol (50 mL) was added, the mixture was stirred for 30 min and sodium borohydride (1.14 g, 30 mmol) was added. After 2.5h was saturated aqueous ammonium chloride added and the mixture was extracted with dichloromethane. The organic phases was pooled, dried over magnesium sulfate and concentrated. The residue was suspended in diethyl ether and filtered, the solid was collected and dried in vacuo to give 0.33 g (10% yield) of the title compound: 1H NMR (DMSO-J6) δ 7.73 (t, J = 1.88 Hz, 1 H), 7.56 - 7.52 (m, 1 H), 7.51 - 7.48 (m, 1 H), 7.37 (t, J= 7.91 Hz, 1 H)5 3.48 (d, J= 9.54 Hz, 1 H), 1.23 - 1.14 (m, 1 H), 0.66 - 0.59 (m, 1 H), 0.54 - 0.44 (m, 2 H), 0.40 - 0.32 (m, 1 H).
Example 2 l-Bromo-3-rcyclopropyl('isothiocyanato)methyl1ben2ene
Figure imgf000037_0001
Thiophosgene (0.122 mL, 1.6 mmol) was added in portions to a stirred solution of l-(3- bromophenyl)-l-cyclopropylmethanamine (0.33 g, 1.46 mmol) and saturated aqueous sodium bicarbonate (30 mL) in dichloromethane (30 mL) at 0 0C. The mixture was stirred for 2 h at 0 0C and the organic phase were collected. The aqueos phase was extracted with dichloromethane and the combined organics was washed with brine, dried over sodium sulfate, filtrated and concentrated to give 0.353 g (90% yield) of the title compound: 1H NMR (DMSO-J6) δ 7.64 - 7.62 (m, 1 H), 7.59 - 7.56 (m, 1 H), 7.47 - 7.44 (m, 1 H), 7.42 - 7.38 (m, 1 H), 4.59 (d, J= 8.78 Hz, 1 H), 1.51 - 1.43 (m, 1 H), 0.72 - 0.66 (m, 1 H), 0.60 - 0.53 (m, 2 H), 0.53 - 0.46 (m, 1 H).
Example 3 4-(3-Bromophenyl)-4-cyclopropyl-1.3-thiazolidine-2,5-dithione
Figure imgf000038_0001
l-Bromo-3-[cyclopropyl(isothiocyanato)methyl]benzene (0.35 g, 1.32 mmol) and carbon disulfide (0.16 mL, 2.63 mmol) in dry tetrahydrofuran (8 mL) was added drop wise to a stirred mixture of potassium tert-butoxide (0.27 g, 2.38 mmol) in dry tetrahydroruran (8 mL) at -78 0C. After the addition, the mixture was allowed to reach room temperature overnight. Water, brine and ethyl acetate was added and the organic phase was collected. The aqueos phase was extracted with ethyl acetate, the organic extracts were pooled, washed with brine, dried over sodium sulfate and evaporated to give 0.47 g (quantitative yield) of the title compound: 1H NMR (DMSCW6) δ 7.62 - 7.59 (m, 1 H), 7.56 - 7.52 (m, 1 H), 7.52 - 7.49 (m, 1 H)5 7.39 - 7.34 (m, 1 H), 0.90 - 0.82 (m, 1 H), 0.76 - 0.63 (m, 2 H), 0.59 - 0.48 (m, 1 H), 0.34 - 0.25 (m, 1 H).
Example 4
8-(3-Bromophenyl)-8-cvclopropyl-3,4J,S-tetrahydroimidazori,5-α1pyrimidine-6('2/i)- thione
Figure imgf000038_0002
4-(3-Bromophenyl)-4-cyclopropyl-l,3-thiazolidine-2,5-dithione (0.47 g, 1.36 mmol) and 1,3-diaminopropane (0.3 g, 4 mmol) was heated to 70 0C in ethanol (10 mL) for 1.5 h. The mixture was cooled to room temperature and concentrated, the residue was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate and brine. The organic extracts were pooled, dried over sodium sulfate and evaporated. Silica chromatography using ethyl acetate from 0 - 30 % in n-heptane afforded 0.3 g (63 % yield) of the title compound: 1H NMR (DMSCW6) 6 10.05 (s, 1 H), 7.72 (t, J= 1.88 Hz, 1 H), 7.59 - 7.56 (m, 1 H)5 7.55 - 7.51 (m, 1 H), 7.36 (t, J= 7.91 Hz5 1 H)5 3.73 - 3.59 (m, 2 H), 3.42 - 3.36 (m, 2 H), 1.75 - 1.68 (m, 2 H)5 1.57 - 1.50 (m, 1 H)5 0.52 - 0.47 (m, 3 H), 0.45 - 0.38 (m, 1 H); MS (ESI) m/∑ 348, 350 (M-I]".
Example 5
8-('3-Bromoρhenyπ-8-cvctopiOpyl-2,3,4,8-tetrahydroiniidazo[l,5-αlpyrimidin-6-amine
Figure imgf000039_0001
8-(3-Bromophenyl)-8-cyclopropyl-354,7,8-tetrahydroimidazo[l,5-ι3]pyriniidiiie-6(2H)- thione (0.3 g, 0.86 mmol) was dissolved in methanol (10 mL) and aqueous ammonia (25%, 3 mL). Tert-butyl hydroperoxide (1.8 mL, 70% in water, 13 mmol) was added and the mixture was left over night at room temperature and then heated to 30 0C for 3 h. Most of the methanol was evaporated, water was added and the mixture was extracted with ethyl acetate. The organic extracts were pooled, washed with water, brine, dried over sodium sulfate and evaporated, drying in vacuo oven afforded 0.3 g (quantitative yield) of the title product: 1H NMR (DMSO-^6) δ 10.73 (br s, 2 H)5 7.80 (t, J= 1.76 Hz, 1 H)5 7.71 - 7.67 (m, 1 H), 7.40 - 7.36 (m, 1 H), 7.25 (t, J= 7.78 Hz, 1 H), 3.51 - 3.37 (m, 4 H), 1.65 - 1.60 (m, 2 H), 1.41 - 1.36 (m, 1 H), 0.36 - 0.31 (m, 1 H), 0.29 - 0.22 (m, 3 H); MS (ESI) m/∑ 333, 335 [M+lf.
Example 6
8-Cvclopropyl-8-C3',S'-dichlorobiphenyl-3-yl)-2,3<4,8-tetrahvdroimidazori,5-α'|pyrimidin- 6-amine acetate
Figure imgf000040_0001
8-(3-Bromophenyl)-8-cyclopropyl-2,3,4,8-tetrahydroimidazo[l,5-σ]pyrimidin-6-amine (80 mg, 0.23 mmol), [l,r-bis(diphenylρhosphino)ferrocene]palladium(II) chloride dichloromethane adduct (19 mg, 0.023 mmol), cesium carbonate (300 mg, 0.92 mmol), and 3,5-dichlorobenzeneboronic acid (53 mg, 0.35 mmol) was dissolved in tetrahydrofUran: water (9: 1) (3 mL) and heated at 130 0C for 30 min in a microwave. When cooled to ambient temperature the mixture was filtered and purified by preparative HPLC to give 24 mg (23% yield) of the title compound: 1H NAdR (DMSO-J6) δ 7.94 - 7.92 (m, 1 H), 7.79 - 7.76 (m, 1 H), 7.65 - 7.63 (m, 2 H), 7.61 - 7.56 (m, 2 H), 7.41 (t, J= 7.78 Hz, 1 H), 3.53 - 3.48 (m, 4 H), 1.88 (s, 3 H, acetate), 1.65 (t, J= 5.65 Hz, 2 H), 1.56 - 1.49 (m, 1 H), 0.43 - 0.36 (m, 1 H), 0.33 - 0.26 (m, 3 H); MS (ESI) m/z 399, 401 [M+l]+.
Example 7
1 -(3-Bromophenyl)-l -cyclohexylmethanamine
Figure imgf000040_0002
The title compound was synthesized as described for example 1 in 11% yield starting from cyclohexylmagnesiumchloride. With the exception that diethyl ether was used as solvent and the product was isolated using silica chromatography, gradient elution from dichloromethane to dichloromethane: methanol 95:5: 1H NMR (DMSO-J6) δ 7.49 - 7.47 (m, 1 H), 7.39 - 7.34 (m, 1 H), 7.27 - 7.21 (m, 2 H), 3.51 (d, J= 6.78 Hz, 1 H), 1.86 - 1.78 (m, 1 H), 1.71 - 1.65 (m, 1 H), 1.63 - 1.54 (m, 2 H), 1.40 - 1.29 (m, 2 H), 1.18 - 1.05 (m, 3 H), 0.95 - 0.78 (m, 2 H). Example 8 l-Bromo-S-l'cvclohexyK'isothiocvanato^ethylibenzene
