AU2003265740B2 - Neurologically-active compounds - Google Patents

Description

WO 2004/031161 PCT/AU2003/001303 NEUROLOGICALLY-ACTIVE COMPOUNDS The present invention relates to neurologically-active compounds, processes for their preparation and their use as pharmaceutical or veterinary agents, in 5 particular for the treatment of neurological conditions, more specifically neurodegenerative conditions such as Alzheimer's disease. BACKGROUND OF THE INVENTION All references, including any patents or patent applications, cited in this 10 specification are hereby incorporated by reference. No admission is made that any reference constitutes prior art. The discussion of the references states what their authors assert, and the applicants reserve the right to challenge the accuracy and pertinency of the cited documents. It will be clearly understood that, although a number of prior art publications are referred to herein, this reference does not constitute an admission that 15 any of these documents forms part of the common general knowledge in the art, in Australia or in any other country. The life span is thought to be biologically fixed for each species, and the length of the human life span is uncertain, but may be up to 120 years. Since life expectancy has risen significantly in this century, the elderly are an increasing segment 20 of our population, and their health care needs will continue to grow for decades. Although normal aging is characterized by modest reductions in the mass and volume of the human brain, which may be due to the atrophy and/or death of brain cells, these changes are far more profound in the brains of patients who succumb to a neurodegenerative condition. Most of these conditions are sporadic (i.e., not due to 25 genetic mutations) and of unknown cause, but hundreds of different mutations in many genes have been shown to cause familial (inherited) variants of several neurodegenerative conditions. Many of the dozen or more genes that harbor these mutations were discovered in the quest to determine the genetic basis of neurodegenerative conditions just in the last ten years. Neurodegenerative conditions 30 evolve gradually after a long period of normal brain function, due to progressive degeneration (i.e., nerve cell dysfunction and death) of specific brain regions. Since symptomatic expression of disease occurs when nerve cell loss exceeds a "threshold" for the continuing function (e.g., memory, movement) performed by the affected brain region, the actual onset of brain degeneration may precede clinical expression by many 35 years. Intellectual and higher integrative cognitive faculties become progressively impaired and interfere with activities of daily living in neurological WO 2004/031161 PCT/AU2003/001303 -2 conditions resulting in dementia. The precise prevalence of dementia in the elderly population is unknown, but may be 15% of people over 65 years old with 5% severely and 10% mildly to moderately demented. The prevalence of severe dementia increases from 1% at 65 years to 45% at 85 years. There are many causes of dementia, but 5 Alzheimer's Disease (AD) accounts for 50% of demented patients over 65 years of age. AD is a primary degenerative disease of the brain. It is characterized by progressive decline of cognitive functions such as memory, thinking, comprehension, calculation, language, learning capacity and judgement. Dementia is diagnosed when these declines are sufficient to impair personal activities of daily living. AD shows an 10 insidious onset with slow deterioration. This disease needs to be clearly differentiated from age-related nonnal decline of cognitive functions. The normal decline is much less, much more gradual and leads to milder disabilities. The onset of AD is usually after 65 years of age, although earlier onset is not uncommon. As age advances, the incidence increases rapidly (it roughly doubles every 5 years). This has obvious 15 implications for the total number of individuals living with this disorder as life expectancy increases in the population. The aetiology of dementia of AD is unclear. There is considerable evidence of a heritable predisposition for some forms of AD (reviewed in St George Hyslop, 2000), and the expression of certain isoforms of ApoE has also been linked to a 20 higher risk of AD (Corder et al, 1993; Czech et al 1994). The toxic accumulation of aluminium has been suggested as a causative agent in AD, although this hypothesis has now been largely superseded. The brains of AD patients display abnormal deposits which include P-amyloid protein (Ap). Ap is known to be present in the brains of individuals with certain 25 neurodegenerative diseases, but it is not known whether it is symptomatic of an underlying disease process, or is actually involved in the aetiology of the disease. For example, some authors believe that the Ap deposits may be indicative of a normal brain defence mechanism, in which the brain attempts to sequester the Ap; such deposits can be present in the brains of normal individuals. There is a mutation of tau protein in 30 which neurofibrillary tangles, but no amyloid plaques are present in the brain; this condition is known as tauopathy. One proposed approach to AD therapy is to inhibit production of As in the brain. Proteolytic cleavage of APP by BACEl and y-secretase generates the full length Ap, which is then released from cells (Nunan and Small, 2000). Therefore 35 inhibitors of either BACE1 or y-secretase may be of therapeutic value. Alternatively, a number of studies have shown that cholesterol can influence Ap release (Simons et al., 1998; Hartmann, 2001; Fassbender et al., 2001; Frears et al., 1999; Friedhoff et al., WO 2004/031161 PCT/AU2003/001303 -3 2001). However, there is some disagreement in the art as to the value of lowering cholesterol levels, and some workers consider that cholesterol is actually beneficial. For example, Ji et al, (2002) have suggested that the binding of AP to cholesterol might prevent As toxicity by inhibiting its oligomerization. 5 In an alternative approach, it has been proposed that by unravelling the proteolytic processing of the amyloid precursor protein (APP), which generates the As amyloid monomer, a number of possible therapeutic targets may be possible (Shearman et al., 2000; Sinha et al., 1999), and this approach is in an early stage of clinical development. Attempts to promote the clearance of AP from the brain through 10 immunization with AP, while efficacious in a transgenic mouse model for AD (Schenk et al 1999), have been found to have significant adverse effects (Brower, 2002). It has also been suggested that deposition of amyloid-like fibrils may also be important in other neurodegenerative diseases. These include Parkinson's disease, dementia with Lewy body formation, multiple system atrophy, Hallerboden 15 Spatz disease, and diffuse Lewy body disease. One of the competing theories of the aetiology of AD is that the causative step(s) lies within the pathway of the intracerebral biogenesis and accumulation of the AP amyloid protein (see recent reviews by Selkoe, 2001; Beyreuther et al., 2001; Bush, 2001). However, to date no drugs or agents which target 20 this pathway have been demonstrated to have a lasting effect on modifying the clinical expression of the disease or in preventing or ameliorating the decline in cognitive function associated with neurodegenerative disorders, including Alzheimer's disease. A further hypothesis is that AD is caused by the toxic accumulation of As amyloid, due in part to excess binding of copper and zinc, metal ions which are 25 abundant in the regions most affected. Moreover, it has been suggested that when Zn 2 + and Cu 2 + ions interact with AP, aggregation of AP into fibrils and plaques occurs (Atwood et al., 1998); confirmed by recent data from animals deficient in synaptic Zn+ (Lee et al., 2002). It has also been suggested that redox-active Cu 2 +-Ap interactions can generate H 2 0 2 from 02 (Huang et al., 1999). Both Cu 2 + and Zn 2 + have been shown to 30 affect Ap-lipid membrane interactions (Curtain et al., 2001). The brain is an organ that concentrates metal ions and recent evidence suggests that a breakdown in metal homeostasis plays a critical role in a variety of age related neurodegenerative diseases. Common features of these diseases include the deposition of misfolded protein (each disease has its own specific amyloid protein) and 35 substantial cellular damage as a result of oxidative stress. Indeed data is now rapidly accumulating that metallochemical reactions could emerge as the common denominator underlying amyloidogenic neurological disorders such as Alzheimer's disease, WO 2004/031161 PCT/AU2003/001303 -4 amylotrophic lateral sclerosis (ALS), prion diseases - including Creutzfeldt-Jakob Disease (CJD), transmissible spongioform encephalopathies (TSE), cataracts, mitochondrial disorders, Parkinson's disease and Huntington's disease. In these instances, the pathological aggregation of a specific protein is promoted by abnormal 5 redox activity in a physiological environment typified by the presence of transition metals and available reducing agents. [Bush, 2000 (Curr Opin Chem Biol. 2000 Apr; 4(2):184-91)]. A method of treatment of AD using iodochlorohydroxyquinoline an antibiotic [also known as clioquinol (CQ)], is disclosed and claimed in US patent 10 Nos. 5,994,323 and 6,001,852 by P.N. Geromylatos S.A. and in US patent application No. 09/972,913 by Bush et al. CQ was withdrawn as an antibiotic in 1970, because of its association with an uncommon neurological syndrome, subacute myelo-optic neuropathy (SMON), which was observed only in Japan in the 1960s, in patients thought to have received the drug over long periods and probably at doses higher than 15 those recommended at the time (Shiraki, 1975). However, recent evidence suggests that SMON was caused by an overuse-related vitamin B 12 deficiency in an exceptionally vulnerable population, and therefore could be rehabilitated for study in a clinical setting (Yassin et al., 2000; Bush and Masters, 2001). However, no in vivo results in animal models or in humans are provided 20 in the Geromylatos and Bush patents. US 5,994,323 discloses a composition comprising CQ and Vitamin B 12, and its use for the treatment of "diseases or disorders responsive to CQ administration while inhibiting detrimental side effects" of CQ. These diseases include AD. US 6,001,852 discloses a method of treatment of AD using CQ, preferably together with Vitamin B12. Both US 5,994,323 and US 6,001,852 suggest a dosage of 25 10-750 mg per day; US 5,994,323 recommends that if treatment is over a long period CQ should be given intermittently, for up to 3 weeks at a time followed by a "wash-out" period of 1-4 weeks. In US application No. 09/972,913 CQ is exclusively referred to in terms of its ability to disaggregate AP deposits. No other mechanism of neurotoxicity is 30 discussed. PCT/US99/05291 by General Hospital Corporation discloses the use of CQ in combination with specific copper and zinc chelators to promote dissolution of amyloid plaques and inhibition of amyloid plaque formation and/or the production of ROS by Ap. US 6,001,852 also suggests that a composition comprising CQ and 35 Vitamin B12 could be used in the treatment of Parkinson's disease; however, in this context it is suggested that CQ acts primarily via clearing iron from the substantia nigra.

- 5 The efficacy of CQ in the treatment of AD rests upon its ability to enter the CNS and then sequester the transition metals Cu, Zn and Fe from various AP entities thereby reducing As toxicity and liberating it for clearance. The effectiveness of CQ is restricted by its poor aqueous solubility which limits its oral bioavailability. CQ is also 5 known to undergo considerable conjugative metabolism and has a history of toxicity as discussed above. The fact that CQ is a bidentate metal ligand makes necessary the commitment of at least two molecules for every metal ion captured. SUMMARY OF THE INVENTION 10 The present invention provides a means of treating neurological conditions including those characterised by the abnormal reaction between proteins and metals. We have now developed heterocyclic compounds having two fused 15 6-membered rings with a nitrogen atom at position 1 and a hydroxy or mercapto group at position 8 with at least one ring being aromatic through the collective optimization of one or more of the following properties: (a) metal chelation (as hereinafter defined); (b) aqueous solubility; 20 (c) reduced cell toxicity; (d) amyloid dispersion properties; (e) membrane permeability appropriate for CNS penetration; and (f) metabolic stability. These compounds include examples of therapeutics which are 25 concentrated in the CNS through active transport, contain antioxidant activity in addition to their metal chelation properties which in some cases leads to enhanced metal chelation properties and demonstrate a prodrug strategy which masks the 8-hydroxy or 8-mercapto moiety to favour CNS penetration and make use of the known esterase activity which resides on the inner surface of the blood brain barrier (BBB). 3 0 According to the present invention there is provided a method for the treatment, amelioration and/or prophylaxis of a neurological condition which comprises the administration of an effective amount of a compound of formula IA: 2114997_1 (GHMatters) 10/11/09 -6 -- 63 WY

