Specification
DtPHENYUMIDAZOPYRtMIDlNE AND -IMIDAZOLE AMINES AS INHIBITORS OF P-SECRETASE
BACKGROUND OF THE INVENTION
Alzheimer''s disease (AD), a progressive degenerative disease of the brain primarily associated with aging, is a serious healthcare problem. Clinically, AD is characterized by the of toss of memory, cognition, reasoning, judgment, and orientation. Also affected, as the disease progresses, are motor, sensory, and linguistic abilities until global impairment of multiple cognitive functions occurs. These cognitive losses take place gradually, but typically lead to severe impairment and eventual death in 4-12 years. Patients with AD display characteristic (3-amyloid deposits in the brain and in cerebral blood vessels (p-amy!oid angiopathy) as well as neuroflbrillary tangles. Amytoidogenic plaques and vascular amyloid angiopathy also characterize the brains of patients with Trisomy 21 (Down''s Syndrome), Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch-type (HCHWA-D), and other neurodegenerative disorders, Neurofibrillary tangles also occur in other dementia-inducing disorders.
The family of proteins known as {3-amyloid are thought to be causal for the pathology and subsequent cognitive decline in Alzheimer''s disease. Proteolytic processing of the amyloid precursor protein (APR) generates amyloid p (A-beta) peptide; specifically, A-beta is produced by the deavage of APP at the N-termtnus by (3-secretase and at the C-terminus by one or more y-secretases. Aspartyl protease enzyme, or {3-secretase enzyme (BACE), activity is correlated directly to the generation of A-beta peptide from APP (Sinha, et al, Nature, 1999. 402, 537-540). Increasingly, studies indicate that the inhibition of the p-secretase enzyme, inhibits
the production of A-beta peptide. The inhibition of p-secretase and consequent lowering of A-beta peptide may lead to the reduction of p-amyloid deposits in the brain and p-arnyloid levels in the cerebral biood vessels and to an effective treatment of a disease or disorder caused thereby.
Therefore, if is an object of this invention to provide compounds which are inhibitors of p-secretase and are useful as therapeutic agents in the treatment, prevention or amelioration.of a disease ordisorder characterized by elevated p-amyloid deposits or p-amy!oid levels in a patient.
It is another object of this invention to provide therapeutic methods and pharmaceutical compositions useful for the treatment, prevention or amelioration of a disease or disorder characterized by elevated p-amyloid deposits or p-amyloid levels in a patient.
It is a feature of this invention that the compounds provided may also be useful to further study and elucidate the p-secretase enzyme.
These and other objects and features of the invention will become more apparent by the detailed description set forth hereinbelow.
SUMMARY OF THE INVENTION
The present invention provides a imidazopyrimidine or imidazoimidazole
(I)
amine of formula
(Figure Remove)
wherein X is N, NO or CRi9; YisN.NOorCRi,;
Z is N, NO or CR-o with the proviso that no more than two of X, Y or Z may be N or NO;
RI and R,2 arc each independently H, CN or an optionally substituted C--C4alkyl group;
R3 and R4 are each independently H, or an optionally substituted C,-C4 alkyl group or R3 and R4 may be taken together to form a 3- to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S or R3 may be taken together with the atom to which it is attached and an adjacent carbon atom to form a double bond;
R5 and R6 are each independently H, halogen, NO2l CN, OR13, NR14R15 or a CrC6alkyl, C,-C6haloaikyl, C2-C6alkenyl, C2-C6alkynyl or C3-C8cycloalkyi group each optionally substituted or when attached to adjacent carbon atoms R5 and R6 may be taken together with the atoms to which they are attached to form an optionally substituted 5-to 7-membered ring optionally containing one or two heteroatoms selected from 0. N or S;
R7, R8, R9, RIO, RII, Rig and R20 are each independently H, halogen, NO2, CN, OR16, NRI7R,a or a C,-C6alky1, CrC6ha!oalkyl, C2-C6alkenyl, Ca-Cealkynyi or C3-C8cycloalkyl group each optionally substituted;
m is 0 or 1;
nisO, 1.2 or 3;
--—- is a single bond or a double bond with the proviso that when m is 0
c
then —-.-.- must be a single bond;
RI2, R,3 and Rie are each independently H or a C1-C6alky1, C1-C6haloalkyt. C2-C6alkenyl, C2-C6alkyny!, C3-C8cycloalkyl or aryl group each optionally substituted; and
R-14, RIS, RIT and R« are each independently H or d-C^alkyJ; or a tautomer thereof, a stereoisomer thereof or a pharmaceutically acceptable salt thereof.
The present invention also provides therapeutic methods and pharmaceutical compositions useful for the treatment, prevention or amelioration of a disease or disorder characterized by increased p-amyloid deposits or increased (3-amyloid levels in a patient.
DETAILED DESCRIPTION OF THE INVENTION
Alzheimer''s disease (AD) is a major degenerative disease of the brain which presents clinically by progressive loss of memory, cognition, reasoning, judgement and emotional stability and gradually leads to profound mental deteoriation and death. The exact cause of AD is unknown, but increasing evidence indicates that amyloid beta peptide (A-beta) plays a central role in the pathogenesis of the disease. (D. B. Schenk; R. E. Rydei et a/, Journal of Medicinal Chemistry, 1995. 21,4141 and D. J. Selkoe, Physiology Review, 2001. 81, 741). Patients with AD exhibit characteristic neuropathological markers such as neuritic plaques (and in p-amyloid angiopathy, deposits in cerebral blood vessels) as well as neurofibrillary tangles detected in the brain at autopsy. A-beta is a major component of neuritic plaques in AD brains. In addition, p-amyloid deposits and vascular (3-amyloid angiopathy also characterize individuals with Downs Syndrome, Hereditary Cerebral Hemmorhage with Amyloidosis of the Dutch type and other neurodegenreative and dementia-inducing disorders. Over expression of the amyloid precursor protein (APR), altered deavage of APR to A-beta or a decrease in the clearance of A-beta from a patient''s brain may increase the levels of soluble or fibrullar forms of A-beta in the brain. The p-site APR cleaving enzyme, BACE1, also called memapsin-2 or Asp-2, was identified in 1999 (R. Vassar. B. D. Bennett, et al, Nature, 1999. 402, 537). BACE1 is a membrane-bound aspartic protease with all the known functional properties and characteristics of p-secretase. Parallel to BACE1, a second homologous asp''artyl protease named BACE2 was found to have p-secretase activity in vitro. Low molecular weight, non-peptide, non-substrate-related inhibitors of BACE1 or p-secretase are earnestly sought both as an aid in the study of the P-secretase enzyme and as potential therapeutic agents.
Surprisingly, it has now been found that diphenylimidazopyrimidine amine or diphenylirrndazoimidazole amine compounds of formula I demonstrate inhibition of p-secretase and the selective inhibition of BACE1. Advantageously, said pyrimidinamine or imidazolamine compounds may be used as effective therapeutic agents for the treatment, prevention or amelioration of a disease or disorder characterized by elevated p-amyloid deposits or p-amyloid levels in a patient.
Accordingly, the present invention provides an imidazopyrimidine or imidazoimidazole amine compound of formula i
(Figure Remove)
wherein X is N, NO or CR19;
Y is N, NO or CR,,;
Z is N, NO or CRso with the proviso that no more than two of X, Y or Z may be N or NO;
RI and R2 are each independently H, CN or an optionally substituted Cr C4alkyl group;
R3 and R4 are each independently H, or an optionally substituted Ct-C4 alkyf group or R3 and R« may be taken together to form a 3- to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S or R3 may be taken together with the atom to which it is attached and an adjacent carbon atom to form a double bond;
R5 and R6 are each independently H, halogen, NO2l CN, OR13, NR,4Ris or a CrCealkyl, CfCehaloalkyl, CrC6aikenyl, C2-C6alkynyl or Ca-Cacycloalkyl group each optionally substituted or when attached to adjacent carbon atoms Rs and RS may be taken together with the atoms to which they are attached to form an optionally substituted 5-to 7-membered ring optionally containing one or two heteroatoms selected from O. N or S;
R?, Rfl, Ra, Rio, Rn, RIS and R20 are each independently H, halogen, NO2, CN, OR1G, NR17R18or a CrC6alkyl, CrCehaloalkyl, C2-CGaikenyl, C2-Ci,alkynyI or C3-Cecycioalkyl group each optionally substituted;
tn is 0 or 1;
n is 0, 1, 2 or 3;
----—- is a single bond or a double bond with the proviso that when rn then ------ must be a single bond;
RI?., Rn and R16 are each independently H or a C|-C6alkyl, C^
C2-C6alkenyl, C2-CGalkynyl, C3-C8cycloa!kyl or aryf group each optionally substituted; and
Ri4, RIS, RI? and R18 are each independently H or C1-C4alkyl; or a tautomer thereof, a stereoisomer thereof or a pharmaceuticaily acceptable salt thereof.
X is N, NO or CR19; preferably N or CR19, more preferably N. Y is N, NO or CRu; preferably CRu. Z is N. NO or CR20; preferably CR20. The symbol m is 0 or 1, preferably 1. R7, Ra, R9, R10, R^, R19 and Rao are each independently H, halogen. NO2l CN, OR16, NR17Ria or a d-Cealkyl, CrC6haloalkyl, CrC6alkenyI, C2-C6alkynyl or C3-C8cycloalkyl group each optionally substituted; preferably H or halogen. —— is a single bond or a double bond, preferably a double bond. The ring containing X, Y and Z substitutes the phenyl group, preferably at the 3-position thereof. The ring member of the ring containing X, Y and Z that is linked to the phenyl group is preferably the carbon atom linked to Y and Z.
RT and R2 are each independently H, -CN or an optionally substituted Cr C4alkyl group; preferably H or an optionally substituted C1-C4alkyl group; more preferably H, a CrC4alkyl group or a substituted CrC^alkyl group substitute^ by halogen; advantageously H.
R3 and R« are each independently H, or an optionally substituted CrC4 alkyl group or R3 and R< may be taken together to form a 3- to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S or R3 may be taken together with the atom to which it is attached and an adjacent carbon atom to form a double bond. Preferably R3 and R4 are each independently H or a Ct-C4 alkyl group or, when taken together with the carbon ato-n to which they are attached, form a 3- to 7-membered ring (advantageously a 3- to 5-membered ring) optionally containing one or two heteroatoms selected from O, N or S. More preferably the ring is carbocycfic or contains one heteroatom selected from O, N or S (advantageously O). Advantageously R3 and R« are each independently H or, when taken together with
the carbon atom to which they are atlached, form the aforesaid ring. The symbol n is 0, 1, 2 or 3, preferably 0, 1 or 2, advantageously 1.
Preferably R5 and Re are each independently H. halogen, CN. OR-3; or a C,-C6aikyi, C,-Cuhaioalkyl, or C3-C8cycloalky! group each optionally substituted or, when attached lo adjacent carbon atoms, R5 and RB may be taken together with said adjacent carbon atoms to form an optionally substituted 5- to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S, the ring preferably containing two oxygen atoms as heteroatoms. More preferably R5 and R6 are each independently H, halogen, CN, QRu, or a Ci-C6alkyl, C-i-Cehaloalkyi, or C3-Cacyc!oalkyJ group or, when attached to adjacent carbon atoms, R5 and Re may be taken together with said adjacent carbon atoms to form a 5- to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S, the ring being optionally substituted by 1, 2 or 3 halogen atoms. Advantageously at least one of RS and R6 is OR^ or RS and RB when taken together represent a divalent radical which is a methylenedtoxy or ethyienedioxy radical optionally substituted by one or more halogen atoms. R13 is H or a CrC6alkyf, CrCBhaloalkyl, C2-C6alkenyl, Cz-Cealkynyl, C3-Cacycloalky] or aryl group each optionally substituted; preferably d-C6alkyl substituted by 0,1,2 or 3 substituents independently selected from halogen atoms, nitro, cyano, thiocyanato, cyanato, hydroxyl, alkyl, haloalkyl, alkoxy, haloalkoxy, amino, alkylamino, dialkylamino, formyl, alkoxycarbonyl, carboxyl, alkoxycarbonyl, alkanoyl, alkanoyloxy, alkylthio, alkylsuphinyl, alkylsulphonyl, carbamoyl, alkylamido, phenyt, phenoxy, benzyl, benzyfoxy, heterocycfyl and cycloalkyl groups.,
As used in the specification and claims, the term halogen designates F, CJ, Br or I and the term cycloheteroaikyl designates a five- to seven-membered cycloalkyl ring system containing 1 or 2 heteroatoms, which may be the same or different, selected from N, O or S and optionally containing one double bond. Exemplary of the cycloheteroaikyl ring systems included in the term as designated herein are the following rings wherein Xi is NR, O or S; and R is H or an optional substituent as described hereinbelow:
Similarly, as used in the specification and claims, the term heteroaryl designates a five- to ten-membered aromatic ring system containing 1, 2 or 3 heteroatoms, which may be the same or different, selected from N, O or S. Such heteroaryl ring systems include pyrrolyl, azolyl, oxazolyl, thiazolyl, imidazolyl, furyl, thienyl, quinolinyl, isoquinolinyl, indolyl, benzothienyl, benzofuranyl, benzisoxazolyl or the like. The term aryl designates a carbocyclic aromatic ring system such as phenyl, naphthyl, anthracenyl or the like. The term aryl{C,-C4)alkyl designates an aryl group as defined hereinabove attached to a CrC^alkyl group which may be straight or branched. Said aryl(CrC4)alkyl groups include benzyl, phenethyl, napthtylmethyl, or the like. The term haloalkyl as used herein designates a CnH2n*i group having from one to 2n-M halogen atoms which may be the same or different and the term haloalkoxy as used herein designates an OCnH2n+i group having from one to 2n+1 halogen atoms which may be the same or different. Preferably the term haloalkyl designates CF3 and the term haloalkoxy designates OCF3.