Figure imgf000041_0001
The title compound was synthesized as described for example 2 in quantitative yield starting from l-(3-bromophenyl)-l-cyclohexylmethanamine: 1H NMR (DMSO-c^) δ 7.57 7.53 (m, 2 H), 7.41 - 7.33 (m, 2 H), 4.99 (d, J= 6.78 Hz, 1 H), 1.88 - 1.78 (m, 1 H), 1.75 - 1.58 (m, 4 H), 1.48 - 1.41 (m, 1 H), 1.21 - 1.00 (m, 5 H).
Example 9 4-(3 -Bromophenyiy 4-cγclohexyl- 1 ,3 -thiazolidine-2, 5-dithione
Figure imgf000041_0002
The title compound was synthesized as described for example 3 in 95% yield starting from l-bromo-3-[cyclohexyl(isothiocyanato)methyl]benzene: 1H NMR (DMSO-J6) δ 7.53 - 7.47 (m, 3 H), 7.38 - 7.33 (m, 1 H), 1.75 - 1.66 (m, 2 H), 1.66 - 1.58 (m, 3 H), 1.39 - 1.33 (m, 1 H), 1.27 - 1.20 (m, 3 H), 1.10 - 1.01 (m, 2 H).
Example 10
8-(3-Bromophenyl)-8-cyclohexyl-3,4,7,8-tetrahvdroimidazori,5-g1pyrimidine-6(2H)- thione
Figure imgf000042_0001
The title compound was synthesized as described for example 4 in 67% yield starting from 4-(3-bromoρhenyl)-4-cyclohexyl-l,3-thiazolidine-2J5-dithione: 1H NMR (DMSO-afe) δ 10.35 (s, 1 H), 7.71 (t, J= 1.S8 Hz, 1 H), 7.60 - 7.57 (m, 1 H), 7.51 - 7.48 (m, 1 H), 7.35 (t, J= 7.91 Hz, 1 H), 3.75 - 3.68 (m, 1 H), 3.58 - 3.51 (m, 1 H), 3.50 - 3.42 (m, 1 H), 3.38 - 3.35 (m, 1 H), 2.02 - 1.94 (m, 1 H), 1.73 - 1.66 (m, 3 H), 1.64 - 1.55 (m, 2 H), 1.53 - 1.46 (m, 1 H), 1.22 - 1.13 (m, 1 H), 1.11 - 0.95 (m, 5 H); MS (ESI) m/z 390, 392 [M-I]".
Example 11
8-f3-Bromophenvπ-8-cvclohexyl-2,3,4,8-tetrahvdroimidazo[l,5-α1pyrimidin-6-amine
Figure imgf000042_0002
The title compound was synthesized as described for example 5 in quantitative yield starting from 8-(3-bromophenyl)-8-cyclohexyl-3,4,7,8-tetrahydroimidazo[l,5- α]pyrimidine-6(2H)-thione: 1H NMR (DMSCW6) δ 10.73 (br s, 2 K), 7.82 - 7.79 (m, 1 H), 7.70 - 7.67 (m, 1 H), 7.38 - 7.34 (m, 1 H), 7.23 (t, J= 7.91 Hz, 1 H), 3.51 - 3.45 (m, 1 H), 3.41 - 3.34 (m, 3 H), 1.77 (m, 1 H) 1.62 - 1.5S (m, 2 H), 1.58 - 1.53 (m, 2 H), 1.48 - 1.41 (m, 1 H), 1.07 - 0.95 (m, 7 H); MS (ESI) m/z 375, 377 [M+l]+.
Example 12 8-Cvclohexyl-S-(3',5'-dichlorobiphenyl-3-yl)-2,3,4,8-tetrahvdroimidazori,5-αlpyrimidin-6- amine acetate
Figure imgf000043_0001
The title compound was synthesized as described for example 6 in 47% yield starting from 8-(3-bromophenyl)-8-cyclohexyl-2,3,4,8-tetrahydroimidazo[l,5-α]pyrimidin-6-amine except that 1 ,2-dimethoxy ethane: ethanol: water (6:3:1) was used as solvent: 1H NMR (DMSO-J6) δ 7.93 - 7.90 (m, 1 H)3 7.78 - 7.74 (m, 1 H), 7.66 - 7.63 (m, 2 H), 7.61 - 7.57 (m, 2 H), 7.42 (t, J= 7.78 Hz, 1 H), 3.55 - 3.51 (m, 4 H), 1.97 - 1.90 (m, 1 H)5 1.88 (s, 3 H, acetate) 1.70 - 1.61 (m, 3 H), 1.59 - 1.50 (m, 3 H), 1.124 - 1.13 (m, 1 H)5 1.10 - 0.96 (m, 5 H); MS (ESI) m/z 441, 443 [M+ 1]+.
Example 13
1 -(3 -Bromophenyl)- 1 -cyclobutylmefhanamine
Figure imgf000043_0002
1,3-dibromobenzene (2.42 mL, 20.0 mmol) was dissolved in dry diethyl ether (30 mL) and cooled to -78 0C. n-Butyllithium (8.0 mL, 20.0 mmol, 2.5 M in hexane) was added drop wise and the mixture was stirred for 70 min. Cyclobutanecarbonitrile (1.62 g, 20.0 mmol) in dry diethyl ether (10 mL) was added and the mixture was stirred at -78 0C for 15 min and at 0 0C for 40 min. Dry methanol (30 mL) was added and after 5 min at 0 °C sodium borohydride (1.51 g, 40.0 mmol) was added. The mixture was stirred over night at room temperature, saturated aqueous ammonium chloride was added and the mixture was extracted with dichloromethane. The organic phases were pooled, dried over magnesium sulfate and concentrated. Silica chromatography, gradient elution from 0 to 8.5 % methanol in dichloromethane afforded 2.54 g (52.9 % yield) of the title compound: 1H NMR (DMS(W6) δ 7.51 (t, J= 1.76 Hz, 1 H), 7.3S - 7.34 (m, 1 H), 7.31 - 7.26 (m, 1 H), 7.25 - 7.19 (m, 1 H), 3.66 (d, J= 8.78 Hz, 1 H), 2.39 - 2.27 (m, 1 H), 1.88 - 1.75 (m, 2 H), 1.75 - 1.65 (m, 3 H), 1.65 - 1.55 (m, 1 H).
Example 14 l-Bromo-3-rcvclobuM(isothiocyanato)meΦvHbenzene
Figure imgf000044_0001
The title compound was synthesized as described for example 2 in quantitative yield starting from l-(3-bromophenyl)-l-cyclobutylmethanamine. With the exception that the temperature was allowed to rise to room temperature 5 min after complete addition and the stirring time was reduced to 1 h: 1H NMR (DMSCMs) δ 7.62 - 7.50 (m, 2 H), 7.44 - 7.31 (m, 2 H), 5.12 (d, J= 8.03 Hz, 1 H), 2.87 - 2.75 (m, 1 H), 2.07 - 1.83 (m, 2 H), 1.83 - 1.61 (m, 4 H).
Example 15 4-f3-BromophenylV4-evclobuM-13-thiazolidine-2,5-dithione
Figure imgf000044_0002
The title compound was synthesized as described for example 3 in 91% yield starting from l-bromo-S-fcyclobutyKisothiocyanato^ethylJbenzene: 1H NMR (DMSO-J6) δ 12.35 (br s, 1 H), 7.62 - 7.54 (m, 1 H), 7.49 - 7.45 (m, 1 H), 7.42 - 7.37 (m, 2 H), 3.72 - 3.60 (m, 1 H), 2.20 - 2.07 (m, 1 H), 1.94 - 1.82 (m, 2 H), 1.82 - 1.67 (m, 3 H); MS (ES) m/z 356, 358 [M- H]-. Example 16
8-0-Bromophenyl)-8-cyclobutyl-3,4,7,8-tetrahvdroimidazo[l,5-a]pyrimidine-6(2H)- thione
Figure imgf000045_0001
The title compound was synthesized as described for example 4 in 65% yield starting from 4-(3-bromophenyl)-4-cyclobutyl-l,3-thiazolidine-2,5-dithione: 1H NMR (DMSO-Ci6) δ 10.50 (s, 1 H), 7.71 - 7.67 (m, 1 H), 7.55 (dd, J= 7.78, 1.00 Hz, 1 H), 7.52 - 7.47 (m, 1 H), 7.36 - 7.30 (m, 1 H), 3.78 - 3.68 (m, 1 H), 3.61 - 3.52 (m, 1 H), 3.45 - 3.36 (m, 1 H), 3.10 - 2.98 (m, 1 H), 2.01 - 1.86 (m, 1 H), 1.80 - 1.67 (m, 5 H), 1.67 - 1.57 (m, 3 H); MS (ES) m/z 362, 364 [M-H]-.
