WO2009152027A1 - 5,7-dihydro-6h-pyrrolo[2,3-d]pyrimidin-6-one derivatives for mark inhibition - Google Patents

Abstract

Compounds of formula I: are potent and selective inhibitors of microtubule affinity regulating kinase (MARK), and hence find use in treatment of Alzheimer's disease and other conditions associated with hyperphosphorylation of tau.

Description

5,7-D1HYDRO-6H-PYRROLO[2,3-d]PYRIMIDIN-6-ONE DERIVATIVES FOR MARK

INHIBITION

This invention relates to methods and materials for the treatment or prevention of neurodegenerative diseases such as Alzheimer's disease. In particular, there is disclosed a particular class of 5,7-dihydro-6H-pyrrolo[2,3-ώT|pyrimidin-6-one derivatives which selectively inhibit microtubule affinity regulating kinase (MARK).

Alzheimer's disease (AD) is the most common cause of dementia in the elderly and is characterised by a decline in cognitive function, that progresses slowly and results in symptoms such as memory loss and disorientation. Death occurs, on average, 9 years after diagnosis. The incidence of AD increases with age, so that while about 5% of people over the age of 70 are sufferers, this figure increases to 20% of those over 80 years old.

Existing treatments exclusively target the primary symptoms of AD. Diseased neurons may release insufficient or excessive amounts of particular neurotransmitters, and so current drugs are aimed at increasing neurotransmitter levels or at reducing the stimulation of nerve cells by neurotransmitters. Although these drugs provide some improvement in the symptoms of AD, they fail to address the underlying cause of the disease.

The classic clinical and neuropathological features of AD consist of senile or neuritic plaques and tangled bundles of fibers (neurofibrillary tangles) [Verdile, G., et al, Pharm. Res, 50:397-409 (2004)]. In addition, there is a severe loss of neurons in the hippocampus and the cerebral cortex. Neuritic plaques are extracellular lesions, consisting mainly of deposits of β- amyloid peptide (Aβ), surrounded by dystrophic (swollen, damaged and degenerating) neurites and glial cells activated by inflammatory processes. In contrast, neurofibrillary tangles (NFTs) are intracellular clusters composed of a hyperphosphorylated form of the protein tau, which are found extensively in the brain (e.g. mainly in cortex and hippocampus in AD). Tau is a soluble cytoplasmic protein which has a role in microtubule stabilisation. Excessive phosphorylation of this protein renders it insoluble and leads to its aggregation into paired helical filaments, which in turn form NFTs.

The amyloid cascade hypothesis proposes that abnormal accumulation of Aβ peptides, particularly Aβ42, initiates a cascade of events leading to the classical symptoms of AD and ultimately, to the death of the patient. There is strong evidence [e.g. Rapoport, M., et al (2002) Proc. Natl. Acad. Sci USA 99:6364-6369] that dysregulation of tau function is a key step in the cascade of Alzheimer's disease pathology leading ultimately to neuronal death. Furthermore, tau mutations and NFTs are found in other dementias in which Aβ pathology is absent, such as frontotemporal dementia, Pick's disease and parkinsonism linked to chromosome 17 (FTDP- 17) [Mizutani, T. (1999) Rinsho Shikeigaku 39; 1262-1263]. Also, in AD the frequency of NFTs correlates to the degree of dementia better than that of senile plaques [Arriagada, P.V., et al (1992) Neurology 42:631-639], while significant numbers of amyloid plaques are often found in the brains of non-demented elderly people, suggesting that amyloid pathology on its own is not sufficient to cause dementia. For these reasons, normalisation of tau function (in particular prevention of hyperphosphorylation) is seen as a desirable therapeutic goal for the treatment of AD and other dementing conditions. Tau is a 352-441 amino acid protein encoded by the Mapt (Microtubule-associated protein tau) gene which is widely expressed in the central nervous system (CNS) with localisation primarily in axons [Binder et al J Cell Biol. 1985, 101(4), 1371-1378]. The major function of tau is regulation of the stability of microtubules (MTs), intracellular structural components comprised of tubulin dimers which are integral in regulating many essential cellular processes such as axonal transport and elongation as well as generation of cell polarity and shape. Tau binding to tubulin is a key factor in determining the rates of polymerisation/depolymerisation (termed dynamic instability) of MTs, and tau is therefore key to the regulation of many essential cellular processes [see, for example, Butner, K. A., Kirschner, M.W. (1991) J.Cell. Biol. 115: 717-730]. Tau is a basic protein with numerous serine and threonine residues, many of which are susceptible to phosphorylation, While normal tau has two to three phosphorylated amino acid residues, hyperphosphorylated tau found in AD and other tauopathies typically has eight or nine phosphorylated residues. A variety of kinases promote phosphorylation of these sites, including proline-directed kinases such as glycogen synthase kinase 3β (GSK3β) and cyclin dependent kinase 5 (cdk5), and non-proline-directed kinases such as protein kinase A (PKA) and calmodulin (CaM) kinase II, which phosphorylate tau at Lys-(Ile/Cys)-Gly-Ser sequences, also known as KXGS motifs. One KXGS motif is found in each of the MT binding repeats. Phosphorylation at these sites is important for the regulation of tau-MT binding and while the degree of phosphorylation is normally low, it has been shown to be increased in brain tissue from AD patients. Phosphorylation of one particular residue within the KXGS motifs, Ser-262 has been shown to be elevated in tau protein extracted from the NFTs in AD [Hasegawa, M. et al (1992) J. Biol. Chem 267:17047-17054] and phosphorylation at this site also appears to dramatically reduce MT binding [Biernat, J. et al. (1993) Neuron 11: 153-163].

Nishimura et al. [Cell 116: 671-682 (2004)] demonstrated that overexpression of the kinase PAR-I in Drosophila led to enhanced tau-mediated toxicity and an increase in the phosphorylation of tau on Ser-262, Ser-356, and other amino acid residues, including sites phosphorylated by GSK3β and Cdk5. Their findings suggest that PAR-I kinase acts as a master kinase during the process of tau hyperphosphorylation, with the phosphorylation of the Ser-262 and Ser-356 sites being a prerequisite for the subsequent phosphorylation at downstream sites by other kinases.

The mammalian ortholog of PAR-I is microtubule affinity-regulating kinase (MARK). There are four MARK isoforms and these form part of the AMP-dependent protein kinase (AMPK) family. Like PAR-I, MARK is thought to phosphorylate tau, perhaps in response to an external insult, such as the disruption of Ca²⁺ homeostasis caused by Aβ, priming it for further phosphorylation events. It is not clear whether the phosphorylation of tau by MARK leads directly to its detachment from MTs or the subsequent phosphorylation events cause detachment. The resulting unbound, hyperphosphorylated tau is delocalised to the somatodendritic compartment and is then cleaved by caspases to form fragments prone to aggregation [Drewes, G. (2004). Trends Biochem. Sci 29:548-555; Gamblin, T.C., et al, (2003) Proc. Natl. Acad. Sci. U.S.A. 100:10032-10037]. These aggregates can grow into filaments, which are potentially toxic, eventually forming the NFTs found in AD.

For these reasons, it is proposed that MARK inhibitors will enable the prevention or amelioration of neurodegeneration in AD and other tauopathies.

WO 2008/012571, WO 2007/141571, WO 2007/085873 and WO 2007/088401 disclose various classes of compounds as MARK inhibitors, but do not disclose or suggest the compounds described herein.

WO 2007/140222 discloses pyrrolopyrimidine compounds as inhibitors of protein kinases, useful in treating proliferative diseases and other conditions, including Alzheimer's disease, The compounds are apparently indiscriminately active against a wide range of kinases, although there is no mention of activity towards MARK.

There is therefore a need for potent and selective MARK inhibitors, in particular compounds showing good biovailability after oral administration and showing good brain penetration.

In accordance with the invention there is provided a compound of formula I:

I or a pharmaceutically acceptable salt or hydrate thereof; wherein: R¹ represents Ar or C_1-4alkyl which is optionally substituted with Ar, where Ar represents phenyl or pyridyl either of which optionally bears up to 3 halogen substituents; R^1a represents methyl; or R' and R^1a together with the carbon atom to which they are attached complete a bicyclic group of formula (a):

R represents H, halogen, OH, C_1-4alkyl, C_1-4alkoxy or benzyloxy;

Z represents CH₂ or O;

R² represents a group selected from:

and

III

X represents CR⁵ or N;

R³, R⁴ and R⁵ independently represent H, halogen, C_1-4alkoxy or C_1-4alky! which is optionally substituted with OH, C_1-4alkoxy, or with up to 3 halogen atoms; or one of R³, R⁴ and R^s represents

and the other two of R³, R⁴ and R⁵ are as defined previously;

R⁶ represents C_1-6alkyl which is optionally substituted with 1-3 halogens or with Ct. ₄alkoxy or CO₂C_1-4alkyl, or R⁶ represents m is 1, 2 or 3; each R^a independently represents H or C_1-4alkyl which is optionally substituted with 1-3 halogens, or two R^a groups attached to the same carbon may represent =0, or one R^a group together with R⁷ and the intervening atoms may complete a ring of 4 to 7 members; with the proviso that when m>1, only one -CRV may be other than -CH₂-; R⁷ represents H, C_1-4alkyl, acetyl or t-butoxycarbonyl, or R⁷ together with R^a and the intervening atoms completes a ring of 4-7 members;

R⁸ represents H or C_1-6alkyl which is optionally substituted with up to 3 halogen atoms, or with OR^b, COR^b, CO₂R^b, NR^b ₂ or C0NR^b ₂; R^b represents H or C_1-4alkyl which is optionally substituted with OH or C_1-4alkoxy; or R^s represents

or R⁸ together with R³ or R⁴ and the intervening atoms may complete a 5- or 6-membered ring; or R⁷ and R⁸ together with the nitrogen to which they are both attached may complete a monocyclic or bicyclic ring system of up to 10 ring atoms in total, of which up to two, in addition to the nitrogen to which R⁷ and R⁸ are bonded, are selected from N, O and S, the remainder being carbon, said ring system optionally bearing up to 3 substituents independently selected from halogen, CF₃, C_1-4alky], C₃^cycloalkyl, phenyl, =0, OH, C_1-4alkoχy, hydroxy C_1-4alkyl and C_{1- 4}alkoxy C_1-4alkyl ; n is O, 1, 2 or 3;

R^c represents H or C_1-4alkyl with the proviso that when n>l, only one -CRV may be other than -CH₂-; and Y represents a monocyclic or bicyclic ring system of up to 10 ring atoms, of which 0- 3 are selected from N, O and S, the remainder being carbon, said ring system optionally bearing up to 3 substituents independently selected from halogen, CF₃, C_1-4alkyl, C_3-6cycloalkyl, phenyl, =O, OH, C_1-4alkoxy, hydroxyC_1-4alkyl, C_1-4alkoxyC_1-4alkyl and C_1-4alkylcarbonyl.

The invention further provides a pharmaceutical composition comprising a compound of formula I as defined above or a pharmaceutically acceptable salt or hydrate thereof and a pharmaceutically acceptable carrier.

The invention further provides a compound of formula I as defined above or a pharmaceutically acceptable salt or hydrate thereof for use in therapeutic treatment of humans or animals.

The invention further provides the use of a compound of formula I as defined above or a pharmaceutically acceptable salt or hydrate thereof for the manufacture of a medicament for treatment or prevention of a neurodegenerative disease associated with hyperphosphorylation of tau in a human patient.

There is also disclosed a method for treatment or prevention of a neurodegenerative disease associated with hyperphosphorylation of tau in a human patient, said method comprising administering to that patient an effective amount of a compound of formula I as defined above, or a pharmaceutically acceptable salt or hydrate thereof, Neurodegenerative diseases associated with hyperphosphorylation of tau include AD, fronto temporal dementia, Pick's disease and parkinsonism linked to chromosome 17 (FTDP-17).

The invention further provides a compound of formula I as defined above, or a pharmaceutically acceptable salt or hydrate thereof, for use in reducing or preventing the hyperphosphorylation of tau in a human patient.

In one embodiment of the invention the compounds of formula I have the stereochemical configuration shown in formula IA:

IA wherein R¹, R^1a and R² are as defined previously, such that when R¹ and R^1a together with the carbon atom to which they are attached complete a bicyclic group of formula (a), it has the stereochemical configuration shown in formula (b):

where R and Z are as defined previously.

In a subembodiment of the compounds of formula IA, R^1a represents methyl;

R⁶ represents C_1-6alkyl which is optionally substituted with 1-3 halogens or with CO₂C_{1- 4}alkyl, or R represents

R⁷ represents H or C_1-4alkyl, or together with R^a and the intervening atoms completes a ring of 4-7 members; Y represents a monocyclic or bicyclic ring system of up to 10 ring atoms, of which 0-3 are selected from N, O and S, the remainder being carbon, said ring system optionally bearing up to 3 substituents independently selected from halogen, CF₃, C_1-4alkyl, C_3-6cycloalkyl, phenyl, =0, OH, C_1-4alkoxy, hydroxy C_1-4alky I and C_1-4alkoxyC_1-4alkyl; and all other variables are as originally defined.

As used herein, the expression "C_1-xalkyl" where x is an integer greater than 1 refers to straight-chained and branched alkyl groups wherein the number of constituent carbon atoms is in the range 1 to x. Particular alkyl groups are methyl, ethyl, n-propyl, isopropyl and t-butyl. Derived expressions such as "C_2-6alkenyl", "hydroxyC_1-6alkyl", "heteroarylC_1-6alkyl", "C_2-6alkynyl" and " C_1-6alkoxy" are to be construed in an analogous manner. Most suitably, the number of carbon atoms in such groups is not more than 6.