(R

1 )m N (R 1 )m 5 RH IA in which R is O or S; R' is independently selected from H, optionally substituted alkyl, optionally 10 substituted alkenyl; optionally substituted alkynyl; optionally substituted aryl; optionally substituted heterocyclyl; an antioxidant; a targeting moiety; CN; halo; CF 3 ;

SO

3 H; and OR 2 , SR 2 , SOR 2 , S0 2

R

2 , NR 2

R

3 , (CH 2 )nNR 2

R

3 , HCNOR 2 , HCNNR 2

R

3 ,

CONR

2

R

3 , CSNR 2

R

3 , NCOR 2 , NCSR 2 , COR 2 , CO 2

R

2 , CSR 2 or SO 2

NR

2

R

3 in which R 2 and R 3 are independently selected from H, optionally substituted alkyl, optionally 15 substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heterocyclyl, an antioxidant or a targeting moiety and n is an integer of I to 10; W is CH, N or NH; U is CH, CO or N; 20 Y is absent or together with the ring to which it is attached forms a 5- or 6 membered optionally substituted aryl or a 5- or 6-membered optionally substituted heterocyclyl; Y' is absent or together with the ring to which it is attached forms a 6-membered N-containing optionally substituted heterocyclyl; and 25 m is an integer from I to 3; salts, hydrates, solvates, derivatives, pro-drugs, tautomers and/or isomers thereof to a subject in need thereof, in which the prodrug is a C 1

-

6 alkyl ester or aryl ester moiety located at position 8; 30 with the provisos that: (i) at least one of W and U is other than CH; (ii) phanquinone or tautomers thereof are excluded i.e., when R is 0, R' at position 7 is OH, W and are CH and Y is absent, then Y' is not N (iii) when R is 0, Y is absent, W is CH other than at position 3 40 - NH ' N NH 2114997_2 (GHMattero) 22/12/09 PUT/AUUU3/OO1303 Received 31 December 2004 - 6a (iii) when R is 0, Y is absent, Z is CH, X is CH other than at position 3 Me where X is N, m is 2 and R is -NH NH 10 N at position 3, then R at position 2 is not -s / NHAc or -s /aNH-SO 2 Me ; and 20 (iv) clioquinol i.e, when R is 0, Y is absent, Z and X are CH and m is 2, then R' at position 5 is not chloro and R' at position 7 is not iodo. Amended Sheet

IPEA/AU

-7 where W is N, m is 2 and R' is 5 at position 3, then R' at position 2 is not -s NHAc or -S /NH-SO 2 Me ; and 10 (iv) clioquinol i.e, when R is 0, Y and Y' are absent, W and U are CH and m is 2, then R' at position 5 is not chloro and R' at position 7 is not iodo. Further according to the present invention there is provided use of the 15 compound of formula IA in the manufacture of a medicament for the treatment, amelioration and/or prophylaxis of a neurological condition. The invention also provides use of the compound of formula IA for the treatment, amelioration and/or prophylaxis of a neurological condition. The invention further provides the compound of formula IA for use in 20 the treatment, amelioration and/or prophylaxis of a neurological condition. The invention still further provides use of the compound of formula IA as a pharmaceutical, preferably a neurotherapeutic or neuroprotective agent, more preferably an antiamyloidogenic agent. Preferably, the neurological condition is a neurodegenerative condition, more preferably neurodegenerative amyloidosis such as 25 Alzheimer's disease or Parkinson's disease. R is preferably 0. R' is preferably halo, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkyl, OR 2 , SR 2 , (CH 2 )nNR 2

R

3 , CONR 2

R

3 and

NCOR

2 in which n, R 2 and R 3 are as defined above. More preferably R' is fluoro; iodo; 30 chloro; optionally substituted phenyl such as 4-halophenyl, for example, 4-fluorophenyl or 4-chlorophenyl; an optionally substituted unsaturated 3 to 6-membered heteromonocyclic group containing I to 4 nitrogen atoms such as imidazolyl or pyridinyl; an optionally substituted saturated 3 to 6-membered heteromonocyclic group containing I to 4 nitrogen atoms such as imidazolidinyl or piperazinyl; an optionally 35 substituted saturated 3 to 6-membered heteromonocyclic group containing I to 2 oxygen atoms and I to 3 nitrogen atoms such as morpholinyl; optionally substituted C,. 4 alkyl such as methyl or ethyl; optionally substituted C 2 .6 cycloalkyl such as cyclopropyl; optionally substituted C 1 - alkoxy; optionally substituted thio; CH 2

NR

4

R

5 in which R 4 and R 5 are independently selected from H and Ci4 alkyl; or 40 CONH(CH 2

)

2

R

6 in which R6 is optionally substituted heterocyclyl. 2114997_1 (GHMattern) 10/11/09 - 8 Y is preferably an optionally substituted phenyl; an optionally substituted unsaturated 5- or 6-membered heteromonocyclic group containing 1 to 4 nitrogen atoms such as imidazolyl or pyridinyl; or an optionally substituted saturated 5 or 6-membered heteromonocyclic group containing I to 2 oxygen atoms and I to 3 nitrogen atoms such 5 as morpholinyl. While not wishing to be bound by theory, it is believed that substituent R' has a limited effect, electronically or sterically, in the chelating properties of the compounds of the present invention. Substitution can therefore be used to modulate other parameters such as cytotoxicity and physicochemical properties including the 10 number of hydrogen bond donors and acceptors, lipophilicity (ClogP, ElogP and LogD), solubility and polar surface area. Modulation of these parameters contribute to the optimisation of the pharmacokinetic profile of the compounds. It is also postulated that when substituent R' is located at positions 2 and/or 7 in addition to modulating cytotoxicity and physicochemical properties could also affect activity if the substituent 15 provides chelating properties. Illustrative classes of compounds of formula I are as follows: (R)m 0 H 8-hydroxy-4(3H)-quinazolinones OH 20 8-hydroxy-quinazoline OH 25 8-hydroxy-quinoxaline OH Nj (R )m [l,6]naphthyridin-8-ol 30 OH 0 - N y N\ 9-hydroxypyrimido[ 1,6-a]pyrimidin-4-one OH (R) 35 2114997_1 (GHMatters) 10/11/09 9 'I~ 8-hydroxy-cinnoline OH 4-hydroxy-acridine OH 10 10 \N R)N, 4,7(4, 1 O)-phenanthrolin-5-oI R' N OH OH (R'). 15 (R) OH 0 N 9-hydroxypyrido[ I ,2-a]pyrim idin-4-one (R')<K - 20 qN\ ) OH (R'), O(0H N(P')q I ) [1,5]naphthyridine-4,8-diol 0OH 25 (R) quinazol-8-oI OH 30 2114991 _2 (CHMater) 22/12/09 - 10 in which R' is as defined above, p is an integer of 1 to 4 and q is an integer of I or 2. The 8-hydroxyl or 8-mercapto group on the compounds of formula I may 5 be blocked to form a prodrug, in particular an ester prodrug. The 8-hydroxy or 8 mercapto represents a principal site of metabolism for the compound of formula I: conjugation with glucuronic acid or sulphate gives a hydrophilic species ready to be excreted. Such conjugates probably do not pass the blood brain barrier. The ester prodrug may protect the compound of formula I from conjugation. Esterases integral to 10 the blood brain barrier may then release the C8-hydroxy or mercapto on passage through that barrier activating the compound for its role in the CNS. 2114997_1 (GHMatter.) 10/11/09 - 11 Preferred compounds of formula IA are as follows: (i) Formula Ia O 5 N

(R

1 )m N (R ) RH 10 Ia in which R, R', m and q are as defined above. Preferably R' is located at positions 2, 3, 5 and/or 7 and is selected from halo, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkyl and (CH 2 )nNR 2

R

3 in which n, R 2 and R 3 are as defined above. More 15 preferably R' is chloro, optionally substituted phenyl, C 3 .6 cycloalkyl, CH 2

NR

4 R' in which R 4 and R 5 are independently selected from H and CI.4 alkyl or optionally substituted pyridinyl. Particularly preferred examples are shown below. 21149971 (GHMattere) 10/11/09 WO 2004/031161 PCT/AU2003/001303 - 12 C1 0 F1 Cl o N F CI N CI N C N OH OH 1055 1061 10 C 0 0 NH NH ~N C1 N-N 15 OH 1067 1049 (ii) Formula lb 20 N

R

1 ) ()m(R 1 )q N RH 25 Ib in which R, R 1 , m and q are as defined above. Preferably R' is located at positions 2, 4, 5 and/or 7 and is selected from halo and optionally substituted heterocyclyl. More preferably, R 1 is chloro and/or morpholinyl.

PCT/AU2003/001303 Received 31 December 2004 - 13 Preferred examples are shown below. 0 0 CI N N 5-N N N CI N CI NO OH OH OH 10 CLogP = 3.087 CLogP = 2.35872 15 (iii) Formula Ic N (R4 )m1 (R)(R )q N 20 RH Ic in which R, R 1 , m and q are as defined above. Preferably R1 is located at positions 2, 5 and/or 7 and is selected from 25 halo and CH 2

NR

4

R

5 in which R 4 and R 5 are independently selected from H and C14 alkyl. Preferred examples are shown below. CI 30 N O NMe 2 CI N CI N OH OH NMe 2 35 1066 CLogP =2.57029 Amended Sheet

IPEA/AU

WO 2004/031161 PCT/AU2003/001303 - 14 C1 CI N N 5 CI N Cl N OH OH 1064 1065 10 (iv) Formula Id

(R

1 )q N

(R

1 )m 15 N RH Id 20 in which R, R', n and q are as defined above. Preferably R 1 is located at positions 2 and/or 7 and is selected from optionally substituted heterocyclyl, C0 2

R

2 , (CH 2 )nNR 2

R

3 and CONR 2

R

3 in which n, R2 and R3 are as defined above.

WO 2004/031161 PCT/AU2003/001303 - 15 Preferred examples are shown below. CI N N 5 N N MeOOC N 0 OH OH 1053 1045 10 H N N) N 15 1070 O OH HN (v) Formula le 0 20 RN (R) N RH 25 in which R, R 1 , m and q are as defined above. Preferably R' is located at positions 2, 3, 6 and/or 7 and is selected from halo, optionally substituted aryl and (CH 2 )nNR 2

R

3 in which n, R 2 and R 3 are as defined above.