Example 17 S-fS-Bromophenyl^-δ-cyclobuM^^^^-teiTahvdroimidazorLS-fltipyrirnidin-β-amine
Figure imgf000045_0002
8-(3-Bromophenyl)-8-cyclobutyl-3,4,758-tetrahydroimidazo[l,5-α]pyrimidine-6(2H)- thione (0.495 g, 1.36 mmol) was dissolved in methanol (15 mL) and heated to 30 0C. Aqueous ammonia (25%, 3 mL) and tert-butyl hydroperoxide (2.8 mL, 70 % in water, 20.4 mmol) were sequentially added and the mixture was stirred over night at 30 0C. Water and saturated aqueous ammonium chloride were added and the mixture was extracted with dichlorome thane. The organic phases were pooled, dried over magnesium sulfate and concentrated. Silica chromatography using ammonia in methanol (3.5 N) from 0-6% in dichloromethane afforded 0.265 g (56%) of the title compound: 1H NMR (DMSO-J6) δ 7.83 - 7.80 (m, 1 H), 7.67 (d, J= 7.78 Hz, 1 H), 7.38 - 7.33 (m, 1 H), 7.22 (t, J= 7.91 Hz, 1 H), 3.53 - 3.45 (m, 1 H), 3.44 - 3.37 (m, 1 H), 3.26 - 3.18 (m, 1 H), 2.90 - 2.79 (m, 1 H), 1.94 - 1.82 (m, 1 H), 1.73 - 1.50 (m, 7 H), 1.50 - 1.40 (m, 1 H); MS (ES) m/z 347, 349 [M+H]+.
Example 18
S-Cvclobu^l-S-rS'.S'-dichlorobiphenyl-S-vn^.S^^-tetrahvdroimidazorLS-fllpyrimidin-o- amine acetate
Figure imgf000046_0001
The title compound was synthesized as described for example 6 in 38% yield starting from 8-(3-bromophenyl)-8-cyclobutyl-2,3,4,8-tetrahydroimidazo[l,5-«]pyrimidin-6-amine: 1H NMR (DMSCW6) δ 7.92 (br s, 1 H), 7.76 (d, J= 7.78 Hz, 1 H), 7.63 (d, J= 1.76 Hz, 2 H)5 7.60 (d, J= 1.76 Hz, 1 H), 7.55 (d, J- 7.78 Hz3 1 H), 7.38 (t, J= 7.78 Hz, 1 H), 3.56 - 3.48 (m, 3 H), 3.04 - 2.94 (m, 1 H)3 1.97 - 1.89 (m, 1 H), 1.88 (s, 3 H), 1.76 - 1.44 (m, 8 H); MS (ES) m/z 411, 413 [M-H]".
Example 19
1 -( 3 -Bromophenyl)- 1 -cyclopentylmethanamine
Figure imgf000046_0002
The title compound was synthesized as described for example 1 in 57% yield starting from 3-bromobenzonitrile and cyclopentylmagnesium chloride (2 M in diethyl ether): 1H NMR (DMSCW6) δ 7.52 (t, J= 1.76 Hz, 1 H), 7.39 - 7.34 (m, 1 H), 7.32 - 7.27 (m, 1 H), 7.26 - 7.20 (m, 1 H), 3.51 (d, J= 8.53 Hz3 1 H), 1.99 - 1.87 (m, 1 H), 1.79 - 1.70 (m, 1 H), 1.62 - 1.28 (m, 5 H), 1.26 - 1.16 (m, 1 H), 1.12 - 1.01 (m, 1 H). Example 20 l-Bromo-3-rcvclopentylfisothiocvanato)methyllbenzene
Figure imgf000047_0001
The title compound was synthesized as described for example 2 in quantitative yield starting from l-(3-bromophenyl)-l-cyclopentylmethanamine: 1H NMR (DMSO-dg) δ 7.61 - 7.58 (m, 1 H), 7.57 - 7.54 (m, 1 H), 7.42 - 7.35 (m, 2 H), 5.01 (d, J= 8.03 Hz, 1 H), 2.48 - 2.39 (m, 1 H), 1.79 - 1.69 (m, I H), 1.69 - 1.38 (m, 6 H), 1.25 - 1.13 (m, I H).
Example 21 4-('3-BromophenyD-4-cvclopentyl- 1 ,3-thiazolidine-2,5-dithione
Figure imgf000047_0002
The title compound was synthesized as described for example 3 in quantitative yield starting from l-bromo-3-[cyclopentyl(isothiocyanato)methyl]benzene: 1H NMR (DMSO- d6) δ 12.18 (br s, 1 H), 7.58 (d, J= 7.78 Hz5 1 H), 7.54 (t, J= 1.76 Hz, 1 H)5 7.51 - 7.45 (m, 1 H)5 7.40 (t, J= 7.91 Hz5 I H), 3.30 - 3.22 (m, I H), 1.71 - 1.58 (m, 3 H), 1.58 - 1.50 (m, 3 H), 1.50 - 1.42 (m, 1 H), 1.29 - 1.20 (m, 1 H); MS (ES) m/z 370, 372 [M-H]".
Example 22
8-f3-Bromophenyl)-8-cvclopentyl-3,4,7,8-tetrahydroimidazo["l,5-Qipyrimidine-6C2/j)- thione
Figure imgf000048_0001
The title compound was synthesized as described for example 4 in 62% yield starting from 4-(3-bromophenyl)-4-cyclopentyl-l53-thiazolidine-2,5-dithione: 1H NMR (DMSO-d6) δ 10.37 (s, 1 H), 7.74 (t, J= 1.76 Hz5 1 H), 7.63 - 7.58 (m, 1 H)5 7.52 - 7.47 (m, 1 H)5 7.34 (t, J= 7.91 Hz5 1 H)5 3.78 - 3.69 (m, 1 H)5 3.60 - 3.51 (m, 1 H)5 3.46 - 3.38 (m, 2 H), 2.69 - 2.59 (m, 1 H)5 1.76 - 1.62 (m, 2 H)5 1.59 - 1.37 (m, 5 H)5 1.35 - 1.04 (m, 3 H); MS (ES) m/z 376, 378 [M-H]-.
Example 23 8-(3-Bromophenyl)-8-cyclopentyl-2,3,4,8-tetrahvdroimidazo[l,5-a;1pyrimidin-6-amine
Figure imgf000048_0002
The title compound was synthesized as described for example 5 in 52% yield starting from 8-(3-bromophenyl)-8-cyclopentyl-354,758-tetrahydroimidazo[l55-α]pyrimidine-6(2H)- thione with the exception that he stirring time was 48 h and the silica chromatography gradient was ammonia in methanol (3.5 N) from 0-10 % in dichloromethane: 1H NMR (DMSO-dβ) δ 7.84 (br S5 1 H)5 7.72 (d, J= 7.78 Hz5 1 H)5 7.36 (d, J= 8.03 Hz5 1 H)5 7.23 (t, J= 7.91 Hz7 1 H), 3.56 - 3.45 (m, 1 H)5 3.44 - 3.36 (m, 1 H)5 3.26 - 3.15 (m, 1 H)5 2.48 - 2.36 (m, 1 H)5 1.68 - 1.55 (m, 2 H)5 1.55 - 1.24 (m, 7 H), 1.13 - 0.95 (m, 2 H); MS (ES) m/z 359, 361 [M-H]-.
Example 24
8-Cyclopentyl-8-π',5'-dichlorobiphenyl-3-yl)-2,3<4,8-tetrahydroimidazofl,5-α]ρyrimidin- 6-amine acetate
Figure imgf000049_0001
The title compound was synthesized as described for example 6 in 28% yield starting from 8-(3-bromophenyl)-8-cyclopentyl-2,3,4,8-tetrahydroimidazo[l,5-α]pyrimidin-6-amine: 1H NMR (DMSO-ύfe) δ 7.9S - 7.94 (m, 1 H)3 7.80 (d, J= 7.78 Hz, 1 H), 7.64 (d, J= 2.01 Hz, 2 H), 7.60 (t, J= 1.88 Hz, 1 H), 7.58 - 7.53 (m, 1 H), 7.39 (t, J- 7.78 Hz, 1 H), 3.56 - 3.48 (m, 4 H), 2.65 - 2.54 (m, 1 H), 1.88 (s, 3 H), 1.71 - 1.57 (m, 2 H), 1.56 - 1.40 (m, 4 H), 1.39 - 1.29 (m, 2 H), 1.19 - 1.05 (m, 2 H); MS (ES) 425, 427 [M-H]".
Assays Compounds were tested in at least one of the following assays:
β-Secretase Enzyme
The enzyme used in the IGEN Cleavage-, Fluorescent-, TR-FRET- and BiaCore assays is described as follows: The soluble part of the human β-Secretase (AA 1 — AA 460) was cloned into the ASP2- Fc 10-1 -IRES-GFP -neoK mammalian expression vector. The gene was fused to the Fc domain of IgGl (affinity tag) and stably cloned into HEK 293 cells. Purified sBACE-Fc is stored in Tris buffer, pH 9.2 and has a purity of 95%.
IGEN Cleavage Assay