The term "halogen" as used herein includes fluorine, chlorine, bromine and iodine of which fluorine and chlorine are preferred.

The expression " C_3-6cycloalkyl" as used herein refers to nonaromatic monocyclic hydrocarbon ring systems comprising from 3 to 6 ring atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl and cyciohexyl.

The term "heterocyclyl" as used herein refers to a ring system in which at least one of the ring atoms is N, O or S. Depending on the maximum number of ring atoms permitted, said ring system may be mono- or bicyclic. Any ring comprised by said system may saturated or unsaturated to any degree, including aromatic unless indicated otherwise. Attachment may be via any available ring atom unless indicated otherwise. "N-heterocyclyl" indicates attachment via a ring nitrogen and "C-heterocyclyl" indicates attachment via a ring carbon.

The term "heteroaryl" refers to heterocyclic groups in which at least one ring comprising a heteroatom is aromatic. For use in medicine, the compounds of formula I may be in the form of pharmaceutically acceptable salts. Other salts may, however, be useful in the preparation of the compounds of formula I or of their pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts of the compounds of this invention include acid addition salts which may, for example, be formed by mixing a solution of the compound according to the invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulphuric acid, methanesulphonic acid, benzenesulphonic acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Alternatively, where the compound of the invention carries an acidic moiety, a pharmaceutically acceptable salt may be formed by neutralisation of said acidic moiety with a suitable base. Examples of pharmaceutically acceptable salts thus formed include alkali metal salts such as sodium or potassium salts; ammonium salts; alkaline earth metal salts such as calcium or magnesium salts; and salts formed with suitable organic bases, such as amine salts (including pyridinium salts) and quaternary ammonium salts. When the compounds useful in the invention have one or more asymmetric centres, they may accordingly exist as enantiomers. Where the compounds according to the invention possess two or more asymmetric centres, they may additionally exist as diastereoisomers. It is to be understood that all such isomers and mixtures thereof in any proportion are encompassed within the scope of the present invention unless expressly indicated otherwise, e.g. by the use of stereochemical identifiers in structural formulae.

When a compound useful in the invention is capable of existing in tautomeric keto and enol forms, both of said forms are considered to be within the scope of the invention.

A nitrogen atom forming part of a heteroaryi ring may be in the form of the N-oxide. A sulphur atom forming part of a nonaromatic heterocycle may be in the form of the S-oxide or S,S-dioxide.

A heteroaryi group may be attached to the remainder of the molecule via a ring carbon or a ring nitrogen, provided that this is consistent with preservation of aromaticity.

In one group of the compounds of formula I, R^1a represents methyl and R¹ represents Ar or a C_1-4alkyl group which is optionally substituted with Ar, where Ar represents phenyl or pyridyl which optionally bears up to 3 halogen substituents. Very suitably, R¹ represents C_{1- 4}alkyl such as methyl, ethyl, n-propyl or isopropyl, in particular ethyl. Other suitable identities for R¹ include 3-fiuoro-2-pyrϊdyl, 4-fluorophenyI, 2-fluorophenyl, 2,4-difluorophenyl, phenyl, 5- fluoro-2-ρyridyl, 2,6-difluorophenyl, 2-pyridyl and 2,5~difluorophenyl, Alternatively, R¹ and R^1a together with the carbon atom to which they are attached complete a bicyclic group of formula (a):

(a); where R represents H, halogen (in particular F, Cl or Br), OH, C_1-4alkyl (in particular methyl), C_1-4alkoxy (in particular methoxy) or benzyloxy, and Z represents CH₂ or O. Most suitably, Z represents CH₂.

In a first embodiment of formula I, R² represents a phenyl or pyridyl group of formula II:

where X represents CR⁵ or N, and R³, R⁴ and R⁵ are as defined previously. Suitable identities for R³, R⁴ and R⁵ include H, halogen (especially F or Cl), C_1-4alkoxy (such as methoxy) and C_1-4alkyl which is optionally substituted with OH or with up to 3 fluorines (such as hydroxymethyl or CF₃). Very suitably, at least one of R³, R⁴ and R⁵ is H. In a particular embodiment, one of R³ and R⁴ (preferably R⁴) is a group represented by the formula IV:

IV where m, R^a, R⁷ and R⁸ are as defined previously. When one of R³ and R⁴ represents a group of formula IV, the other one of R³ and R⁴, together with the intervening atoms, optionally completes a 5- or 6-membered ring with R⁸. Said ring typically comprises only carbon atoms apart from the nitrogen to which R⁸ is bonded. Very suitably, the ring is 6-membered. Thus, in particular examples of this embodiment, R² is a group of formula:

or

where X and R⁷ are as defined previously.

In formula IV, m is 1, 2 or 3; preferably 1 or 2, and in a particular embodiment m is 1.

Each R^a is independently H or C_1-4alkyl which is optionally substituted with 1-3 halogens; or two R^a groups attached to the same carbon may complete a carbonyl group; or one R^a group together with the R⁷ and the intervening atoms may complete a ring of 4-7 members; provided that when m>l, only one -CRV maybe other than CH₂. A ring completed by R^a and R⁷ together typically comprises only carbon atoms apart from the nitrogen to which R⁷ is attached. Examples of groups in accordance with formula IV therefore include:

In formula IV R⁷ represents H, C_1-4aUcyl, acetyl or t-butoxycarbonyl, in particular H or C_1-4alkyl, or together with R^a completes a ring of 4-7 members as described above, Alternatively, R⁷ may complete a ring in combination with R⁸, as described hereinafter.

In one embodiment of formula IV, R⁸ represents H or C_1-6alkyl which is optionally substituted with up to 3 halogens or with OR^b, COR^b, CO₂R^b, NR^b ₂ or CONR^b ₂, where R^b represents H or C_1-4 alkyl which is optionally substituted with OH or C_1-4alkoxy. Within this embodiment, examples of groups represented by R⁸ include H, 2-methoxy-l -methyl ethyl, 2- hydroxyethyl, 3,3,3-trifluoropropyl, isopropyl, N,N-dimethylcarbamoylmethyl, 2-(2- hydroxyethoxy)ethyl, 2-hydroxy-3,3-dimethylbutyl, 2-methoxyethyl, 2-hydroxy-2-methylproρyl, 2-fluoroethyl, and 2,2-difluoroethyl.

In an alternative embodiment of formula IV, R⁸ together with R³ or R⁴ and the intervening atoms completes a 5- or 6-membered ring as described previously, In a further embodiment of formula IV, R⁸ together with R⁷ and the nitrogen to which they are both attached complete a monocyclic or bicyclic ring system of up to 10 ring atoms in total. Said ring atoms are selected from C, N, O and S, but not more than two are other than carbon, apart from the nitrogen to which R and R are both attached. The monocyclic or bicyclic ring system optionally bears up to 3 substituents independently selected from halogen, CF₃, C]-₄alkyl_; C₃-₆cycloalkyl, phenyl, =0, OH, C_1-4alkoxy, hydroxy C_1-4alkyl and C_1-4alkoxyC_{1- 4}alkyl. When more than one substituent is present, preferably at least one of the substituents is halogen or C_1-4alkyl. The monocyclic or bicyclic ring system may comprise folly saturated, unsaturated or aromatic rings, but in a particular embodiment it is saturated. Examples of suitable monocyclic systems include azetidine, pyrrolidine, piperidine, piperazine, morpholine, homopiperidine, pyrrole, pyrazole, imidazole and triazole. Examples of suitable bicyclic systems include azabicyclo[3.1,0]hexyl, azabicyclo[2.2.1]heptyl, 5,6,7,8-tetrahydroirnidazo[1,2- a]pyrazine, tetrahydropyridine and telrahydroquinoline. Within this embodiment, specific examples of groups represented by -NR⁷R⁸ include 3-hydroxy-3-phenylpyrrolidin-1-yl, 6- hydroxymethyl-3-azabicyclo[3, 1.0]hex-3-yl, piperidin-1-yl, 2-azabicyclo[2.2.1]hept-2-yl, 2,6- dimethylmorpholin-4-yl, morpholin-4-yl, hompiperidin-1-yl, pyrrolidin-1-yl, 5,6- dihydroimidazo[1,2-a]pyrazin-7(8H)-yl, 3-fluoropiperidin-1-yl and 4-methylpiperazin-1-yl. In a yet further embodiment of formula IV, R⁸ is a group of formula:

where n, R^c and Y are as defined previously.

Within this embodiment of formula IV, R⁷ is very suitably H. n is very suitably 0, 1 or 2. Each R^c is preferably H. Specific examples of the moiety

therefore include a bond, -CH₂- and -CH₂CH₂-. Y represents a monocyclic or bicyclic ring system of up to 10 ring atoms, 0-3 of which are selected from O, N and S, the remainder being carbon. The monocyclic or bicyclic ring system optionally bears up to 3 substituents independently selected from halogen, CF₃, Ci-4alkyl, C_3-6cycloalkyl, phenyl, =0, OH, C_1-4alkoxy, hydroxyC_1-4alkyl, C_1-4alkoxyC_1-4alkyl and C_1-4alkylcarbonyl. When more than one substituent is present, preferably at least one of the substituents is halogen or C_]-4alkyl. The monocyclic or bicyclic ring system may comprise fully saturated, unsaturated or aromatic rings. Examples of monocyclic rings included in the definition of Y include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, azetidinyl, pyrrolidinyl, piperidinyl, hompiperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, phenyl, pyridyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furyl, thienyl and isoxazolyl. Examples of bicyclic systems included in the definition of Y include pyrazolo[1,5-a]pyridinyl, imidazo[1,2-a]pyridinyl, and tetrahydronaphthalenyl.

Within this embodiment of formula IV, specific identities of R⁸ include: (pyrazolo[1,5-a]pyridin-3-yl)methyl; (5-oxopyrrolidin-2-yl)methyl; 2-(l-hydroxycyclopentyl)ethyl; 1 -phenylpiperidin-4-yl; 2-( 1 H-pyrazol- 1 -yl)ethyl ; cyclopentyl; (3 -methy loxetan-3 -y l)methy 1 ; (imidazo[ 1 ,2-a]pyridine-3-yl)methyl; 2-(l H-1 ,2,3-triazol- 1 -yl)ethyl;

(5-methylisoxazol-3-yl)methyl;

2-(methoxymethyl)cyclopentyl;

(2-methyltetrahydrofuran-2-yl)methyl; cyclopropylmethyl;

1,2,3 ,4-tetrahydronaphthal en-2-yl ;

1 -cycl opropy lethyl ;

3 ,3 -difluorocyclobutyl ; tetrahydro-2H-pyran~3-yl; (1 -methyl- 1 H-pyrazol- 3 -y l)methy 1 ;

(pyrazolo[1,5-a]pyridin-7-yl)methyl; oxetan-3-yl;

1 -methyl- 1 H-pyrazol-4-yl;

1 -methylpiperϊdin-4-yl; and l-acetylpiperidin-4-yl,

In a second embodiment of formula ϊ, R² represents a pyrazolyl group of formula III:

III where R⁶ represents Cj.galkyl which is optionally substituted with up to 3 halogens or with C_1-4alkoxy or Cθ₂C_1-4alkyl, or R⁶ represents:

where n, R° and Y are as described above.

Within this embodiment of formula I, suitable identities for R⁶ include: benzyl, isopropyl; (piperidin-4-yl)methyl;

2-(piperidin-4-yl)ethyl; piperidin-4-yl;

2-hydroxy-2-methylcyclopentyl ;

2-aminoethyl; pyrrolidin-3-yl;

2-(4-methylpiρerazin- 1 -yl)ethyl;

2,2-difiuoroethyl; methyl; azetidin-3-yl;

1 ,1 -dimethyl-2-methoxy-2-oxoethyl;

2-hydroxycyclopentyl ; ethyl;

2-(morpholin-4-yl)ethyl;

2-methoxy-2-oxoethyl;

( 1 -methyl- 1 H-imidazol-2-yl)methyl ;

2-methoxyethyl; (tetrahydrofuran-2-yl)methyl;

2-(l H-imidazol- 1 -yl)ethyl;

2-(pyrid-2-yl)ethyl; and 2-methoxyethyl.

A subset of the compounds of the invention consists of the compounds of formula V:

and pharmaceutically acceptable salts and hydrates thereof; wherein X, R¹, R⁷ and R⁸ have the same definitions and specific identities as described above.

In a particular embodiment of this subset, R¹ is ethyl.

A second subset of the compounds of the invention consists of the compounds of formula VI: VI

and pharmaceutically acceptable salts and hydrates thereof: wherein R¹ and R⁶ have the same definitions and specific identities as described above.

In a particular embodiment of this subset, R¹ is ethyl.

A third subset of the compounds of the invention consists of the compounds of formula VII:

VII and pharmaceutically acceptable salts and hydrates thereof; wherein R, X, R³ and R⁴ have the same definitions and specific identities as described above.

In one embodiment of this subset, R³ and R⁴ are both H.

In an alternative embodiment of this subset, R³ and R⁴ complete a fused-ring structure represented by:

or

where X and R⁷ have the same definitions and specific identities as described above. In specific examples of this embodiment, R⁷ represents H, methyl, acetyl or t-butoxycarbonyl and X is H. In the compounds of formula VII, is suitably (but not necessarily) OH which is attached to the 4-position of the indane ring.

Specific examples of compounds in accordance with the invention are provided in the Examples section hereinbelow.