In the specification and claims, when the terms CrC6alkyl, d-C6haloalkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C8cycloalkyl, cycloheteroalkyf, aryl, aryl(CrC^alkyl or heteroaryl are designated as being optionally substituted and where a 5- to 7-membered ring formed by taking R5 and Re together with the atoms to which R5 and Re are attached is designated as optionally substituted, the substituent groups which are optionally present may be one or more of those customarily employed in the development of pharmaceutical compounds or the modification of such compounds to influence their structure/activity, persistence, absorption, stability or other beneficial property. Specific examples of such substituents include halogen atoms, nitro, cyano, thiocyanato, cyanato, hydroxyl, alkyl, haloalkyl, alkoxy, haloalkoxy, amino, alkylamino, dialkylamino. formyl, alkoxycarbonyl, carboxyl, alkanoyl, alkylthio, alkylsuphinyl, alkylsulphor.yl, carbamoyl, alkylamido, phenyl. phenoxy, benzyl, henzyloxy, heterocyclyl or cycloalkyl groups, preferably halogen atoms or lower alkyl
or lower alkoxy groups. Typically, 0-3 substituents may be present. When any of the foregoing substituents represents or contains an alkyl substituent group, this may be linear or branched and may contain up lo 12, preferably up to 6. more preferably up to 4 carbon atoms.
Pharmaceutically acceptable salts may be any acid addition salt formed by a compound of formula ! and a pharmaceutical!y acceptable acid such as phosphoric, sutfuric, hydrochloric, hydrobromic, citric, maleic, malonic, mandeiic, succinic, fumaric, acetic, lactic, nitric, sulfonic, p-toluene sulfonic, methane sulfonic acid or the like.
Compounds of the invention include esters, carbamates or other conventional prodrug forms, which in general, are functional derivatives of the compounds of the invention and which are readily converted to the inventive active moiety in vivo. Correspondingly, the method of the invention embraces the treatment of the various conditions described hereinabove with a compound of formula I or with a compound which is not specifically disclosed but which, upon administration, converts to a compound of formula I in vivo. Also included are metabolites of the compounds of the present invention defined as active species produced upon introduction of these compounds into a biological system.
Compounds of the invention may exist as one or more tautomers. One skilled in the art will recognize that compounds of formula I may also exist as the tautomer It as shown below.
(Figure Remove)
Tautomers often exist in equilibrium with each other. As these tautomers interconvert under environmental and physiological conditions, they provide the same
useful biological effects. The present invention includes mixtures of such tautomers as well as the individual tautomers of Formula I and Formula It.
The compounds of the invention may contain one or more asymmetric carbon atoms or one or more asymmetric (chiral) centers and may thus give rise to optical isomers and diastereomers. Thus, the invention includes such optical isomers and disastereomers; as well as the racemic and resolved, enantiomerically pure stereoisomers; as well as other mixtures of the R and S stereoisomers. One skilled in the art will appreciate that one stereoisomer may be more active or may exhibit beneficial effects when enriched relative to the other stereoisomer(s) or when separated from the other stereoisomer(s). Additionally, the skilled artisan knows how to separate, enrich or selectively prepare said stereoisomers. Accordingly, the present invention comprises compounds of Formula I, the stereoisomers thereof, the tautomers thereof and the pharmaceutically acceptable salts thereof. The compounds of the invention may be present as a mixture of stereoisomers, individual stereoisomers, or as an optically active or enantiomerically pure form.
The stereoisomers of the invention include compounds in which formula I is formula IA and compounds in which formula I is formula IB.
(Figure Remove)
Preferred compounds of the invention are those compounds of formula I wherein Ri and R2 are H. Another group of preferred compounds of the invention are those compounds of formula I wherein m and n are 1. Also preferred are those Formula I compounds wherein X is N. A further group of preferred compounds of the invention are those compounds of formula I wherein the nitrogen-containing 5-
(Figure Remove)
rnembered or 6-membered heteroaryl ring is attached to the phenyl ring in the 3-position of the phenyl ring; this preferred group of formula I compounds is designated in the specification and claims as formula la. The formula la compound is shown beiow.
More preferred compounds of the invention are those compounds of formula la wherein the nitrogen-containing heteroaryl ring is a 6-membered ring and is attached to the phenyl ring in the 3-position of said heteroaryl ring; this more preferred group of formula I compounds is designated in the specification and claims as formula Ib. Formula Jb is shown below.
(Figure Remove)
Another group of more preferred compounds of the invention are those compounds of formula Ib wherein Rt and R2 are H. A further group of more preferred compounds
of the invention are those compounds of formula Ib wherein Y is CRn and R, and R? are H.
Examples of preferred compounds of formula I include; 8-(3-pyrimidin-5-ylphenyl)-8-[4-(trifluofomethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrirnidin-6-amine;
7-(3-pyrimidin-5-ylphenyl)-7-[4-(trifluoromethoxy)phenyl]-7H-imidazo[1,5-alimida2ol-5-amine;
8-[4-f!uoro-3-(4-fluoropyridin-3-yi)phenyI]-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[3-(5-f]uoropyridin-3-y!)phenyl]-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[3-(5-chloropyridin-3-yl}phenyl]-8-[4-(trifluoromethoxy)pheny!]-2,3,4,8-
tetrahydroimfdazo[1,5-a]pyrimidin-6-amine; (8S)-8-[3-{2-fluoropyridin-3-yl)pheny!l-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1 , 5-ajpyrimidin-6-amine; {8R)-8-[3-(2-fluoropyridin-3-yl)pnenyl3-8-[4-{trifluoromethoxy)phenyl]-2,3)4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; (8R)-8-(3-pyrimidin-5-yJphenyl)-8-[4-(trifIuoromethoxy}phenyl]-2,3,4,8-
{efrahydroimidazo[1,5-a]pyrimidin-6-amine;
tetrahydroirnidazo[1,5-ajpyrimidin-6-amine; 8-[3-(4-fIuoropyridin-3-yl)ph€ny!]-8-[4-(trifluoromethoxy)phenyI]-2I3,4I8-
(etrahydroimidazo[1 , 5-aJpyrimidin-6-amine; 8-[3-(2-fluoropyridin-3-yl)phenyI3-8-(4-methoxyphenyI]-2,3I4,8-{etrahydroimidazo[1,5-
a]pyrimidin-6-amine; 8-(4-methoxyphenyl)-8-(3-pyrimidin-5-yIpheny!)-2,3,4I8-tefrahydroimidazo[1,5-
a]pyrimidin-6-amine; 8-(4-fluoro-3-pyrimidin-5-ylphenyl)-8-(4-methoxyphenyl)-2)3,4l8-
tetrahydroimidazo[1,5-a]pyrimWin-6-amine; 8-[4-fluoro-3-(2-fluoropyridin-3-yl)phenyl]-8-(4-methoxyphenyl}-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-{4-fluoro-3-pyrimidin-5-ylphenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3)4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine;
8-{3-(2-fluoropyridJn-3-yf)phenyl]-8-[4-(trifluoroiiiethoxy)phenyl]''2,3.4,8-!etrahydroirnidazo[1,5-a]pyrimidin-6-amine;
8--f4-fluoro-3-(5-fluoropyridin-3-yl)phenyl]-8-[4-(trifluorornethox\/)pheny!J-2,3,4,S-tetrahydroimidazo[1,5-a]pyrimidin-6-amine;
8-[4-fluoro-3-{2-fluoropyridin-3-yl)phenyl]-8-[4-(lrJfluoromethoxy)pheny!]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine;
the tautomers thereof; the stereoisomers thereof; or the pharmaceutically acceptable salts thereof.
Advantageously, the present invention provides a process for the preparation of a compound of formula I which comprises reacting a compound of formula II wherein Hal is Cl or Brwith a compound of formula III wherein W is B(OH)2, Sn(Bu)3 or Sn(CH3)3 in the presence of a palladium catalyst and an inorganic base optionally in the presence of a solvent. The process is shown in flow diargram I, wherein Hal and W are as defined hereinabove.
Ftow Diagram I
** *
(II)
R,o(Figure Remove)
X—
•^™-f
W
(III)
Palladium catalysts suitable for use in the process of the invention include Pd{0) or Pd(ll) catalysts such as dichlorobis(tri-o-toiy(phosphine)palladium(ll), Pd(OCOCH3)2/tri-o-tolylphosphine. tetrakis(triphenylphosphine)palladium(0), tris(dibenzylideneacetone)dipalladium(0)triphenylphosphine, or the like.
Inorganic bases suitable for use in the inventive process include Na or K hydroxides, carbonates or bicarbonates, preferably Na2CO3or K2CO3.
Solvents suitable for use in the inventive process include polar or non-polar organic solvents such as toluene, diethoxy ethyl ether, dioxane, ethyleneglycol
dimethyl ether or any non-reactive organic solvent which is capable of solubilizing the formula II or formula III compounds.
Compounds of forrv.uis !! m3u be nren3red usin^1 convsntiona! s^nlh^lic methods and, if required, standard separation or isolation techniques. For example, compounds of formula II wherein R, and R2 are H (Ha), may be prepared by reacting a ketone of formula IV with a phenyi magnesium bromide of formula V in the presence of a catalyst such as Cul to give a 1 ,1 ,1-trisubstituted methanol compound of formula VI; reacting said formula VI meJhanol sequentially with thionyl chloride and ammonia to give the corresponding methylamine of formula VII; and reacting said formula VII amine with cyanogen bromide in the presence of acetonitrile to give the desired formula Ha product. The reaction is shown in flow diagram II wherein Hal is Cl or Br.
Flow Diagram II
MgBr
!| "S
(Figure Remove)
1) SOCI2 2)NH3
BrCN(Figure Remove)
CH3CN
(Ha)
Ketones of formula IV may be prepared using conventional techniques, for example, by reacting a benzoyl halide of formula VIM with an imidazole or tetrahydropyrirnidine of formula IX in the presence of a base such as NaOH, or by
oxidizing the appropriate meihanol compound of formula X with an oxidizing agent such as MnO2, The reactions are shown in flow diagram III.
(Figure Remove)
Flow Diagram 111
o
(VIII)
(Figure Remove)
Compounds of formula X may be prepared by reacting a benzaJdehyde of formula XI with NaHSO3 and NaCN to give the corresponding cyanomethanol of
t
formula Xlf; reacting said formula Xll compound with ethanol and HCI to give the irnidate of formula XIII; and reacting said formula Xlil compound with a diamine of formula XIV to give the desired methanol derivative of formula X. The reaction is shown in flow diagram IV wherein Ha! is Cl or Br.
Flow Diagram IV
Hal
NaSO3
''
) NaCN
OH
CN
(XI)
(XII)
(Figure Remove)
(XIII)
Compounds of formula lla may be converted to the corresponding compounds of formula I wherein R, and R2are H using the procedure described hereinabove in flow diagram I.
Compounds of formula I wherein RI and R2 are other than H may be prepared using standard alkylation techniques such as reacting the compound of formula I wherein R, and R2 are H with an alkyl halide, RrHal, to give the compound of formula I wherein R2 is H (Id) and optionally reacting said formula Id compound with a second alkyl halide, R2-Hal, to give the desired formula I compound wherein Rt and R2 are other than H.
Advantageously, the compounds of the invention are useful for the treatment, prevention or amelioration of a disease or disorder characterized by elevated p-amyloid deposits or p-amyloid levels in a patient, including Alzheimer''s disease, Downs Syndrome, Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch type or other neurodegenerative or dementia-inducing disorders. Accordingly, the present invention provides a method for the treatment, prevention or amelioration of a disease or disorder characterized by elevated p-amyloid deposits or p-amyloid
levels in a patient which comprises providing said patient with a therapeutically effective amount of a compound of formula I as described hereinabove. The compound may be provided by oral or parenteral adminis(ra!ion or In any common manner known to be an effective administration of a therapeutic agent to a patient in need thereof.
The term "providing" as used herein with respect to providing a compound or substance embraced by the invention, designates either directly administering such a compound or substance, or administering a prodrug, derivative or analog which forms an equivalent amount of the compound or substance within the body.
The therapeutically effective amount provided in the treatment of a specific CMS disorder may vary according to the specific condition(s) being treated, the size, age and response pattern of the patient, the severity of the disorder, the judgment of the attending physician and the like. In general, effective amounts for daily oral administration may be about 0.01 to 1,000 mg/kg, preferably about 0.5 to 500 mg/kg and effective amounts for parenteral administration may be about 0.1 to 100 mg/kg, preferably about 0.5 to 50 mg/kg.
In actual practice, the compounds of the invention are provided by administering the compound or a precursor thereof in a solid or liquid form, either neat or in combination with one or more conventional pharmaceutical carriers or excipients. Accordingly, the present invention provides a pharmaceutical composition which comprises a pharmaceutically acceptable carrier and an effective amount of a compound of formula I as described hereinabove.
Solid carriers suitable for use in the composition of the invention incfude one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aides, binders, tablet-disintegrating agents or encapsulating materials. In powders, the carrier may be a finely divided solid which is in admixture with a finely divided compound of formula I. In tablets, tine formula I compound may be mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. Said powders and tablets may contain up to 99% by weight of the formula t compound. Solid carriers suitable for use in the composition of the invention include calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin,
cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrroiidine, low melting waxes and ton exchange resins.
Any pharmaceutical!" 3cc8nt3b!s !!n|jid carrier suitable for rirerv3r!nrj solutions, suspensions, emulsions, syrups and elixirs may be employed in the composition of the invention. Compounds of formula 1 may be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, or a pharmaceutically acceptable oil or fat, or a mixture thereof. Said liquid composition may contain other suitable pharmaceutical additives such as solubtlizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, coloring agents, viscosity regulators, stabilizers, osmo-regulators, or the like. Examples of liquid carriers suitable for oral and parenteral administration include water (particularly containing additives as above, e.g., cellulose derivatives, preferably sodium carboxymethyi cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g., glycols) or their derivatives, or oils (e.g., fractionated coconut oil and arachis oil). For parenteral administration the earner may also be an oily ester such as ethyf oleate or isopropy! myristate.
Compositions of the invention which are sterile solutions or suspensions are suitable for intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions may also be administered intravenously. Inventive compositions suitable for oral administration may be in either liquid or solid composition form.