The enzyme was diluted to 43 μg/ml in 40 mM MES pH 5.0. The IGEN substrate was diluted to 12 μM in 40 mM MES pH 5.0. Compounds were diluted to the desired concentration in dimethyl sulfoxide (final dimethyl sulfoxide concentration in assay is 5%). The assay was performed in a 96 well PCR plate from Greiner (#650201). Compound in dimethyl sulfoxide (3 μL) and enzyme (27 μL) were added to the plate, and pre- incubated for 10 min. The reaction was started with substrate (30 μL). The final dilution of enzyme was 20 μg/ml and the final concentration of substrate was 6 μM. After 20 minutes reaction at room temperature (RT), the reaction was stopped by removing 10 μL of the reaction mix and diluting it 1 :25 in 0.2 M Trizma-HCl, pH 8.0. The product was quantified by adding 50 μL of a 1:5000 dilution of the neoepitope antibody to 50 μL of the 1:25 dilution of the reaction mix (all antibodies and the strep tavidin coated beads were diluted in PBS containing 0.5% BSA and 0.5% Tween20). Then, 100 μL of 0.2 mg/mL streptavidin coated beads (Dynabeads M-280) and a 1:5000 dilution of ruthenylated goat anti-rabbit (Ru-GaR) antibody was added. The mixture was measured for electro- chemiluminescence in a BioVeris M8 Analyzer after 2 hours of incubation with shaking at RT. The dimethyl sulfoxide control defined 100% activity level and 0% activity was defined by exclusion of the enzyme (using 40 mM MES pH 5.0 buffer instead).
Fluorescent Assay
The enzyme was diluted to 52 μg/ml in 40 mM MES pH 5.0. The substrate (Dabcyl-Edans) was diluted to 30 μM in 40 mM MES pH 5.0. Compounds were diluted to the desired concentration in dimethyl sulfoxide (final dimethyl sulfoxide concentration in assay is 5%). The assay is done in a Corning 384 well round bottom, low volume, non-binding surface plate (Corning #3676). Enzyme (9 μL) together with 1 μL of compound in dimethyl sulfoxide were added to the plate and pre-incubated for 10 min. Substrate (10 μL) was added and the reaction proceeded in the dark at RT for 25 min. The final dilution of enzyme was 23 μg/ml, and the final concentration of substrate was 15 μM (Km of 25 μM). The fluorescence of the product was measured on a Victor II plate reader with an excitation wavelength of 360 nm and an emission wavelength of 485 nm using a protocol for labelled Edans peptide. The dimethyl sulfoxide control defined 100% activity level and 0% activity was defined by exclusion of the enzyme (using 40 mM MES pH 5.0 buffer instead).
TR-FRET Assay
Enzyme was diluted to 6 μg/mL and the substrate (Europium)CEVNLDAEFK(Qsy7) to 200 nM in reaction buffer (NaAcetate, chaps, triton x-100, EDTA pH 4.5). Compounds were diluted to the desired concentration in dimethyl sulfoxide (final dimethyl sulfoxide concentration in assay is 5%). The assay was done in a Costar 384 well round bottom, low volume, non-binding surface plate (Corning #3676). Enzyme (9 μL) and 1 μL of compound in dimethyl sulfoxide was added to the plate, mixed and pre-incubated for 10 min. Substrate (10 μL) was added and the reaction proceeded in the dark for 15 min at RT. The reaction was stopped with the addition of 7 μL NaAcetate, pH 9. The fluorescence of the product was measured on a Victor II plate reader with an excitation wavelength of 340 5 nm and an emission wavelength of 615 nm. The final concentration of the enzyme was 2.7 μg/ml and the final concentration of the substrate was 100 nM (Km of 290 nM). The dimethyl sulfoxide control defined the 100% activity level and 0% activity was defined by exclusion of the enzyme (using reaction buffer instead).
i o BA CE Biacore Sensor Chip Preparation
BACE was assayed on a Biacore3000 instrument by attaching either a peptidic transition state isostere (TSI) or a scrambled version of the peptidic TSI to the surface of a Biacore CM5 sensor chip. The surface of a CM5 sensor chip has 4 distinct channels that can be used to couple the peptides. The scrambled peptide KPES-statine-ETIAEVENV was is coupled to channel 1 and the TSI inhibitor KTEEISEVN-statine-VAEF was coupled to channel 2 of the same chip. The two peptides were dissolved at 0.2 mg/mL in 20 mM sodium acetate pH 4.5, and then the solutions were centrifuged at 14K rpm to remove any particulates. Carboxyl groups on the dextran layer were activated by injecting a one to one mixture of 0.5 M N-ethyl-N' (3-dimethylaminopropyl)-carbodiimide and 0.5 M N-
20 hydroxysuccinimide at 5 μL/min for 7 min. Then the stock solution of the control peptide was injected in channel 1 for 7 min at 5 μL/min., and then the remaining activated carboxyl groups were blocked by injecting 1 M ethanolamine for 7 min at 5 μL/min.
BA CE Biacore Assay Protocol
25 The BACE Biacore assay was done by diluting BACE to 0.5 μM in sodium acetate buffer at pH 4.5 (running buffer minus dimethyl sulfoxide). The diluted BACE was mixed with dimethyl sulfoxide or compound diluted in dimethyl sulfoxide at a final concentration of 5% dimethyl sulfoxide. The BACE/inhibitor mixture was incubated for 30 minutes at RT before being injected over channel 1 and 2 of the CM5 Biacore chip at a rate of 20 μL/min.
30 As BACE bound to the chip the signal was measured in response units (RU). BACE binding to the TSI inhibitor on channel 2 gave a certain signal. The presence of a BACE inhibitor reduced the signal by binding to BACE and inhibiting the interaction with the peptidic TSI on the chip. Any binding to channel 1 was non-specific and was subtracted from the channel 2 responses. The dimethyl sulfoxide control was defined as 100% and the effect of the compound was reported as percent inhibition of the dimethyl sulfoxide control.
Beta-Secretase TIfyole Cell Assays Generation ofHEK293-APP695