Compounds of formula I may be prepared by reacting a sulfone (1) with R²NH₂:

(1) where R¹, R^1a and R² have the same meanings as before. The reaction may be carried out in an ethereal solvent such as THF in the presence of strong base such as sodium bis(trimethylsilyl)amide. When R² is an electron-rich heteroaryl group (such as 1-alkyl-1H- pyrazol-4-yl), the reaction can be carried out at higher temperature in the absence of base, e.g. in dioxan. In an alternative method of performing the same transformation, R²-NH-CHO is treated with NaH in DMF, then reacted with the sulfones (1).

Sulfones (1) are obtainable by reaction of compounds (2) with (R¹)(R^1a)CH-NH₂, followed by oxidation of the thioether functionality

(2)

Reaction with (Rⁱ)(R^1a)CH-NH₂ takes place in ethanol in the presence of a tertiary amine. Details of the preparation of (2) and its conversion to (1) are provided in the Examples section.

Alternatively, the thioether (2) may be oxidised to the sulfones, then reacted with R²NH₂ or R²-NH-CHO in the manner described above, then reacted with (R¹)(R^1a)CH-NH₂ to provide the compounds of formula I.

Chiral compounds of formula IA are obtainable via use of (R¹)(R^1a)CH-NH2 in the appropriate chiral form.

An alternative route, suitable for preparing compounds of formula V and analogs thereof, includes reductive alkylation of R⁷R⁸NH with aldehydes of formula (3a):

(3) where R¹, R^1a, R³, R⁷, R⁸ and X have the same meanings as before. The reaction takes place in the presence of sodium triacetoxyborohydride. The aldehydes (3a) are available via oxidation of vinyl derivatives (3b). The vinyl derivatives are obtainable by reaction of sulfones (1) with the appropriate amine. Details of these procedures are provided in the Examples section.

Where they are not themselves commercially available, the starting materials and reagents described above may be obtained from commercially available precursors by means of well known synthetic procedures and/or the methods disclosed in the Examples section herein.

Where the above-described processes for the preparation of the compounds of use in the invention give rise to mixtures of stereoisomers, these isomers may be separated by conventional techniques such as preparative chromatography. The compounds may be prepared in racemic form, or individual enantiomers may be prepared either by enantiospecific synthesis or by resolution. The compounds may, for example, be resolved into their component enantiomers by standard techniques such as preparative HPLC, or the formation of diastereomeric pairs by salt formation with an optically active acid, such as di-/?-toluoyl-D-tartaric acid and/or di-/?-toluoyl- L-tartaric acid, followed by fractional crystallization and regeneration of the free base, The compounds may also be resolved by formation of diastereomeric esters or amides, followed by chromatographic separation and removal of the chiral auxiliary.

During any of the above synthetic sequences it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T. W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley Sc Sons, 1991. The protecting groups may be removed at a convenient subsequent stage using methods known from the art.

The compounds of formula I are suitably administered to patients in the form a pharmaceutical composition comprising the active ingredient (i.e. the compound of formula I or pharmaceutically acceptable salt or hydrate thereof) and a pharmaceutically acceptable carrier. Preferably these compositions are in unit dosage forms such as tablets, pills, capsules, powders, granules, sterile parenteral solutions or suspensions, metered aerosol or liquid sprays, drops, ampoules, transdermal patches, auto-injector devices or suppositories; for oral, parenteral, intranasal, sublingual or rectal administration, or for administration by inhalation or insufflation. The principal active ingredient typically is mixed with a pharmaceutical carrier, e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate and dicalcium phosphate, or gums, dispersing agents, suspending agents or surfactants such as sorbitan monooleate and polyethylene glycol, and other pharmaceutical diluents, e.g. water, to form a homogeneous preformulation composition containing a compound of the present invention, or a pharmaceutically acceptable salt thereof. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This preformulation composition is then subdivided into unit dosage forms of the type described above containing from 0.1 to about 500 mg of the active ingredient of the present invention. Typical unit dosage forms contain from 1 to 100 mg, for example 1, 2, 5, 10, 25, 50 or 100 rng, of the active ingredient. Tablets or pills of the composition can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.

The liquid forms in which the compositions useful in the present invention may be incorporated for administration orally or by injection include aqueous solutions, liquid- or gel- filled capsules, suitably flavoured syrups, aqueous or oil suspensions, and flavoured emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles. Suitable dispersing or suspending agents for aqueous suspensions include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylceliulose, methylcellulose, poly(ethylene glycol), ρoly(vinylpyrrolidone) or gelatin.

In one embodiment of the invention, the compound of formula I is administered to a patient suffering from AD, FTDP- 17, Pick's disease or frontotemporal dementia, preferably AD.

In an alternative embodiment of the invention, the compound of formula I is administered to a patient suffering from mild cognitive impairment or age-related cognitive decline. A favourable outcome of such treatment is prevention or delay of the onset of AD. Age-related cognitive decline and mild cognitive impairment (MCI) are conditions in which a memory deficit is present, but other diagnostic criteria for dementia are absent (Santacruz and Swagerty, American Family Physician, 63 (2001), 703-13). (See also "The ICD-10 Classification of Mental and Behavioural Disorders", Geneva: World Health Organisation, 1992, 64-5). As used herein, "age-related cognitive decline" implies a decline of at least six months' duration in at least one of: memory and learning; attention and concentration; thinking; language; and visuospatial functioning and a score of more than one standard deviation below the norm on standardized neuropsychologic testing such as the MMSE. In particular, there may be a progressive decline in memory. In the more severe condition MCI, the degree of memory impairment is outside the range considered normal for the age of the patient but AD is not present. The differential diagnosis of MCI and mild AD is described by Petersen et at,, Arch. Neurol, 56 (1999), 303-8. Further information on the differential diagnosis of MCI is provided by Knopman et al, Mayo Clinic Proceedings, 78 (2003), 1290- 1308. In a study of elderly subjects, Tuokko et al {Arch, Neurol, 60 (2003) 577-82) found that those exhibiting MCI at the outset had a three-fold increased risk of developing dementia within 5 years.

Grundman et al (J, MoI Neurosci., 19 (2002), 23-28) report that lower baseline hippocampal volume in MCI patients is a prognostic indicator for subsequent AD. Similarly, Andreasen et al {Acta Neurol. Scand, 107 (2003) 47-51 ) report that high CSF levels of total tau, high CSF levels of phospho-tau and lowered CSF levels of Aβ42 are all associated with increased risk of progression from MCI to AD.

Within this embodiment, the compound of formula I is advantageously administered to patients who suffer impaired memory function but do not exhibit symptoms of dementia. Such impairment of memory function typically is not attributable to systemic or cerebral disease, such as stroke or metabolic disorders caused by pituitary dysfunction. Such patients may be in particular people aged 55 or over, especially people aged 60 or over, and preferably people aged 65 or over. Such patients may have normal patterns and levels of growth hormone secretion for their age. However, such patients may possess one or more additional risk factors for developing Alzheimer's disease. Such factors include a family history of the disease; a genetic predisposition to the disease; elevated serum cholesterol; and adult-onset diabetes mellitus.

In a particular embodiment of the invention, the compound of formula I is administered to a patient suffering from age-related cognitive decline or MCI who additionally possesses one or more risk factors for developing AD selected from: a family history of the disease; a genetic predisposition to the disease; elevated serum cholesterol; adult-onset diabetes mellitus; elevated baseline hippocampal volume; elevated CSF levels of total tau; elevated CSF levels of phospho- tau; and lowered CSF levels of Aβ(l-42).

A genetic predisposition (especially towards early onset AD) can arise from point mutations in one or more of a number of genes, including the APP, presenilin-1 and presenilin-2 genes. Also, subjects who are homozygous for the ε4 isoform of the apolipoprotein E gene are at greater risk of developing AD.

The patient's degree of cognitive decline or impairment is advantageously assessed at regular intervals before, during and/or after a course of treatment in accordance with the invention, so that changes therein may be detected, e.g. the slowing or halting of cognitive decline. A variety of neuropsychological tests are known in the art for this purpose, such as the Mini-Mental State Examination (MMSE) with norms adjusted for age and education (Folstein et al.J. Psych. Res,, 12 (1975), 196-198, Anthony el al, Psychological Med, 12 (1982), 397-408; Cockrell et al, , Psychopharmacology, 24 (1988), 689-692; Crum et al. , J. Am. Med. Assoc'n, 18 (1993), 2386-2391). The MMSE is a brief, quantitative measure of cognitive status in adults. It can be used to screen for cognitive decline or impairment, to estimate the severity of cognitive decline or impairment at a given point in time, to follow the course of cognitive changes in an individual over time, and to document an individual's response to treatment. Another suitable test is the Alzheimer Disease Assessment Scale (ADAS), in particular the cognitive element thereof (AD AS-cog) (See Rosen et al , Am. J. Psychiatry, 141 (1984), 1356-64).

For treating or preventing Alzheimer's disease, a suitable dosage level is about 0.01 to 250 mg/kg per day, preferably about 0.01 to 100 mg/kg per day, and more preferably about 0.05 to 50 mg/kg of body weight per day, of the active compound. The compounds may be administered on a regimen of 1 to 4 times per day. In some cases, however, a dosage outside these limits may be used.

The compound of formula I optionally may be administered in combination with one or more additional compounds known to be useful in the treatment or prevention of AD or the symptoms thereof. Such additional compounds thus include cognition-enhancing drugs such as acetylcholinesterase inhibitors (e.g. donepezil and galanthamine), NMDA antagonists (e.g. memantine) or PDE4 inhibitors (e.g. Ariflo™ and the classes of compounds disclosed in WO 03/018579, WO 01/46151, WO 02/074726 and WO 02/098878). Such additional compounds also include cholesterol-lowering drugs such as the statins, e.g. simvastatin. Such additional compounds similarly include compounds known to modify the production or processing of Aβ in the brain ("amyloid modifiers"), such as compounds which modulate the secretion of Aβ

(including γ-secretase inhibitors, γ-secretase modulators and β-secretase inhibitors), compounds which inhibit the aggregation of Aβ, and antibodies which selectively bind to Aβ. Such additional compounds further include growth hormone secretagogues, e.g. as described in WO 2004/080459. In this embodiment of the invention, the amyloid modifier may be a compound which inhibits the secretion of Aβ, for example an inhibitor of γ-secretase (such as those disclosed in WO 01/53255, WO 01/66564, WO 01/70677, WO 01/90084, WO 01/77144, WO 02/30912, WO 02/36555, WO 02/081435, WO 02/081433, WO 03/018543, WO 03/013506, WO 03/013527, WO 03/014075, WO 03/093251, WO 03/093252, WO 03/093253, WO 03/093264, WO 2004/031137, WO 2004/031 138, WO 2004/031 139, WO 2004/039370, WO 2004/039800, WO 2004/101538, WO 2004/101539 and WO 2005/030731), or a β-secretase inhibitor (such as those disclosed in WO 03/037325, WO 03/030886, WO 03/006013, WO 03/006021 , WO 03/006423, WO 03/006453, WO 02/002122, WO 01/70672, WO 02/02505, WO 02/02506, WO 02/02512, WO 02/02520, WO 02/098849 and WO 02/100820), or any other compound which inhibits the formation or release of Aβ including those disclosed in WO 98/28268, WO 02/47671, WO 99/67221 , WO 01/34639, WO 01/34571, WO 00/07995, WO 00/38618, WO 01/92235, WO 01/77086, WO 01/74784, WO 01/74796, WO 01/74783, WO 01/60826, WO 01/19797, WO 01/27108, WO 01/27091, WO 00/50391, WO 02/057252, US 2002/0025955 and US2002/0022621,

Alternatively, the amyloid modifier may be a compound which modulates the action of γ- secretase so as to selectively attenuate the production of Aβ(l-42). Compounds reported to show this effect include certain non-steroidal antiinflammatory drugs (NSAIDs) and their analogues (see WO 01/78721 and US 2002/0128319 and Weggen et al Nature, 414 (2001) 212-16; Morihara et al, J. Neurochem., 83 (2002), 1009-12; and Takahashi et al, J. Biol Chem., 278 (2003), 18644-70), and compounds which modulate the activity of PPARα and/or PPARδ (WO 02/100836), Further examples of γ-secretase modulators are disclosed in WO 2006/008558, WO 2006/043064, WO 2005/054193, WO 2005/013985 and WO 2005/108362. Alternatively, the amyloid modifier may be a compound which inhibits the aggregation of

Aβ. Suitable examples include chelating agents such as clioquinol (Gouras and Beal, Neuron, 30 (2001), 641-2) and the compounds disclosed in WO 99/16741, in particular that known as DP- 109 (Kalendarev et al, J. Pharm. Biomed. Anal, 24 (2001), 967-75). Other inhibitors of Aβ aggregation suitable for use in the invention include the compounds disclosed in WO 96/28471, WO 98/08868 and WO 00/052048, including the compound known as Apan™ (Praecis); WO 00/064420, WO 03/017994, WO 99/59571 and the compound known as Alzhemed™ (Neurochem); WO 00/149281 and the compositions known as PTI-777 and PTI-00703 (ProteoTech); WO 96/39834, WO 01/83425, WO 01/55093, WO 00/76988, WO 00/76987, WO 00/76969, WO 00/76489, WO 97/26919, WO 97/16194, and WO 97/16191. Further examples include phytic acid derivatives as dsiclosed in US 4,847,082 and inositol derivatives as taught in US 2004/0204387.