Alternatively, the use of sustained delivery devices may be desirable, in order to avoid the necessity for the patient to take medications on a daily basis. "Sustained delivery" is defined as delaying the release of an active agent, i.e., a compound of the invention, until after placement in a delivery environment, followed by a sustained release of the agent at a later time. Those of skill in the art know suitable sustained delivery devices. Examples of suitable sustained delivery devices include, e.g., hydrogels (see, e.g., US Patent Nos. 5,266,325; 4,959,217; and 5,292,515), an osmotic pump, such as described by Alza (US Patent Nos. 4,295,987 and 5,273,752) or Merck (European Patent No. 314,206), among others; frydrophobic membrane materials, such as ethylenemethacrylate (EMA) and ethyfenevinylacetate (EVA); bioresorbable polymer systems (see, e.g., International Patent Publication No. WO 98/44964. Bioxid and Celiomeda; US Patent Nos. 5,756.127 and 5.854.388V other
bioresorbable implant devices have been described as being composed of, for example, polyesters, polyanhydrides, or lactic acid/glycolic acid copolymers (see, e.g., US Patent No. 5,817.343 (Alkermes Inc.)). For use in such sustained delivery devices, ihe compounds of the invention may be formulated as described herein.
In another aspect, the invention provides a pharmaceutical kit for delivery of a product. Suitably, the kit contains packaging or a container with the compound formulated for the desired delivery route. For example, if the kit is designed for administration by inhalation, it may contain a suspension containing a compound of the invention formulated for aerosol or spray delivery of a predetermined dose by inhalation. Suitably, the kit contains instructions on dosing and an insert regarding the active agent. Optionally, the kit may further contain instructions for monitoring circulating levels of product and materials for performing such assays including, e.g., reagents, well plates, containers, markers or labels, and the like. Such kits are readily packaged in a manner suitable for treatment of a desired indication. For example, the kit may also contain instructions for use of the spray purnp or other delivery device.
Other suitable components to such kits will be readily apparent to one of skill in the art, taking into consideration the desired indication and the delivery route. The doses may be repeated daily, weekly, or monthly, for a predetermined length of time or as prescribed.
For a more clear understanding, and in order to illustrate the invention more clearly, specific examples thereof are set forth hereinbelow. The following examples are merely illustrative and are not to be understood as limiting the scope and underlying principles of the invention in any way. Indeed, various modifications of the invention, in addition to those shown and described herein, will become apparent to those skilled in the art from the examples set forth hereinbelow and the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.
Unless otherwise noted, all parts are parts by weight. The term NMR designates nuclear magnetic resonance. The terms THF and DMFdesignate tetrahydrofuran and dimethyl formamide, respectively.
EXAMPLE 1 Preparation of 2-(3-Bromobenzoyl)-1 H-imidazole
(Figure Remove)
A solution of imidazole (2.66 g, 39.1 mmol) and triethylamine (Et3N) 8.03g, 79.4 mmol) in pyridine at 0°C is treated with 3-bromobenzoyl chloride (17.5 g, 79.9 mmol), stirred for 5 min, allowed to warm to room temperature for 45 min, treated with an aqueous sodium hydroxide solution (7.5 N, 20 ml, 150 mmol), heated at reflux temperature for 2 h, cooled to room temperature, diluted with water, further cooled with an ice bath for 1 h and filtered.. The filtercake is washed with water and dried under vacuum at 50 °C overnight to afford the title compound as a light tan solid, 4.89 g (50%yield), identified by NMR and mass spectral analyses. 1H NMR (500 MHz, CDCI3) 8 10.58 (br s, 1H), 8.73 (t, J= 1.8 Hz, 1H), 8.61 (dt, J = 7.8,1.2 Hz, 1H), 7.75-7.72 (m, 1H), 7.42-7.38 (m, 2H), 7.32 (s, 1H); ESI MS m/z25Q [C10H7BrN2O + H]*.
EXAMPLE 2 Preparation of 1-(3-Bromophenvl)-1H''^dazot"2"V^"''^''f4^tr''f1''-lol''omethoxv)-
K
phenvnmethanol
(Figure Remove)
A mixture of magnesium (0.644 g, 87.7 mmol) in THF (13 mL) at 50 °C is treated dropwise with a solution of 1-bromo-/l-(trifluoromethoxy)benzene (6.32 g,
2.G. 2 mmoi) in THF over a period of 5 min, stirred at 50 °C for an additional 1 .5 h. cooled to room temperature, treated with copper(l) iodide (0.041 g, 0.215 mmol) and a solution of 2~(3-bromobenzoyl}-1H-irnfdazo!e (2.6Q g. 1Q.4 ITVTIO!) in THP, heaieu ai 65 °C overnight, cooled to room temperature and diluted with a 1:1:1 mixture of ethyl acetate, water and saturated aqueous ammonium chloride. The phases are separated and the aqueous phase is extracted with ethyl acetate. The combined organic extracts are dried over sodium sulfate, filtered and concentrated. The resultant residue is purified by flash chromatography (silica, 85:15 to 75:25 hexanes/ethyl acetate as eluent) affords the title product as a yellow solid, 3.47 g (81% yield), identified by NMR and mass spectral analyses. 1H NMR (500 MHz, CDCI3) 5 8.98 (br s, 1 H), 7.55 (t. J = 1 .6 Hz, 1 H), 7.45 (dt, J = 8. 1 , 1 .9 Hz, 1 H), 7.37-7.34 (m, 2H), 7.24-7.20 (m, 2H), 7.18 (d, J= 8.1 Hz, 2H), 7.07 (brs, 1H), 6.98 (br s. 1H), 4.30 (br s, 1H); ESI MS m/z 412 [Ct7H12BrF3N2O2 + H]*.
EXAMPLE 3 Preparation of 1-13-B^^ (trifluoromethoxytehenynmethylarnme
(Figure Remove)
OCF3
OCF3
A mixture of 1-(3-bromophenyl)-1-(imidazo!-2-yl>-1-[4-(trifluorometho)cy)-phenyl]methanol (1.89 g, 4.57 mmol) and thionyl chloride (2.13 g, 17.9 mmol) in benzene is heated at 80 "C for 3 h, cooled to room temperature and concentrated in vacua to dryness. This orange solid residue is dispersed in isopropanol, bubbled with ammonia gas at ice-bath temperature until the solution is saturated. This ammonia saturated solution is heated in a sealed tube at 35 °C overnight, cooled io room temperature, concentrated and partitioned between chloroform (50 mL) and 1 N HCi (50 ml). The phases are separated and the organic phase is washed with
additional 1 N HCI. The combined HCI washes are cooled to 0 °C, basified to pH >10 by the addition of solid sodium hydroxide and extracted with chloroform. The combined chloroform extracts are dried over sodium suifaie. filtered and concentrated to afford the title product as a light yellow solid, 1.68 g (89% yield), identified by NMR and mass spectral analyses. 1H NMR (500 MHz, CDCI3) 8 9.12 (br s, 1H), 7.51 (t, J = 1.2 Hz, 1H), 7.44-7.41 (m, 1H), 7.35-7.31 (m, 2H), 7.22-7.19 (m, 2H),.7.18-7.15 (m, 2H), 7.15 (t, J= 1.5 Hz, 1H), 6.97 (t, J= 1.7 Hz, 1H), 2.48 (brs. 2H); ESI MS m//412 [Cl/H13BrF3N3O + H]+.
EXAMPLE 4
Preparation of 7-(3-(Bromophenyl)-7-r4-(trifluoromethoxv)phenvn-7H-imidazori.5-a1imidazol-5-ylamine
(Figure Remove)
OCF3
BrCN CH3CN ,
''OCF,
A mixture of 1-(3-bromophenyl)-1-(imidazol-2-yl)-1-[4-(trifluoromethoxy)-phenyljmethylamine (1.67 g, 4.05 mmol) and cyanogen bromide (1.75 g, 16.5 mmol) in acetonitrile is heated at 100 "C in a sealed tube overnight, cooled to room temperature and concentrated. The resultant residue is purified twice by flash chromatography (silica, 96:4:0.5 methylene chloride/methanol/concentrated ammonium hydroxide, then 85:15 to 50:50 hexanes/ethyl acetate as eluent) to afford the title compound as a yellow solid 0.671 g (38%yield), identified by NMR and mass spectral analyses. 1H NMR (500 MHz, CDCI3) 8 7.61 (t, J = 1.6 Hz, 1H), 7.53-7.48 (m, 2H), 7.46-7.41 (m, 3H), 7.27-7.22 (m, 4H); ESI MS m/z437 [C,8H12BrF3N4O +
EXAMPLE 5
Preparation of 7-ff3-(Pvrimidin-5-vl)phenvn-7-f4-(triflaoromethoxy)phenvn-7H-iiTjjdazcrTJ ^-
(Figure Remove)
"OCF3
''OCF-,
A mixture of 7-{3-(bromophenyl)-7-[4-(trifluoromethoxy)pheny!]-7H-imidazo[1,5-a]imidazol-5-ylamine (0.201 g, 0.460 mmol), 5-pyrimidine boronic acid (0.073 g, 0.587 mmol), bis(triphenylphosphino)palladium(ll) chloride (0.016 g, 0.0232 mmol}, triphenylphosphine (0.012 g, 0.047 mmol) and potassium carbonate (0.139 g, 1.37 rnmol) in 5:1 dioxane/water is heated at 100 °C for 3.5 h, cooled to room temperature and concentrated. The resultant residue is purified by flash chromatography (silica, 96:4:0.5 methylene chloride/methanoi/concentrated ammonium hydroxide as eluent) to afford the title compound as an off-white solid, 0.037 g (19%yieid), mp 120-130 "C; identified by NMR and mass spectral analyses. 1H NMR (500 MHz, CD3OD) Q9.12 (s, 1H), 9.02 (s, 2H), 7.81 (s, 1H), 7.67-7.62 (m, 2H), 7.56-7.51 (m, 3H), 7.47 (d, J = 1.1 Hz, 1H), 7.27 (d, J = 1.5 Hz, 1H), 7.23 (d, J = 8.3 Hz, 2H); IR (ATR) 3143,1672, 1505,1443, 1414, 1253, 1216, 1159, 79*1, 723 cm-1; ESI MS m/z437 [C22H15F3N6O + Hf
EXAMPLE 6 Preparation of 1-(3-Bromo-4-fluorophenv[)-1-cvanomethanol
(Figure Remove)
2} NaCN
A solution of sodium bisulfite (28.3 g, 271 mmol) in water at 50 °C is treated with 3-bromo-4-fluorobenzaldehyde (45.8 g, 225 mmol), stirred at 50 °C for 2 h, cooled with an ice bath, diluted with ether, treated dropwise with an aqueous solution of sodium cyanide (12.2 g, 248 mmol) over a period of 30 min and stirred at room temperature overnight. The reaction mixture is separated, and the aqueous phase is extracted with ether. The extracts are combined with the organic phase, washed with brine, dried over magnesium sulfate and concentrated to dryness to afford the title compound as a clear oil, 47.6 g (92%yield), identified by NMR analysis. 1H NMR (300 MHz, CDCI3) 5 7.75 (m, 1H), 7.47 (m, 1H), 7.23 (m, Hi), 5.54 (m, 1H), 3.19 (m, Hi).
EXAMPLE 7
Preparation of Ethyl 2-(3-Bromo-4-fluorophenvn-2-hvdroxvethanimidoate Hvdrochloride
H'' HCI
OC2H5
C2H5OH HCI
(Figure Remove)
A solution of 1-(3-bromo-4-fluorophenyi)-1-cyanomethanol (47.5 g, 206 mmol) and ethanol (10.9 g, 237 mmol) in ether is cooled with an ice bath and treated dropwise with HCI (258 ml_ of a 1.0 M solution in diethyl ether, 258 mmol) over a period of 40 min., stirred at ice-bath temperature for 2 h, stored at 0°C for 6 days, warmed to room temperature, diluted with hexanes and filtered. The filtercake is dried to afford the title compound as a white solid, 39.8 g (62% yield), identified by NMR and mass spectral analyses. The title compound is a mixture of E and Z isomers. 1H NMR (300 MHz, CD3OD) 5 7.78 (m, 1H), 7.49 (m, 1H), 7.28 (m, 1H), 5.54 and 5.17 (2m. 1H), 4.45 and 4.15 (2m, 2H), 1.38 and 1.20 (2t, 3H); ESI MS m/z 261 [C10H11BrFNO2 + H]*
EXAMPLE 8 Preparation of 1-(3-Bromo-4-fluorophenvn-1-(2-tetrahvdropy^
(Figure Remove)
OC2H5:
-NH2
A mixture of ethyl 2-(3-bromo-4-fluorophenyl)-2-hydroxyethanimidoate Hydrochloride (39.8 g, 127 mmo!} and 1,3-diaminopropane (9.43 g, 127 mmol) in ethanoi is heated at 120°C in a sealed tube overnight, cooled to room temperature, concentrated to remove the solvent, diluted with water, stirred vigorously for 1 h and filtered. The filtrate is cooled with an ice bath, made strongly basic with 1 N NaOH, cooled for 1 h in an ice bath and filtered. The filtercake is dried to afford th title compound as a white solid 23.4 g (64% yield), identified by NMR and mass spectral analyses. ''H NMR (500 MHz, CDCI3) 5 7.59 (m, 1H), 7.30 (m, 1H), 7.10 (t, J = 8.4 Hz, 1H), 4.79 (s, 1H), 3.35 (m, 4H), 1.76 (m, 2H); ESI MS m/z 287 [CnH
HI*.
EXAMPLE 9 Preparation of 2-(3-Bromo-4-fluorobenzovl)-2,3,4,5-tetrahvdropvrimidine
MnO2
(Figure Remove)
A mixture of 1-(3-bromo-4-fluorophenyl)-1-{2-tetrahydropyrimidinyl)methanol (23.4 g, 81.5 mmol) and manganese dioxide (70.8 g, 815 mmol) in methylene chloride is stirred at room temperature for 3 days and filtered through diatomaceous earth. The filtercake is washed with chloroform. The filtrates are combined and concentrated to dryness to afford the title compound as a yellow-green solid, 18.9 g (81% yield), identified by NMR and mass spectral analyses. 1H NMR (500 MHz,
CDCI3) 58.51 (m, 1H), 8.23 (m, 1H), 7.15 (t, J = 8.4 Hz, 1H), 6.10 (br s, lH),3.65(br s, 2H), 3.41 (brs, 2H), 1.83(m, 2H); ESI MS m/z284 [C,,H10BrFN2O+ Hf.