The pcDNA3.1 plasmid encoding the cDNA of human full-length APP695 was stably transfected into HEK-293 cells using the Lipofectamine transfection reagent according to manufacture's protocol (Invitrogen). Colonies were selected with 0.1-0.5 mg/mL of zeocin. Limited dilution cloning was performed to generate homogeneous cell lines. Clones were characterized by levels of APP expression and Aβ secreted in the conditioned media using an ELISA assay developed in-house.
Cell culture for HEK293-APP695
HEK293 cells stably expressing human wild-type APP (HEK293-APP695) were grown at 37 0C, 5% CO2 in DMEM containing 4500 g/L glucose, GlutaMAX and sodium pyruvate supplemented with 10% FBS, 1% non-essential amino acids and 0.1 mg/mL of the selection antibiotic zeocin.
Aβ40 release assay
HEK293-APP695 cells were harvested at 80-90% confluence and seeded at a concentration of 0.2x106 cells/mL, 100 mL cell suspension/well, onto a black clear bottom 96-well poly-D-lysine coated plate. After over night incubation at 37 0C, 5% CO2, the cell medium was replaced with cell culture medium with penicillin and streptomycin (100 LVmL, 100 μg/mL, respectively) containing test compounds in a final dimethyl sulfoxide concentration of 1%. Cells were exposed to the test compounds for 24 h at 37 0C, 5% CO2. To quantify the amount of released Aβ, 100 μL cell medium was transferred to a round bottom polypropylene 96-well plate (assay plate). The cell plate was saved for the ATP assay, as described below. To the assay plate, 50 μL of primary detection solution containing 0.5 μg/mL of the rabbit anti-Aβ40 antibody and 0.5 μg/mL of the biotinylated monoclonal mouse 6E10 antibody in DPBS with 0.5 %BSA and 0.5% Tween-20 was added per well and incubated over night at 4 0C. Then, 50 μL of secondary detection solution containing 0.5 μg/mL of a ruthenylated goat anti-rabbit antibody and 0.2 mg/mL of streptavidin coated beads (Dynabeads M-280) was added per well. The plate was vigorously shaken at RT for 1-2 hours. The plate was then measured for electro- chemiluminescence in a BioVeris M8 Analyzer.
Cell culture for SH-SY5Y
SH-SY5Y cells were grown 37 0C with 5% CO2 in DMEM/F-12 1:1 containing GlutaMAX supplemented with 1 mM HEPES3 10% FBS and 1% non-essential amino acids.
sAPPβ release assay
SH-SY5Y cells were harvested at 80-90% confluence and seeded at a concentration of 1.5xlO6 cells/mL, 100 mL cell suspension/well, onto a black clear flat bottom 96-well tissue culture plate. After 7 hours of incubation at 37 0C, 5% CO2, the cell medium was replaced with 90 μl cell culture medium with penicillin and streptomycin (100 XJ /mL, 100 μg/mL, respectively) containing test compounds in a final dimethyl sulfoxide concentration of 1%. Cells were exposed to the test compounds for IS h at 370C, 5% CO2. To measure sAPPβ released into the cell medium, sAPPβ microplates from Meso Scale Discovery (MSD) were used and the assay was performed according to the manufacture's protocol. Briefly, 25 μL cell medium was transferred to a previously blocked MSD sAPPβ microplate. The cell plate was saved for the ATP assay, as described below. The sAPPβ was captured during shaking at RT for 1 hour, by antibodies spotted in the wells of the microplate. After multiple washes, SULFO-TAG labeled detection antibody was added (25μL/well, final concentration InM) to the assay plate and the plate was incubated with shaking at RT for 1 hour. Following multiple washes, 150 μl/well of Read Buffer T was added to the plate. After 10 minutes at RT the plate was read in the SECTOR™ Imager for electro-chemiluminescence.
ATP assay As indicated above, after transferring medium for analysis of Aβ40 or sAPPβ from the cell plate, the plate was used to analyze cytotoxicity using the ViaLight™ Plus cell proliferation/cytotoxicity kit from Cambrex BioScience that measures total cellular ATP. The assay was performed according to the manufacture's protocol. Briefly, 50 μL cell lysis reagent was added per well. The plates were incubated at RT for 10 min. Two min after addition of 100 μL reconstituted ViaLight™ Plus ATP reagent, the luminescence was measured in a Wallac Victor2 1420 multilabel counter.
hERG Assay
Cell culture
The hERG-expressing Chinese hamster ovary Kl (CHO) cells described by (Persson, Carlsson, Duker, & Jacobson, 2005) were grown to semi-confluence at 37 0C in a humidified environment (5% CO2) in F- 12 Ham medium containing L-glutamine, 10% foetal calf serum (FCS) and 0.6 mg/ml hygromycin (all Sigma-Aldrich). Prior to use, the monolayer was washed using a pre-warmed (37°C) 3 ml aliquot of Versene 1:5,000 (Invitrogen). After aspiration of this solution the flask was incubated at 37 0C in an incubator with a further 2 ml of Versene 1 :5,000 for a period of 6 minutes. Cells were then detached from the bottom of the flask by gentle tapping and 10 ml of Dulbecco's Phosphate-Buffered Saline containing calcium (0.9 mM) and magnesium (0.5 mM) (PBS; Invitrogen) was then added to the flask and aspirated into a 15 ml centrifuge tube prior to centrifugation (50 g, for 4 mins). The resulting supernatant was discarded and the pellet gently re-suspended in 3 ml of PBS. A 0.5 ml aliquot of cell suspension was removed and the number of viable cells (based on trypan blue exclusion) was determined in an automated reader (Cedex; Innovatis) so that the cell re-suspension volume could be adjusted with PBS to give the desired final cell concentration. It is the cell concentration at this point in the assay that is quoted when referring to this parameter. CHO-KvI.5 cells, which were used to adjust the voltage offset on Ion Works™ HT, were maintained and prepared for use in the same way.
Elecfrophysiology
The principles and operation of this device have been described by (Schroeder, Neagle, Trezise, & Worley, 2003). Briefly, the technology is based on a 384-well plate