Alternatively, the amyloid modifier may be an antibody which binds selectively to Aβ. Said antibody may be polyclonal or monoclonal, but is preferably monoclonal, and is preferably human or humanized. Preferably, the antibody is capable of sequestering soluble Aβ from biological fluids, as described in WO 03/016466, WO 03/016467, WO 03/015691 and WO

01/62801. Suitable antibodies include humanized antibody 266 (described in WO 01/62801) and the modified version thereof described in WO 03/016466. Suitable antibodies also include those specific to Aβ-derived diffusible ligands (ADDLS), as disclosed in WO 2004/031400.

As used herein, the expression "in combination with" requires that therapeutically effective amounts of both the compound of formula I and the additional compound are administered to the subject, but places no restriction on the manner in which this is achieved. Thus, the two species may be combined in a single dosage form for simultaneous administration to the subject, or may be provided in separate dosage forms for simultaneous or sequential administration to the subject. Sequential administration may be close in time or remote in time, e.g. one species administered in the morning and the other in the evening. The separate species may be administered at the same frequency or at different frequencies, e.g. one species once a day and the other two or more times a day. The separate species may be administered by the same route or by different routes, e.g. one species orally and the other parenterally, although oral administration of both species is preferred, where possible. When the additional compound is an antibody, it will typically be administered parenterally and separately from the compound of formula I.

EXAMPLES MARK 3 Assay

MARK3 activity was assayed in vitro using a Cdc25C biotinylated peptide substrate (Cell Signalling Technologies). The phosphopeptide product was quantitated using a Homogenous Time-Resolved Fluorescence (HTRF) assay system (Park et al., 1999, Anal. Biochem. 269:94- 104). The reaction mixture contained 50 mM HEPES/Tris-HCl, pH 7.4; 10 mM NaCl, 5 mM MgCl₂, 0.2 mM NaVO₄, 5 mM β-glycerol phosphate, 0.1 % Tween-20, 2 mM dithiothreitol, 0.1% BSA, 10 μM ATP, 1 μM peptide substrate, and 10 nM recombinant MARK3 enzyme (University of Dundee) in a final volume of 12 μl. The buffer additionally contained protease inhibitor cocktail (Roche EDTA-free, 1 tab per 50 ml). The kinase reaction was incubated for 2 hours at 25°C, and then terminated with 3 μl Stop/Detection Buffer (50 mM HEPES, pH 7.0, 16,6 mM EDTA, 0.5M KF, 0.1 % Tween-20, 0.1 % BSA, 2 μg/ml SLX^ent 665 (CISBIO), and 2 μg/ml Eu³⁺ cryptate label antibody (CISBIO)). The reaction was allowed to equilibrate overnight at 0°C, and relative fluorescent units were read on an HTRF enabled plate reader (e.g. TECAN GENios Pro). Inhibitor compounds were assayed in the reaction described above to determine compound ICSOs, Aliquots of compound dissolved in DMSO were added to the reaction wells in a third-log dilution series covering a range of 1 nM to 10 μM. Relative phospho substrate formation, read as HTRF fluorescence units, was measured over the range of compound concentrations and a titration curve generated.

The compounds described below gave IC50 values of 10OnM or less, typically 5OnM or less, in the above assay.

METHODS OF SYNTHESIS

Method A

General procedures to prepare compounds of the instant invention are described in Scheme A. Treatment of diethyl succinate with ethyl formate under basic conditions gave diethyl 2-formylsuccinate I, which upon condensation with S-methylthiourea semi-sulfate and exposure to acetic acid afforded ethyl [2-(methylthio)-4-oxo-1,4-dihydropyrimidin-5-yl]acetate II. Transformation of the 4-hydroxypyrimidine II to ethyl [4-chloro-2-(methyIthio)pyrimidin-5- yl] acetate III was accomplished using phosphorous(V) oxychloride. Treatment of 4- chloropyrimidine III with methyl iodide in a solution of DMF followed by exposure to sodium hydride leads to ethyl 2-[4-chloro-2-(methylthio)pyrimidin-5-yl]-2-methylpropanoate IV. (S)- sec-(-)-butyl amine can be coupled to IV in the presence of l,4-diazabicyclo[2.2.2]octane and ethanol at high temperature and pressure to afford the bicyclic 5,5-dimethyl-7-[(1S)-1- methyIpropyl]-2-(methylthio)-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one V. Exposure of V to meta-perchlorobenzoic acid provides the oxidized product 5,5-dimethyl-7-[(1S )-1- methylpropyl]-2-(methylsulfonyl)-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one VI.

Scheme A

General procedures to prepare compounds of the instant invention are also described in Scheme B. Coupling 5,5-dimethyl-7-[(15)-1-methylpropyl]-2-(methylsulfonyl)-5,7-dihydro-6H- pyrrolo[2_;3-_<^pyrimidin-6-one VI with iV-(4-vinylphenyl)formamide VII, which was prepared by treatment of 4-vinylaniline with ethyl formate, using sodium hydride gives N-{5,5-dimethyl-7- [(1S)"l-methylpropyl]-6-oxo-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-2-yl}-N-(4- vinylphenyl)formamide VIIL Removal of the formyl group under mildly basic conditions leads to 5,5-dimethyl-7-[(liS)-1-methylpropyl]-2-[(4-vinylphenyl)amino]-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one IX which is followed by osmium tetroxide-mediated oxidative cleaved of the vinyl group to give 4-({5,5-dimethyl-7-[(1S)-1-methylpropyl]-6-oxo-6,7-dihydro-5-pyrrolo[2,3-d]pyrimidin-2-yl}amino)benzaldehyde X. The aldehyde X can be coupled with various primary or secondary amines XI using standard reductive amination conditions to provide benzylic amines of the type XII.

Scheme B

XII IX

General procedures to prepare compounds of the instant invention are also described in Scheme C. The sulfone of the 2-sulfonylpyrimidine VI can be displaced using the anion of various aryl and heteroaryl amines XIII, which are generated in situ through deprotonation with sodium bis(trimethylsilyl)amide, to give substituted 2-aminopyrimidine products of the class XIV.

Scheme C

Vl XIV

General procedures to prepare compounds of the instant invention are also described in Scheme D. The sulfone of the 2~sulfonylpyrimidine VI can be displaced with electron rich heteroaryl amines such as l-alkyl-lH-pyrazol-4-amines XV under high temperature conditions to afford substituted 2-aminopyrimidines of type XVI.

Scheme D

VI XVI

In the following non-limiting Examples, unless otherwise stated:

1. All the end products of the formula XII, XIV, and XVI were analyzed by NMR, LCMS.

2. Intermediates were analyzed by NMR and/or TLC and/or LCMS. 3. Most compounds were purified by flash chromatography on silica gel, reverse phase HPLC, recrystallization and/or swish (suspension in a solvent followed by filtration of the solid). 4. The course of reactions was followed by thin layer chromatography (TLC) and/or LCMS and reaction times are given for illustration only.

Intermediate 1

5,5-dimethyl-7-[(1S' )-1-methylpropyI]-2-(methylsulfonyl)-5,7-dihydro-6H-pyrrolo[2,3- d] pyrimidin-6-one

Step 1 : Diethyl 2-formylsuccmate

Ethanol (27.0 g, 0.574 mol) was added dropwise to a suspension of sodium hydride (60 wt%, 23.0 g, 0.574 mol) in tetrahydrofuran (300 mL); the mixture was stirred at room temperature for 1 hour. Upon cooling to 10 °C (cold water bath), solutions of diethyl succinate (100 g, 0.574 mol) in tetrahydrofuran (100 mL) followed by ethyl formate (42.5 g, 0.574 mol) in tetrahydrofuran (100 mL) were added dropwise. After stirring for 16 h at room temperature, water (500 mL) was added and the resulting solution was washed with diethyl ether (2 x 250 mL). The water phase was acidified with 50% aq. H2SO4 and the resulting suspension was extracted with diethyl ether (2 x 250 mL). The combined organic layer was washed with water (3 x 200 mL), dried with sodium sulfate, filtrated through Celite, and concentrated under reduced pressure. The residue was distilled in vacuo (5 mm Hg) to afford 62.1 g (53% yield) of the desired diethyl 2-formylsuccinate, b.p. 90 °C. ¹H NMR (400 MHz, CDCl₃) δ: 11.40 (d, J= 11.0 Hz, 0.5 H), 9.80 (s, 0.5 H), 7.00 (d, J= 11.0 Hz, 0.5 H), 4.30-4.00 (m, 4 H), 3.70-3.61 (m, 0.5 H), 3.00 (s, 1 H), 2.85-2.70 (m, 1 H), 1.28-1.10 (m, 6 H).

Step 2: Ethyl [2-(methylthio)-4-oxo-L4-dihydropyrimidin-5-yl]acetate

To a solution of diethyl 2-formylsuccinate (53.4 g, 0.264 mol) and S- methylthiourea semi-sulfate (36.8 g, 0.264 mol) in 240 mL of water, a solution of sodium hydroxide (15.8 g, 0.396 mol) water (60 mL) was added, The reaction mixture was stirred at 100 °C for 1.5 hours, cooled to room temperature, and acidified with slow addition of acetic acid. After stirring at room temperature for 15 min, the resulting precipitate was filtered, thoroughly washed with water, and finally dried by evaporation with acetonitrile (3 times) to afford ethyl [2- (methylthio)-4-oxo-1,4-dihydropyrimidin-5-yl]acetate (32.6 g, 50% yield), 1H NMR (400 MHz, CDCl₃) δ: 7.81 (s, 1 H), 4.18 (q, J = S.I Hz, 2 H), 3.42 (s, 2 H), 2.55 (s, 3 H), 1.25 (t, J= 5.7 Hz, 3 H).

Step 3: Ethyl [4-chl oro-2-(methylthio)pyrimidin-5-yl 1 acetate

A mixture of ethyl [2-(methylthio)-4-oxo-1,4-dihydropyrimidin-5-yl]acetate (28.8 g, 0.128 mol) in POCl₃ (240 mL) was refluxed for 4 hours and then concentrated under reduced pressure. The residue was twice re-evaporated with benzene and purified by column chromatography on silica gel (ethyl acetate-hexanes, 1 :3) to afford ethyl [4-chloro-2-

(methylthio)pyrimidin-5-yl]acetate (29.72 g, 95.5% yield).

¹HNMR (400 MHz, CDCl₃) δ: 8.32 (s, 1 H), 4.13 (q, J- 5.7 Hz, 2 H), 3.62 (s, 2 H), 2.52 (s, 3 H), 1.21 (t, J= 5.7, 3 H).

Step 4: Ethyl 2-[4-chloro-2-(methylthio)pyrimidin-5-yl]-2-methylpropanoate

A solution of ethyl [4-chloro-2-(methylthio)pyrimidin-5-yl]acetate (150 g, 608 mmol) and methyl iodide (259 g, 1824 mmol) in N,N-dimethylformamide (2 L) was cooled to 0 °C and sodium hydride (60 wt%, 60.8 g, 1520 mmol) was added over ninety minutes. The cooling bath was removed upon completion of the addition and the reaction mixture was allowed to warm to room temperature overnight. Excess base was quenched by the addition of 7% aqueous citric acid solution (150 mL); the resulting mixture was poured into an extractor containing water (5 L) and extracted with diethyl ether 4 times. The combined organic layers were washed with water, dried over sodium sulfate, filtered, and concentrated to give a dark reddish oil. This crude oil was purified by chromatography using a 1.5 kg silica column (1: 19 ethyl acetate-heptane, grading to 1 :3 ethyl acetate-heptane) to give 2~[4-chloro-2-(methylthio)pyrirnidin~5-yl]-2- methylpropanoate (151.8 g, 91% yield) Calcd for C₁₁H₁₅ClN₂O₂S [M+l]⁺: 275, Found: 275

Step 5,5-Dimethyl-7-[(1 S )-1-methylpropyl ]-2-(methylthio)-5,7-dihvdro-6H-pyrrolo [2,3-d]pyrimidin~6-one

A solution of 2-[4-chloro-2-(methylthio)pyrimidin-5-yl]-2-methylpropanoate (28.4 g, 103 mmol), (S)-sec-(-)-butyl amine (9.07 g, 124 mmol), and l,4-diazabicyclo[2.2.2]octane (23.19 g, 207 mmol) in ethanol (500 mL) was heated to 140 °C at high pressure (110 psi) for 32 hours. The mixture was concentrated to afford a crude brown oil, which was purified by silica gel chromatography (ethyl acetate-heptanes, 1:4) to give 5,5-dimethyl-7-[(lS)-1-methylpropyl]-2- (methylthio)-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one (24.0 g, 87%). Calcd for CnHi₉N₃OS [M+l]⁺: 266, Found: 266

Step 6: 5,5-Dimethyl-7-[(1 S )-1-methylpropyl]-2-(methylsulfonyl)-5,7-dihvdro-6H-pyrrolo[2,3-d]pyrimidin-6-one

To a solution of 5,5-dimethyl-7-[(1S)-1-methylpropyl]-2-(methylthio)-5J~dihydro-6H- pyrrolo[2,3-d]pyrimidin-6-one (24.3 g, 92 mmol) in dichloromethane (500 mL) was added meta- chloroperoxybenzoic acid (77 wt%, 47.2 g, 211 mmol), After being stirred overnight, the reaction was diluted with dichloromethane and the resulting organic layer was washed with aqueous saturated sodium bicarbonate solution, aqueous 1 M sodium hydroxide solution twice, water, and finally brine. The organic layer was dried over sodium sulfate, filtered, and concentrated. The resulting off-white solid was triturated with methyl tert-butyl ether and the solids were collected by filtration to yield 5,5-dimethyl-7-[( 1S)-I -methylpropyl]-2- (methylsulfonyl)-5,7-dihydro-6H-pyrrolo[2_}3-d]pyrimidm-6-one (19.4g, 71% yield) as a white solid,