EXAMPLE 10
Preparation of 1-(3-Bromo-4-fluorophenvl)-1-itetrahvdropyrimidin-2-vl)-1-r4-{trifluoromethoxv)phenvl]methanol
MgBr
OCF3
Cul
(Figure Remove)
OCF-,
A mixture of magnesium (2.13 g, 87.7 mmol) in THF at 50 °C is treated dropwise with a solution of 1-bromo-4-(trifluoromethoxy)benzene (21.1 g, 87.7 mmol) in THF over a period of 20 min., stirred at 50 °C for an additional 1.5 h, cooled to room temperature, treated with copper(l) iodide (0.13 g, 0.70 mmol) and a solution of 2-(3-bromo-4-fluorobenzoyl)-2,3,4,5-tetrahydropyrimidine (10.0 g, 14.9 mmol) in THF, heated at 65 °C overnight, cooled to room temperature and diluted with ethyl acetate and saturated aqueous ammonium chloride. The phases are separated. The organic phase is washed sequentially with water and brine, dried over sodium sulfate. and concentrated to afford 18.5 g of a brown oil. The oil is purified by flash chromatography (silica, 90:10:0.5 methylene chloride/methanol/concentrated ammonium hydroxide as eluent) to afford the title compound as a yellow oil, 10.0 g (64% yield), identified by NMR and mass spectral analyses. 1H NMR (300 MHz, CDCI3) 5 7.60 (dd, J = 6.4, 2.3 Hz, 1H), 7.41 (dd, J - 6.9, 2.0 Hz, 2H), 7.28-7.20 (m, 3H), 7.10 (t, J = 8.3 Hz, 1H), 3.50 (m, 6H), 1.96 (m, 2H); ESI MS m/z447 [C18H15BrF4N2O2 + H]*.
EXAMPLE 11 -4-f luorophenyl).1 -
rim idi n-2-y|) -1 -[4.
(trifluoromethoxv)phenvnrnethvlaminp.
(Figure Remove)
OCF-i
1)SOCI2
.—— to,
2)NH3
OCF-,
A mixture of 1-(3-bromo-4-fluorophenyl)-l-(tetrahydropyrimidin-2-yl}-1-[4-(trifluoromethoxy)phenyl]methanol (4.60 g, 10.3 mmol) and thionyl chloride (12.2 g, 103 mmol) in toluene is heated at 110 °C overnight, cooled to room temperature and concentrated to dryness to give a tan solid residue. The residue is dispersed in isopropanol and bubbled through with ammonia gas until the mixture is saturated with ammonia. The saturated mixture is heated in a sealed tube at 45 °C overnight, cooled to room temperature and concentrated. The resultant residue is partitioned '' between chloroform and 1 N NaOH. The aqueous phase is separated and extracted with chloroform. The extracts are combined with the organic phase, dried over potassium carbonate and concentrated to afford the title compound as a dark oil, 0.89 g (>100%, -80% purity), identified by NMR and mass spectral analyses. 1H NMR (300 MHz, CDCI3) 5 7.65 (m, 1H), 7.41-7.303 (m, 36H), 3.41 (br s, 4H), 2.32 (br s, 2H), 1.77 (m, 2H); ESI MS m/z446 [C,8H16BrF4N3O + Hf.
EXAMPLE 12
Preparation of 8-13-Brcmo-4-fluorophenvl)-8-f4-(tnTluoromethoxy)pheiQyj]-2J3,4,8-t_etr_a.hydrqlrnidazpf1,5-alpvrirnidin-6-ylarni_ne
OCF-,
OCF3
BrCN CH3CN
(Figure Remove)
A mixture of 1-(3-romo-4-fluorophenyl)-1-(tetrahydropyrimidin-2-yl)-1-{4-(trifluoromethoxy)phenyl]methylamine (0.85 g, ~80% purity, -1.52 mmol) and cyanogen bromide (0.81 g, 7.62 mmol) in acetonitrile is stirred at room temperature for 45 rnin, heated at 100 "C in a sealed tube overnight, cooled to room temperature and concentrated. The resultant residue is purified by flash chromatography (silica, 95:5:0.25 methylene chloride/methanol/concentrated ammonium hydroxide as eluent) to afford the title compound as a tan solid, 0.26 g (36% yield), identified by NMR and mass spectral analyses. 1H NMR (300 MHz, CDCI3) 5 7.74 (dd, J = 6.7, 2.3 Hz, 1H), 7.49 (dd. J= 6.7, 2.1 Hz, 2H), 7.38 (m, 1H), 7.13 (d, J- 8.1 Hz, 2H). 7.03 (t, J= 8.5 Hz, 1H), 3.58 (m, 4H), 1.86 (m, 2H); ESI MS m/z471 [C19H15BrF4N4O + HT
EXAMPLE 13
Preparation of 8-(f(4-Fluoro-3-pvrimidln-5-y()phenyn-8-f4-(trifluorornethoxv)phenvn-2,3.4.8-tetrahydroimidazof1.5-a1pyrimidin-6-vl>amine
OCF3
(Figure Remove)
A mixture of 8-(3-bromo-4-fluorophenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-tetrahydroimida2o[1,5-a]pyrimidin-6-ylamine (0.26 g, 0.552 mmol), 5-pyrimidine boronic acid (0.082 g, 0,662 mmol), ietra(kistriphenylphosphino)-palladium(O) (0.032 g. 0.0276 mmo!) and potassium carbonate (0.23 g, 1.65 mrnol) in 5:1 dioxane/water is heated at 100 °C for 1 h, treated with additional tetra(kistriphenylphosphino)palladium{0) (0.032 g, 0.0276 mmol), heated at 100 QC for 3.5 h, cooled to room temperature and concentrated. The resultant residue is purified by flash chromatography (silica, 95:5:0.25 methylene chloride/methanol/ concentrated ammonium hydroxide as eluent) to afford the title compound as an off-white solid, 0.079 g (30% yield), mp 105-115 °C, identified by NMR and mass spectral analyses. 1H NMR (300 MHz, CD3OD) Q9.14 (s, 1H), 8.97 (s, 2H), 7.55 (m, 2H), 7.43 (dd. J = 6.8, 2.1 Hz, 2H), 7.21-7.31 (m, 3H), 3.69 (m, 2H), 3.50 (m, 2H), 1.87 (m, 2H); ESI MS m/z471 [C23H18 F4NeO+ H]*;
EXAMPLE 14
Preparation of (8-f4-Fiuoro-3-(5-fluoropvrim}din-3-vl)phenvn-8-f4-(trifluoromethoxv)phenvn-2.3.4.8-tetrahvctroimidazof1.5-alDvrimidin-6-Yl}amtng
''OCF3
OCF3
H2(Figure Remove)
A mixture of 8-(3-bromo-4-fluorophenyl)-8-{4-(trifluorometho)Qr)phenyl]-2l3,4,8-tetrahydroimidazo[1,5-a]pyrimidin-6-ylamine (0.075 g, 0.159 mmol), 3-fluoro-5-{tributy!stannyl)pyridine (0.092 g, 0.239 mrnol), and dichlorobis(tripheny1phos-phine)palladium(ll) (0.006 g, 0.008 mmol) in DMF is degassed, heated at 150 °C in a sealed tube for 1.5 h, cooled to room temperature and diluted with ethyl acetate (50 ml) and 5% aqueous LiCI. The reaction mixture is separated. The organic phase is washed with 5% aqueous Lid, dried over sodium sulfate and concentrated. The resultant residue is purified by flash chromatography (silica, 95.5:0.25 methylene
chloride/methanol/ concentrated ammonium hydroxide as e!uent) to afford the title product as a white solid, 0.043 g (56% yield), mp 94-105 °C; identified by NMR and
1,/t-no ''u MI\/ID conn IUU-T ,-^n orv\ no c« //-< f — -t o i_i— -< < n
M IOOO oi''^*-''l*tJ
,jro-j. . . . «i..i> ,w,v.v, ,,.. ./.. .^^v.-L.,; ..^._._.^ Vu, j - , .0 , i^, niy,
(d, J = 2.6 Hz, 1H), 7.83 (d, J - 9.7 Hz, 1H), 7.54-7.44 (m, 4H), 7.41-7.21 (m. 3H), 3.69 (m, 2H), 3.50 (m, 2H), 1.87 (m, 2H); ESI MS m/z488 [C24H1S F5N5O ^ H]*;
EXAMPLES 15-28 Preparation of Diphenylimidazopyrimidinvlarnine Derivatives
(Figure Remove)
Using essentially the same procedures described in Examples 5, 13 and14 hereinabove and employing the appropriate azacyclic reagent wherein W is B(OH)2 or Sn(n-Bu}3, the compounds on Table I were obtained and identified by NMR and mass spectral analyses.
(Figure Remove)Table Remove
EXAMPLE 29
Preparation of 8-(3-Pvrimidin-5-vl-phenvO-8-(4-trifluoromethoxY-phenyl)-2,3.4.8-tetr3hvdro-Sm!d3Zo[1,5-alpyrJmidin-6-vl-cvanamide
(Figure Remove)
"OCF3 -OCF3
A mixture of ethylene glycol dimethyl ether, tris(dibenzylideneacetone)-dipalladium (0) (0.014 g, 16.0 pmol), triphenylphosphine (0.008 g, 32.0 ^mol) under a nitrogen atmosphere is stirred for 5 min., treated with 2 (0.153 g, 0.320 mmol), pyrimidine-5-boronic acid (0.047 g, 0.380 mmof), sodium carbonate (0.101 g, 0.96 mmol) and water (2 ml), heated at 85 °C for 1 hr, cooled to room temperature and concentrated, the resultant residue is purified by flash chromatography (silica, 97.5:2.5:0.5 methylene chloride/methanol/concentrated ammonium hydroxide as eluent) to afford the title compound as a white solid, 0.130 g (85% yield), mp 227-231 °C; identified by NMR and mass spectral analyses. 1H NMR (500 MHz, CD3OD) S 9.13 (s, 1H), 9.02 (s, 2H), 7.77 (d, J = 7.5 Hz, 1H), 7.71 (s, 1H), 7.62 (dd, J= 7.9, 7.6 Hz, 1 H) 7.57 (d, J = 8.0 Hz, 1H), 7.50 (d, J = 8.9 Hz, 2H), 7.33 (d, J = 8.3 Hz, 2H), 3.70 (m, 2H), 3.58 (m, 2H), 1.89 (m, 2H); IR (ATR) 3106, 2187, 1622, 1500, 1412, 1256. 1218, 1159cm''1; ESI MS m/z478 [C24H18F3N7O + H
EXAMPLE 30
Preparation of (8R)-8-f3-(2-fluoropvridin-3-vQphenvn-8-f4-(trifIuoronTethoxv)phenvn-2.3.4.8-tetrahvdroirnidazof1.5-a1pyrim?din-6-amine(A) and(8S)-8-f3-(2-fluoropvrid?n-3-vnphenvn-8-f4-ftrifluoromethoxy)phenvn-2.3.4.8-tetrahvdroimidazof1.5-alpvrirnidin-6-amine (B)
(Figure Remove)
OCF3
Chiral separation
HiN
OCF3
(A)
(Figure Remove)
N
''OCF,
A racemic mixture of 8-[3-(2-fluoropyridin-3-yl)phenyl]-8-[4-
(trif1uoromethoxy)phenyl]-2,3,4,8-tetrahydroimidazo[1,5-a]pyrimidin-6-amine.(0.740g, 1.57 mmol) is placed on a Chiralpak AD 5 x 50 cm column (90:10:0.1 heptane/ethanol/diethylamine as eluent). The second eiuting peak (fR = 26 min) is collected and concentrated to a pale yellow oil. The oil residue is re-dissolved in a minimal amount of methylene chloride, triturated with hexanes and filtered. The filtercake is dried under vacuum for 24 h to afford the title product A as an off-white solid, 0.299 g, mp 126-131 °C; [a]25^ -11.6° (c = 0.5 in MeOH); identified by NMR, infrared and mass spectral analyses. ''H NMR (500 MHz, CD3OD) 8 8.16 (dd, J = 4.8,1.1 Hz, 1H), 8.03-7.98 (m. 1H), 7.58 (s, 1H), 7.55-7.52 (m, 1H), 7.49-7.45 (m, 3H), 7.44-7.41 (m, 1 H), 7.40-7.37 (m, 1 H), 7.23 (d, J = 8.2 Hz, 2H), 3.70 (t, J = 6.0 Hz, 2H), 3.49 (t, J = 5.4 Hz, 2H), 1.90-1.85 (m, 2H); IR (ATR) 3062,2954,1654, 1602, 1504,1434,1251,1216,1159, 791 cm''1; ESI MS m/z470 [C24H19F4N5O + H]*;