(PatchPlate ) in which a recording is attempted in each well by using suction to position and hold a cell on a small hole separating two isolated fluid chambers. Once sealing has taken place, the solution on the underside of the PatchPlate is changed to one containing amphotericin B. This permeablises the patch of cell membrane covering the hole in each well and, in effect, allows a perforated, whole-cell patch clamp recording to be made.
A β-test IonWorks™ HT from Essen Instrument was used. There is no capability to warm solutions in this device hence it was operated at room temperature (~21°C), as follows. The reservoir in the "Buffer" position was loaded with 4 ml of PBS and that in the "Cells" position with the CHO-hERG cell suspension described above. A 96-well plate (V-bottom, Greiner Bio-one) containing the compounds to be tested (at 3-fold above their final test concentration) was placed in the "Plate 1" position and a PatchPlate™ was clamped into the PatchPlate™ station. Each compound plate was laid-out in 12 columns to enable ten, 8- point concentration-effect curves to be constructed; the remaining two columns on the plate were taken up with vehicle (final concentration 0.33% DMSO), to define the assay baseline, and a supra-maximal blocking concentration of cisapride (final concentration 10 mM) to define the 100% inhibition level. The fiuidics-head (F-Head) of IonWorks™ HT then added 3.5 μl of PBS to each well of the PatchPlate™ and its underside was perfused with "internal" solution that had the following composition (in mM): K-Gluconate 100, KCl 40, MgCl2 3.2, EGTA 3 and HEPES 5 (all Sigma-Aldrich; pH 7.25-7.30 using 10 M KOH). After priming and de-bubbling, the electronics-head (E-head) then moved round the PatchPlate™ performing a hole test (i.e. applying a voltage pulse to determine whether the hole in each well was open). The F-head then dispensed 3.5 μl of the cell suspension described above into each well of the PatchPlate™ and the cells were given 200 seconds to reach and seal to the hole in each well. Following this, the E-head moved round the PatchPlate™ to determine the seal resistance obtained in each well. Next, the solution on the underside of the PatchPlate™ was changed to "access" solution that had the following composition (in mM): KCl 140, EGTA 1, MgCl2 1 and HEPES 20 (pH 7.25-7.30 using 10 M KOH) plus 100 μg/ml of amphotericin B (Sigma-Aldrich). After allowing 9 minutes for patch perforation to take place, the E-head moved round the PatchPlate™ 48 wells at a time to obtain pre-compound hERG current measurements. The F-head then added 3.5 Dl of solution from each well of the compound plate to 4 wells on the PatchPlate™ (the final DMSO concentration was 0.33% in every well). This was achieved by moving from the most dilute to the most concentrated well of the compound plate to minimise the impact of any compound carry-over. After approximately 3.5 mins incubation, the E-head then moved around all 384-wells of the PatchPlate™ to obtain post-compound hERG current measurements. In this way, non-cumulative concentration-effect curves could be produced where, providing the acceptance criteria were achieved in a sufficient percentage of wells (see below), the effect of each concentration of test compound was based on recording from between 1 and 4 cells.
The pre- and post-compound hERG current was evoked by a single voltage pulse consisting of a 20 s period holding at -70 mV, a 160 ms step to -60 mV (to obtain an estimate of leak), a 100 ms step back to -70 mV, a 1 s step to + 40 mV, a 2 s step to -30 mV and finally a 500 ms step to -7OmV. In between the pre- and post-compound voltage pulses there was no clamping of the membrane potential. Currents were leak-subtracted based on the estimate of current evoked during the +1OmV step at the start of the voltage pulse protocol. Any voltage offsets in Ion Works™ HT were adjusted in one of two ways. When determining compound potency, a depolarising voltage ramp was applied to CHO- KvI.5 cells and the voltage noted at which there was an inflection point in the current trace (i.e. the point at which channel activation was seen with a ramp protocol). The voltage at which this occurred had previously been determined using the same voltage command in conventional electrophysiology and found to be -15 mV (data not shown); thus an offset potential could be entered into the Ion Works™ HT software using this value as a reference point. When determining the basic electrophysiological properties of hERG, any offset was adjusted by determining the hERG tail current reversal potential in IonWorks™ HT, comparing it with that found in conventional electrophysiology (-82 mV) and then making the necessary offset adjustment in the IonWorks™ HT software. The current signal was sampled at 2.5 kHz.
Pre- and post-scan hERG current magnitude was measured automatically from the leak subtracted traces by the IonWorks™ HT software by taking a 40 ms average of the current during the initial holding period at -70 mV (baseline current) and subtracting this from the peak of the tail current response. The acceptance criteria for the currents evoked in each well were: pre-scan seal resistance >60 MΩ, pre-scan hERG tail current amplitude >150 pA; post-scan seal resistance >60 MΩ. The degree of inhibition of the hERG current was assessed by dividing the post-scan hERG current by the respective pre-scan hERG current for each well.
Results
Typical IC50 values for the compounds of the present invention are in the range of about 1 to about 10,000 nM. Biological data on exemplified final compounds is given below in Table 1.
TABLE 1.
Figure imgf000057_0001