PCT/AU2003/001303 Received 31 December 2004 - 16 Preferred examples are shown below. 0ci N Cl 50 1 NN NN OH 1063 N 10 0 OH 1069 N 15 OH 1048 20 (vi) Formula If

(R

1 )m N 25 (R)m R1 N OH 30 in which R' and m are as defined above. Preferably R' is located at positions 2 and/or 6 and is selected from halo and (CH 2 )nNR2R 3 in which n, R2 and R 3 are as defined above. Amended Sheet

TPEA/AU

- 17 Preferred examples are shown below. N NN 'N N cl N cl N OH OH OH NMe 2 1026 CLogP = 2.77788 CLogP = 2.61188 10 In a further aspect, the invention provides a pharmaceutical or veterinary composition comprising the compound of formula IA as defined above, together with a pharmaceutically or veterinarily acceptable carrier. Some of the compounds of formula IA are novel per se. Accordingly, the invention provides a compound of formula IA with the 15 proviso that at least one R' is other than H. Preferred compounds of formula IA are compounds of the formulae la, Ib, Ic, Id and le defined above, more preferably 1045, 1061, 1066, 1053, 1063, 1064, 1065, 1067, 1069 and 1070. The compound of formula IA defined above may be prepared using the 20 processes described in detail hereinafter. DETAILED DESCRIPTION OF THE INVENTION In the claims of this application and in the description of the invention, except where the context requires otherwise due to express language or necessary 25 implication, the words "comprise" or variations such as "comprises" or "comprising" are used in an inclusive sense, i.e. to specify the presence of the stated features but not to preclude the presence or addition of further features in various embodiments of the invention. The term "alkyl" used either alone or in compound words such as 30 "optionally substituted alkyl" or "alkylamino" refers to straight chain, branched chain or cyclic hydrocarbon groups having from I to 10 carbon atoms, preferably I to 6 carbon atoms, more preferably I to 4 carbon atoms. Illustrative of such alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, neopentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. Preferred alkyl 35 groups are C1 alkyl such as methyl or ethyl and C 2 - cycloalkyl such as cyclopropyl. 2114997_1 (GHatters) 10/11/09 WO 2004/031161 PCT/AU2003/001303 - 18 The term "alkenyl" used either alone or in compound words such as "optionally substituted alkenyl", denotes linear, branched or mono- or poly-cyclic radicals having at least one carbon-carbon double bond of 2 to 20 carbon atoms, preferably 2 to 14 carbon atoms, more preferably 2 to 6 carbon atoms. Examples of 5 alkenyl radicals include allyl, ethenyl, propenyl, butenyl, iso-butenyl, 3-methyl-2 butenyl, 1-pentenyl, cyclopentenyl, 1-methyl-cyclopentenyl, 1-hexenyl, 3-hexenyl, cyclohexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, cyclooctenyl, 1-nonenyl, 2-nonenyl, 3 nonenyl, 1-decenyl, 3-decenyl, 1,3-butadienyl, 1,4-pentadienyl, 1,3-cyclopentadienyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl,1,3 10 cycloheptadienyl, 1,3,5-cycloheptatrienyl, 1,3,5,7-cycloocta-tetraenyl and the like. The tenn "alkynyl" used either alone or in compound words such as "optionally substituted alkynyl" refers to straight chain or branched chain radicals having at least one carbon-carbon triple bond of 2 to 20 carbon atoms, preferably 2 to 14 carbon atoms, more preferably 2 to 6 carbon atoms. Examples include ethynyl, 1 15 propynyl, 1- and 2-butynyl, 2-methyl-2-propynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 10-undecynyl, 4-ethyl-1-octyn-3-yl, 7 dodecynyl, 9-dodecynyl, 10-dodecynyl, 3-methyl-1-dodecyn-3-yl, 2-tridecynyl, 11 tridecynyl, 3-tetradecynyl, 7-hexadecynyl, 3-octadecynyl and the like. The term "heterocyclyl group" used either alone or in compound words 20 such as "optionally substituted heterocyclyl" refers to monocyclic or polycyclic heterocyclic groups containing at least one heteroatom atom selected from nitrogen, sulphur and oxygen. Suitable heterocyclic groups include N-containing heterocyclic groups, such as, unsaturated 3 to 6-membered heteromonocyclic groups containing 1 to 4 25 nitrogen atoms, for example, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl; saturated 3 to 6-membered heteromonocyclic groups containing 1 to 4 nitrogen atoms, such as, pyrrolidinyl, imidazolidinyl, piperidino or piperazinyl; unsaturated condensed heterocyclic groups containing 1 to 5 nitrogen 30 atoms, such as indolyl, isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl or tetrazolopyridazinyl; unsaturated 3 to 6-membered heteromonocyclic group containing an oxygen atom, such as, pyranyl or furyl; unsaturated 3 to 6-membered heteromonocyclic group containing I to 2 35 sulphur atoms, such as, thienyl; unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, oxazolyl, isoxazolyl or oxadiazolyl; WO 2004/031161 PCT/AU2003/001303 - 19 saturated 3 to 6-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, morpholinyl; unsaturated condensed heterocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, such as, benzoxazolyl or benzoxadiazolyl; 5 unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as, thiazolyl or thiadiazolyl; saturated 3 to 6-membered heteromonocyclic group containing 1 to 2 sulphur atoms and 1 to 3 nitrogen atoms, such as, thiazolidinyl; and unsaturated condensed heterocyclic group containing 1 to 2 sulphur 10 atoms and 1 to 3 nitrogen atoms, such as, benzothiazolyl or benzothiadiazolyl. Preferably the heterocyclyl is an unsaturated 5 or 6-membered heteromonocyclic group containing 1 to 3 nitrogen atoms such as imidazolyl or pyridinyl; a saturated 5 or 6-membered heteromonocyclic group containing 1 to 4 nitrogen atoms such as imidazolidinyl or piperazinyl; or a saturated 5 or 6-membered 15 heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as morpholinyl. The term "aryl" used either alone or in compound words such as "optionally substituted aryl" denotes a carbocyclic aromatic system containing one, two or three rings wherein such rings may be attached together in a pendent manner or may 20 be fused. The term "aryl" embraces aromatic radicals such as phenyl, naphthyl, tetrahydronaphthyl, indane and biphenyl. Preferably, the aryl is optionally substituted phenyl such as 4-halophenyl, more preferably 4-fluorophenyl or 4-chlorophenyl. The term "halo" refers to fluorine, chlorine, bromine or iodine, preferably fluorine, iodine or chlorine. 25 The term "alkoxy" refers to straight chain or branched oxy-containing radicals preferably each having alkyl portions of 1 to about 6 carbon atoms. Examples of alkoxy include methoxy, ethoxy, propoxy, butoxy and tert-butoxy. The term "optionally substituted thio" refers to optional substituents such as radicals containing a linear or branched alkyl of 1 to 10 carbon atoms, 30 preferably 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms, attached to a divalent sulphur atom. Examples of alkylthio radicals include methylthio, ethylthio, propylthio, butylthio and hexylthio. The term "optionally substituted" refers to a group which may or may not be further substituted with one or more groups selected from alkyl, alkenyl, alkynyl, 35 aryl, aldehyde, halo, haloalkyl, haloalkenyl, haloalkynyl, haloaryl, hydroxy, alkoxy, alkenyloxy, aryloxy, benzyloxy, haloalkoxy, haloalkenyloxy, haloaryloxy, nitro, nitroalkyl, nitroalkenyl, nitroalkynyl, nitroaryl, nitroheterocyclyl, amino, alkylamino, WO 2004/031161 PCT/AU2003/001303 - 20 dialkylamino, alkenylamino, alkynylamino, arylamino, diarylamino, benzylamino, dibenzylamino, acyl, alkenylacyl, alkynylacyl, arylacyl, acylamino, diacylamino, acyloxy, alkylsulphonyloxy, arylsulphenyloxy, heterocyclyl, heterocycloxy, heterocyclamino, haloheterocyclyl, alkylsulphenyl, arylsulphenyl, carboalkoxy, 5 carboaryloxy, mercapto, alkylthio, benzylthio, acylthio, phosphorus-containing groups and the like. Preferably, the optional substituent is C1.

6 alkyl, more preferably C 1

.

4 alkyl; CF 3 ; fluorine; chlorine; iodine; cyano; C 1

.

6 alkoxy, more preferably C 1

.