The first eiuting peak (/R = 19 min) is collected and concentrated to a pale yellow oil. The oil residue is re-dissolved in a minimal amount of methylene chloride, triturated with hexanes and filtered. The filtercake is dried under vacuum for 24 h to afford the title product B as an off-white solid, 0.315 g, mp 124-128 °C; [ctj250: + 12.6° (c = 0.5 in MeOH); identified by NMR, infrared and mass spectral analyses. ''H NMR (500 MHz, CD3OD) 5 8,16 (dd, J = 4.8. 1.1 Hz, 1H), 8.03-7.98 (m, 1H), 7.58
(s, 111), 7.55-7.52 (m, 1H), 7.49-7.45 (m. 3H), 7.44--7.41 (m, 1H), 7.40-7.37 (m. 1H), 7.23 (d, J= 8.2 Hz, 2H), 3.70 (t, J= 6.0 Hz, 2H), 3.49 (t, J = 5.4 Hz, 2H), 1.90-1.85 (m, 2H); !R (AIR) 3064, 2947, 1653, 1602, 1504, 1434. 1251. 1216 1155 791 an"1-ESI MS m/z470 ^H^NsO + H]
EXAMPLE 31
Preparation of (8R)-8-(3-pvrimidin-5-vlphenvl)-8-[4-(trifluoromethoxv)ph6nvl1-2.3,4,8-tetrahvdroirnidazof1,5-a1pvrimidin-6-amine (A) and (8S)-8-(3-pvrimidin-5-vlphenvl)-8-f4-(trifluoromethpxv)phenvll-2.3.4.8-tetrahvdrQimidazori,5-a]pyrimidin-6-arnine (B)
OCF3
N
(Figure Remove)
''OCF3
Chira! separation
OCF3
(B)
A racemic mixture of 8-(3-pyrimidin-5-ylphenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3l4l8-tetrahydroimidazo[1,5-a]pyrimidin-6-amine_(2.0 g, 1.57 mmol) is placed on a Chiralpak AD 5 x 50 cm column (90:10:0.1 heptane/ethanol/ diethylamine as eluent). The first eluting peak (fo = 34 min) is collected and concentrated to give a pale yellow oil. The oil is re-dissolved in a minimal amount of methylene chloride, triturated with hexanes and filtered. The filtercake is dried under vacuum for 24 h to afford the title compound A as an off-white solid, 0.815 g, mp 178-186 °C; [a]25D: -14.7° (c = 0.50
in MeOH); identified by NMR, infrared and mass spectial analyses. 1H NMR (500 MHz, CD3OD) 5 9.12 (s, 1H). 9.02 (s, 2H). 7.70-7.67 (m, 2H), 7.57-7.51 (m, 2H), 7.48-7.44 (m, 2H), 7.26 (d, J = 8.7 Hz, 2H). 3.73 (t, J = 6.3 Hz, 2H). 3.55-3.52 (m. 2H), 1.92-1.88 (m, 2H); IR (ATR) 3040, 2956, 2859, 1655, 1504, 1413, 1253. 1160, 786 cm''1; ESI MS m/z453 [C23H19F3N5O + H]+;
The second eluting peak (tR = 46 min) is collected and concentrated to give a pale yellow oil. The oil residue is re-dissolved in a minimal amount of methylene chloride, and then triturated with hexanes and filtered. The filtercake is dried under vacuum for 24 h to afford the title compound B as an off-white solid, 0.798 g, mp 180-186 °C; [a}25D: +9.7° (c = 0.51 in MeOH), identified by NMR, infrared and mass spectral analyses. 1H NMR (500 MHz, CD3OD) 5 9.13 (s, 1H), 9.02 (s, 2H), 7.73-7.69 (m, 2H), 7.58-7.51 (m, 2H), 7.49-7.46 (m, 2H), 7.29-7.26 (m, 2H), 3.76 (t, J = 6.3 Hz, 2H), 3.57-3.55 (m, 2H), 1.93-1.90 (m, 2H); IR (ATR) 3040, 2955,1655, 1553, 1505, 1413, 1253, 1201, 1162, 786 cm''1; ESI MS m/z453 [C^M^NgO + Hf
EXAMPLE 32
Preparation of (8S)-8-f3-(5-ChloroDvridm-3-vnphenvn-8.r4-ftrifluoromethoxY)-phenvH-2.3.4.8-tetrahvdroim{dazori.5-a1pvrim8din-6-amine FA1 and r8ffl-8-f3^5-Chloropvridin"3-vl)phenvn-8-r4-(trifluoromethoxv^phenvn-2.3.4.8-tetrahydroimidazofl ,5-a1pyrimidin-6-amine TB1
.N
(Figure Remove)
H,N
OCF3
OCF3
(Figure Remove)
(Figure Remove)
A racemie rnixtue of 8-[3-(5-chloropyridin-3-yl)phenyl]-8-[4-
irifluoromethoxy)phenyl]-2,3,4,8-tctrahydroirnidazo[1 ,5-a]pyrimidin-6-amine (1 .73 g) vas separated into its enantiomfir.s usinn a Cnirainak AD 5 x 50 cm column 90:10:0. 1 heptarue/ethanol/diethylamine) to afford the title S-isomer (A) as an off-A/hite solid, mp 104-1 16°C; [a]25D: -8.6° (c - 0.51% in MeOH); 1H NMR (500 MHz, 3D3OD) 8 8.69 (d, J=1.9Hz, 1H), 8.53 (d, J= 2.2 Hz, 1H), 8.11 (t, J=2.1 Hz, 1H), 7.69-7.67 (m, 1H), 7.65 (t, J - 1.7 Hz, 1H), 7.54 (t, J = 7.6 Hz, 1H), 7.51-7.45 (m, 3H), 7.29 (d, J = 8.3 Hz, 2H), 3.77 (t. J = 5.9 Hz, 2H), 3.58-3.55 (m, 2H), 1.97-1.92 (m, 2H); ESI MS m/z486 ^H^CIFsNsO + H]+; and the title R-isomer (B) as an off-white solid, mp 114-118°C; [a]25D: +13.3° (c= 0.53% in MeOH); 1H NMR (500 MHz, CD3OD) O 8.69 (d, J = 1.9 Hz, 1H), 8.54 (d, J = 2.3 Hz, 1 H), 8.1 1 (t, J = 2.1 Hz, 1H), 7.70-7.67 (m, 1H), 7.65 (t, J= 1.6 Hz, 1H), 7.55 (t, J= 7.7 Hz, 1H), 7.52-7.46 (m. 3H), 7.29 (d, J = 8.2 Hz, 2H), 3.77 (t, J- 6.0 Hz. 2H), 3.59-3.56 (m, 2H), 1.95-1.90 (m, 2H); ESI MS m/z486 [C24H19CIF3N5O + H]+.
EXAMPLE 33
Preparation of (8S)-8-r3-(4-fluoropvridin-3-yt)phenyn-8-r4-(trifjuoromethoxv)-phenvll-2.3.4.o-tetrahydroim{dazof1.5-a]pvrimi''din-8-amine [A] and (8f?)-8-r3-f4-fluoropvridin-3-vl)phenvn-8-f 4-(trifluoromethoxy)phenvn-2. 3.4.8-tetrahvdroimidazof 1 ,5-alpyrimidin-6-amtne [Bl
OCF3
H2N.
"OCF,
,N
(Figure Remove)
OCF,
A racemic mixture of 8-[3-(4-fIuoropyridin-3-yl)phenyl]-8-[4-
(lrifluoromethoxy)phenyl]-2,3,4,8-tetrahydroimidazo[1,5-a]pyrimidti>6-amine (1.89 g) was separated into its enantiomers using a Chiraipak AD 5 x 50 cm column (93:7:0.1 heptane/ell >anoi/ diethyiamine) to give the title S-isomer (A) as an off-white solid (0.755g),mp 171 °C; [a]25D: +10.6° (c = 0.5% in MeOH); 1H NMR (500 MHz, CD3OD) 5 8.63 (d, J = 10.0 Hz, 1H), 8.52 (dd, J = 7.3, 5.7 Hz, 1H), 7.57-7.52 (m, 2H), 7.51-7.45 (m, 4H), 7.32 (dd, J = 10.3, 5.7 Hz, 1H), 7.24 (d, J = 8.1 Hz, 2H), 3.71 (t, J = 5.9 Hz, 2H), 3.50 (t, J = 5.5 Hz, 2H), 1.90-1.85 (m, 2H); ESI MS m/z470 [C24H13F4N5O + H]*; and the title R-isomer (B) as an off-white solid (0.675 g), mp 115-116 °C; [a]250: -10.9° (c= 0.5 % in MeOH); 1H NMR (500 MHz, CD3OD) Q 8.63 (d, J = 10.1 Hz, 1H), 8.53 (dd, J- 7.3, 5.7 Hz, 1H), 7.57-7.54 (m, 2H), 7.53-7.45 (m, 4H), 7.32 (dd. J = 10.3, 5.7 Hz, 1H), 7.26 (d, J= 8.2 Hz, 2H), 3.73 (t, J = 6.0 Hz, 2H), 3.52 (t, J = 5.7 Hz, 2H), 1.90 (quintet, J = 6.3 Hz, 2H; ESI MS m/z 470 [C24H19F4N50 + Hf.
EXAMPLE 34
Preparation of 8-f3-(2-Ruoro 1-oxy-pvridin-3-vn-phenvl|-8-(4-trif(uoromethoxv-phenvl)-2.3.4,8-tetrahvdro-im»dazoI1,5-alpyrimidin-6-vfamme
''OCF3
(Boc)jO, DMA?
THF
''OCF3
(Figure Remove)
urea-hydrogen peroxide con^lcx
TFAA CH2a2,40°C
HC1
''OCF3
-OCF3
Step a) Preparation of compound 2.
A mixture of 1 (0.580 g, 1.24 mmol), di-terf-butyl dicarbonale (0.670 g, 3.10 mmol) and 4-dimethylaminopyridine (0.151 g, 1.24 mmol) in tetrahydrofuran (15 ml) was stirred at room temperature for 1 h. The mixture was then diluted with melhylene chloride (75 ml_), washed with 1 M citric acid (25 mL), water (25 ml) anc
orine (25 ml), dried over sodium sulfate, filtered and concentrated to afford 2 (0.74 g, 89%) as a colorless oil: 1H NMR (500 MHz, CDC!3) 8 8.17 (d, J = 7.1 Hz, 1H), 7.84 (m, 1H), 7.73 (s, 1H), 7.60-7.55 (m, 3H), 7.51-7.47 (m, 1H), 7.42 (t, J = 7.8 Hz, 1H), 7.26-7.22 (in, 1H), 7.17 (d, J= 8.1 Hz, 2H), 3.66-3.61 (m, 2H), 3.50 (t, J = 6.0 Hz, 2H), 1.88-1.83 (m, 2H), 1.33 (s, 18H); ESI MS m/zQ70 [C34H35F4N5O5 + Hf. Step b) Preparation of compound 3.
Trifluoroacetic anhydride (0.365 g, 1.74 mmol) was added dropwise to a stirred suspension of urea-hydrogen peroxide complex (0.169 g, 1.80 mmol) in methylene chloride (15 rr»L) at 0 °C. The mixture was stirred for 5 min and then a solution of 2 (0.200 g, 0.29 mmol) in methylene chloride (10 mL) was added dropwise. The reaction was warmed to room temperature and then heated at 40 °C for 45 min. After this time, the reaction was cooied to room temperature, diluted with methylene chloride (50 ml) and washed with saturated aqueous sodium bicarbonate (2 * 20 ml) and brine (20 mL), dried over sodium sulfate, filtered and concentrated. Purification by flash chromatography (silica, 97.5:2.5 to 95:5 methylene chloride/methanol) afforded 3 (0,086 g, 41%) as a colorless oil: ''H NMR (300 MHz, CDCI3) 8 8.25 (dt, J = 6.3, 1.5 Hz, 1H), 7.73 (s, 1H), 7.68-7.62 (m, 1H), 7.57 (d, J = 6.9 Hz, 2H), 7.49-7.43 (m, 2H), 7.40-7.33 (m, 1H), 7.20-7.13 (m, 3H), 3.64 (t, J = 5.4 Hz, 2H), 3.50 (t, J = 5.8 Hz, 2H), 1.90-1.81 (m, 2H), 1.45 (s, 18H); ESI MS m/z 686 [C^Hg^NgOe + Hf. Step c Preparation of 8-[3-(2-FIuoro 1-oxy-pyridin-3-yl)-phenyIl-8-(4-
*
trifluoromethoxy-phenyl)-2,3,4,8-tetrahydro-imidazo[1,5-a3pyrimidin-6-ylamine
A mixture of 3 (0.08 g, 0.11 mmol) and 4 M HCI/dioxane (5 mL) was stirred at room temperature for 20 h. The solvents were evaporated and the residue diluted with saturated aqueous sodium bicarbonate (15 mL) and methylene chloride (20 mL). The layers were separated and the organic layer washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated. The crude product was purified by preparative HPLC. The appropriate fractions were combined, concentrated, then neutralized with saturated aqueous sodium bicarbonate (10 mL) and extracted with methylene chloride (3*10 mL). The combined organic layers were washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated. The residue was freeze dried from acetonitrile/water (8 mL, 1:1) to afford the title product as a white solid, 0.018 g (32% yield), 1H NMR (300 MHz, CDCI3) o 8.24 (dt, J = 6.3,1.6 Hz, 1H),
7.74 (s, 1H), 7.62-7.53 (m, 3H), 7.48-7.32 (m, 3H), 7.20-7.12 (m, 3H), 3.66 (t, J = 5.9 Hz, 2H), 3.60 (t, J = 5.5 Hz, 2H), 1.88 (t, J - 5.7 Hz, 2H); ESI MS m/z486 [C;14H19F4N5O2 + H]+.