Calcd for C₁₃Hi₉N₃O₂S [M-H]⁺: 298, Found: 298

Example 1

2-[(4-{ [(2-Methoxy- 1 -methylethyl)amino3methyϊ}phenyl)amino]-5,5-dimethyl-7-[(1S)-I - methylpropyl]-5,7-dihydro-6H-pyrrol o [2,3-d] pyrimidin-6-one

Step 1 : N-(4-VinylphenvI)formamide

4- Amino styrene (2.0Og, 16,78 mmol) was treated with ethyl formate (4,10 mL, 50.4 mmol) and stirred at 48 °C for 26 hours. The reaction was concentrated to remove ethanol and additional ethyl formate (4.10 mL) was added. The reaction was stirred at 64 °C for 19 hours and then concentrated under reduced pressure to afford N-(4-vinylphenyl)formamide (2.26 g, 91% yield). Calcd for C₉H₉NO [M+ 1]⁺: 148, Found: 148

Step 2 : 5,5-Dimethyl-7-r(1S)-1-methvlpropvll-2-[f4-vinylphenyl)amino1-5,7-dihydro-6H- pyrrolof2,3-(f|pyrimidin-6-one

To a stirred solution of 5,5<limethyl-7-[(lS)-1-memylρropyl]-2-(methylsuIfonyl)-5,7- dihydro-6H-pyrrolo[2,3-(f|pyrimidin-6-one (6.9 g, 23.20 mmol), and N-(4- vinylphenyl)formamide (3.93 g, 26.7 mmol) in N,N-dimethylforrnamide (186 ml) was added sodium hydride (60 wt%, 1.485 g, 37.1 mmol) over 5 minutes. The reaction mixture was stirred at room temperature for 50 minutes and then partitioned between ethyl acetate and water. The organic layer was washed with one portion of water and one portion of brine, dried over sodium sulfate, filtered, and concentrated to afford an orange oil. The oil was dissolved in methanol (41 ml), tetrahydrofuran (41 ml) and saturated aqueous sodium bicarbonate solution (41 ml). The reaction was stirred at room temperature for 40 minutes and then partitioned between ethyl acetate and water. The organic layer was washed with one portion of brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel flash chromatography (1: 19 ethyl acetate-hexanes, grading to 3:5 ethyl acetate-hexanes) to afford 5,5-dimethyl-7-[(lS)- 1 -methylpropyl]-2-[(4-vinylphenyl)amino]-5,7-dihydro-6H-pyrroio[2,3-tir|pyrimidin-6-one (4.8 g, 62% yield). Calcd for C₂₀H₂₄N₄O [M+l]⁺: 337, Found: 337

Step 3: 4-( { 5 ,5 -Dimelhy 1-7- IY 1 S)- 1 -methγlpropγl1-6-oxo-6 J-dihydro-5 -pyrrolof 2,3 - Jlpyrimidin- 2-yl I ammo)benzaldehyde

A stirred solution of 5,5-dimethyl-7-[(lS)-1-methylproρyl]-2-[(4-vinylphenyl)amino]-5,7- dihydro-6H-pyrrolo[2,3-c/jpyrimidin-6-one (4.8 g, 14.27 mmol) in 1,4-dioxane (86 mL) was treated dropwise with water (57 mL), then sodium periodate (6.41 g, 30.0 mmol) and osmium tetroxide (4% in water, 1.1 mL, 0.143 mmol). The reaction mixture was stirred at room temperature for 35 minutes and then concentrated. The residue was partitioned between water and ethyl acetate. The organic layer was washed with one portion of saturated aqueous sodium thiosulfate solution, one portion of brine, dried over sodium sulfate, filtered, and concentrated to afford 4-( { 5 , 5 -dimethyl-7- [( 1 S)- 1 -methy lpropy 1] -6-oxo-6 , 7-dihydro-5 -pyrrolo [2,3 -d\ pyr imidin- 2-yl}amino)benzaldehyde (4.7 g, 97% yield). Calcd for Cj₉H₂₂N₄O₂ [M+l]⁺: 339, Found: 339

Step 4: 2-f⁽4-([(2-Metho^χy-1-methyleth^yl⁾amino1meth^yUphen^yl^')aminol-5,5-dimethyl-7-Kiy⁾-1- methy lpropyl] - 5 ,7-dihydro -6H-pyrrolo [2,3 -d\ pyr imidin-6 -one

To a stirred solution of 4-({5,5-dimethyl-7-[(1S)-1-methylpropyl]-6-oxo-6,7-dihydro-5- pyrrolo[2,3-(f|pyrimidin-2-yl}amino)benzaldehyde (300 mg, 0.887 mmol) in 5:95 acetic acid / 1,2-dichloroethane (8.9 mL), l-methoxy-N-methylpropan^-amine (95 mg, 1.064 mmol) and 4A molecular sieves were added. The reaction mixture was stirred at room temperature for 16 hours and then sodium triacetoxyborohydride (263 mg, 1.241 mmol) was added. The reaction mixture was stirred for 1 hour and then partitioned between ethyl acetate and saturated aqueous sodium bicarbonate solution. The organic layer was washed with one portion of brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel flash chromatography (dichloromethane, grading to 1 :13 methanol-dichloromethane) to afford 2-[(4-{[(2-Methoxy-1- methy lethyl)amino] methyl } phenyl)amino] -5,5 -dimethyl-7- [( 1S)- 1 -methylpropyl] -5 , 7-dihydro- 6H-pyrrolo[2,3-d]pyrimidin-6-one (132 mg, 36% yield). Calcd for C₂₃H₃₃NsO₂ [M+lf: 412, Found: 412

Using the procedures in Example 1, the compounds listed in Table 1 were also prepared.

Example 40

2-[(6-Methoxypyridin-3-yl)amino]-5,5-dimethyl-7-[(liS)-1-methylpropyl3-5,7-dihydro-6H- pyrrolo[2,3-(fipyrimidin-6-one

To a stirred solution of 5,5-dimethyl-y-Jfl^-1-melhylpropylj^-tmethylsulfonyl)-5-,7- dihydro-6/f-pyrrolo[2,3-_<i]pyrirnidin-6-one (75 mg, 0.252 mmol) and 6-methoxyρyridin-3-amine (34.4 mg, 0.277 mmol) in tetrahydrofuran (3 mL) under an argon atmosphere was added sodium bis(trimethylsilyl)amide (1 M in tetrahydrofuran, 0.328 mL, 0.328 mmol) over 30 seconds. After 15 minutes acetic acid (0.025 mL, 0.433 mmol) was added and the reaction mixture was concentrated. The residue was purified by reverse phase HPLC (1 :9 acetonitri Ie- water with 0.05% trifluoroacetic acid, grading to 7:3 acetonitrile-water with 0.05% trifluoroacetic acid) to provide 2-[(6-methoxypyridin-3-yl)amino]-5,5-dimethyl-7-[(1S)- 1 -methylpropyl]-5,7~dihydro- 6H-pyrrolo[2,3-t/]pyrimidin-6-one (22.7 mg, 0.066 mmol), Calcd for C₁₈H₂₃N₅O₂ [M+l]⁺: 342, Found: 342

Example 41

2-[(l-Benzyl-17f-pyrazol-4-yl)amino]-5,5-dimethyl-7-t(1S)-1-methylpropyl]-5,7-dihydro-6H- pyrrolo[2,3-<i]pyrimidin-6-one

A solution of 5,5-dimethyl-7-[(1S)-1-methylpropyl]-2-(methylsulfonyl)-5,7-dihydro-6H- pyrrolo[2,3-_</|pyrirnidin-6~one (49.4 mg, 0.166 mmol) and 1 -benzyl- l/i-pyrazol-4-amine (29.1 mg, 0.168 mmol) in 1,4-dioxane (1.5 mL) was heated at 130 °C for 16 hours. The reaction mixture was allowed to cool and then it was concentrated. The resulting residue was purified by reverse phase ΗPLC (1 :9 acetonitrile-water with 0.05% trifluoroacetic acid, grading to 9:1 acetonitrile-water with 0.05% trifluoroacetic acid) to afford 2-[(l-benzyl~1H-pyrazol-4- yl)amino]-5,5-dimethyl-7-[(1S)-1-methylpropyl]-5,7-dihydro-6H-pyrrolo[2,3-<|pyrimidin-6-one (24.4 mg, 0.062 mmol). Calcd for C₂₂H₂₆N₆O [M+l]⁺: 391, Found: 391

Examples 42 - 67 The following were prepared by the methods described for Intermediate 1 and Example 1, substituting the appropriate benzylic amine for seobutylamine in step 5 of Intermediate 1. Any benzyl ic amine intermediates that are not commercially available were made according to the methods of Cogan et al. Tetrahedron 1999, 55, 8883-8904, Any N-I alkyl 4-amino pyrazole intermediates that are not commercially available were made according to the methods of Zabierck et al. Tetrahedron Lett. 2008, 49, 2996-2998. When a protecting group for nitrogen was needed, a tert-butyl carbamate was employed and later cleaved using conditions familiar to those skilled in the art (e.g. trifluoroacetic acid and dichloromethane).

Intermediate 2

( 1.S)-4-hydroxy-2,3-dihydro- 1 /i-inden- 1 -aminium trifluoro acetate

Step 1 : tert-butγl (4-hydrρxy-2,3-dihvdro- 1 H-inden- 1 -ypcarbamate

Ammonium acetate (128 g, 1660 mmol) was suspended in MeOH and concentrated in vacuo three times, then combined with 4-hydroxy-1-indanone (16.4 g, 1 1 1 mmol) and sodium cyanoborohydride (11.1 g, 177 mmol) and MeOH (250 mL) in a IL pressure vessel. The vessel was sealed and the reaction mixture was heated to 70 °C overnight. The reaction was cooled down to ambient temperature, and then concentrated HCl was carefully added until the pH = 1. The mixture was stirred for an additional 20 minutes and the solvent was concentrated to about 25% initial volume under reduced pressure. Aqueous ammonia was added until the pH = 10, then the mixture was diluted with ethyl acetate. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered, and concentrated to afford crude l-amino-2,3-dihydro-l/i-inden-4- ol.

To a solution of crude l-amino-^-dihydro-l/f-inden^-ol in THF (400 ml) was added triethylamine (21.7 ml, 156 mmol) and di-tert-butyl dicarbonate (22.7 g, 104 mmol). The reaction mixture was allowed to stir at room temperature for 1 hour, then diluted with ethyl acetate, washed with saturated aqueous sodium bicarbonate and brine. The organic layer was separated, and the aqueous layer was back extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (0-50% EtOAc/hexanes gradient to give tert-buty\ (4- hydroxy-2,3 -dihydro- 1 H-inden- 1 -yl)carbamate.

Racemic tert-butyl (4~hydroxy-2,3-dihydro-1H-inden-1-yl)carbamate was purified by chiral chromatography on a Berger Multigram SFC (Chiral Technology AD-H 2.1X25cm, 5uM; 25%/75% lsopropanol/CO₂; 70 mL/min, 5min run time; 220 πm detection) to deliver enantiomer peak 1 (elution time = 2 min) and enantiomer peak 2 (elution time = 3.5 min).

Step 2: ( 1 SM-hydroxy-2,3 -dihydro- 1 H-inden- 1 -amim^'um trifluoroacetate

Enantiomer peak 2 (5 g, 20.06 mmol) from Step 1 was dissolved in dichloromethane (120 ml) and cooled to 0 °C. Trifluoroacetic acid (40 ml, 519 mmol). Was quickly added and the reaction was allowed to warm to ambient temperature. After 30 minutes, the reaction mixture was concentrated to an oil, then dissolved in 100 mL of heptane and concentrated in vacuo to afford (1S)-4-hydroxy-2,3-dihydro-lH-inden-1-aminium trifluoroacetate. The absolute stereochemistry of this salt was determined via analysis of an X-ray crystal structure of the derivative 2-anilino-7-[(l S)-4-hydroxy-2,3-dihydro-l H-inden-1-yl]-5,5-dimethyl-5,7-dihydro- 6H-ρyrrolo[2,3-d]pyrimidin-6-one (Example 42).

Other l-amino-2,3-dihydro-1H-indenes having different (or no) substitution on the aromatic ring, were prepared by the same method, starting from the appropriately substituted indanone.

Example 68

2-aniIino-7-[(l S)-4-hydroxy-2,3-dihydro-1H-inden-1-yl]-5,5-dimethyl-5,7-dihydro-6H- pyrro Io [2 ,3 -d] pyrimidin-6-one

Step 1: ethyl 244-chloro-2-(methylsulfonyl)pyrimidin-5-yll-2-methylpropanoate

rø-CPBA (30.1 g, 131 mmol) was added over a period of 5 minutes to a stirred solution of ethyl 2-[4-chloro-2-(methylthio)pyrimidin-5-yl]~2-methylpropanoate (18 g, 65.5 mmol) in dichloromethane (250 ml) at 0 °C. The reaction mixture was stirred at 0 °C for 10 minutes, then allowed to warm to ambient temperature. After 2 hours, additional rø-CPBA (3.77 g, 16.38 mmol) was added and the reaction mixture was stirred overnight at ambient temperature. The mixture was poured into ethyl acetate (IL), washed 1 x 250 mL saturated sodium bicarbonate solution, 1 x 300 mL 0.5N NaOH, then 2 x 100 mL brine. The organics were dried over magnesium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (0-100% ethyl acetate/hexanes gradient) to afford 2-[4-chloro-2- (methylsulfonyl)ρyrimidin-5-yl]-2-methylpropanoate (20.62 g). Calcd for C_HH_I6CIN₂O₄S [M+l]⁺: 307.0, Found: 307.0.