4 alkoxy; aryl; heterocyclyl; amino; or alkylamino. The term "antioxidant" is used herein in its broadest sense and refers to a 10 group which has the capacity to react with a reactive oxygen species such as a hydroxyl radical in such a way as to generate a non toxic product. Examples include phenols such as 3,4,5-trimethoxyphenyl and 3,5-di-t-butyl-4-hydroxyphenyl, indole amines such as melatonin and flavonoids. Other examples may be found the literature (Wright, 2001; Karbownik, 2001; Gilgun-Sherki, 2001). 15 The term "targeting moiety" is used herein in its broadest sense and refers to a group which will facilitate the brain delivery of the drug by way of an active transport mechanism. The targeting moiety is recognised by specific transporter enzymes integral to the blood brain barrier and these transporter enzymes then provide a mechanism for the drug to be imported into the brain. Typically such transporters are 20 sodium dependant and their substrates contain carboxylic acids such as ascorbic acid and L-glutamate. Conjugation of the targeting moiety to the drug is enacted so as to retain the acid moiety. Examples can be found in the literature (Manfredini, 2002, Sakaedu, 2001). The term "metal chelator" is used herein in its broadest sense and refers 25 to compounds having two or more donor atoms capable of binding to a metal atom, preferably Cu, Zn or Fe wherein at least two of the donor atoms are capable of simultaneous binding to the metal atom and the resultant metal complex has a thermodynamic stability greater than or equal to that of the metal ion: biological ligand complex. The use of metal chelators as treatments for neurological disorders in the 30 present invention is distinguished from the previously known concept of "chelation therapy". "Chelation therapy" is a term associated clinically with the removal of bulk metals such as in Wilson's disease, B-thallesemia and haemochromatosis. The break down in metal homeostasis in these diseases can be described as a catastrophic event much like a dam bursting leading to overwhelming flooding of the problem metal. The 35 mechanism of action of such compounds is that bulk metal is sequestered by the chelators and cleared by excretion. By way of comparison the breakdown in metal homeostasis associated with neurological conditions of the present invention is more - 21 akin to the constant drip of a leaky tap, which if left long enough will eventually cause local damage over a long period of time. The intention of the "metal chelator" of the present invention is to disrupt an abnormal metal-protein interaction to achieve a subtle repartitioning of metals and a subsequent normalization of metal distribution with the 5 aim that once the toxic cycle is short-circuited, endogenous clearance processes can cope more effectively with the accumulating amyloidogenic protein. The salts of the compound of Formula I or II are preferably pharmaceutically acceptable, but it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the present invention, since these are useful 10 as intermediates in the preparation of pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts include salts of pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonium and alkylammonium; acid addition salts of pharmaceutically acceptable inorganic acids such as hydrochloric, orthophosphoric, sulphuric, phosphoric, nitric, carbonic, boric, 15 sulfamic and hydrobromic acids; or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, trihalomethanesulphonic, toluenesulphonic, benzenesulphonic, salicylic, sulphanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic 20 and valeric acids. In addition, some of the compounds of the present invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention. By "pharmaceutically acceptable derivative" is meant any 25 pharmaceutically acceptable salt, hydrate, ester, amide, active metabolite, analogue, residue or any other compound which is not biologically or otherwise undesirable and induces the desired pharmacological and/or physiological effect. The term "pro-drug" is used herein in its broadest sense to include those compounds which are converted in vivo to compounds of Formula IA. Use of the pro 30 drug strategy optimises the delivery of the drug to its site of action, for example, the brain. In one aspect, the term refers to the presence of a C 1 _ alkyl or arylester moiety which is designed to resist hydrolysis until the pro-drug has crossed the BBB, where esterases on the inner surface of the BBB act to hydrolyse the ester and liberate the C8 hydroxyl of the compounds of formula IA. In a second aspect, the term refers to the 35 attachment at position 2 of an antioxidant group, in particular the 3,4, 5trimethoxyphenyl moiety or derivatives thereof. Exposure to the prooxidative environment of the brain will then lead to hydroxylation of the 3,4,5-trimethoxyphenyl 21149971 (GHMattero) 10/11/09 - 22 group to give a 2-hydroxy-3,4,5-trimethoxyphenyl substituent, the hydroxyl group of which acts to enhance the chelation properties of the compounds of formula I or 11. The term "tautomer" is used herein in its broadest sense to include compounds of Formula IA which are capable of existing in a state of equilibrium 5 between two isomeric forms. Such compounds may differ in the bond connecting two atoms or groups and the position of these atoms or groups in the compound. The term "isomer" is used herein in its broadest sense and includes structural, geometric and stereo isomers. As the compound of Formula IA may have one or more chiral centres, it is capable of existing in enantiomeric forms. 10 The compositions of the present invention comprise at least one compound of Formula IA together with one or more pharmaceutically acceptable carriers and optionally other therapeutic agents. Each carrier, diluent, adjuvant and/or excipient must be pharmaceutically "acceptable" in the sense of being compatible with the other ingredients of the composition and not injurious to the subject. Compositions is include those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. The compositions may conveniently be presented in unit dosage form and may be prepared by methods well known in the art of pharmacy. Such methods include the step of bringing into association the active ingredient with the 20 carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers, diluents, adjuvants and/or excipients or finely divided solid carriers or both, and then if necessary shaping the product. The term "neurological condition" is used herein in its broadest sense 25 and refers to conditions in which various cell types of the nervous system are degenerated and/or have been damaged as a result of neurodegenerative disorders or injuries or exposures. In particular, compounds of formula IA can be used for the treatment of resulting conditions, in which damage to cells of the nervous system has occurred due to surgical interventions, infections, exposure to toxic agents, tumours, 30 nutritional deficits or metabolic disorders. In addition, compounds of the formula IA can be used for the treatment of the sequelae of neurodegenerative disorders, such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, amylotrophic lateral sclerosis, epilepsy, drug abuse or drug addiction (alcohol, cocaine, heroin, amphetamine or the like), spinal cord disorders and/or injuries, dystrophy or degeneration of the 35 neural retina (retinopathies) and peripheral neuropathies, such as diabetic neuropathy and/or the peripheral neuropathies induced by toxins. 2114997_1 (GHMatteral 10/11/09 WO 2004/031161 PCT/AU2003/001303 - 23 The term "neurodegenerative disorder" as used herein refers to an abnormality in which neuronal integrity is threatened. Neuronal integrity can be threatened when neuronal cells display decreased survival or when the neurons can no longer propagate a signal. 5 Neurological disorders that can be treated with the compounds of the present invention include acute intermittent porphyria; adriamycin-induced cardiomyopathy; AIDS dementia and HIV-1 induced neurotoxicity; Alzheimer's disease; amylotrophic lateral sclerosis; atherosclerosis; cateract; cerebral ischaemia; cerebral palsy; cerebral tumour; chemotherapy-induced organ damage; cisplatin 10 induced nephrotoxicity; coronary artery bypass surgery; Creutzfeldt-Jacob disease and its new variant associated with "mad cow" disease; diabetic neuropathy; Down's syndrome; drowning; epilepsy and post-traumatic epilepsy; Friedrich's ataxia; frontotemporal dementia; glaucoma; glomerulopathy; haemochromatosis; haemodialysis; haemolysis; haemolytic uraemic syndrome (Weil's disease); 15 haemorrhagic stroke; Hallerboden-Spatz disease; heart attack and reperfusion injury; Huntington's disease; Lewy body disease; intermittent claudication; ischaemic stroke; inflammatory bowel disease; macular degeneration; malaria; methanol-induced toxicity; meningitis (aseptic and tuberculous); motor neuron disease; multiple sclerosis; multiple system atrophy; myocardial ischaemia; neoplasia; Parkinson's disease; peri-natal 20 asphyxia; Pick's disease; progressive supra-nuclear palsy; radiotherapy-induced organ damage; restenosis after angioplasty; retinopathy; senile dementia; schizophrenia; sepsis; septic shock; spongiform encephalopathies; subharrachnoid haemorrage/cerebral vasospasm; subdural haematoma; surgical trauma, including neurosurgery; thalassemia; transient ischaemic attack (TIA); traumatic brain injury (TBI); traumatic spinal injury; 25 transplantation; vascular dementia; viral meningitis; and viral encephalitis. Additionally, compounds of the present invention may also be used to potentiate the effects of other treatments, for example to potentiate the neuroprotective effects of brain derived nerve growth factor. The invention is particularly directed to conditions which induce 30 oxidative damage of the central nervous system, including acute and chronic neurological disorders such as traumatic brain injury, spinal cord injury, cerebral ischaemia, stroke (ischaemic and haemorragic), subharrachnoid haemorrage/cerebral vasospasm, cerebral tumour, Alzheimer's disease, Creutzfeldt-Jacob disease and its new variant associated with "mad cow" disease, Huntington's disease, Parkinson's disease, 35 Friedrich's ataxia, cataract, dementia with Lewy body formation, multiple system atrophy, Hallerboden-Spatz disease, diffuse Lewy body disease, amylotrophic lateral sclerosis, motor neuron disease, multiple sclerosis, fatal familial insomnia, Gertsmann WO 2004/031161 PCT/AU2003/001303 - 24 Straussler Sheinker disease and hereditary cerebral haemorrhage with amyoidoisis Dutch type. More particularly, the invention is directed to the treatment of neurodegenerative amyloidosis. The neurodegenerative amyloidosis may be any 5 condition in which neurological damage results from the deposition of amyloid. The amyloid may be formed from a variety of protein or polypeptide precursors, including but not limited to AP, synuclein, huntingtin, or prion protein. Thus the condition is preferably selected from the group consisting of sporadic or familial Alzheimer's disease, amyotrophic lateral sclerosis, motor neuron 10 disease, cataract, Parkinson's disease, Creutzfeldt-Jacob disease and its new variant associated with "mad cow" disease, Huntington's disease, dementia with Lewy body formation, multiple system atrophy, Hallerboden-Spatz disease, and diffuse Lewy body disease. More preferably the neurodegenerative amyloidosis is an Ap-related 15 condition, such as Alzheimer's disease or dementia associated with Down syndrome or one of several forms of autosomal dominant forms of familial Alzheimer's disease (reviewed in St George-Hyslop, 2000). Most preferably the Ap-related condition is Alzheimer's disease. In a particularly preferred embodiment of all aspects of the invention, 2 0 prior to treatment the subject has moderately or severely impaired cognitive function, as assessed by the Alzheimer's Disease Assessment Scale (ADAS)-cog test, for example an ADAS-cog value of 25 or greater. In addition to slowing or arresting the cognitive decline of a subject, the methods and compounds of the invention may also be suitable for use in the treatment 25 or prevention of neurodegenerative conditions, or may be suitable for use in alleviating the symptoms of neurodegenerative conditions. The compounds may be able to provide at least a partial reversal of the cognitive decline experienced by patients. If administered to a subject who has been identified as having an increased risk of a predisposition to neurodegenerative conditions, or to a subject exhibiting pre-clinical 3 0 manifestations of cognitive decline, such as Mild Cognitive Impairment or minimal progressive cognitive impairment, these methods and compounds may be able to prevent or delay the onset of clinical symptoms, in addition to the effect of slowing or reducing the rate of cognitive decline. Currently Alzheimer's disease and other dementias are usually not 35 diagnosed until one or more warning symptoms have appeared. These symptoms constitute a syndrome known as Mild Cognitive Impairment (MCI), which was recently defined by the American Academy of Neurology, and refers to the clinical state of WO 2004/031161 PCT/AU2003/001303 - 25 individuals who have memory impairment, but who are otherwise functioning well, and who do not meet clinical criteria for dementia (Petersen et al., 2001). Symptoms of MCI include: (1) Memory loss which affects job skills 5 (2) Difficulty performing familiar tasks (3) Problems with language (4) Disorientation as to time and place (getting lost) (5) Poor or decreased judgement (6) Problems with abstract thinking 10 (7) Misplacing things (8) Changes in mood or behaviour (9) Changes in personality (10) Loss of initiative MCI can be detected using conventional cognitive screening tests, such 15 as the Mini Mental Status Exam, and the Memory Impairment Screen, and neuropsychological screening batteries. The term "subject" as used herein refers to any animal having a disease or condition which requires treatment with a pharmaceutically-active agent. The subject may be a mammal, preferably a human, or may be a domestic or companion 20 animal. While it is particularly contemplated that the compounds of the invention are suitable for use in medical treatment of humans, it is also applicable to veterinary treatment, including treatment of companion animals such as dogs and cats, and domestic animals such as horses, ponies, donkeys, mules, llama, alpaca, pigs, cattle and sheep, or zoo animals such as primates, felids, canids, bovids, and ungulates. 25 Suitable mammals include members of the Orders Primates, Rodentia, Lagomorpha, Cetacea, Carnivora, Perissodactyla and Artiodactyla. Members of the Orders Perissodactyla and Artiodactyla are particularly preferred because of their similar biology and economic importance. For example, Artiodactyla comprises approximately 150 living species 30 distributed through nine families: pigs (Suidae), peccaries (Tayassuidae), hippopotamuses (Hippopotamidae), camels (Camelidae), chevrotains (Tragulidae), giraffes and okapi (Giraffidae), deer (Cervidae), pronghorn (Antilocapridae), and cattle, sheep, goats and antelope (Bovidae). Many of these animals are used as feed animals in various countries. More importantly, many of the economically important animals such 35 as goats, sheep, cattle and pigs have very similar biology and share high degrees of genomic homology.