EXAMPLE 35
Preparation of 7-I3-(2-Fluoro-pvridin-3-vl)-phenvl]-7-(4-trifluoromethoxv-phenYr 2.7-dihvdro-3H-lmidazof1,5-a1imidazol-5-vlamine
MgBr
CN F3CO
2. NH4C1, NH4OH
OCF,
NaBH4
MeOH
OCF3
CSC12
sat. aq. NaHCOj CH2C12
NCS
OCF3
t-BuOK, €82
THF
H?N
,NH,
EtOH,70°C ''OCF overnight
OCF-,
t-BuOOH
NHjOH, MeOH rt, overnight
,N
OCF3
cc
Na2CO3 (Figure Remove)
:1DME/H2O, reflux
OCF,
Step a) Preparation of compound 2
A mixture of magnesium (0.60 g, 24.7 mmol) in THF (6 ml_) was heated to 5 °C and treated dropwise with a solution of 1-bromo-4-(trifluoromethoxy)-benzene (5.96 g, 24.7 mmol) in THF (18 ml) over a period of 10 min. After stirring at 50 °C I an additional 1.5 h, the mixture was cooled to room temperature and treated with a solution of 1 (3.0 g, 16.5 mmol) in THF (12 ml). The mixture was then reheated to
65 "C for 1 h. After this time, the reaction mixture was cooled to room temperature 3rid poured onto a solution of saturated aqueous ammonium chloride (20 ml) and cone, ammonium hydroxide (20 mL) at -15 °C and stirred for 5 min. This mixture was then filtered through a pad of celite 521 with ether (100 ml). The organic layer in Ihe filtrate was separated, washed with brine (50 ml), dried over magnesium sulfate, filtered and concentrated to afford the crude tmine (3.90 g, 68%) as an amber oil. A solution of this crude imine (3.90 g, 11.3 mmol) in MeOH (20 rnL) was cooled with an ice bath and treated with sodium borohydride (0.86 g, 22.7 mmol). The cooling bath was removed and the mixture stirred at room temperature for 3 h. After this time the mixture was concentrated and partitioned between 1 N NaOH (100 mL) and methylene chloride (100 ml). The organic layer was separated and washed with brine (100 ml), dried over potassium carbonate, filtered and concentrated. Purification by flash chromatography (silica, 1:4 ethyl acetate/hexanes) afforded 2 (1.98 g, 34% over 2 steps): 1H NMR (300 MHz, CDC13) 5 7.56 (t, J = 1.7 Hz, 1H), 7.40-7.15 (m, 7H), 5.19 (s, 1H); ESI MS m/z 329 [CieH15BrF4N2O2 - NH2+ Hf. Step b) Preparation of compound 3
A mixture of 2 (0.66 g, 1.91 mmol) in methylene chloride (2 mL) and saturated aqueous sodium bicarbonate (2 ml) was cooled with an ice bath, treated with thiophosgene (0.24 g, 2,10 mmol) and stirred vigorously for 30 min.. The organic layer was separated, washed with brine (2 ml), dried over sodium sulfate and concentrated to afford 3 (0.74 g, 100%) as a yellow oil: 1H NMR (300 MHz, CDCI3) 6 7.49-7.22 (m, 8H), 5.97 (s, 1H). Step c) Preparation of compound 4
To a mixture of potassium f-butoxide (0.070 g, 0.623 mrnol) in tetrahydrofuran (2 mL) at -78 °C was added dropwise a solution of 3 (0.220 g, 0.567 mmol) and carbon disulfide (0.065 g, 0.850 mmol) in tetrahydrofuran (3 ml). The reaction was stirred at -78 °C for 0.5 h, then warmed to room temperature slowly and stirred overnight at room temperature. The reaction was then diluted with ethyl acetate (50 mL) and water (10 mL). The organic layer was separated, washed with brine (10 mL}, dried over sodium sulfate and concentrated, to afford 4 (0.26 g, 99%) as a clear oil: "H NMR (300 MHz, CDCI3) 5 7.86-7.10 (m, 8H), 3.70 (s, br, 1H). Step d) Preparation of compound 5
A solution of 4 (0.850 g, 1.83 mmol) and ethylenediamine (0.330 g, 5.49 mmol) in ethanol (15 ml) was heated overnight at 70 °C. The reaction was cooled to room temperature and concentrated. Purification by flash chromatography (silica, 1:4 ethyl acetate/hexanes) afforded 5 (0.45 g, 54%) as a white solid: 1H NMR (300 MHz, CDCI3) 5 8.50 (s, 1H), 7.59-7.19 (m, 8H), 4.46 (t, J = 8.5 Hz, 2H), 3.87 (t, J = 8.5 Hz, 2H), ESI MS m/z456 [C,aH13BrF3N3OS + H]*. Step e) Preparation of compound 6
A mixture of 5 (0.200 g, 0.438 mmo!) and f-butyl hydroperoxide (0.79 g of a 70% solution in water, 8,80 mmol) in methanol (20 mL) and concentrated aqueous ammonium hydroxide (4 ml) was stirred overnight at room temperature. The reaction was then concentrated. Purification by flash chromatography (silica, 95:5.0.25 methylene chloride/methanot/concentrated ammonium hydroxide) afforded 6 (0.156 g, 81%) as a white solid: 1H NMR (300 MHz, CD3OD) 5 7.58 (t, J = 1.5 Hz, 1H), 7.49-7.44 (m, 3H), 7.36 (dt, J= 7.8,1.5 Hz, 1H), 7.29-7.23 (m, 3H), 4.41 (t, J = 8.7 Hz, 2H), 3.75 (t, J = 8.7 Hz, 2H); ESI MS m/z440 [C18H14BrF3N4O + Hf. Step f) Preparation of 7-{3-(2-f!uoro-pyridin-3-yl)-phenyl]-7-(4-trifluoromethoxy-phenyi)-2,7-dihydro-3H-im(dazo[1,5-a3imidazoN5-y!amme
A mixture of 6 (0.070 g, 0.159 mmo!), triphenylphosphine (0.004 g, 0.016 mmol), bis(dibenzylideneacetone)paJladium{0) (0.007 g, 0.008 mmol), sodium carbonate (0.051 g, 0.478 mmol) and 2-fluoro-3-boronic acid (0.040 g, 0.287 mmol) in ethylene glycol dimethyl ether (6 ml) and water (2 mL) was degassed and heated at 80 °C for 2.5 h. The mixture was cooled to room temperature and diluted with ethyl acetate (100 ml) and water (50 ml). The organic layer was separated, washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated. Purification by flash chromatography (silica, 93:7:0.25 methylene chloride/methanol/concentrated ammonium hydroxide) and then by preparative HPLC afforded a mixture of starting material and product A mixture of this material (0.032 g, 0.073 mmol), triphenylphosphine (0.002 g, 0.007 mmol), bis(dibenzylideneacetone)pa!ladium(0) (0.003 g, 0.004 mmol), sodium carbonate (0.023 g, 0.220 mmol) and 2-fluoro-3-boronic acid (0.019 g, 0.131 mrnol) in ethylene glycol dimethyl ether (6 mL) and water (2 mL) was degassed and heated at 80 °C overnight. The organic layer was separated, washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated. Purification by flash chromatography (silica, 95:5:0.25 methylene
chlonde/methanol/concentrated ammonium hydroxide) afforded the title product as a white solid, 0.030 cj (41% yielctt, nip 95-100 "C; ''H NMR (500 MHz, CD3OD) 8.17 (dt, .7 -- 4 !r 1.0 Hz: 1H), 8.02 (ddd: J= 7.5. 2 0. 2.0 Hz, 1H).. 7.64-7.38 (m. 9HV 7.2R M. J = 8.0 Hz, 2H), 4.43 (d, J = 9.0 Hz, 2H), 3.78 (d, J = 9.0 Hz, 2H); IR (ATR) 2925, 1644, 1601, 1504, 1450, 1434, 1400, 1250, 1211, 1 157, 1010, 966, 791 cm''1; ESI MS m/z456 [C?3H17F4N5O+ H]*.
EXAMPLE 36
Preparation of 9-f3-(2-Fluoro-pyridin-3-yn-phenvn-9-(4-trifluoromethoxy-phenyn:2A5^-tetrahydrQr3H-irnida2Q[1,5-alf1,3ldiazepin-7-vlaminie
H-,N
OCF,
OCFi
/-BuOOH
NH4OH,
H2N
.N
(Figure Remove)
(Figure Remove)
OCF3
B(OH)2
OCF3
Step a) Preparation of compound 2,
A solution of 1 (0.50 g, 1.08 mmo!) and 1,4-diarnino propane (0.28 g, 3.23 mmol) in ethanol was heated at 70 °C for 18 h, cooled to room temperature and concentrated in vacuo. The concentrate was partitioned between ethyl acetate and water. The organic layer was separated, washed with brine, dried over magnesium sulfate and concentrated. Purification of the resultant residue by flash chromatography (silica, 1:4 ethyl acetate/hexanes) afforded 2 (0.22 g, 42%) as a white foam: ''H NMR (300 MHz, CDCI3) 5 7.47 (m, 2H), 7.33 (d, J = 8.9 Hz, 2H), 7.25-7.18 (m, 4H), 4.15 (m, 2H), 3.81 (m. 2H), 1.99 (m. 4H); ESI MS m/z484 [C?0Hv/BrF3N3OS + H]''.
Stop b) Preparation of compound 3
A mixture of 2 (0.22 g, 0.454 mmol) and ^-butyl hydroperoxide (1.17 g of a 70% solution in water, 9.08 mmol) in methanol and concentrated aqueous ammonium hydroxide (4.4 ml) was stirred overnight at room temperature, treated with 10% aqueous sodium thiosulfate (30 ml_) and concentrated to remove most of the methanol. Tiie remaining aqueous mixture was extracted with methyiene chloride. The methyiene chloride extracts were combined, washed with brine, dried over magnesium sulfate, and concentrated. Purification of the resultant residue by flash chromatography (silica, 95:5:0.25 methyiene chloride/methanoi/concentrated ammonium hydroxide) afforded 3 (0.136 g, 65%) as a white foam: 1H NMR (300 MHz, CDCI3) 5 7.63 (t, J= 1.7 Hz, 1H), 7.46 (dd, J = 6.8, 2.0 Hz, 2H), 7.39-7.33 (m, 2H), 7.19-7.12 (m, 3H), 3.73 (m, 2H), 3.57 (m, 2H), 1.93 {m. 4H); ESI MS m/z467 [C20HiaBrF3N40 + Hf.
Step c) Preparation of 9-[3-(2-FIuoro-pyridin-3-yl}-phenyl]-9-{4-trifluorometh-oxyphenyl)-2A5,9-tetrahydro-3H-imidazo[1,5-a][1,3]diazepin-7-ylamine
A mixture of 3 (0.065 g, 0.139 rnmoi), 2-fluoropyridine-3-boronfc acid (0.035 g, 0.250 mmol), bis(triphenytphosphino)pa!ladium(ll) chloride (0.0049 g, 0.007 mmol), triphenylphosphine (0.0036 g, 0.014 mmol) and sodium carbonate (0.044 g, 0.417 mmol) in 3:1 DME/waterwas heated at reflux temperature for 1 h, cooled to room temperature and diluted with ethyl acetate and water. The organic layer was separated, washed with brine, dried over magnesium sulfate and concentrated. Purification of the resultant residue by flash chromatography (silica, 95*:5:0.25 methyiene chloride/methanol/concentrated ammonium hydroxide) afforded 0.049 g of a white foam. This material was freeze dried from 2:1 acetonitrile/water to afford the title product as a white solid, 0.041 g (62% yield), mp 88-97 °C; 1H NMR (500 MHz, CD3OD) 5 8.16 (d, J = 4.3 Hz, 1H), 8.00 (m, 1H), 7.59-7.37 (m, 7H), 7.22 (d, J - 8.7 Hz, 2H), 3.73-3.64 (m, 4H), 1.96 (m, 4H); ESI MS m/z 484 [C25H2iF4N5O + Hf;
EXAMPLE 37
Preparation of^^1-f3-(2-Fluoro-pvridin-3-vO-phenvn-1-(4-trifluoromeUioxv-phenvl)-1.4.5,6r7,8-hexahvdro-2,3a.9-triaza-cvclopentacvcloocten-3-vlamine
HI-
(Figure Remove)
MiuOOH NH4OH
OCF3
,N
OCF3
B(OH)2
OCFi
Step a) Preparation of compound 2
A solution of 1 (0.50 g, 1.08 mrnol) and 1,5-diarnino pentane (0.33g, 3.23 mmol) in ethanol was heated at 70 °C for 5 h, then at 100 °C for 17 h and finally at 120 °C for 6 h. The reaction was cooled to room temperature, concentrated and the concentrate was partitioned between ethyl acetate and water. The organic layer was separated, washed with, dried over magnesium sulfate and concentrated. Purification of the resultant residue by flash chromatography (silica, 1:4 ethyl acetate/hexanes) afforded 2 (0.153 g, 28%) as a white foam: 1H NMR (300 MHz, CDCI3) 8 7.54 (t, J = 1.7 Hz, 1H)), 7.50-7.19 (m, 7H), 4.45 (t, J = 6.6 Hz, 2H), 4.01 (t. J = 6.6 Hz, 2H), 1.96-1.86 (m, 4H), 1.48-1.44 (m, 2H); ESI MS m/z 49R [C21H19BrF3N3OS + H]+. Step b) Preparation of compound 3
A mixture of 2 (0.15 g, 0.301 mmol) and f-butyl hydroperoxide (0.77 g of a 70% solution in water, 6.02 mmol) in methanol and concentrated aqueous ammonium hydroxide (3 mL) was stirred overnight at room temperature treated with 10% aqueous sodium thiosulfate (20 ml) and concentrated to remove most of the methanol. The remaining aqueous mixture was extracted with methylene chloride. The methylene chloride extracts were combined, washed with brine, dried over magnesium sulfate and concentrated. Purification of the resultant residue by flash chromatography (silica, 95:5:0.25 methylene chloride/methanol/concentrated ammonium hydroxide) afforded 3 (0.073 g. 52%) as a white foam: \\ NMR (300
MHz, CDC!3) S 7.63 (I, J = 1 .3 Hz, 1H), 7 49 (dd, J = 6.8, 2.0 Hz, 2H), 7.49-7.37 (m,
2H), 7.20-7.13 (m, 3H), 3.96 (m, 4H), 1.90 (m, 4H), 1.56 (m, 2H); ESI MS m/z482
!C3iH2nBrF,NN
N
"N
O O"
3 N HC1
(Figure Remove)
CHC1F-,
FCOH, /-PrOH
OCHF2
A mixture of 8-[3-(2-fiuoropyridin-3-yl)phenyl]-8-(4-methoxymethoxyphenyl)-2,3,4,8-tetrahydroimida2o[1,5-a]pyrimidin-6-ylamine (1.10 g, 2.46 mmol) and 3 N hydrochloric acid (60 ml) in methanol was stirred at room temperature for 16 h, neutralized with aqueous sodium hydroxide and concentrated in vacuo. The residue obtained was triturated with ethanol, the solids removed by filtration and the filtrate concentrated. Purification of the concentrate by flash chromatography (silica, 90:10 methylene chloride/methanol) afforded a white solid (0.70 g, 70%). A 0.040 g sample of this solid was further purified by semi-preparative liquid chromatcgraphy to afford 4-{6-amino-8-[3-(2-fluoro-pyridin-3-yl)-phenyl]-2,3,4,8-tetrahydro-imidazo[1,5-a]pyrimidin-8-yl}-phenol as a white solid (0.0073 g), mp 164-181 °C; 1H NMR (500 MHz, CD3OD) 8 8.15 (d, J= 5.0 Hz, 1H), 8.03-7.97 (m, 1H), 7.57 (br s,«1H), 7.54-7.48 (m, 1H), 7.46-7.41 (m, 2H), 7.40-7.36 (m, 1H), 7.16 (d, J = 8.7 Hz, 2H), 6.73 (d, J = 8.7 Hz, 2H), 3.69 (t, J~ 6.0 Hz, 2H), 3.50-3.44 (m, 2H), 1.90-1.83 (m, 2H); ESI MS m/z 402 [CaHzoFNgO + H]*.