Step 2: ethyl 2-f4-chloro-2-fphenylamino)pyrimidin-5-vn-2-methylpropanoate

NaH (3.03 g, 76 mmol) was added to a solution of formanilide (8.04 g, 66.3 mmol) in DMF (100 ml) at 0 °C. The mixture was allowed to warm to ambient temperature and stirred for 30 minutes. This mixture was added via cannula to a cooled solution of 2-[4-chloro-2- (methylsulfonyl)pyrimidin-5-yl]-2-methylpropanoate (19.38 g, 63.2 mmol) in DMF (120 ml) at 0 °C over period of 10 minutes. The mixture was allowed to warm to ambient temperature and stirred for 1 additional hour. The reaction mixture was cooled to 0 °C and slowly quenched with 100 mL of saturated sodium bicarbonate solution, followed by 50 mL of methanol. The resulting slurry was stirred overnight at ambient temperature, then diluted with an additional 750 mL water and 500 mL ethyl acetate. The organic layer was separated, and the aqueous layer was washed with ethyl acetate (2 x 250 mL). The combined organic layers were washed 1 x 300 mL water, then 2 x 100 mL brine, dried over magnesium sulfate, filtered, and concentrated. The resulting semi-solids were dissolved in 100 mL hot ethyl acetate and partially concentrated to approximately 25 mL of solution. The resulting solution was stirred vigorously for several minutes, during which time significant precipitation of white solids began. Hexanes (450 mL) were added over a period of several minutes, during which time significant precipitation of colorless solids continued as the suspension was stirred. The mixture was allowed to stir for 2 hours. The colorless solids were filtered and dried in vacuo to afford ethyl 2-[4-chloro-2- (phenylamino)pyrimidin-5-yl]-2-methylpropanoate (15.1 g). The mother liquor was concentrated and purified by flash chromatograhpy (0-40% ethyl acetate/hexanes gradient) to afford an additional 2.2 g of ethyl 2-[4-chloro-2-(phenylamino)pyrimidin-5-yl]-2-methylpropanoate. Calcd for Ci₆Hi₉ClN₃O₂ [M+l]^÷: 319.1, Found: 320.1.

Step 3: 2-anilino-7-^iS)-4-hvdroxv-2,3-dihvdro-1H-inden-1-yll-5,5-dimethyl-5J-dihvdro-6H- pyrrolo[2,3-d1pyrimidin-6-one.

(l£)-4-hydroxy-2,3-dihydro-l/f~inden-1-aminium trifluoroacetate (4.94 g, 20.06 mmol) was dissolved with 2-ethoxyethanol (10 ml) in a pressure vessel. Diisopropylethylamine (7.51 ml, 43.0 mmol) was added, followed ethyl 2-[4~chloro-2-(phenylamino)pyrimidin-5-yl]-2- methylpropanoate (2.75 g, 8.60 mmol), The reaction vessel was sealed and the mixture was heated to 160 °C for two days. The reaction mixture was cooled to ambient temperature, diluted with ethylacetate and washed with saturated aqueous sodium bicarbonate. The organic layer was separated, and the aqueous layer was back extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography on silica gel (EtOAc/hexanes) to afford 2.5 g of 2-anilino-7-[(l S)- 4-hydroxy-2,3-dihydro-1H-inden-l -yl]~5,5-dimethyl-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6- one. The absolute stereochemistry was determined by X-Ray analysis. ¹H NMR (500 MHz, DMSO-d₆) δ 9.49 (s_} 1H); 9.46 (s, 1H); 8.22 (s, 1H); 7.25 (br s, 2H); 7.15 (app t, 2H); 6.92 (dd, 1H); 6.83 (dd, 1H); 6.66 (d, 1H); 6.40 (d, 1H); 5.85 (t, 1H); 3.01-3.09 (m, 1H); 2.73-2.82 (m, 1 H); 2.30-2.64 (m, 2H); 1.39 (s, 3H); 1.33 (s, 3H). LRMS (ESI) calculated for C₂₃H₂₃N₄O₂ [M+Hf, 387.2; found 387.2.

Example 69 7-[(l₁S)-4-hydroxy-2,3-dihydro-1H-inden-1-yl]-5,5-dimethyl-2-(1,2,3,4-tetrahydroisoquinolin-6- ylamino)-5,7~dihydro-6H-ρyrrolo[2,3~(f]pyrimidin-6-one

Step 1 : 5,5-dimethyl-2-fmethylthioV7-jϊl^-4-αetrahvdro-2/j^r-pyran-2-yloxyV2,3-dihvdro-1H- inden- 1 -yl] - 5 ,7 -dihydro-6H-pyrrolo \23-d] pyr imidin-6-one

7-[(1S)-4-hydroxy-2,3-dihydro-1H-inden-1-yl]-5,5-dimethyl-2-(methylthio)-5,7-dihydro- 6H-pyrrolo[2,3-ύ?]pyrimidin-6-one was prepared as described for Intermediate 1 steps 1-5, substituting (1S)-4-hydroxy-2,3-dihydro-lH-inden-l -aminium trifluoroacetate for ^ec-butylamine in step 5. To a solution of this compound (250 mg, 0.732 mmol) in dichloromethane (10 ml) at 20

°C was added 3,4-dihydro-2H-pyran (0.669 ml, 7.32 mmol) and p-TsOH pyridinium salt (3 mg, 0.012 mmol). After 16 hours, the reaction mixture was concentrated and purified by column chromatography on silica gel, eluting (ethyl acetate/hexanes gradient) to afford 5, 5 -dimethyl -2- (methylthio)-7 - [( 1S)-4-(tetrahydro-2H~ρyran-2-yloxy>2, 3 -dihydro- 1 H-inden- 1 -yl] - 5 ,7-dihydro- 6H-pyrrolo[2,3-f/]pyrimidin-6-one.

¹H NMR (500 MHz, DMSOd₆) δ 8.38 (d, 1H); 7.06-7.03 (m, 1H); 6.92 (dd, 1H); 6.61 (dd, 1H); 5.90-5.84 (ra, 1H); 5.51 (dt, 1H); 3.79-3.72 (m, 1H); 3.56-3.53 (m, 1H); 3.22-3.14 (m, 1H); 2.92-2.82 (m, 1H); 2.51-2.42 (m, 2H); 2.20 (d, 3H); 1.92-1.84 (m, 1H); 1.84-1.72 (m, 2H); 1.66-1.48 (m, 3H); 1.35 (dd, 6H).

Step 2: 7-r(iy)-4-hvdroxy-2,3-dihvdro-1H-inden-1-yll-5,5-dimethyl-2-fmethylsulfonyl)-5J- dihydro-6H-pyrroϊoj-2,3-<i1pyrimidin-6-one

Oxone (1835 mg, 2.98 mmol) in water (2388 μl) was added to a mixture of SjS-dimethyl^- tmethylthioH-JOSH-ttetrahydro^H-pYran^^ 6H-pyrrolo[2,3-d]pyrimidin~6-one (254 mg, 0.597 mmol), saturated sodium bicarbonate (1.2 mL), THF (1.2 ml), and methanol (1.2 ml) at 20 °C. After 16 hours, the reaction mixture was filtered and washed with dichloromethane. Filtrate was concentrated to afford 7-[(1S)-4- hydroxy-2,3-dihydro-lH-inden-1-yl]-5,5-dimethyl-2-(methylsulfonyl)-5,7-dihydro-6H- pyrrolo[2,3-</)pyrimidin-6-one. Material was used crude without purification. LRMS (ESI) calculated for C₁₈H₂₀N₃O₄S [M+H]⁺, 374.1 ; found 374.1.

Step 3 : 5 ,5 ~dimethyl-2-fmethv!sulfonγD- 7- iϊ 1 ₎S^r)-4-Ctetrahvdro-2/f-pyran-2-yloxy^')-2,3 -dihydro-

1 /i-inden- 1 -y 1] - 5 ,7-dihydro-6H-pyrrolo [2,3 -<^]pyrimidin-6-one To a solution of 7-[(1S)-4-hydroxy-2,3-dihydro-lf/-inden-1-yl]-5,5-dimethyl-2-(methylsιιlfonyl)- 5,7-dihydro-6H-pyrrolo[2,3-tf]pyrimidin-6-one (223 mg, 0.597 mmol) in dichloroniethane (10 ml) at 20 ^QC was added 3,4-dihydro-2H-pyran (1.092 ml, 1 1.94 mmol) and p-TsOH pyridinium salt (3 mg, 0.012 mmol). After 16 hours, the reaction mixture was concentrated and purified by column chromatography on silica gel, eluting (ethyl acetate/hexanes gradient) to afford 5,5- dimethyl-2-(methylsulfonyl)-7-[(1S)-4-(tetrahydro-2H-pyran-2-yloxy)-2,3-dihydro--1H-inden-1- yl]-5,7-dihydro-6H-pyrrolo[2,3-^]pyrimidin-6-one.

¹H NMR (500 MHz, DMSOd₆) δ 8.74 (d, 1H); 7.08-7.02 (m, 1H); 6.93 (dd, 1H); 6.70 (t, 1H); 5.94-5.89 (m, 1H); 5.48 (dt, 1H); 3.79-3.72 (m, 1H); 3.56-3.50 (m, 1H); 3.30-3.19 (m, 1H); 3.11 (d, 3H); 2.95-2.85 (m, 1H); 2.50-2.38 (m, 2H); 1.95-1.85 (m, 1H); 1.85-1.72 (m, 2H); 1.68- 1.50 (m, 3H); 1.40 (dd, 6H). LRMS (ESI) calculated for C₂₃H₂₈N₃O₅S [M+H]⁺, 458.2; found 458.2.

Step 4: tert-butyl 6-({5,5-dimethyl-6-oxo-7-IY iSV4-ftetrahvdro-2ff-pyran-2-yloxyV2,3-dihydro~ 1 /f-inden- 1 -yl] -6,7-dihydro-5H-pyrrolo [2,3 -d] pyrimidin-2-yl } amino)-3 ,4-dihγdroisoquinoline- 2( I /JVcarboxylate

To a solution of tert-b\xty\ 6-(formylamino)-3,4-dihydroisoquinoline-2(l/i)-carboxylate (93 mg, 0.337 mmol) in DMF (5 ml) at 0 °C was added sodium hydride (15.91 mg, 0.398 mmol). After 30 minutes, the reaction mixture was added via syringe to a solution of 5,5-dimethyl-2- (methylsulfonyl)-7-[(1S)-4-(tetrahydro-2/f-pyran-2-yloxy)-2,3-dihydro- 1 /f-inden- 1 -yl]-5,7- dihydro-6/f-pyrrolo[2,3-<|pyrimidin-6-one (140 mg, 0.306 mmol) in DMF (5 ml) at 0 °C. Allowed reaction mixture to warm to ambient temperature. After 30 minutes, the reaction mixture was quenched with saturated sodium bicarbonate solution, diluted with ethyl acetate. Organics were separated, washed with brine, dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was purified by column chromatography on silica gel, eluting (ethyl acetate/hexanes gradient) to afford tert-bxxtyl 6-({5,5-dimethyl-6-oxo-7-[(lS)- 4-(tetrahydro-2H-pyran-2-yloxy)-2,3-dihydro- l//-inden- 1 -yl]-6,7-dihydro-5/f-pyrrolo[2,3- t/jρyrimidin-2-yl } amino)-3,4-dihydroisoquinoline-2(l H)-carboxylate.

LRMS (ESI) calculated for C₃₆H₄₄N₅O₅ [M+H]⁺, 626.3; found 626.4.

Step 5: 7-[fl5^f)-4-hvdroxy-23-dihvdro-lH-inden-1-yll-5,5-dimethyl-2-(1.23,4- tetrahydroisoquinolin-6-ylamino)-5,7-dihydro-6H-ρyrrolor2,3-<f1pyrimidin-6-one

To a solution of tert-butyl 6-({5,5-dimethyl-6-oxo-7-[(1S)-4-(tetrahydro-2H-pyran-2-yloxy)~2,3- dihydro-lH-inden-1-yl]-6,7-dihydro-5H-pyrrolo[2_J3-(f|pyrimidin-2-yl}amino)-3,4- dihydroisoquinoline-2(1H)-carboxylate (206 mg, 0.329 mmol) in Dichloromethane (2 ml) at 20 °C was added TFA (2 mL, 26.0 mmol). After 30 minutes, the reaction mixture was concentrated and purified by preparative ΗPLC Reverse phase (C- 18), eluting with acetonitrile/water + 0.05% TFA to give 7-t(1S)-4-hydroxy-2,3-dihydro-l/Mnden-1-yl]-5,5-dimethyl-2-(1,2,3,4- ietrahydroisoquinolin-6-ylamino)-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one.

¹H NMR (500 MHz, DMSOd₆) δ 9.50-9.40 (bs, 1H); 9.35 (s, 1H); 8.20 (s, 1H); 7.16- 7.06 (bs, 2H); 6.91 (t, 1H); 6.85 (d, 1H); 6.64 (d, 1H); 6.39 (d, 1H); 5.82 (t, 1H); 3.82 (s, 2H); 3.10-3.02 (m, 1H); 3.00-2.94 (m, 2H); 2.80-2.72 (m, 1H); 2.68-2.55 (m, 3H); 2.38-2.28 (m, 1H); 1.33 (d, 6H). LRMS (ESI) calculated for C₂₆H₂₈N₅O₂ [M+H]⁺, 442.2; found 442.2.