- 26 The Order Perissodactyla comprises horses and donkeys, which are both economically important and closely related. Indeed, it is well known that horses and donkeys interbreed. As used herein, the term "therapeutically effective amount" is meant an 5 amount of a compound of the present invention effective to yield a desired therapeutic response, for example, to treat, ameliorate or prevent a neurological condition. The specific "therapeutically effective amount" will, obviously, vary with such factors as the particular condition being treated, the physical condition of the subject, the type of subject being treated, the duration of the treatment, the nature of 10 concurrent therapy (if any), and the specific formulations employed and the structure of the compound or its derivatives. The compounds of the present invention may additionally be combined with other medicaments to provide an operative combination. It is intended to include any chemically compatible combination of pharmaceutically-active agents, as long as 15 the combination does not eliminate the activity of the compound of formula IA. It will be appreciated that the compound of the invention and the other medicament may be administered separately, sequentially or simultaneously. Other medicaments may include, for example, where the condition is a p-amyloid related condition, particularly Alzheimer's disease, an inhibitor of the 20 acetylcholinesterase active site, for example phenserine, galantamine, or tacrine; an antioxidant, such as Vitamin E or Vitamin C; an anti-inflammatory agent such as flurbiprofen or ibuprofen optionally modified to release nitric oxide (for example NCX 2216, produced by NicOx) or an oestrogenic agent such as 17-p-oestradiol. Methods and pharmaceutical carriers for preparation of pharmaceutical 25 compositions are well known in the art, as set out in textbooks such as Remington's Pharmaceutical Sciences, 20th Edition, Williams & Wilkins, Pennsylvania, USA. As used herein, a "pharmaceutical carrier" is a pharimaceutically acceptable solvent, suspending agent or vehicle for delivering the compound of formula IA to the subject. The carrier may be liquid or solid and is selected with the planned 30 manner of administration in mind. Each carrier must be pharmaceutically "acceptable" in the sense of being compatible with other ingredients of the composition and non injurious to the subject. The compound of formula IA may be administered orally, topically, or parenterally in dosage unit formulations containing conventional non-toxic 35 pharmaceutically acceptable carriers, adjuvants, and vehicles. The term parenteral as used herein includes subcutaneous injections, aerosol for administration to lungs or 2114997_1 (GKMatters) 10/11/09 - 27 nasal cavity, intravenous, intramuscular, intrathecal, intracranial, injection or infusion techniques. The present invention also provides suitable topical, oral, and parenteral pharmaceutical formulations for use in the novel methods of treatment of the present 5 invention. The compounds of the present invention may be administered orally as tablets, aqueous or oily suspensions, lozenges, troches, powders, granules, emulsions, capsules, syrups or elixirs. The composition for oral use may contain one or more agents selected from the group of sweetening agents, flavouring agents, colouring agents and preserving agents in order to produce pharmaceutically elegant and palatable 10 preparations. Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharin. Suitable disintegrating agents include corn starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar. Suitable flavouring agents include peppermint oil, oil of wintergreen, cherry, orange or raspberry flavouring. Suitable preservatives include sodium benzoate, vitamin E, 15 alphatocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate. The tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of 20 tablets. These excipients may be, for example, (1) inert diluents, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating and disintegrating agents, such as corn starch or alginic acid; (3) binding agents, such as starch, gelatin or acacia; and (4) lubricating agents, such as magnesium stearate, stearic 25 acid or talc. These tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. Coating may also be performed using techniques described in the U.S. Pat. Nos. 4,256,108; 4,160,452; and 30 4,265,874 to form osmotic therapeutic tablets for control release. The compound of formula IA as well as the pharmaceutically-active agent useful in the method of the invention can be administered, for in vivo application, parenterally by injection or by gradual perfusion over time independently or together. Administration may be intravenously, intraarterial, intraperitoneally, intramuscularly, 35 subcutaneously, intracavity, transdermally or infusion by, for example, osmotic pump. For in vitro studies the agents may be added or dissolved in an appropriate biologically acceptable buffer and added to a cell or tissue. 21149971 (GHMatter.) 10/11/09 - 28 Preparations for parenteral administration include sterile aqueous or non aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous 5 solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as, for example, anti 10 microbials, anti-oxidants, chelating agents, growth factors and inert gases and the like. Generally, the terms "treating", "treatment" and the like are used herein to mean affecting a subject, tissue or cell to obtain a desired pharmacologic and/or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or sign or symptom thereof, and/or may be therapeutic in terms of 15 a partial or complete cure of a disease. "Treating" as used herein covers any treatment of, or prevention of disease in a vertebrate, a mammal, particularly a human, and includes: (a) preventing the disease from occurring in a subject that may be predisposed to the disease, but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; or (c) relieving or ameliorating the effects of the disease, 20 i.e., cause regression of the effects of the disease. The invention includes various pharmaceutical compositions useful for ameliorating disease. The pharmaceutical compositions according to one embodiment of the invention are prepared by bringing a compound of formula IA, analogues, derivatives or salts thereof, or combinations of compound of formula IA and one or 25 more pharmaceutically-active agents into a form suitable for administration to a subject using carriers, excipients and additives or auxiliaries. Frequently used carriers or auxiliaries include magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatin, starch, vitamins, cellulose and its derivatives, animal and vegetable oils, polyethylene glycols and solvents, such as sterile water, alcohols, 30 glycerol and polyhydric alcohols. Intravenous vehicles include fluid and nutrient replenishers. Preservatives include antimicrobial, anti-oxidants, chelating agents and inert gases. Other pharmaceutically acceptable carriers include aqueous solutions, non toxic excipients, including salts, preservatives, buffers and the like, as described, for instance, in Remington's Pharmaceutical Sciences, 20th ed. Williams and Wilkins 35 (2000) and The British National Formulary 43rd ed. (British Medical Association and Royal Pharmaceutical Society of Great Britain, 2002; http://bnf.rhn.net), the contents of which are hereby incorporated by reference. The pH and exact concentration of the 2114997 1 (GHMatrera) 10/11/09 WO 2004/031161 PCT/AU2003/001303 - 29 various components of the pharmaceutical composition are adjusted according to routine skills in the art. See Goodman and Gilman's The Pharmacological Basis for Therapeutics (7th ed., 1985). The pharmaceutical compositions are preferably prepared and 5 administered in dose units. Solid dose units may be tablets, capsules and suppositories. For treatment of a subject, depending on activity of the compound, manner of administration, nature and severity of the disorder, age and body weight of the subject, different daily doses can be used. Under certain circumstances, however, higher or lower daily doses may be appropriate. The administration of the daily dose can be 10 carried out both by single administration in the form of an individual dose unit or else several smaller dose units and also by multiple administration of subdivided doses at specific intervals. The pharmaceutical compositions according to the invention may be administered locally or systemically in a therapeutically effective dose. Amounts 15 effective for this use will, of course, depend on the severity of the disease and the weight and general state of the subject. Typically, dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of the pharmaceutical composition, and animal models may be used to determine effective dosages for treatment of the cytotoxic side effects. Various considerations are described, e.g., in 20 Langer, Science, 249: 1527, (1990). Formulations for oral use may be in the form of hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil. 25 Aqueous suspensions normally contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspension. Such excipients may be (1) suspending agent such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; (2) dispersing or wetting agents which may be (a) naturally occurring 30 phosphatide such as lecithin; (b) a condensation product of an alkylene oxide with a fatty acid, for example, polyoxyethylene stearate; (c) a condensation product of ethylene oxide with a long chain aliphatic alcohol, for example, heptadecaethylenoxycetanol; (d) a condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol such as polyoxyethylene sorbitol monooleate, 35 or (e) a condensation product of ethylene oxide with a partial ester derived from fatty acids and hexitol anhydrides, for example polyoxyethylene sorbitan monooleate.

- 30 The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension may be formulated according to known methods using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation 5 may also a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil 10 may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Compounds of formula IA may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of 15 phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines. The compounds of formula IA may also be presented for use in the form of veterinary compositions, which may be prepared, for example, by methods that are conventional in the art. Examples of such veterinary compositions include those adapted for: 20 (a) oral administration, external application, for example drenches (e.g. aqueous or non-aqueous solutions or suspensions); tablets or boluses; powders, granules or pellets for admixture with feed stuffs; pastes for application to the tongue; (b) parenteral administration for example by subcutaneous, intramuscular or intravenous injection, e.g. as a sterile solution or suspension; or (when 25 appropriate) by intramammary injection where a suspension or solution is introduced in the udder via the teat; (c) topical applications, e.g. as a cream, ointment or spray applied to the skin; or (d) intravaginally, e.g. as a pessary, cream or foam. 30 Dosage levels of the compound of formula I or II of the present invention are of the order of about 0.5 mg to about 20 mg per kilogram body weight, with a preferred dosage range between about 0.5 mg to about 10 mg per kilogram body weight per day (from about 0.5 gms to about 3 gms per patient per day). The amount of active ingredient that may be combined with the carrier materials to produce a single 35 dosage will vary depending upon the host treated and the particular mode of administration. For example, a formulation intended for oral administration to humans may contain about 5 mg to Ig of an active compound with an appropriate and 2114997 1 (GHMatterO) 10/11/09 WO 2004/031161 PCT/AU2003/001303 - 31 convenient amount of carrier material which may vary from about 5 to 95 percent of the total composition. Dosage unit forms will generally contain between from about 5 mg to 500 mg of active ingredient. Optionally the compounds of the invention are administered in a divided 5 dose schedule, such that there are at least two administrations in total in the schedule. Administrations are given preferably at least every two hours for up to four hours or longer; for example the compound may be administered every hour or every half hour. In one preferred embodiment, the divided-dose regimen comprises a second administration of the compound of the invention after an interval from the first 10 administration sufficiently long that the level of active compound in the blood has decreased to approximately from 5-30% of the maximum plasma level reached after the first administration, so as to maintain an effective content of active agent in the blood. Optionally one or more subsequent administrations may be given at a corresponding interval from each preceding administration, preferably when the plasma level has 15 decreased to approximately from 10-50% of the immediately-preceding maximum. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the 20 severity of the particular disease undergoing therapy. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a flow chart of subjects studied; Fig. 2 are graphs showing mean change (±SE) over time from baseline in 25 cognitive abilities (as assessed with ADAS-cog) in (A) two arms of CQ vs placebo and (B) stratification by severity within treatment arms [less-severely affected (ADAS-cog < 25), more-severely affected (ADAS-cog >25) (*p 5 0.05; ** p 5 0.01); Fig. 3 are graphs showing mean change (±SE) over time from baseline in plasma Ap 42 levels in (A) the arms of CQ vs placebo and (B) stratification by severity 30 as in Fig 8. (***p< 0.001); Fig. 4 are graphs showing mean change (±SE) over time from baseline in (A) plasma Zn (B) plasma Cu in the two arms of CQ vs placebo; and Fig. 5 is a graph showing relative changes in behavioral (ADAS-cog) and biochemical (plasma/CSF Ap) levels over the course of AD. 35 WO 2004/031161 PCT/AU2003/001303 - 32 EXAMPLES The invention will now be described in detail by way of reference only to the following non-limiting examples. 5 (1) PREPARATION OF 8-HYDROXY-QUINOXALINES AND 8 HYDROXY-3H-QUINAZOLIN-4-ONES 8-Hydroxy-quinoxalines and 8-hydroxy-3H-quinazolin-4-ones) can be prepared 10 by processes known in the literature. For example, condensation of an appropriately substituted 1,2-phenylenediamine with 1,2-dicarbonyl compounds such as pyruvic aldehyde provides a range of 2- or 3-substituted 8-hydroxyquinoxalines (see for example: Abe and coworkers, J Med Chem., 1998, 41, 4062) (Scheme 1). An alternative synthesis of such compounds via condensation of the 1,2-phenylenediamine 15 with epoxides in the presence of Bismuth(0) is shown in Scheme 2 (see Antoniotti and coworkers, Tetrahedron Letters, 2002, 4, 3971). Further elaboration of the 2- and/or 3 position(s) can be achieved using known methods. Scheme 1 20 R2 R3

NH

2 N R 2 N R3 (Ri)n o O , (R1)n + (R)n qNH 2 N 2'N R OH OH OH 25 Scheme 2

NH,

2 N R2N R 3 (RI)n-- R3 (R ')n + (RI)n qNH, N R' N R 2 OH OH OH 30 8-Hydroxy-3H-quinazolin-4-one itself was prepared according to a literature procedure (see Iyer and Dhar, J. Sci. Ind. Res., 1956, 15C, 1). Derivatives of 8-hydroxy 3H-quinazolin-4-one can be synthesised by numerous methods known in the literature. One of the most common approaches to 3H-quinazolin-4-one derivatives is via 35 substituted 2-aminobenzamides. The 2-aminobenzamides may be accessed via two different approaches (Scheme 3). For example, a 2-nitrobenzoic acid 3A is first converted into the corresponding anthranilic acid 3B. In the presence of amine and an WO 2004/031161 PCT/AU2003/001303 - 33 activating agent such as CDI, 3B gives the 2-aminobenzamide 3C (Path A). Alternatively, 3A and an amine in the presence of CDI can first be converted into the 2 nitrobenzamide 3D. Subsequent reduction of 3D gives 3C (Path B). 5 Scheme 3 R COOH Path A R COOH R CONHR

NO

2

NH

2

NH

2 3A 3B 3C 10 Path B R CONHR

NO

2 3D 15 The compound, 4,6-dichloro-3-hydroxy-2-nitrobenzoic (4F) (Scheme 4) is a key intermediate used for the provision of a range of 8-hydroxy-3H-quinazolin-4-ones and specifically for the synthesis of 5,7-dichloro-substituted derivatives. Hence, according to Golstein and Schaaf (Helv. Chim. Acta, 1957, 57(23), 132), commercially available 20 2,4-dichlorobenzoic acid (4A) is nitrated to give 2,4-dichloro-5-nitrobenzoic acid (4B). Compound 4B is converted, via the amine 4C and the acetamide 4D, into 3-acetamide 4,6-dichloro-2-nitrobenzoic acid (4E). Subsequent base hydrolysis of 4E gives the 2 nitrobenzoic acid 4F. All steps proceed in high yields (about 90%). 25 Scheme 4 ci Cl CI CI C COOH C COOH C COOH C COOH 4A