A mixture of 4-{6-amino-8-[3-(2-fluoro-pyridin-3-yl)-phenylJ-2,3,4,8-tetrahydro-imidazo[1,5-a]pyrimidin-8-yl}-phenol (0.285 g, 0.710 mmol) and potassium hydroxide (0.396 g, 7.10 mmol) in 2-propanol was stirred for 10 min at room temperature, cooled to -45 °C, bubbled with FREON 22 (6.7 g. 77.0 mmol) and sealed. The sealed reaction mixture was warmed to room temperature, gradually warmed to 50 °C, stirred for 90 min at 50 °C. cooled to room temperature, unsealed and quenched by carefully adding the reaction mixture to water. The aqueous mixture was diluted with methylene chloride and the layers separated. The organic layer was washed with brine, dried aver sodium sulfate and concentrated. Purification of this residue by flash
chromatography (silica, 95:5:0.25 methylene chloride/methanol/concentrated ammonium hydroxide) afforded an off-white solid (0.055 g, 17%). This solid was further purified by semi-preparative LC to afford (he title product as a white solid, (0.033 g), mp 104-114 °C; 1H NMR (500 MHz, CDCI3) 5 8.16 (dt, J - 4.7, 1.3 Hz, 1H), 7.85-7.80 (m, 1H), 7.69(d, J= 1.4 Hz, 1H), 7.53-7.45 (m, 4H), 7.40 (t, J = 7.7 Hz, 1H), 7.25-7.21 (m, 1H), 7.05 (d, J= 8.7 Hz, 2H),6.47 (t, J= 74.0 Hz, 1H), 3.63-3.55 (m, 4H), 1.91-1.84 (m, 2H); ESI MS m/z 452 [C24H,oF3N5O •*• H]*.
EXAMPLE 42
Preparation of 7-f3-<2-Fluoropyridin-3-vOph6nvll-2,2-dirnethyf-7-f4-trifluoromethoxvphenv})-2.7-dihvdro-3H-imldazQF1,5--alirnjdazol-5-vlamme
(Figure Remove)
OCF3
H
,CH3 NH2
EtOH,0-40°C
OCF,
f-BuOOH
5:1 MeOH/NH4OH
OCFi
Na2CO3 (Figure Remove)
DME,H2O
OCF3
Step a) Preparation of compound 2
A solution of 1 (0.250 g, 0.54 mmol) in ethanol at 0 °C was treated with 2-methylpropane-1,2-diamine (0.144 g, 1.62 mmol), stirred at 0 °C for 3 h , then at room temperature for 45 minutes, heated to 40 °C for 2h and concentrated in vacua. The resultant residue was partitioned between ethyl acetate and water. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated. This residue was purified by flash chromatography (silica, gradient 100% hexanes to 1:9 ethyl acetate/hexanes) to afford 2 as a white solid. 1H NMR (300 MHz. CDCI3) 5 7.73 (s, 1H). 7.56 (t. J = 1.8 Hz, 1H ). 7.49 {dt, J = 7.7. 1.3 Hz, 1H), 7.43 (t, J = 3,0
Hz. 1H), 7.40 (t, J = 2.2 Hz, 1H), 7.34-7.21 (m. 4H), 3.60 (s, 2H), 1.44 (s, 3H), 1.43 (s, 3H); ESI MS m/z484 [C30H17BrF3N3OS + H]+. Step b) Preparation of compound 3.
A mixture of 2 (0.201 g, 0.42 mmol) and f-butyl hydroperoxide (0.75 g of a 70% solution in water, 8.30 mmol) in methanol and concentrated aqueous ammonium hydroxide (5 mL) was stirred overnight at room temperature, treated 10% aqueous sodium thiosulfate (30 ml) and concentrated to remove most of-the methanol. The remaining aqueous mixture was extracted with methylene chloride. The methylene chloride extracts were combined, washed with brine, dried over sodium sulfate and concentrated. Purification of the residue by flash chromatography (silica, 97:2.5:0.5 methylene chloride/methanol/concentrated ammonium hydroxide) afforded 3 (0.134 g. 68%) as a white solid: 1H NMR (300 MHz, CDCI3) 87.71 (t, J= 1.7 Hz, 1H ), 7.54 (d, J= 8.8 Hz, 2H), 7.43-7.37 (m, 2H), 7.19-7.13 (m, 3H), 3.36 (s, 2H), 1.42 (s, 6H); ESI MS m/z467 [C2oH18BrF3N4O + H]+. Step c) Preparation of 7-[3-(2-fluoropyridin-3-yl)phenyJ]-2,2-dimethyI-7-(4-triffuoromethoxyphenyl)-2,7-dihydro-3H-imidazo[1,5-a]imidazol-5-ylamine
A mixture of 3 (0.134 g, 0.29 mmol), 2-fluoropyridine-3-boronic acid (0.081 g, 0.57 mmol), bis(triphenylphosphino)palladium(l!) chloride (0.010 g, 0.015 mmol), triphenylphosphine (0.008 g, 0.029 mmol) and sodium carbonate (0.092 g, 0.87 mmol) in 3:1 DME/water (8.0 mL) was heated at 80 °C for 5 h, cooled to room temperature and diluted with ethyl acetate and water. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated. Purification of the resultant residue by flash chromatography (silica, 97:2.5:0.5 methylene chloride/methanof/ concentrated ammonium hydroxide) afforded 0.020 g of an off-white solid. This material was freeze dried from 2:1 acetonitrile/water (6 mL) to afford the title product as a white solid, 0.0189 g (14% yield), mp 89-97 °C; ''H NMR (300 MHz, CDCI3) 8 8.18-8.17 (m, 1H), 7.84-7.78 (m, 2H), 7.62-7.54 (m, 3H), 7.48-7.42 (m, 2H), 7.27 (m, 1H), 7.16 (d, J « 8.2 Hz, 2H), 3.39 (d, J= 2.2 Hz, 2H), 1.44 (s, 3H), 1.42 (s, 3H); ESI MS m/z 484 [CasH^NsOz* Hf.
EXAMPLE 43
Preparation of 8-(2.2-Difluoro-benzof1.3ldioxol-5-vl)-8-r3-{2-fluoro-pyridin-3-vn-phenYn-2,3,4,8-tetrahydro-imidazof1.5-a1pvrimidin-6-vlamine
1. /-BuLi, Et2O, -78 °C Rr
" CH2C!2 o sat. NaCO, (aq) --F
2 Brx^x-^CN li i •v^- J-
3. NaBH4, MeOH
CICSCI
Br
NCS
/-BuOK, CS: THF, -7S °C
/-BuOOH 5:1 MeOH/NH,OH
'' °~h
DME, H2O
(Figure Remove)
Step a) Preparation of Compound 3
«
A solution of 1 (2.00 g, 8.44 mmol) in diethyl ether (16 mL) was added dropwise to a mixture of f-butyl lithium (4.96 mL of a 1.7 M solution in pentane, 8.44 mmol) in diethyJ ether (20 mL) at-78 °C and stirred at this temperature for 25 min. To this was added 3-bromobenzonitrile (0.731 g, 4.02 mmol) in diethyl ether (20 mL) at -78 °C, and the reaction mixture was stirred at -78 °C for an additional 1.5 h. The reaction was then warmed to 0 °C and methanol (60 mL) was added, followed by the portion-wise addition of sodium borohydride (0.319 g, 8.43 mmol). The cooling bath was removed and then the mixture was stirred at room temperature for 2 hr. Saturated aqueous ammonium chloride (10 mL) was added, most of the methanol and diethyl ether was removed under reduced pressure and the aqueous residue obtained was diluted with methylene chloride (200 mL) and saturated aqueous sodium bicarbonate solution (30 mL). The organic layer was separated and washed
with brine, dried over sodium sulfate, filtered and concentrated. Purification by flash chromatography (silica, 9:1 hexanes/ethyl acetate) afforded 3 (0.237 g, 18%) as a colorless syrup: 1H NMR (300 MHz, CDC!:,} S 7.54 (t, J - 1.6 Hz, 1HA 7 3Q. >/
OCHF,
Step a) Preparation of Compound 2
A mixture of 1 (1.0 g, 5.22 mmol) and potassium hydroxide (0.345 g of 85%, 5.22 mmol) in ethanol (2 ml) was heated at reflux for 3 h. The reaction was cooled to room temperature, diluted with ethyl acetate (10 ml) and the solids that formed removed by filtration. The filtrate was concentrated to afford 2 (0.65 g, 80%) as a colorless liquid: 1H NMR (300 MHz, CDCI3) 5 4.47 (s, 4H), 3.95 (s, 4H). Step b) Preparation of Compound 3
A mixture of 2 (0.63 g, 4.06 mmo!) and sodium azide (0.66 g, 10.2 mmol) in DMSO was heated at 65 °C overnight. The reaction was cooled to room temperature, diluted with water and extracted into methylene chloride. The combined methylene chloride extracts were washed with water, dried over sodium sulfate, filtered and concentrated at room temperature to a volume of 2 mL and diluted with THF. This solution was then treated with triphenylphospine (2.56 g, 9.75 mmol)) in THF and stirred at room temperature for 10 min, treated with water (0.29 g, 16.2 mmol) and stirred overnight at room temperature. The mixture was then concentrated, diluted with methylene chloride and extracted with 10% aqueous HCI. The combined aqueous extracts were washed with methylene chloride and
concentrated to afford 3 (0.51 g, 66%) as a white solid: ''H NMR (500 MHz, D2O) o
4.49 (s, 4H), 3.42 (s, 4H).
Step c) Preparation of Compound 5
A mixture of 4 (0.50 g, 1.12 mmol), 3 (0.50 g, 2.64 mmo!) and triethylamine (0.57g, 5.60 mmol) in ethano! was heated at 70 °C for 3 h, cooled to room temperature, concentrated and partitioned between ethyl acetate and water. The organic layer was separated, washed with brine, dried over sodium sulfate, filtered and concentrated. Purification by flash chromatography (silica, 1:4 ethyl acetate/hexanes) afforded 5 (0.389 g, 70%) as a white foam: 1H NMR (500 MHz, COCI3) 6 7.61 (s, 1H). 7.49 (dt, J= 7.5,1.5 Hz, 1H), 7.47-7.44 (m, 1H), 7.33-7.19 (m, 4H), 7.11 (d, J= 8.8 Hz, 2H), 6.52 (t, J = 73.4 Hz, 1H), 4.52-4.47 (m, 4H), 4.14-4.06 (m, 2H), 3.87-3.77 (m, 2H); ESI MS m/z494 [C21H)eBrF2N3O2S + HJ*. Step c) Preparation of Compound 6
A mixture of 5 (0.389 g, 0.79 mmol) and f-butyl hydroperoxide (1.42 g of a 70% solution in water, 15.8 mmol) in methanoi and concentrated aqueous ammonium hydroxide (5.0 ml) was stirred at room temperature for 7 h, treated with 10% aqueous sodium thiosulfate (30 mL)and concentrated to remove most of the methanol. The resultant aqueous mixture was extracted with methylene chloride. The methylene chloride extracts were combined and washed with brine, dried over sodium sulfate, filtered and concentrated. Purification of this residue by flash chromatography (silica, 95:5:0.25 melhytene chloride/methanol/concentrated ammonium hydroxide) afforded 6 (0.179 g, 47%) as a white foam: *H NMR (300 MHz, CDCI3) 5 7.56 (t. J = 1.7 Hz, 1H), 7.40-7.37 (m, 1H), 7.38 (d, J= 8.8 Hz, 2H), 7.29-7.26(m, 1H), 7.15 (t, J= 7.9 Hz, 1H), 7.04 (d, J = 8.7 Hz, 2H), 6.48 (t, J = 73.9 Hz, 1 H), 4.57-4.53 (m, 2H), 4.46-4.42 (m, 2H), 3.79 (s, 4H); ESI MS m/z 477 [C2,H19BrF2N402 + HJStep d) Preparation of of 8-[4-(difluoromethoxy)phenyl]-8-[3-{2-fluoropyridin-3-yl)phenyl]-2,8-dihydrospiro[lmidazo[1,5-a]pyrimidine-3,3*-oxetan]-6-amIne
A mixture of 6 (0.10 g, 0.21 mmol), 2-fluoropyridine-3-boronic acid (0.035 g, 0.250 mmol), bis(triphenylphosphfno)palladium(ll) chloride (0.0073 g, 0.11 mmol), triphenylphosphine (0.0055 g, 0.021 mmol) and sodium carbonate (0.066 g, 0.63 mmol) in 3:1 DME/water was heated at reflux temperature for 2 h, cooled to room temperature and diluted with ethyl acetate and water. The organic layer was
Separated and washed with brine, dried over sodium sulfate, filtered and concentrated. Purification of this residue by flash chromatography (silica, 97:2.5:0.5 melhylene chloride/metnanoi/concentrated ammonium hydroxide) afforded a yellow foam (0.089 g, 85%). This foam was further purified by semi-preparative LC to afford the title product as a white solid; mp 96-107 °C; ''H NMR (500 MHz, CD3OD) D8.17-8.16(m, 1H), 8.02-7.98 (m, 1H), 7.52-7.51 (m, 2H), 7.45 (t, J= 7.9 Hz, 1H),7.40-7.34 (m, 4H), 7.09 (d, J = 8.8 Hz, 2H), 6.01 (t. J = 74.1 Hz, 1H), 4.50-4.47 (m, 4H), 3.97-3.89 (m, 2H), 3.76-3.69 (m, 2H); ESI MS m/z496 [C26H22F3N5O2 + Hf;
EXAMPLE 47
Preparation of 8''-r4-(difluoromethoxv)phenvl1-8''-r3-(2-fluoropvridin-3-vl)phenyl|-^S''-dihydrospirofcvclobutane-I^Mrriidazofl.S-alpvrimldlnl-S''-arnine
(Figure Remove)
•2Hd
/-BuOOH
NH4OH, McOH
r\/-tre EtOH, triethyiamine
OCHF O
OCHF,
a:
,N^F
^B(OH)j
(Figure Remove)
H2N ^4-J
OCHF2
,N_F VN y
3: IDME/H2O, reflux
, OCHF2
Using essentially the same procedure described in Example 44 and employing 1,1-di(arninomethylcyclobutane as starting reactant, the title product was obtained as a white solid, mp 220-226 °C; 1H NMR (500 MHz, CD3OD) Q8.16-8.15 (m, 1H), 8.01-7.97 (m, 1H), 7.54-7.51 (m, 2H), 7.44 (t, J - 7.8 Hz, 1H)t 7.40-7.37 (m, 4H), 7.08 (d, J = 8.8 Hz, 2H), 6.80 (t, J= 74.1 Hz, 1H), 3.65-3.58 (m, 2H), 3.46-3.39 (m, 2H), 2.09-2.01 (m, 2H), 1.92-1.87 (m, 4H); ESi MS m/z492 [C2rH2JF3N5O + HJ*.