Example 70

7-[(1S)-4-hydroxy-2_;3-dihydro-lH-inden-1-yl]-5_J5-dimelhyl-2-[(2-methyl-l ,2,3,4- tetrahydroisoquinolin-6-yl)amino]-5,7-dihydro-6H-pyrrolo[2,3-βr|pyrimidin-6-one

To a solution of 7-[(1S)-4-hydroxy-2,3-dihydro-l/f-inden-1-yl3-5,5-dimethyl~2-(1,2,3,4- tetrahydroisoquinolin-6-ylamino)-5,7-dihydro-6H-pyiτolo[2,3-d]pyrimidin-6-one (10 mg, 0.023 mraol) in 1 ,2-dichloroethane (1 ml) at 20 °C was added Formaldehyde (3.37 μl, 0.045 mmol). After 10 minutes, the reaction mixture was cooled to OC and sodium Iriacetoxyborohydride (5,28 mg, 0.025 mmol) added. After 20 minutes, the reaction mixture was quenched with aqueous ammonium chloride and diluted with ethyl acetate. Organics were separated, washed with water, dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was purified by preparative ΗPLC Reverse phase (C-18), eluting with acetonitrile/water + 0.05% TFA to give 7-[(1S)-4-hydroxy-2_J3-dihydro-1H-inden-1-yl]-5_J5~dimethyl-2-[(2-methyl-l , 2,3,4- tetrahydroisoquinolin-6-yl)amino]-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one.

¹H NMR (500 MHz, Methanol-d₄) δ 8.05 (s, 1H); 7.06-7.00 (m, 2H); 6.98-6.90 (m, 2H); 6.64 (d, 1H); 6.40 (d, 1H); 5.94 (ι, 1H); 3.54 (s, 2H); 3.20-3.15 (m, 1H); 2.88-2.82 (m, 3H); 2.74- 2.66 (m, 3H); 2.43 (s, 3H); 2.42-2.35 (s, 1 H); 1.42 (s, 6H). LRMS (ESI) calculated for C₂₇H₃₀N₅O₂ [M+H]⁺, 456.2; found 456.2.

Example 71

2-[(2-acetyl-1,2,3,4-tetrahydroisoquinolin-6-yl)amino]-7-[(1S)-4-hydroxy-2,3-dihydro-1H-inden- l-yl]-5,5-dimethyl-5,7-dihydro-6H-pyrrolo[2,3-(i]pyrimidin-6-one

To a solution of 7~[(lS)-4-hydroxy-2_J3-dihydro-lH-inden-1-yl]-5,5-dimelhyl-2-(1,2,3,4- tetrahydroisoquinolin-6-ylamino)-5,7-dihydro-6H-pyrrolo[2,3-(/3pynmidin-6-one (25 mg, 0.057 mmol) in dichloromethane (1 ml) at 0 °C was added pyridine (0.024 ml, 0.297 mmol). After 5 minutes, acetic anhydride (5.34 μl, 0.057 mmol) was added to the reaction mixture. After 20 minutes, the reaction mixture was quenched with saturated sodium bicarbonate and diluted with ethyl acetate. Organics were separated, dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was purified by preparative HPLC Reverse phase (C-18), eluting with acetonitrile/water + 0.05% TFA to give 2-[(2-acetyl- 1,2,3, 4-tetrahydroisoquinolin-6- yl)amino]-7-[( lS)-4-hydroxy-2,3~dihydro- 1H-inden- 1 -yl]-5,5-dimethyl-5,7-dihydro-6H- pyrrolo[2,3-<i]pyrimidin-6-one.

¹H NMR (500 MHz, Methanol-d₄) δ 8.06 (s, 1H); 7.12-6.96 (m, 4H); 6.66 (d, 1H); 6.44 (d, 1H); 5.95 (t, 1H); 4.64-4.56 (m, 2H); 3.76-3.66 (m, 2H); 3.20-3.12 (m, 1H); 2.90-2.81 (m, 2H); 2.78-2.64 (m, 2H); 2.44-2.36 (m, 1H); 2.18 (d, 3H); 1.43 (d, 6H). LRMS (ESI) calculated for C₂₈H₃₀N₅O₃ [M+H]⁺, 484.2; found 484.2.

Example 72

7.[(l,S).4-hydroxy-2,3-dihydro-l/f-inden-1-yl3-5_J5-dimethyl-2-(1,2,3,4-tetrahydroisoquinolin-7- ylamino)-5 ,7-dihydro-6/i-pyrrolo [2,3 -ώf]pyrimidin-6-one

Step 1: fert-butyl 7-(formylamino)-3,4-dihvdroisoquinoline-2f1H)-carboxylate

To ethyl formate (20 ml, 246 mmol) was added tert-butyl 7-amino-3,4- dihydroisoquinoline-2(l//)-carboxylate (2.0 g, 8.05 mmol). The reaction mixture was heated to 60 °C. After three days, the reaction mixture was concentrated in vacuo, then diluted with ethyl formate (20 ml, 246 mmol) and heated to 60 °C. After 24 hours, the reaction mixture was concentrated and then purified by column chromatography on silica gel, eluting (ethyl acetate/hexanes gradient) to afford tert-butyl 7-(formylamino)-3,4-dihydroisoquinoline-2(l/f)- carboxylate. ¹H NMR (500 MHz, DMSOd₆) δ 10.10 (s, 1H); 8.22 (s, 1H); 7,44-7.36 (bs, 1H); 7.35- 7.25 (bs, 1H); 7.08 (d, 1H); 4.43 (s, 2 H); 3.54-3.48 (m, 2H); 2.71-2.67 (m, 2H); 1.41 (s, 9H). LRMS (ESI) calculated for Ci₅H₂]N₂O₃ [M+H]^÷, 277.2; found 277.2.

Step 2 : tert-butyl 7- { [4-chloro-5-C2-ethoxy- 1 , 1 -dimemyl-2-oxoethyl)pyrimidin-2~yl] amino } -3 ,4- dihydroisoquinoline-2fl//)-carboxylate

To a solution of tert-bntyl 7-(formylamino)-3,4-dihydroisoquinoline~2(l/i)-carboxylate (0.991 g, 3.59 mmol) in DMF (10 ml) at 0 °C was added sodium hydride (0.169 g, 4.24 mmol). The reaction mixture was allowed to warm to ambient temperature, then after 30 minutes was added via syringe to a solution of ethyl 2-[4-chloro-2-(methylsulfonyl)pyrimidin-5-yl3-2- methylpropanoate (1 g, 3.26 mmol) in DMF (5 ml) at 0 °C. The reaction mixture was allowed to warm to ambient temperature and after 30 minutes was quenched with saturated sodium bicarbonate solution and diluted with ethyl acetate. Organics were separated, dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was dissolved in THF (60 ml), methanol (50 ml), acetonitrile (10 ml), and saturated sodium bicarbonate solution (15 ml) and stirred. After 3 days, the reaction mixture was partially concentrated, then diluted with ethyl acetate. Organics were washed with water and then brine, dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was purified by column chromatography on silica gel, eluting (ethyl acetate/hexanes gradient) to afford tert-butyl 7-{[4- chloro-5-(2-ethoxy-l,l-dimethyl~2-oxoethyl)pyrimidin-2-yl]amino}-3,4-dihydroisoquinoline- 2(l//)-carboxylate.

¹H NMR (500 MHz, DMSO-d₆) δ 9.95 (s, 1H); 8.53 (s, 1H); 7.50 (s, 1H); 7.45 (d, 1H); 7.07 (d, 1H); 4.48-4.40 (bs, 2H); 4.07 (q, 2H); 3.55-3.50 (m, 2H); 2.72-2.66 (m, 2H); 1.53 (s, 6H); 1.41 (s, 9H); 1.12 (t, 3H). LRMS (ESI) calculated for C₂₄H₃₂ClN₄O₄ [M+Hf, 475.2; found 475.2.

Step 3: tert-butyl 7-r{7-[(iy)-4-hvdroxy-23-dihvdro-l/f-inden-1-yl1-5.5-dimethyl-6-oxo-6.7- dihvdro-5H-pyn:olo[2J-(f|pyrimidin-2-yl}amino)-3,4-dihvdroisoquinoline-2(l/j^r)-carboxylate To a suspension of tert-bntyl 7-{[4-chloro-5-(2-ethoxy-l,l-dimethyl-2- oxoethyl)pyrimidin-2-yl] amino }~3,4-dihydroisoquinoline-2(l/-/)-carboxylate (183 mg, 0.385 mmol) and (1S)-4-hydroxy-2,3-dihydro-l//"inden-1-aminium trifluoroacetate (291 mg, 0.771 mmol) in 2-ethoxyethanol (1 ml) was added Hunig's base (0.404 ml, 2.312 mmol). The reaction mixture was heated to 130 °C. After two days, the reaction mixture was diluted with ethyl acetate. The organics were washed with brine, dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was purified by column chromatography on silica gel, eluting (ethyl acetate/hexanes gradient) to afford tert-butyl 7-({7-[(l.S)-4-hydroxy-2,3- dihydro- 1 H-inden- 1 -yl] - 5 , 5 -dimethy l-6-oxo-6, 7-dihydro-5 H-pyrroIo [2,3 -d] pyr imidin-2 - yl} amino)-3 ,4-dihydroisoquinoline-2( 1 H)-carboxylate. LRMS (ESI) calculated for C₃₁H₃₆N₅O₄ [M+H]⁺, 542.3; found 542.3.

Step 4: 7-[fl₁S^f)-4-hvdroxy-2,3-dihvdro-lH-inden-1-vn-5.5-dimethyl-2-(L2.3.4- tetrahydroisoquinolin-7-ylamino)-5,7-dihvdro-6H-pyrrolor23-(^1pyrimidin-6-one To a solution of tert-bulyl 7-({7-[(lS)-4-hydroxy-2,3-dihydro-lH-inden-1-yl]-5,5- dimelhyl-6-oxo-6,7-dihydro-5H-pyrrolo[2,3-(f|pyrimidin-2-yl}amino)-3,4-dihydroisoquinoline- 2(1H)-carboxylate (162 mg, 0.299 mmol) in Dichloromethane (10 ml) was added TFA (2.304 ml, 29.9 mmol). After 1 hour the reaction mixture was concentrated and then purified by preparative ΗPLC Reverse phase (C- 18), eluting with acetonitrile/water + 0.05% TFA to afford 7-f(1S)-4-hydroxy-2,3-dihydro-lH-inden-l~yl]-5,5-dimethyl-2-(1,2,3,4-tetrahydroisoquinolin-7- ylamino)-5j7-dihydro-6/f-pyrrolo[2j3-tflpyrimidin-6-one.

¹H NMR (SOO MHz, DMSOd₆) δ 9.50-9.40 (bs, 1H); 9.33 (s, 1H); 8.19 (s, 1H); 7.15- 7.00 (m, 2H); 6.94-6.86 (m, 2H); 6.64 (d, 1H); 6.39 (d, 1H); 5.82 (t, 1H); 3.76 (s, 2H); 3.09-3.02 (m, 1H); 2.96-2.91 (m, 2H); 2.80-2.70 (m, 1H); 2.65-2.58 (m, 3H); 2.37-2.30 (m, 1H); 1.33 (d, 6H). LRMS (ESl) calculated for C₂₆H₂₈N₅O₂ [M+H]⁺, 442.2; found 442.2.

Example 73

7-[(15)-4-hydroxy-2,3-dihydro-lH-inden-1-yl]-5,5-dimethyl-2-[(2-methyl-l_J2,3,4- tetrahydroisoquinolin-7-yl)amino]-5,7-dihydro-6H~pyrrolo[2,3-ti]pyrimidin-6-one

Step 1 : ethyl 2-[4-chloro-2-f1,2,3,4-tetrahvdroisoquinolin-7-ylamino)pyrimidin-5-yl]-2- methylpropanoate trifluoroacetate

To a solution of tert-butyl 7-{[4-chloro-5-(2-ethoxy-l,l-dimethyl-2-oxoethyl)ρyrimidin- 2-yl]amino}-3,4-dihydroisoquinoline~2(l//)-carboxylate (1.469 g, 3.09 mmol) in dichloromethane (10 ml) was added TFA (5 ml, 64,9 mmol) at 0 °C. After 2 hours, the reaction mixture at 0 °C was quenched with saturated sodium bicarbonate and diluted with into ethyl acetate. The organics were washed with brine, dried over magnesium sulfate, filtered, and concentrated to afford ethyl 2-[4-chloro-2-(1,2,3,4-tetrahydroisoquinolin-7-ylamino)pyrimidin-5- yl]-2-methylpropanoate trifluoroacetate.