NO

2

NH

2 NHAc 4B 4C 4D CI Cl COOH COOH CI NO, C NO 2 NHAc OH 4E 4F *prepared according to GoIstein and Schaaf, HeIv. Chim. Acta, 1957, 57(23), 132 WO 2004/031161 PCT/AU2003/001303 - 34 The intermediate of formula 4E above is novel and therefore forms part of the present invention. The use of this novel intermediate results in the process of Scheme 4 being high yielding and amenable to scale-up. 5 The conversion of a 2-nitrobenzoic acid such as compound 4F into 8-hydroxy 3H-quinazolin-4-ones (5C) is shown in Scheme 5. Hence, treatment of 4F with an amine in the presence of CDI produces the N-(substituted) benzamide 5A. Reduction of the nitro group and coupling of the resultant amine 5B with formic acid/CDI provides the desired N-alkylated derivative 5C. 10 Scheme 5 ci cl ci cl o COOH CONHR CONHR R C1 NO, C O NO, C O NH, Cl N 15OH OH OH OH 4F 5A 5B 5C N-alkylated quinazolin-4-ones may also be prepared via the route shown in Scheme 6. Hence, the appropriately substituted 2-nitrobenzoic acid such as 4F is first 20 treated with an amine in the presence of an activating agent such as CDI. This provides the corresponding nitrobenzamide which, under reducing conditions, gives the aminobenzamide. Subsequent treatment of the aminobenzamide with refluxing formamide produces the 3H-quinazolin-4-one. The 3H-quinazolin-4-one is then deprotonated with a base such as NaH and treated with an appropriate alkyl halide. 25 Scheme 6 Ci Ci Cl cl 0 C COOH C CONH2 CONH, NH CiNO, Ci1J NO, ci J NH, CI N 30 OH OH OH OH 4F ci 0 C1 N 35 OH WO 2004/031161 PCT/AU2003/001303 - 35 Scheme 7 shows a route to the synthesis of 2-substituted 8-hydroxy-3H quinazolin-4-ones. Hence, treatment of an anthranilic acid such as 2-amino-4,6 dichloro-3-hydroxybenzoic acid with thionyl chloride and subsequent reaction of the acid chloride with ammonia gives the corresponding benzamide. This, in turn, is treated 5 with chloroacetyl acetic acid to provide 5,7-dichloro-2-chloromethyl-8-hydroxy-3H quinazolin-4-one (see for example: Tani and coworkers, J Med. Chem., 1979, 22, 95). Further elaboration of the 2-chloromethyl derivative into a range of 2-substituted methyl derivatives, e.g., 2-(methylamino)methyl derivatives, can be achieved using standard literature methods. N-alkylation of these derivatives using the conditions 10 earlier described produces 2,3-disubstituted derivatives. Scheme 7 CI Cl ci o 15 Cl COOH C CONH2 C7 N CiNH, ci NH, CiN OH OH OH cl ci 0 CI 0 20 CNH N R OH R1 OH RI 2,3-Disubstituted-8-hydroxy-3H-quinazolin-4-ones may also be synthesised via 25 the route shown in Scheme 8. Hence, a 2-amino-3N-(substituted)benzamide is treated with the appropriate carboxylic acid such as Boc-protected glycine. Subsequent dehydration with sodium methoxide (see Lee and coworkers, J. Med. Chem., 1995, 38, 3547) produces the desired analog. 30 Scheme 8 ci o ci o ci o C2NHR N NHR 1 C ci NH, ci Ni clN R OH OH R2 OH 35 WO 2004/031161 PCT/AU2003/001303 - 36 (2 PREPARATION OF 8-METHOXY-4(3H)-QUINAZOLINONE, 8 HYDROXY-CINNOLINE, 5,8-DIHYDROXY-OUINOXALINE, 4,5 DIHYDROXY-PHENAZINE, 4,8-DIHYDROXY-PHENAZINE, 4 HYDROXY-ACRIDINE AND 5-HYDROXY-3-METHYL-2(1H) 5 QUINOXALINONE General The following compounds were prepared by the methods described in 10 the literature: 8-methoxy-4(3H)-quinazolinone (3),5 8-hydroxy-cinnoline (C1), 2 5,8 dihydroxy-quinoxaline (B39), 1 4,5-dihydroxy-phenazine (F5), 3 4,8-dihydroxy phenazine (F2), 3 4-hydroxy-acridine (El), 4 and 5-hydroxy-3-methyl-2(lH) quinoxalinone (B12) 19 . The following compounds were sourced commercially: 8 Hydroxy-quinazoline (Al), 4-hydroxy-phenazine (Fl), 4,10-phenanthrolin-5-ol (D2), 15 4,7-phenanthrolin-5,6-diol (D3) and 8-hydroxy-2-methyl-4(3H)-quinazolinone (1). Amines: ethylamine, histamine, 2-(2-aminoethyl)pyridine, 2-(2 methylaminoethyl)pyridine; aldehydes: 4-imidazolecarboxaldehyde, 2 thiazolecarboxaldehyde and 2-pyridinecarboxaldehyde, azoles: pyrazole, imidazole, methylimidazole and 1H-1,2,3-triazole, boronic acids: phenylboronic acid, 2 20 (trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o-tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid and 4-fluorophenylboronic acid; and organozinc reagents: 2-pyridylzinc bromide, 2 25 (methylthio)phenylzinc iodide, 2-(ethoxycarbonyl)phenylzinc iodide and 6 methylpyridylzinc bromide (0.5 M solution in THF) were commercially available (Aldrich). 3-Pyridylboronic acid was purchased from Frontier Scientific. 2 Aminomethylthiazole was prepared according to the literature.

13 Solvents were analytical grade and used as supplied. THF was distilled from sodium and 30 benzophenone under argon. 1H NMR spectra (8, relative to TMS) were recorded on a Varian Unity 300 spectrometer unless otherwise indicated; J-Values are given in hertz. Mass spectral data were recorded on a Micromass Quattro II mass spectrometer. The synthesis of derivatives of 6 classes of compounds: 8-hydroxy quinazoline, 8-hydroxy-quinoxaline, 8-hydroxy-cinnoline, 4,7(4,10)-phenanthrolin-5 35 ol, 4-hydroxy-acridine and 6-hydroxy-phenazine, is described in Part A, B, C, D, E and F, respectively.

WO 2004/031161 PCT/AU2003/001303 - 37 PART A: SYNTHESIS OF 8-HYDROXY-QUINAZOLINE DERIVATIVES A2 - A138 The preparation of a series of 8-hydroxy-quinazoline derivatives is 5 summarized in Charts A1 - A5. 4 NN OH OH OH Class A Class B Class C N19 1 4 N5 N N N4 4 OH OH OH OH Class D Class E Class F WO 2004/031161 PCT/AU2003/001303 - 38 CHART A1 -R N N Me OH A24. A37 -R

-

N N Me 0 36-49 Cl N Me N O OH NR 1

R

2 A142 - A153 0 ci 35 NH "NN "N N "N l '"' N Me N Me N Me N CHO OH OH OH OH OH NHR 1 A3* 2 A4 N "N *- and 5-chloro; 5-chloro-7-iodo; 7-chloro-5-iodo; 5,7-dichloro OH N, and 5-bromo OR derivatives thereof N N OH NH 2 OH NHCOR AIO* A130* WO 2004/031161 PCT/AU2003/001303 - 39 CHART A2 0 OMe OMe OMe OH N H NN CN COOH OMe 3 OMe OMe 0 OMe OH 11 CI OMe OMe OH ON Me O OMe OMe OMe 0 OH NHR 1 so 95 120 A12- AA17 R OMe OH N N N Rf -N c C Me OMe OH 0OH NR 96 A120 OH 7-20 NHR1 OMe 51 -64 OMe NH NH.SO 2

.CH

4 Me RR Me O I -R OH 0H N OMe7A121* OH NHR 1 97-110 21-34 OMe A38 -A51* OH 7 - Me N OMe 12 1 S 0 N 'N, jOH NHR 1 OH R Al8 - A23* AB - A98 OH NNNMe -R NHH A2 OH S A122 OH a R nd 5-chloro; A99 -A112 I 5-chloro-7-lodo; 7-chloro-5-iodo; 5, 7-dictoro end 5-bromo derivatives 'N N thereof 7 - Me OH S A123* WO 2004/031161 PCT/AU2003/001303 - 40 CHARTA3 N N '-R m OH OH R = 5-membered R =aryl, heteroaryl heterocyclic 'N N A135 -A138* A131 -A134* N I~~" OMeNNeN Ne 129 eN e ""N OMe R OH R t 115-118 All13 -A111S* OMe 114 N ~ N"NH OWe 0 OMe 113 ' "N OH Br ~ >~NH A126 - A127* 'N OMe K-N~ K-1126-1126 N NHY I I-a. 1We 1 1B~R H S" OHA1A139 12 -12 A128: ~qCN O NH OH Al OH A52 C1123 0 NH, 'NNN 'N ~NH C' ~OH kNH ON "N OH A53 N N H A124- 0'NH 02N-I N" N ' OMe 124 N OASS OH ':( N- 2NSN1 65N"N ' N NH 2 'N "N O OH OH OH~~r A11 6-7 " - 'N HH OH O H C 66-7 OOH A7015 A1-8 WO 2004/031161 PCT/AU2003/001303 - 41 CHART A4 OlMe OMe OH- OH 1~N'S NS qNIS C(N 'L H H;( OMe H OMe OH OH R 95 130 134 A142 R akyl, ary, heteroaryl OMe NN S e OOH N R A143* OMe R = alkyl, aryl, N heteroaryl OMe c 1132 OMe OH 0, 0 OMe NHR OH NHR 133 A140 0 -and 5-chioro; 5-chloro-7-iodo; ( N-% 1 7-chloro-5-iodo; 5,7-dichioro I and 5-bromo OH NHR derivatives therreof Al * WO 2004/031161 PCT/AU2003/001303 - 42 CHART A5 F N F N OH A155 F F OH Al OH OH A154 A156 CI Fj N F 4 ::'N OH OH A157 A158 HOOC N RHN N OH OH A160 Al 59 N z "N "" "N RHN CN RHN N C O OH A161 OH A162 DER DER DER: further derivatives via substitution at the 2- (chloro/cyano) position to a subject in need thereof, as shown in CHARTS Al -A4. 5-Position was also halogen-substituted (as shown on this CHART) WO 2004/031161 PCT/AU2003/001303 - 43 Preparation of 4-Chloro-8-hydroxy-2-methyl-quinazoline (A2) (Scheme Al) 0 Cl NHp - -N 0 C "r-N Me MN'Me OH OH OH A2 A3 Scheme Al 5 8-Hydroxy-2-methyl-4(3H)-quinazolinone (0.01 mol) and phosphorus oxychloride (10 mL) were heated under reflux for 30 min. The excess phosphorus oxychloride was removed under reduce pressure and the residue was added to a mixture of ice (50 g) and water (50ml), and the pH adjusted to 6 (aqueous ammonia). 4-Chloro 8-hydroxy-2-methyl-quinazoline (A2) was isolated via filtration. 10 Preparation of 8-Hydroxy-2-methyl-quinazoline (A3) (Scheme Al) 4-Chloro-8-hydroxy-2-methyl-quinazoline (0.01 mol) was treated with hydriodic acid (100 mL; freshly distilled from red phosphorus) according to the method described 6 in the literature. This provided 8-hydroxy-2-methyl-quinazoline (A3) as a 15 solid. Preparation of 8-Hydroxy-quinazoline-2-carboxaldehyde (2) (Scheme A2) NMe NCHO N OH OH OH NHR A3 2 A4 -A7 Scheme A2 20 A solution of 8-hydroxy-2-methyl-quinazoline (A3) (5 mmol) in dioxane (10 mL) was added dropwise over 3 h into a stirred mixture of SeO2 (8.8 mmol) in dioxane (30 mL) at 50'C. The resulting mixture was then heated at 80*C for 16 h, allowed to cool, and the solids filtered off. The filtrate was concentrated and purified 25 via column chromatography on silica (dichloromethane/MeOH, 40:1). This afforded 8 hydroxy-quinazoline-2-carboxaldehyde (2) as a solid.