Claims

Claims
1. A compound of formula I:
Figure imgf000058_0001
wherein
A is independently selected from a 5, 6 or 7 membered heterocyclic ring optionally substituted with one or more R1;
B is independently selected from a 4, 5, 6 or 7 membered heterocyclic ring, C1-balkyl, C2.6alkenyl, C2-6alkynyl, Co-6alkylaryl, Co-ealkylheteroaryl, Co-6alkylC3_8cycloalkyl, Co- 6alkylC3.6cycloalkenyl, Co-6alkylC3.6cycloalkynyl, Co-6alkylC3-6heterocyclyl optionally substituted with one or more R";
C is independently selected from phenyl or a 5 or 6 membered he tero aromatic ring optionally substituted with one or more R3;
R1 is independently selected from halogen, cyano, nitro, OR6, C^aUcyl, C2_6alkenyl, C2-6alkynyl, Co-βalkylaryl, Co-ealkylheteroaryl, C0.6alkylC3.6cycloalkyl, Co-όalkylC3- βcycloalkenyl, C0-6alkylC3-6cycloalkynyl, Co-όalkylC3-6heterocyclyl, NR6R7, CONR6R7, NR6(CO)R7, 0(CO)R6, CO2R6, COR6, (SO2)NR6R7, NR6(SO2)R7, SOR6, SO2R6, OSO2R6 and SO3R6 wherein said Ci-6alkyl, C2.6alkenyl, C2.6alkynyl, Co-βalkylaryl, C0- 6alkylheteroaryl, Co.6alkylC3.6cycloalkyl, Co-6alkylC3.6cycloalkenyl, C0-6alkylC3- gcycloalkynyl and Co-6alkylC3-6heterocyclyl may be optionally substituted with one or more D; or two R1 substituents may together with the atom to which they are attached form a cyclic or heterocyclic ring optionally substituted with one or more D;
R2, R3 and R4 are independently selected from halogen, cyano, nitro, OR6, Chalky!, C2-6alkenyl, Qj.ealkynyl, Co-βalkylaryl, Co-βalkylheteroaryl, C0-6alkylC3.6cycloa.kyl, Co- 6alkylC3-6cycloalkenyl, Co.6alkylC3-6cycloalkynyl, Co-6alkylC3-6heterocyclyl, NR6R7, CONR6R7, NR6(CO)R7, 0(CO)R6, CO2R6, COR6, (SO2)NR6R7, NR6(SO2)R7, SO2R6, SOR6, OSO2R6 and SO3R6 wherein said Ci.6alkyl, C2-6alkenyl, C2.6alkynyl, Co-galkylaiyl, Co-όalkylheteroaryl, Co-<5alkylC3-6cycloalkyl, Co-ealkylCs-βcycloalkenyl, Co-6alkylC3- βcycloalkynyl and C0-6alkylC3.6heterocyc.yl is optionally substituted with one or more D; or two R2, R3 or R4 substituents may together with the atoms to which they are attached form a cyclic or heterocyclic ring optionally substituted with one or more D;
R5 is independently selected from hydrogen, cyano, 0R&, Q^alkyl, C2-6alkenyl, C2-6alkynyl, Co-βalkylaryl, Co-ealkylheteroaryl, C0-6alkylC3.6cycloa.kyl, Co-6alkylC3- ecycloalkenyl, C0.6alkylC3.6cycloalkynyl, Co.6alkylC3^heterocyclyl, CONR6R7, CO2R6, COR6, SO2R6 and SO3R6 wherein said Q.ealkyl, C2-6alkenyl, C2-6alkynyl, C0-6alkylaryl, C0. δalkylheteroaryl, Co.6alkylC3.6cycloalkyl, Co.6alkylC3-6cycloalkenyl, Co-όalkylC3. βcycloalkynyl, Co-6alkylC3-6heterocyclyl may be optionally substituted with one or more D;
D is independently selected from halogen, nitro, CN, OR6, Ci.6alkyl, C^alkenyl,
C2-6alkynyl, Co-βalkylaryl, Co-ealkylheteroaryl, Co-6alkylC3-6cycloalkyl, Co-6alkylC3. δcycloalkenyl, Co.6alkylC3.6cycloalkynyl, Co-βalkylheterocyclyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
NR6R7, CONR6R7, NR6(CO)R7, 0(CO)R6, CO2R6, COR6, (SO2)NR6R7, NR6SO2R7,
SO2R6, SOR6, OSO2R6 and SO3R6, wherein said Ci.6alkyl. C2.όalkenyl, C2.6alkynyl, C0. δalkylaryl. Co-βalkylheteroaryl, Co-6alkylC3-6cycloalkyl, C0-6alkylC3-6cycloalkenyl, C0.
6alkylC3.6cycloalk}'nyl or Co-galkylheterocyclyl may be optionally substituted with one or more substituents independently selected from halo, nitro, cyano, OR6, C^alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoromethoxy, difluoromethoxy and trifluoromethoxy; R6 and R7 are independently selected from hydrogen, C^aUcyl, fluoromethyl, difluoromethyl, trifluoromethyl, C2-6alkenyl, C2-6alkynyl, Co-βalkylaryl, Co- 6alkylheteroaryl, C0-6alkylC3.6cycloalkyl, C0-6alkylC3-6cycloalkenyl, C0-OaIkVlC3- βcycloalkynyl, Co-δalkylheterocyclyl; or
R6 and R7 may together form a 5 or 6 membered heterocyclic ring containing one or more heteroatoms selected from N, O or S;
m = 0, 1, 2 or 3; n = 0, 1, 2 or 3; p = 0, 1, 2 or 3; q = 0, 1, 2 or 3;
as a free base or a pharmaceutically acceptable salt, solvate or solvate of a salt thereof.
2. A compound according to claim 1, wherein A is a 6 membered heterocyclic ring.
3. A compound according to claim 1 or 2, wherein B is a Co-6alkylC3-8cycloalkyl.
4. A compound according to claim 3, wherein Co-6alkylC3_scycloalkyl is represented by C3. gcycloalkyl.
5. A compound according to claim 4, wherein B is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
6. A compound according to any one of claims 1 to 5, wherein R5 is hydrogen.
7. A compound according to any one of claims 1 to 6, wherein m is 0.
8. A compound according to any one of claims 1 to 7, wherein n is 0.
9. A compound according to any one of claims 1 to 8, wherein q is 0.
10. A compound according to any one of claims 1 to 9, wherein C is phenyl.
11. A compound according to any one of claims 1 to 9, wherein C is phenyl substituted with one or more R3.
12. A compound according to claim 11, wherein R3 is halogen.
13. A compound according claim 12, wherein said halogen is chlorine.
14. A compound according to claim 1, wherein A is a 6 membered heterocyclic ring;
B is Co-6alkylC3-6cycloalkyl;
C is phenyl substituted with one or more R3; R3 is halogen;
R5 is hydrogen; m = 0; n = 0;
P = 2; q = 0.
15. A compound selected from:
8-Cyclopropyl-8-(3',5'-dichlorobiphenyl-3-yl)-2,3,4,8-tetrahydroimidazo[l,5-fi!]pyrimidin- 6-amine acetate; 8-Cyclohexyl-8-(3^5'-dichlorobiphenyl-3-yl)-2,3,4,8-te1τahyαϊoimidazo[l,5-α]pyrimidin-6- amine acetate;
8-Cyclobutyl-8-(3',5'-dichlorobiphenyl-3-yl)-2,3,4,8-tetrahydroimidazo[l,5-α]pyrimidin-6- amine acetate and
8-Cyclopent}d-8-(3',5'-dichlorobiphenyl-3-yl)-2,3,4,8-tetrahydroimidazo[l,5-α]pyrimidin- 6-amine acetate;
as a free base or a pharmaceutically acceptable salt, solvate or solvate of a salt thereof.
16. A pharmaceutical composition comprising as active ingredient a therapeutically effective amount of a compound according to any one of claims 1 to 15 in association with pharmaceutically acceptable excipients, carriers or diluents. s
17. A compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, for use as a medicament.
18. Use of a compound of any one of claims 1 to 15 as a medicament for treating oro preventing an Aβ-related pathology.
19. Use of a compound of any one of claims 1 to 15 as a medicament for treating or preventing an Aβ-related pathology, wherein said Aβ-related pathology is Downs syndrome, a β-amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebrals hemorrhage, a disorder associated with cognitive impairment, MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with Alzheimer disease, dementia of mixed vascular origin, dementia of degenerative origin, pre-senile dementia, senile dementia, dementia associated with Parkinson's disease, progressive supranuclear palsy or0 cortical basal degeneration.
20. Use of a compound of any one of claims 1 to 15 in the manufacture of a medicament for treating or preventing an Aβ-related pathology. 5
21. Use of a compound of any one of claims 1 to 15 in the manufacture of a medicament for treating or preventing an Aβ-related pathology, wherein said Aβ-related pathology is Downs syndrome, a β-amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, a disorder associated with cognitive impairment, MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms0 associated with Alzheimer disease, neurodegeneration associated with Alzheimer disease, dementia of mixed vascular origin, dementia of degenerative origin, pre-senile dementia, senile dementia, dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
22. A method of inhibiting activity of BACE comprising contacting said BACE with a 5 compound of any one of claims 1 to 15.
23. A method of treating or preventing an Aβ-related pathology in a mammal, comprising administering to said patient a therapeutically effective amount of a compound of any one of claims 1 to 15. 0
24. The method of claim 23, wherein said Aβ-related pathology is Downs syndrome, a β- amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, a disorder associated with cognitive impairment, MCI ("mild cognitive impairment"), Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimers disease, neurodegeneration associated with Alzheimer disease, dementia of mixed vascular origin, dementia of degenerative origin, pre-senile dementia, senile dementia, dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration. o
25. The method of claim 23, wherein said mammal is a human.
26. A method of treating or preventing an Aβ-related pathology in a mammal, comprising administering to said patient a therapeutically effective amount of a compound of any one of claims 1 to 15 and at least one cognitive enhancing agent, memory enhancing agent, or5 choline esterase inhibitor.
27. The method of claim 26, wherein said Aβ-related pathology is Downs syndrome, a β- amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, a disorder associated with cognitive impairment, MCI ("mild cognitive impairment"),0 Alzheimer Disease, memory loss, attention deficit symptoms associated with Alzheimer disease, neurodegeneration associated with Alzheimer disease, dementia of mixed vascular origin, dementia of degenerative origin, pre-senile dementia, senile dementia, dementia associated with Parkinson's disease, progressive supranuclear palsy or cortical basal degeneration.
28. The method of claim 26, wherein said mammal is a human
PCT/SE2007/001018 2006-11-20 2007-11-19 Amino- imidazolones and their use as medicament for treating cognitive impairment alzheimer disease, neurodegeneration and dementia WO2008063114A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US86645206P 2006-11-20 2006-11-20
US60/866,452 2006-11-20