EXAMPLE 48
Evaluation of the Enzyme Activity of Test Compounds and the Inhibition of hBACEI. !ViuBACE1 and hBACE2 by Test Compounds
Assay Conditions: 10 nM human 8ACE1 (or 10 nM Murine BACE1, 1.5 nM human BACE2) 25 uM substrate (WABC-6, MW 1549.6, from AnaSpec); final buffer conditions:50 mM Na-Acetate, pH 4.5, 0.05% CHAPS, 25% PBS; temperature: room temperature; reagent information:Na-Acetate: Aldrich, Cat.# 24,124-5 CHAPS: Research Organics, Cat. # 1304C 1X PBS: Mediatech (Cellgro), Cat* 21-031-CV; peptide substrate AbzSEVNLDAEFRDpa: AnaSpec. Peptide Name: WABC-6; determination of stock substrate (AbzSEVNLDAEFRDpa) concentration: a 25 mM stock solution in dimethyl suifoxide (DMSO) is prepared using the peptide weight and MW and diluted to 25 uM. The concentration is determined by absorbance at 354 nm using an extinction coefficient z of 18172 M^cm"1, The substrate stock is stored in small aliquots at -80° C. [Substrate Stock] = ABS 354nm * 106 /18172 (in mM)
Determination of Stock Enzyme Concentration: The stock concentration of each enzyme by ABS at 280 nm using D of 64150 M''1cm''1 for hBACEI and MuBACEl, 62870 M''W1 for hBACE2 in 6 M Guanidinium Hydrochtoride (from Research Organics, Cat f 5134G-2), pH 6.
(The extinction coefficient e280™" for each enzyme was calculated based on known amino acid composition and published extinction coefficients for Trp (5.69 M"1 cm*1) and Tyr (1.28 M"1 cm"1) residues (Anal. Btochem. 182,319-326). Dilution and mixing steps: total reaction volume: 100 uL
1. 2X inhibitor dilutions in buffer A(66.7 mM Na-Acetate, pH 4.5,0.0667% CHAPS)
are prepared,
2. 4X enzyme dilution In buffer A(66.7 mM Na-Acetate, pH 4.5,0.0667% CHAPS)
are prepared,
3. 100 uM substrate dilution in 1X PBS is prepared,
4. 50 uL 2X Inhibitor and 25 pL 100 pM substrate are added to each well of 96-well
plate (from DYNEX Technologies, VWR #: 11311-046), the immediately 25 pL 4X
enzyme are added to the inhibitor and substrate mixer, the fluorescence readings
are initiated.
Fluorescence Readings: Readings at Xox 320 nm and ?^m 420 nm are taken every 40-sec for 30 min at room temperature to determine the linear slope for substrate cleavage rate (Vi).
Calculation of % Inhibition: % Inhibition = 100 * (1- Vj / v0) (Vj* substrate cleavage rate in the presence of inhibitor, v0 * substrate cleavage rate in the absence of inhibitor) IC50 Determination: % Inhibition = [(B * ICso") + (100 * I0n)] / (ICsc" + I0n),
Fluorescent Kinetic Assay for human recombinant BACE 2
This assay is used to provide kinetic and selectivity parameters for the analyses of the tested compounds.
Materials and methods: final assay conditions: 10 nM human BACE1 (or 10 nM Murine BACE1,1,5 nM human BACE2) 25 uM Substrate (WABC-6, MW 1549.6, from AnaSpec). Final buffer conditions: 50 mM Na-Acetate, pH 4.5,0.05% CHAPS, 25% PBS. Temperature: room temperature. Reagent information: Na-Acetate: Aldrich, Cat* 24,124-5 CHAPS: Research Organics, Cat # 1304C 1X PBS: Mediatech (Cellgra), Cat* 21-031-CV Peptide Substrate AbzSEVNLDAEFRDpa: AnaSpec, Peptide Name: WABC-6
Determination of stock substrate (AbzSEVNLDAEFRDpa) concentration: A 25 mM stock solution in DMSO is prepared using the peptide weight and MW, and diluted to 25 MM. The concentration is determined by absorbance at 354 nm using an extinction coefficient e of 18172 M''1cm''1. The substrate stock is stored in small aliquots at -80° C. [Substrate Stock] = ABS ** nm * 10* /18172 (in mWI)
Determination of stock enzvme concentration: The stock concentration of each enzyme is determined by ABS at 280 nm using e of 64150 M^cm"4 for hBACEl and MuBACEl, 62870 M''1cnY1 for hBACE2 in 6 M guanWinium hydrochloride (from Research Organic®, Cat * 5134G-2), pH 6. (The extinction coefficient e*0™" for each enzyme is calculated based on known amlno add composition and published extinction coefficients for Trp (5.69 M"1 cm''1) and Tyr (1.28 M"1 cm"1) residues (Anal. Biochem. 182. 319-326).)
Dilution and Mixing Steps: Total Reaction Voume.: 100 uL 1. 2X inhibitor dilutions in buffer A(66.7 mM Na-Acetate, pH 4.5,0.0667% CHAPS) are prepared,
2. 4X enzyme dilution In buffer A(66.7 mM Na-Acetate, pH 4.5. 0.0667%
CHAPS) are prepared,
3. 100 MM substrate difution in 1X PBS, 50 pL 2X Inhibitor and 26 pL 100 uM
substrate are added to each well of 96-wefl plate (from DYNEX Technologies, VWR
#: 1 131 1-046), then immediately 25 yL 4X enzyme is added to the inhibitor and
substrate mixer and the fluorescence readings are initiated.
Fluorescence Readings: Readings at Xe» 320 nm. ?„„,, 420 nm are taken every 40-sec for 30 mln at room temperature and to determine the linear slope for substrate cleavage rate (v,}.
Analysis of calculation of % inhibition: % Inhibition = 100 * (1- v-, I vfl) v, = substrate cleavage rate in the presence of inhibitor. v0 = substrate cleavage rate in the absence of inhibitor) IC8o Determination: % Inhibition = ((B * KV) + (100 * I0n)) / (IC«n + I0"),
The data obtained are shown in Table II below. Unless otherwise noted, the ICto value represents me value obtained at 100% inhibition.
Table Remove
Ex. No.
MO.HllMk.MMl
5
13
14
15
16
17
18
19
Table Remove
Table Remove
Results and Discussion:
As can be seen from the data shown on Table II hereinabove, the compounds of the invention are potent and selective inhibitors of BACE1.
A compound of formula I
(Figure Remove)
wherein X \s N, NO or CRi9; Y is N, NO or CRu; Z is N, NO or CR2o with the proviso that no more than two of X,y or Z may be
N or NO; R, and R2 are each independently H, CN or an optionally substituted d-
C4alkyl group;
R3 and R4 are each independently H, or an optionally substituted (VC4 alkyl group or R3 and R* may be taken together to form a 3- to 7-membered ring optionally containing one or two heteroatoms selected from O, N
t
or S or R3 may be taken together with the atom to which it is attached and an adjacent carbon atom to form a double bond; R5 and Re are each independently H, halogen, NO2, CN, OR13, NR14R15 or a CrC6alkyl, CrC6haloatkyl, C2-C6aIkenyl, Cz-Cealkynyl or C3-C8cycloalkyl group each optionally substituted or when attached to adjacent carbon atoms Rs and R6 may be taken together with the atoms to which they are attached to form an optionally substituted 5-to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S;
R?, RB, RS, RIO, Rn, Rig and R20 are each independently H, halogen, NO2, CN, OR16, NR17R,a or a CrC6a!kyl, C,-C6haloalkyl, C2-CGalkenyl, Cr-Cr.alkynyl or Cs-Cscycloalkyl group each optionally substituted;
m is 0 or 1;
n is 0, 1, 2 or 3;
- ---- is a single bond or a double bond with the proviso that when m is 0 then --— -- must be a single bond;
R12, Ri3 and Ri6 are each independently H or a CrC6alkyl, C^Cehaloalkyl, C2-C6aIkenyl, C2-C6alkynyl, C3-C8cycloalkyl or aryl group each optionally substituted; and
Ri4, RIS, RI/ and Ri8 are each independently H or C,-C4alkyl; or a tautomer thereof, a stereoisomer thereof or a pharmaceutically acceptable salt thereof.
2. The compound according to claim 1 wherein Rt and R2 are H.
3. The compound according to either one of claims 1 and 2 wherein m
and n are 1 .
4. The compound according to any one of claims 1 , 2 and 3, wherein
formula I has the structure la
(Figure Remove)
R4
R8
(la)
The compound according to claim 4, wherein formula I has the ;tructure Ib
(Figure Remove)
6. The compound according to claim 5 wherein RI and R2 are H.
7. The compound according to claim 6 wherein X is N.
8. The compound according to cfaim 7 wherein n is 1.
9. The compound according to claim 1 selected from the group
consisting of:
8-(3-pyrimidin-5-ylphenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimtdazo[1,5-a]pyrimidin-6-amine; 7-(3^yrimidin-5-ylphenyf)-7-[4-(trifluoromethoxy)phenyl]-7H-imidazo[1,5-aJimida23Dl-
5-amine; 8-[4-fluoro-3-(4-fluoropyridin-3-yl)phenyfj-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimJdazo[1,5-a]pyrimidin-6-amine; 8-[3-(5-fluoropyridin-3-yl)phenyl]-8-[4-(trifluoromethoxy)phenyl]-2,3,418-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[3-(5-chloropyridin-3-yl)phenyl]-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazoII.S-aJpyrimidin-S-amine; (8S)-8-[3-(2-fluoropyridin-3-yl)phenyl]-8-[4-(trifluoromethoxy)phenyl]-2,3I4,8-
telrahydroimidazo[1,5-a]pyrimidin-6-amine;
(8R)-8-[3-(2-fluoropyridin-3-yl)phenyl]-8-[4-(trifluoromethoxy)phenyl]-2)3,4,8-tetrahydroimidazo[1,5-a]pyrimidin-6-amine;
(8R)-8-(3-pyrimidin-5-ylphenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; (8S)-8-(3-pyrimidin-5-ylphenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[3-(4-fIuoropyridin-3-yl)phenyl]-8-[4-(trifIuoromethoxy)phenyl]r2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[3-(2-fluoropyridin-3-yl)phenyl]-8-(4-methoxyphenyl]-2,3,4,8-tetrahydroimidazo[1,5-
a]pyrimidin-6-arnine; 8-(4-methoxyphenyl)-8-(3-pyrimidin-5-ylphenyl)-2>3,4,8-tetrahydroimidazo[1,5-
a]pyrimidin-6-amine; 8-(4-fluoro-3-pyrimidin-5-ylphenyl)-8-(4-methoxyphenyl)-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[4-fluoro-3-(2-fluoropyridin-3-yl)phenyl]-8-(4-methoxyphenyl)-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-(4-fluoro-3-pyrimidin-5-ylphenyl)-8-[4-(trifluoromethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[3-(2-fluoropyridin-3-yl)phenyl3-8-[4-(trifluorornethoxy)phenyl]-2,3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[4-fluoro-3-(5-fluoropyridin-3-yl)phenyl]-8-[4-(trif1uoromethoxy)phenyl]-2tJ3,4,8-
tetrahydroimidazo[1,5-a]pyrimidin-6-amine; 8-[4-fluoro-3-(2-fluoropyridJn-3-yl)phenvl]-8-[4-(trifluoromethoxy)phenyl]-213,4,8-
tetrahydroimidazo[1,5-a]pyrimid»n-6-amine; a tautomer thereof; a stereoisomer thereof; and a pharmaceutically acceptable salt thereof
10. Use of a compound as claimed in any one of claims 1 to 9 for the manufacture of a medicament for the treatment, prevention or amelioration of a disease or disorder characterized by elevated B-amyloid deposits or p-amyloid levels in a patient..
11. The use according to claim 10 wherein said disease or disorder is
selected from the group consisting of Alzheimer''s disease; mild cognitive impairment,
Down''s syndrome; hereditary cerebral hemorrhage with amyloidosis of the Dutch
type; cerebral amyloid angiopathy; and degenerative dementia.
12, The use according to claim 11 wherein said disease is Alzheimer''s
disease.
13 A pharmaceutical composition which comprises a pharmaceutically acceptable carrier and an effective amount of a compound as claimed in any one of claims 1 to 9.
14. A process for the preparation of a compound of formula I
(Figure Remove)
wherein X is N, NO or CR19;
Y is N, NO or CR,,;
Z is N, NO or CR^ with the proviso that no more than two of X, Y or Z may be N or NO;
Rn and R2 are each independently H or an optionally substituted Ci-C4alkyl group;
R3 and R4 are each independently H, or an optionally substituted CrC4 alkyl group or R3 and R4 may be taken together to form a 3- to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S or R3 may be taken together with the atom to which it is attached and an adjacent carbon atom to form a double bond;
R;> and R6 are each independently H, halogen, NO2, CN, OR13, NR14R15 or a CrCealkyl, d-Cehaloalkyl. C2-C6alkenyl, C2-C6alkynyl or C3-C8cycloalkyl group each optionally substituted or when attached to adjacent carbon atoms R5 and R6 may be taken together with the atoms to which they are attached to form an optionally substituted 5-to 7-membered ring optionally containing one or two heteroatoms selected from O, N or S;
R/, RS, Rg, RIO, RII, Ri9 and R20 are each independently H, halogen, NO2, CN, OR,6l NR17R18 or a d-C6alkyl, CrC6haloalkyl, C2-C6alkenyl, C2-C6alkynyl or C3-C8cycloalkyl group each optionally substituted;
m is 0 or 1;
n is 0,1,2 or 3;
—— is a single bond or a double bond with the proviso that when m is 0 then -——— must be a single bond;
Ri2, Ri3 and R16 are each independently H or a d-C6alkyl, d-C6haloaIkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C8cycloalkyl or aryl group each optionally substituted; and
(n)
wherein Hal is Cl or Br and R,, R2. R3, R*, RS, Re. R?, RB and n are as described for formula I hereinabove with a compound of formula III
R«. RIS, RI? and R™ are each independently H or d-dalkyl; which process comprises reacting a compound of formula II
(Figure Remove)
(Figure Remove)
(III)
wherein W is B(OH)2, Sn(n Bu)3 or Sn(CH3)3 and X, Y, Z, R9, R10, m and are as
described for formula I hereinabove in the presence of a palladium catalyst optionally in the presence of a solvent.
15. The invention substantially such as herein described