WO 2004/031161 PCT/AU2003/001303 - 44 Preparation of 2-[Alkylamino-methyl]-8-hydroxy-quinoxaline (A4 - A7) (Scheme A2) Sodium triacetoxyborohydride (1 mmol) was added to a stirred solution 5 of 8-hydroxy-quinoxaline-2-carboxaldehyde (1 mmol) and ethylamine (1 mmol) in dichloromethane (10 mL). The mixture was left to stir at RT for 16 h, neutralized (aqueous NaHCO 3 ), and concentrated. The residue, after column chromatography on silica, afforded A4. In a similar fashion, reductive amination of 2 with amines: histamine 10 gave A5, 2-(2-aminoethyl)pyridine gave A6, 2-(2-methylaminoethyl)pyridine gave A7. Preparation of 8-hydroxy-quinazoline-2-carboxaldehyde Oximes (A8 - A9) (Scheme A3) -~"N CHO N OH OH NOR 2 A8 -A9 15 Scheme A3 A mixture of 8-hydroxy-quinazoline-2-carboxaldehyde (2) (1 mmol), NaOAc (2 mmol), hydroxylamine hydrochloride (1.5 mmol) and water (10 mL) was heated at 100 'C for 15 min. The precipitate was isolated by filtration. This provided 8 20 hydroxy-quinazoline-2-carboxaldehyde oxime (A8) as a solid. The reaction, repeated using 2 with methoxylamine hydrochloride in pyridine, gave A9. Preparation of 8-Hydroxy-2-methylamino-quinazoline (A10) (Scheme A4) 25 OH N OH OH NH, AB A10 Scheme A4 A solution of the 2-carboxylic acid oxime (1 mmol) in MeOH (20 mL) was treated under hydrogenolysis conditions (atmospheric H 2 , 10% Pd/C) for 16 h. The WO 2004/031161 PCT/AU2003/001303 - 45 solids were filtered off and the filtrate concentrated to provide 8-hydroxy-2 methylamino-quinazoline (Al0). Preparation of 4,8-Dimethoxy-quinazoline-2-carboxylic acid (All) (Scheme A5) 5 0 OMe OMe OMe NNH CN OMe OMe OMe 0 OMe 3 4 56

NH.NH.SO

2

.C

6

H

4 Me Cl OH OH N O O NNCOOH OH NHR, OH NHR 1 OH NHR 1 OH 21-34 7-20 A12 -A17 All OH NHR1 A18 - A23 Scheme A5 To a stirring mixture of 8-methoxy-4(3H)-quinazolinone 5 (3) (0.05 mol) and THF (100 mL) was added iodomethane (0.1 mol), tetrabutylammonium bromide 10 (100 mg) and aqueous NaOH (prepared from 7.55 g of NaOH in 20 mL H 2 0). After 16 h at 40 C, the mixture was concentrated and the remaining residue partitioned between

H

2 0 and dichloromethane (1:1, 200 mL). The organic layer was washed with brine, dried and concentrated. Column purification gave 4,8-dimethoxy-quinazoline (4). To a stirred solution of 4 (40 mmol) in CHC1 3 (200 mL) at 0 "C was 15 added m-chloroperbenzoic acid (44 mmol) portionwise over 10 min. After a further 30 min at 0*C, the mixture was allowed to warm to RT over 30 min and then concentrated to dryness. To the remaining residue was added ethyl acetate and 1 N NaHCO 3 (1:1, 200 mL); the layers were separated and the organic layer was dried (Na 2

SO

4 ), and concentrated. This provided the N-oxide 5.

WO 2004/031161 PCT/AU2003/001303 - 46 A mixture of 5 (30 mmol), benzene (80 mL) and dimethyl sulphate (35 mmol) was stirred under reflux for 16 h, allowed to cool, and concentrated in vacuo. To the remaining residue in H 2 0 (100 mL) at 0 0 C was added NaCN (90 mmol). After 3 h, the reaction mixture was neutralised (HOAc) and extracted with dichloromethane, the 5 extracts combined and dried. Solvent removal gave the 2-cyano-compound 6. A mixture of 6 (20 mmol) and NaOH (40 mmol) in H 2 0 (20 mL) was heated at 100 C for 4 h, and cooled. The pH of the solution was adjusted to 4 (glacial HOAc) and the mixture extracted with ethyl acetate (50 mL x 4). The combined extracts were dried and the volatiles removed. This provided 4,8-dimethoxy 10 quinazoline-2-carboxylic acid as a solid. Subsequent de-O-methylation with BBr 3 gave 4,8-dihydroxy-quinazoline-2-carboxylic acid (Al1). Preparation of 4,8-dihydroxy-quinazoline-2-carboxylic acid amides (A12 - A17) (Scheme A5) 15 1,3-Dicyclohexylcarbodiimide (1 mmol) was added to a stirred solution of 1-hydroxybenzotriazole hydrate (1 mmol) and 4,8-dihydroxy-quinazoline-2 carboxylic acid (All) (1 mmol) in DMF and dichloromethane (1:1, 10 mL). After 30 min, histamine (1 nnrnol) was added and the mixture stirred at RT for a further 16 h. 20 The volatiles were then removed in vacuo and the remaining residue gave, after purification by column chromatography on silica (ethyl acetate/i-PrOH/2 N NH 4 0H, 6:2:1), 4,8-dihydroxy-quinazoline-2-carboxylic acid[2-(1H-imidazol-4-yl)-ethyl-amide (A12). The above reaction was repeated using amines with All: 2-(2 25 aminoethyl)pyridine gave A13, 2-(aminomethyl)pyridine gave A14, 2-aminothiazole gave A15, 2-aminophenol gave A16, 1,2-phenylenediamine gave A17. Preparation of 8-Hydroxy-quinazoline-2-carboxylic acid amides (A18 - A23) (Scheme 5) 30 The 4-hydroxy-compound (A12) (1.2 mmol) and phosphorus oxychloride (4 mL) were heated under reflux for 15 min and allowed to cool. Ice (50 g) was added and the mixture basified with aqueous ammonia. The mixture was extracted with dichloromethane (20 mL x 3), the extracts combined, dried, and concentrated. This 35 provided the corresponding 4-chloro-compound 7. To a stirred mixture ofp-toluenesulfonhydrazide (Aldrich, 2 mmol) in CHCl 3 (10 mL) at 50 0 C was added portionwise over 10 min, the 4-chloro-compound 7 WO 2004/031161 PCT/AU2003/001303 - 47 (1 mmol). After 16 h, the solid was isolated via filtration, washed with H20, and dried. This provided the 4-N'-(p-toluenesulfonhydrazino)-compound 21. The 4-N'-(p-toluenesulfonhydrazino)-compound 21 (0.8 mmol) was added to Na 2

CO

3 (10 mmol) and H 2 0 (10 mL) at 95 C, and the mixture heated under 5 reflux for 15 min, cooled, filtered, and the filtrate extracted with CHCl 3 . The extracts were combined and dried. Subsequent removal of volatiles afforded the 8-hydroxy quinazoline-2-carboxylic acid amide (A18). In a similar fashion, the remaining 4(3H)-quinazolinones A13 - A17 were converted into 8-hydroxy-quinazoline-2-carboxylic acid amides (A19 - A23). 10 Preparation of 4-Aryl(or heterocyclic)-8-hydroxy-2-methyl-quinoxaline (A24 A37) (Scheme A6) R -R ci ci 5 N N ~ N N Me N Me N Me NMe OH 0 0 OH A2 11 A24 - A37 35 36-49 15 Scheme A6 2-Bromopropane (9 mmol) was added to a stirred mixture of 4-chloro-8 hydroxy-2-methyl-quinazoline (A2) (6 mmol), K 2 C0 3 (24 mmol) and DMSO (20 mL). After 16 h at RT, saturated NH 4 Cl (20 mL) was added and the mixture extracted with 20 dichloromethane (20 mL x 3). The extracts were combined and concentrated. Diethyl ether (100 mL) was added to the residue and the resulting mixture washed successively with 2 N NaOH, H20 and brine, and dried (Na 2 SO4). Solvent removal afforded 4 chloro-8-isopropoxy-2-methyl-quinazoline (35) as a solid. To a stirred mixture of 4-chloro-8-isopropoxy-2-methyl-quinazoline (35) 25 (0.58 mmol), phenylboronic acid (0.62 mmol), 2 N Na 2 C0 3 (7.2 mL), EtOH (1.2 mL) and benzene (6 mL) was added, under a blanket of argon, Pd(PPh 3

)

4 (20 mg). The mixture was stirred under reflux for 16 h, cooled, and concentrated. Subsequent column chromatography (ethyl acetate/hexane) provided 8-isopropoxy-2-methyl-4-phenyl quinazoline (36) as a solid. 30 To a stirred solution of 8-isopropoxy-2-methyl-4-phenyl-quinazoline (36) (0.34 mmol) in dichloromethane (2 mL) at -78 C was added BC1 3 (1.36 mL of a 1 WO 2004/031161 PCT/AU2003/001303 - 48 M solution in dichloromethane, 1.36 mmol). The reaction mixture was allowed to warm to RT (over 2 h) and stirred for a further 2 h. MeOH (5 mL) was added and the mixture was concentrated to dryness. The process was repeated four times. Further washing of the remaining residue with diethyl ether (2 mL x 3) provided 8-hydroxy-2-methyl-4 5 phenyl-quinazoline (A24). In a similar fashion, treatment of 4-chloro-8-isopropoxy-2-methyl-quinazoline (35) with boronic acids: 2-(trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 10 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid, 4 fluorophenylboronic acid and 3-pyridylboronic acid; and isopropoxy cleavage with BC1 3 gave 4-aryl(or heterocyclic)-8-hydroxy-2-methyl-quinazolines (A25 - A37). Preparation of 4-Aryl(or heterocyclic)-8-hydroxy-quinazoline (A38 - A51) 15 (Scheme A7) -R -R 0 c NH NN N OMe OMe OMe OH 3 50 51-64 A38 -A51 Scheme A7 Using the procedure as previously described in Example 9, 8-methoxy 20 4(3H)-quinazolone 5 (3) (10 mmol) and phosphorus oxychloride provided 4-chloro-8 methoxy-quinazoline 50. Treatment of the 4-chloride 50 with phenylboronic acid as described in Example 9 gave, after de-0-methylation with BBr 3 , the 4-phenyl derivative A38. The coupling reaction of 50 was repeated using a range of boronic acids: 2 (trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o-tolylboronic 25 acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid, 4 fluorophenylboronic acid and 3-pyridylboronic acid gave 51 - 64; and subsequent cleavage of the methyl ether with BBr 3 gave the 4-aryl(or heterocyclic)-8-hydroxy 30 quinazolines A39 - A51.

WO 2004/031161 PCT/AU2003/001303 - 49 Preparation of 5-Chloro-8-hydroxy-quinazoline (A52) (Scheme A8) CI CI Cl N N OH OH OH 0 Al A52 A53 65 CI Cl N N N) N) OH R A54 - A67 66-79 Scheme A8 5 Chlorine (12 mmol) was added into a stirred solution of 8-hydroxy quinazoline (Al) (10 mmol) in 93% H2SO4 following a previously published 7 procedure. After 3 h, ice (100 g) and H 2 0 (100 mL) was added, the mixture basified with aqueous ammonia, extracted with dichloromethane, and the extracts dried. Solvent removal gave the 5-chloro-compound A52. 10 Preparation of 5-Chloro-8-hydroxy-7-iodo-quinazoline (A53) (Scheme A8) 5-Chloro-8-hydroxy-quinazoline (10 mmol) was added to a stirred solution of ICl (12 nmol) in concentrated HC1 (10 mL).