Publications (2)

Publication Number Publication Date
WO2008063114A1 WO2008063114A1 (en) 2008-05-29
WO2008063114A9 true WO2008063114A9 (en) 2008-09-04

Family

ID=39429958

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE2007/001018 WO2008063114A1 (en) 2006-11-20 2007-11-19 Amino- imidazolones and their use as medicament for treating cognitive impairment alzheimer disease, neurodegeneration and dementia

Country Status (1)

Country Link
WO (1) WO2008063114A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8829036B2 (en) 2007-02-23 2014-09-09 Merck Sharp & Dohme Corp. Heterocyclic aspartyl protease inhibitors

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7592348B2 (en) 2003-12-15 2009-09-22 Schering Corporation Heterocyclic aspartyl protease inhibitors
US7700603B2 (en) 2003-12-15 2010-04-20 Schering Corporation Heterocyclic aspartyl protease inhibitors
AR056865A1 (en) 2005-06-14 2007-10-31 Schering Corp NITROGEN HETEROCICLES AND ITS USE AS INHIBITORS OF PROTEASES, PHARMACEUTICAL COMPOSITIONS
EP1896478B1 (en) 2005-06-14 2014-05-21 Merck Sharp & Dohme Corp. Aspartyl protease inhibitors
TW200815349A (en) 2006-06-22 2008-04-01 Astrazeneca Ab New compounds
TW201020244A (en) 2008-11-14 2010-06-01 Astrazeneca Ab New compounds
UA108363C2 (en) 2009-10-08 2015-04-27 IMINOTIADIASIADIOXIDE OXIDES AS BACE INHIBITORS, COMPOSITIONS THEREOF AND THEIR APPLICATIONS
EP2694521B1 (en) 2011-04-07 2015-11-25 Merck Sharp & Dohme Corp. Pyrrolidine-fused thiadiazine dioxide compounds as bace inhibitors, compositions, and their use
WO2012138734A1 (en) 2011-04-07 2012-10-11 Merck Sharp & Dohme Corp. C5-c6 oxacyclic-fused thiadiazine dioxide compounds as bace inhibitors, compositions, and their use
EP2747769B1 (en) 2011-08-22 2017-08-02 Merck Sharp & Dohme Corp. 2-spiro-substituted iminothiazines and their mono-and dioxides as bace inhibitors, compositions and their use

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SG163508A1 (en) * 2003-12-15 2010-08-30 Schering Corp Heterocyclic aspartyl protease inhibitors
EP1758907B1 (en) * 2004-06-16 2009-02-18 Wyeth Diphenylimidazopyrimidine and -imidazole amines as inhibitors of b-secretase
BRPI0512213A (en) * 2004-06-16 2008-02-19 Wyeth Corp method for treating, preventing or ameliorating a disease or disorder; pharmaceutical composition; process for the preparation of a compound; and use of a compound
JP2008526966A (en) * 2005-01-14 2008-07-24 ワイス Amino-imidazolones for the inhibition of β-secretase
TW200815447A (en) * 2006-06-14 2008-04-01 Astrazeneca Ab Novel compounds IV
TW200815443A (en) * 2006-06-14 2008-04-01 Astrazeneca Ab Novel compounds I
TW200815449A (en) * 2006-06-14 2008-04-01 Astrazeneca Ab Novel compounds II

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8829036B2 (en) 2007-02-23 2014-09-09 Merck Sharp & Dohme Corp. Heterocyclic aspartyl protease inhibitors

Also Published As

Publication number Publication date
WO2008063114A1 (en) 2008-05-29

Similar Documents

Publication Publication Date Title
WO2007145568A1 (en) Amino-imidazolones and their use as a medicament for treating cognitive impairment, alzheimer disease, neurodegeneration and dementia
US7629356B2 (en) Substituted pyrrolo[3,4-b]pyridinamines and pharmaceutical compositions
WO2008063114A9 (en) Amino- imidazolones and their use as medicament for treating cognitive impairment alzheimer disease, neurodegeneration and dementia
US20070299087A1 (en) New Compounds 319
US20080058349A1 (en) New Compounds 318
US20080214577A1 (en) New Compounds 320
US20080051420A1 (en) New Compounds 317
WO2008150217A1 (en) 2-amino-5, 5-diaryl-imidazol-4-one analogs for the inhibition of beta-secretase
US20080161269A1 (en) Compounds 620
WO2009005470A1 (en) Aryl and heteroaryl substituted isoindole derivatives as bace inhibitors
US20090099217A1 (en) 2-Aminopyrimidin-4-Ones And Their Use For Treating Or Preventing Alpha Beta-Related Pathologies
US20090023762A1 (en) Substituted 2-Aminopyrimidine-4-Ones, Their Pharmaceutical Compositions And Their Use In The Treatment And/Or Prevention Of Ab-Related Pathologies
KR20090031585A (en) Substituted isoindoles as bace inhibitors and their use
WO2008076043A1 (en) Novel 2-amino-5,5-diaryl-imidazol-4-ones
WO2009005471A1 (en) Aryl and heteroaryl substituted isoindole derivatives as bace inhibitors

Legal Events

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

Ref document number: 07835216

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 07835216

Country of ref document: EP

Kind code of ref document: A1