¹H NMR (500 MHz, DMSO-d₆) D 10.04 (s, 1H); 8.51 (s, 1H); 7.54-7.50 (m, 2H); 7.12 (d, 1H); 4.17 (s, 2H); 4.07 (q, 2H); 3.32-3.27 (m, 2H); 2.90-2.85 (m, 2H); 1.53 (s, 6H); 1.12 (t, 3H). LRMS (ESI) calculated for Ci₉H₂₄ClN₄O₂ [M+Hf, 375.2; found 375.1. Step 2; ethyl 2-{4-chloro-2-[T2-methyl-1,2,3,4-tetrahydroisoquiκolin-7-vl)amino]pyrimidm-5- yl } -2-methylpropanoate

To a solution of ethyl 2~[4-chloro-2-(1,2,3,4-tetrahydroisoquinolin-7-ylamino)pyrimidin- 5-yl]-2-methylproρanoate trifluoroacetate (500 mg, 1.023 mmol) in 1 ,2-DichIoroethane (10 ml) at 20 °C was added formaldehyde (0.152 ml, 2.045 mmol). After 10 minutes, the reaction mixture was cooled to 0 °C. Sodium triacetoxyborohydride (271 mg, 1.278 mmol) was added and after 20 minutes the reaction mixture was quenched with ammonium chloride and diluted with ethyl acetate. The organics were washed with water, dried over magnesium sulfate, filtered, and concentrated to afford ethyl 2-{4-chloro-2-[(2-methyl- 1 ,2,3,4-tetrahydroisoquinolin-7- y3)amino]pyrimidin-5-yl } -2-methylpropanoate.

¹H NMR (500 MHz, DMSOd₆) δ 9.87 (s, 1H); 8.50 (s, 1H); 7.42-7.36 (m, 2H); 7.00 (d,

1H); 4.07 (q, 2H); 3.41 (s, 2H); 2.76-2.70 (m, 2H); 2.58-2,52 (m, 2H); 2.31 (s, 3H); 1,52 (s, 6H);

1.12 (t, 3H). LRMS (ESI) calculated for C₂₀H₂₆ClN₄O₂ [M+H]⁺, 389.2; found 389.2.

Step 3: 7-fαSV4-hvdroχy-2,3-dihvdro-l#-inden-l -yll-5.5-dimethyl-2-r(2-methyl-l .2.3.4- tetrahydroisoquinolin-7-yl)aminol-5,7-dihydro-6H-pyrrolo|^'2,3-ιf|pyrimidin-6-ρne

To a solution of ethyl 2-{4-chloro-2-[(2-methyl-1,2,3,4-tetrahydroisoqumolin-7- yl)amino]pyrimidin-5-yl}-2-methylpropanoate (50 mg, 0.129 mmol) and (lS)-4~hydroxy-2,3- dihydro-1H-inden-1-aminium trifluoroacetate (121 mg, 0.321 mmol) in 2-ethoxyethanol (0.3 ml) was added Hunig's base (0.180 ml, 1.029 mmol). The reaction mixture was heated to 16O°C.

After 16 hours, the reaction mixture was cooled and purified directly by preparative HPLC

Reverse phase (C-18), eluting with acetonitrile/water + 0.05% TFA to afford 7-[(liS)-4-hydroxy-

2,3-dihydro-1H-inden-1-yl3-5_J5-dimethyl-2-[(2-methyl-1,2,3_J4-tetrahydroisoquinolin-7- yl)amino]-5,7-dihydro-6H-pyrrolo[2,3-ύr|pyrimidin-6-one.

¹H NMR (500 MHz, DMSO-d₆) δ 9.45 (s, 1H); 9.33 (s, 1H); 8.19 (s, 1H); 7,16-7.00 (m,

2H); 6.94-6.89 (m, 2H); 6.64 (d, 1H); 6.38 (d, 1H); 5.82 (t, 1H); 3.35 (s, 2H); 3.16-3.02 (m, 2H);

2.80-2.72 (m, 1H); 2.72-2.50 (m, 4H); 2.38-2.28 (m, 4H); 1.33 (d, 6H). LRMS (ESI) calculated for C₂₇H₃₀N₅O₂ [M+H]⁺, 456.2; found 456.3.

Example 74

2-[(2-acetyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino]-7-[(lS)-4-hydroxy-2,3-dihydro-1H-inden- l-yl]-5,5-dimethyl-5,7-dihydro-6/f-pyrrolo[2,3-d]pyrimidin-6-one

Step 1 : ethyl 2-{2-[(2-acetyl-1,2,3,4-tetrahvdroisoquinolin-7-yl)amino-|-4-chloropyrimidin-5-yl}-

2-methylpropanoate

To a solution of ethyl 2-[4-chloro-2-(1,2,3,4-tetrahydroisoquinolin-7-ylamino)pyrimidin- 5-y]]-2-methyIpropanoate trifluoroacetate (600 rag, 1.227 mmol) in dichloromethane (20 ml) at 0

°C was added pyridine (0.521 ml, 6.44 mmol). After 5 minutes, acetic anhydride (0.116 ml,

1.227 mmol) was added and after 20 minutes at 0 °C the reaction mixture was quenched with saturated sodium bicarbonate and diluted with ethyl acetate. The organics were dried over magnesium sulfate, filtered, and concentrated to afford residue. The residue was purified by column chromatography on silica gel, eluting (ethyl acetate/ttexanes gradient followed by methanol/ethyl acetate gradient) to afford ethyl 2-{2-[(2-acetyl-1,2,3,4-tetrahydroisoquinolin-7- yl)amino]-4-chloropyrimidin-5-yl}-2-methylpropanoate.

^!H NMR (500 MHz, Methanol-d*) δ 9.98 (d, 1H); 8.41 (s, 1H); 7.54 (d, 1H); 7.48-7.42

(m, 1H); 7.11 (d, 1H); 4.68 (d, 2H); 4.15 (q, 2H); 3.79-3.72 (m, 2H); 2.89 (t, 1H); 2.80 (t, 1H); 2.19 (d, 3H); 1.60 (s, 6H); 1.22 (t, 3H). LRMS (ESI) calculated for C₂₁H₂₆ClN₄O₃ [M+H]⁺,

417.2; found 417.2.

Step 2 : 2- [(2-acetyl- 1 ,2,3.4-tetrahydroi$oquinolin-7-yl)ammo1 -7- f f 1 ^)-4-hvdroχy-2.3 -dihydro-

1 H-inden- 1 -yl]-5,5-dimethyl-5J-dihydro-6H-ρyrrolo[2,3-<^]pyrimidin-6-one To a solution of ethyl 2~{2-[(2-acetyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino]-4- chloropyrimidin-5-yl}-2-methy]propanoate (50 mg, 0.120 mmol) and (ljS)-4-hydroxy-2,3- dihydro-lH-inden-1-aminium trifluoroacetate (113 mg, 0.300 mmol) in 2-ethoxyethanol (0.3 ml) was added Hunig's base (0.168 ml, 0.959 mmol). The reaction mixture was heated to 160 °C.

After 16 hours, the reaction mixture was cooled and purified directly by preparative HPLC Reverse phase (C- 18), eluting with acetonitrile/water + 0.05% TFA to afford 2-[(2-acetyl-

1 ,2,3 ,4-tetrahydroisoquinolin-7-yl)amino] -1- [( 1 S)-4-hydroxy-2 , 3 -dihydro- 1 H-inden- 1 -y 1] -5 ,5 - dimethyl-5,7-dihydro-6H-pyrrolo[2,3-^pyrimidin-6-one.

¹H NMR (500 MHz, DMSOd₆) δ 9.49-9.38 (m, 2H); 8.20 (d, 1H); 7.22-7.12 (bs, 2H);

7.00-6.95 (m, 1H); 6.92 (t, 1H); 6.65 (d, 1H); 6.40 (d, 1H); 5.83 (t, 1H); 4.46 (d, 2H); 3.63-3.58 (m, 2H); 3.10-3.02 (m, 1H); 2.80-2.72 (m, 2H); 2.65-2.55 (m, 2H); 2.37-2.30 (m, 1H); 2.05 (d,

3H); 1.33 (dd, 6H). LRMS (ESI) calculated for C₂₈H₃₀N₅O₃ [M+H]⁺, 484.2; found 484.2. Examples 75 - 94

The following were prepared by analogous methods:

Selectivity for MARK over other kinases

Compounds according to the invention were screened against a variety of kinases in order to assess the degree of selectivity towards MARK. The following table records the percentage inhibition observed for each kinase when exposed to test compound at a concentration of 0.1 uM:

The compounds of the invention gave essentially complete inhibition of MARK (in all three isoforms tested), but inhibited other kinases to a much lesser extent, consistent with a high degree of selectivity for the desired target.

Claims

1. A compound of formula I:

or a pharmaceutically acceptable salt or hydrate thereof; wherein:

R¹ represents Ar or C|_-4alkyl which is optionally substituted with Ar, where Ar represents phenyl or pyridyl either of which optionally bears up to 3 halogen substituents; R^1a represents methyl; or R¹ and R^1a together with the carbon atom to which they are attached complete a bicyclic group of formula (a):

(a);

R represents H, halogen, OH, C_1-4alkyl, C_1-4alkoxy or benzyloxy; Z represents CH₂ or O; R² represents a group selected from:

and

III X represents CR⁵ or N;

R³, R⁴ and R⁵ independently represent H, halogen, C_3-4alkoxy or C_1-4alkyl which is optionally substituted with OH, Q^alkoxy, or with up to 3 halogen atoms; or one of R³, R⁴ and R⁵ represents

and the other two of R³, R⁴ and R⁵ are as defined previously;

R⁶ represents C^aUcyl which is optionally substituted with 1-3 halogens or with C_{1- 4}alkoxy or CO₂C_1-4aIkVl, or R⁶ represents

m is 1, 2 or 3; each R^a independently represents H or C_1-4alkyl which is optionally substituted with 1-3 halogens, or two R^a groups attached to the same carbon may represent -O, or one R^a group together with R⁷ and the intervening atoms may complete a ring of 4 to 7 members; with the proviso that when m>l, only one -CRV may be other than -CH₂-;

R⁷ represents H, Q^alkyl, acetyl or t-butoxycarbonyl, or R⁷ together with R^a and the intervening atoms completes a ring of 4-7 members;

R⁸ represents H or C_halky! which is optionally substituted with up to 3 halogen atoms, or with OR^b, COR^b, CO₂R^b, NR^b ₂ or CONR^b ₂;

R^b represents H or C_1-4alkyl which is optionally substituted with OH or C_1-4alkoxy; or R⁸ represents

or R⁸ together with R³ or R⁴ and the intervening atoms may complete a 5- or 6-membered ring; or R⁷ and R⁸ together with the nitrogen to which they are both attached may complete a monocyclic or bicyclic ring system of up to 10 ring atoms in total, of which up to two, in addition to the nitrogen to which R⁷ and R⁸ are bonded, are selected from N, O and S, the remainder being carbon, said ring system optionally bearing up to 3 substituents independently selected from halogen, CF₃, C_1-4alkyl, C_3-6CyClOaIkVl, phenyl, -O, OH, C_1-4alkoxy, hydroxyC_1-4alkyl and Ci- ₄alkoxyC_1-4alkyl; n is 0, 1 , 2 or 3; R^c represents H or C_1-4alkyl with the proviso that when n>l, only one -CR^C ₂- may be other than -CH₂-; and Y represents a monocyclic or bicycUc ring system of up to 10 ring atoms, of which 0- 3 are selected from N, O and S, the remainder being carbon, said ring system optionally bearing up to 3 substituents independently selected from halogen, CF₃, C_1-4alkyl, C^cycloalkyl, phenyl, =O, OH, C_1-4alkoxy, hydroxyC_1-4alkyl, C_1-4alkoxyC^₄alkyI and C_1-4alkylcarbonyl.

2. A compound according to claim 1 having the stereochemical configuration shown in formula IA:

IA wherein R¹, R^1a and R² are as defined in claim 1, such that when R¹ and R^1a together with the carbon atom to which they are attached complete a bicyclic group of formula (a), it has the stereochemical configuration shown in formula (b):

R

where R and Z are as defined in claim 1.

3, A compound according to claim 1 wherein R ^a represents methyl and R¹ represents ethyl,

4. A compound according to claim 1 wherein R¹ and R^1a together with the carbon atom to which they are attached complete a bicyclic group of formula (a) in which Z represents CH₂ and R is selected from H, OH, F, Cl, Br, methyl, methoxy and benzyloxy.

5. A compound according to claim 1 wherein R represents a phenyl or pyridyl group of formula II:

6. A compound according to claims 5 wherein R⁴ is a group represented by formula

IV:

IV

7. A compound according to claim 5 wherein R represents

or

8. A compound according to claim 6 wherein R represents H or Ci-galkyl which is optionally substituted with up to 3 halogens or with OR^b, COR^b, CO₂R^b, NR^b ₂ or CONR^b ₂, where R^b represents H or Cj_-4 alkyl which is optionally substituted with OH or C_1-4alkoxy.

9. A compound according to claim 6 wherein R⁸ together with R⁷ and the nitrogen to which they are both attached complete a monocyclic or bicyclic ring system of up to 10 ring atoms in total, of which up to two, in addition to the nitrogen to which R⁷ and R⁸ are bonded, are selected from N, O and S, the remainder being carbon, said ring system optionally bearing up to 3 substituents independently selected from halogen, CF₃, C_1-4alkyl, C^cycloalkyl, phenyl, -O, OH, C_1-4alkoxy, hydroxyC_1-4alkyl and C_1-4alkoxy C_1-4alky!..

10. A compound according to claim 6 wherein R⁸ represents

1 1. A compound according to claim 6 which is of formula V:

V

or a pharmaceutically acceptable salt or hydrate thereof.

12. A compound according to claim 1 which is of formula VI:

VI

or a pharmaceutically acceptable salt or hydrate thereof.

13. A compound according to claim 1 which is of formula VII;

VII or a pharmaceutically acceptable salt or hydrate thereof.

14. A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutical carrier.

15. A method for treatment or prevention of a neurodegenerative disease associated with hyperphosphorylation of tau in a human patient, said method comprising administering to that patient an effective amount of a compound of formula I as defined in claim 1 , or a pharmaceutically acceptable salt or hydrate thereof.