8 After 5 min, the precipitate 15 was isolated via filtration, washed successively with H20, saturated sodium thiosulfate and H20, and dried. This provided 5-chloro-8-hydroxy-7-iodo-quinazoline (A53) as a solid. Preparation of 7-Aryl(or heterocyclic)-5-chloro-8-hydroxy-quinazolines (A54 20 A67) (Scheme A8) The 8-hydroxy-compound A53 was converted into the corresponding isopropyl ether 65 using 2-bromopropane according to the method as described in Example 9.

WO 2004/031161 PCT/AU2003/001303 - 50 Treatment of 5-chloro-7-iodo-8-isopropoxy-quinazoline (65), according to the method described in Example 9, with a range of boronic acids: phenylboronic acid, 2-(trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 5 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid, 4 fluorophenylboronic acid and 3-pyridylboronic acid gave 66 - 79; and subsequent cleavage of the 8-isopropoxy group with BC1 3 , gave A54 - A79. 10 Preparation of 7-Chloro-8-hydroxy-quinazoline (A69) (Scheme A9) N )CI N3 CI r OH OH OH OH Al A68 A69 A70 Scheme A9 15 8-Hydroxy-quinazoline (Al) (0.1 mol) and concentrated sulfuric acid (50 mL) were heated at 100 "C for 5 h, and allowed to cool. The solution was then carefully added to 300 mL of cold H 2 0. The resulting precipitate was isolated via filtration, washed with H20, and dried. This provided the 5-sulfonic acid A68 as a solid. To a stirred mixture of 5-sulfonic acid A68 (0.09 mol) and H20 (225 20 mL) was added KOH (0.25 mol) and NaOC1 (230 mL of a solution containing 10-13% available chlorine).

9 After 1.5 h at RT, the solution was passed through a column of Amberlite IR-120(H*) resin. The effluent concentrated to 20 mL. Acetone (20 mL) was then added and the precipitate isolated by filtration. Subsequent washing with acetone and drying gave 7-chloro-8-hydroxy-quinazoline (A69). 25 Preparation of 7-Chloro-8-hydroxy-5-iodo-quinazoline (A70) (Scheme A9) To a solution of 7-chloro-8-hydroxy-quinazoline (A69) (0.1 mol) and potassium acetate (0.15 mol) in MeOH and H 2 0 (19:1, 250 mL) was added, over 30 30 min, a solution of iodine (0.095 mol) in MeOH and H20 (19:1, 350 mL). The mixture was then heated under reflux for 15 min, cooled and H 2 0 (400 mL) was added. The precipitate was isolated by filtration, washed with saturated sodium thiosulfate and H20, and dried. This provided 7-chloro-8-hydroxy-5-iodo-quinazoline (A70) as a solid.

WO 2004/031161 PCT/AU2003/001303 - 51 Preparation of 4-Aryl(or heterocyclic)-7-chloro-8-hydroxy-quinazolines (A71 A84) (Scheme A10) R R C CN N CI N CI N' CI"" N CI1 N OH 0 0 OH A70 A71 - A84 80 81-94 5 Scheme Al0 According to the method described in Example 9, treatment of 5-chloro 7-iodo-8-isopropoxy-quinazoline (80), with a range of boronic acids: phenylboronic acid, 2-(trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o 10 tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid, 4 fluorophenylboronic acid and 3-pyridylboronic acid gave 81 - 94; and subsequent cleavage of the 8-isopropoxy group with BC1 3 , gave A71 - A84. 15 Preparation of 2-Chloro-4,8-dimethoxy-quinoxaline (96) (Scheme Al1) OMe OMe OMe OMe 0 OMe OMe 5 95 96 Scheme All 20 Ac 2 0 (6 mL) was added to a stirred mixture of the N-oxide 5 (10 mmol) and dichloromethane (10 mL). The solvent was then removed in vacuo and the resulting solution heated under reflux for 1 h, cooled, and concentrated. The remaining residue was washed with diethyl ether (10 mL x 2). This provided 4,8-dimethoxy-2(lH) 25 quinazolinone (95) as a solid.

WO 2004/031161 PCT/AU2003/001303 - 52 Treatment of 4,8-dimethoxy-2(lH)-quinazolinone (95) (5 mmol) with phosphorus oxychloride (20 mL) and standard workup according to Example 8 provided the 2-chloride 96. 5 Preparation of 2-Aryl(or heterocyclic)-4,8-dihydroxy-quinazolines (A85 - A98) (Scheme A12) OMe OMe OH c N '--N OMe OMe OH 'O 96 97-110 A85-A98 A99 -A112 Scheme A12 10 According to the procedure previously described in Example 9, coupling of the 2-chloride 96 (0.1 mmol) with boronic acids: phenylboronic acid, 2 (trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o-tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 15 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid, 4 fluorophenylboronic acid and 3-pyridylboronic acid gave 97 - 110; and subsequent cleavage of the 8-0-methyl ether with BBr 3 , gave A85 - A98. Preparation of 2-Aryl(or heterocyclic)-8-hydroxy-quinazoline (Scheme A12) 20 Sequential treatment of the 4-hydroxy-compounds A85 - A98 with phosphorus oxychloride, p-toluenesulfonhydrazide and Na 2

CO

3 , according to the method previously described in Example 6, provided the 2-aryl(or heterocyclic)-8 hydroxy-quinazolines A99 - A112.

WO 2004/031161 PCT/AU2003/001303 -- 5 3 Preparation of 2-Chloro-8-methoxy-quinazoline (113) (Scheme A13) OH OMe OMe 0 OMe Al 111 112 113 I -~~ N - N N R = heterocyclic NMe NMe NHMe OH R OMe R OMe A113-A116 115-118 114 5 Scheme A13 According to the procedure previously described in Example 6, 8 hydroxy-quinazoline (Al) (20 mmol) and iodomethane gave 8-methoxy-quinazoline (111). 8-Methoxy-quinazoline (111) (10 mmol) was then treated with m 10 chloroperbenzoic acid which gave the N-oxide 112. Subsequent treatment of the N oxide 112 with Ac 2 0 and phosphorus oxychloride according to Example 14 afforded 2 chloro-8-methoxy-quinazoline (113). Preparation of 2-[(N'-methyl)-heterocyclic]-8-hydroxy-quinazolines (Al 13 - Al 16) 15 (Scheme A13) A solution of 2-chloro-8-methoxy-quinazoline (113) (10 mmol) in pyridine (10 mL) was added methylamine hydrochloride (Aldrich, 15 mmol). After 16 h at RT, the mixture was concentrated in vacuo. Subsequent column purification of the 20 residue gave the 2-(N'-methyl)-compound 114. Amination of 114 (1 mmol) with 2-bromopyridine (1.2 mmol) in the presence of [Pd 2 (dba) 3 ] and DPPP according to the method described" in the literature, provided 8-methoxy-2-[(N'-pyridyl)methyl]-quinazoline (115). Subsequent treatment of 25 115 with BBr 3 (Example 6) afforded the 8-hydroxy-quinazoline derivative A113. Amination of 114 was repeated using a range of 2-bromo-substituted heterocyclics: 2-bromothiazole, 4-bromo-1H-imidazole and 4-bromo-1- WO 2004/031161 PCT/AU2003/001303 - 54 methylimidazole which provided 116 - 118; and subsequent cleavage of the 0-methyl ether with BBr 3 , gave A114 - A116. Preparation of 5,7-Diamino-8-hydroxy-quinazoline (A117) (Scheme A14) 5 NO,

NH

2 NHCOR ON 2 H'N N I)-NROCHN OH OH OH OH Al 119 A117 A118 - A119 Scheme A14 To a solution of 8-hydroxy-quinazoline (Al) (0.05 mol) in acetic acid (175 mL) was added a solution of nitric acid (0.16 mol) in acetic acid (25 mL), keeping 10 the temperature below 30 "C. After 2 h, the 5,7-dinitro-compound 119 was isolated by filtration, washed with H 2 0, and dried. Hydrogenolysis of the 5,7-dinitro-compound 119 (0.045 mol) in MeOH (200 mL) in the presence of platinum oxide gave, after filtration to remove solids and concentration, 5,7-diamino-8-hydroxy-quinazoline (A117). 15 Preparation of 5,7-Diacylamino-8-hydroxy-quinazolines (A118 - A119) (Scheme A14) Acetic acid (2 mmol) and CDI (2.2 mmol) were heated under reflux in 20 dry THF (10 mL) for 1 h. 5,7-Diamino-8-hydroxy-quinazoline (A117) (2 mmol) was added and the mixture heated under reflux for 16 h. Removal of volatiles in vacuo and subsequent column chromatography of the resulting residue provided 5,7 desacetamido-8-hydroxy-quinazoline (A118). In a similar fashion, treatment of 5,7-diamino-8-hydroxy-quinazoline 25 (A117) with benzoic acid provided 5,7-dibenzoylamido-8-hydroxy-quinazoline (A119).

WO 2004/031161 PCT/AU2003/001303 - 55 Preparation of 2-Acetyl-4,8-dihydroxy-quinazoline (A120) (Scheme A15) OMe OMe OMe N CN Me N Me OMe OMe O OMe S 6 120 121 OH OH Me Me OH 0 OH S A120 A122 Me Me N NY OH 0 OH S A121 A123 Scheme A15 5 Methylmagnesium bromide (1.2 mL of a 3 M solution in diethyl ether, 3.5 mmol) was added dropwise into a stirred solution of 6 (0.6 mmol) in diethyl ether (10 ,L) at -15 C. The resulting solution was allowed to warm to RT over 2 h and stirred at RT for a further 4 h. The reaction mixture was then quenched with saturated

NH

4 Cl and extracted with ethyl acetate (10 mL x 3), the extracts combined, dried and 10 concentrated to provide 120. Treatment of 120 with BBr 3 (Example 6) gave A120. Preparation of 2-Acetyl-8-hydroxy-quinazoline (A121) (Scheme A15) Successive treatment of A120 with phosphorus oxychloride, p 15 toluenesulphonhydrazide and Na 2

CO

3 (Example 6) gave A121. Preparation of 2-S-Acetyl-4,8-dihydroxy-quinazoline (A122) (Scheme A15) A solution of 120 (1 mmol) and Lawesson's reagent (0.7 mmol) in THF 20 (10 mL) was heated under reflux for 16 h and allowed to cool. Concentration and subsequent column chromatography of the residue gave 121. Treatment of 121 with BBr 3 (Example 6) gave A122.

WO 2004/031161 PCT/AU2003/001303 - 56 Preparation of 2-S-Acetyl-8-hydroxy-quinazoline (A123) (Scheme A15) Successive treatment of A122 with phosphorus oxychloride, p toluenesulphonhydrazide and Na 2

CO

3 (Example 6) gave A123. 5 Preparation of 8-Hydroxy-quinazoline-2-urea (A124) (Scheme A16) C NH 2 N N OMe OMe OMe 0 e NH, OMe NH2 113 122 123 124 ""N I' -"N q NH ( NN<NH OH diNH, OH s>'NH2 A124 A125 Scheme A16 10 The 2-chloride 113 was converted into the amine 122 according to standard Chichibabin reaction conditions. The 2-amine 122 (1 mmol) and isocyanate (1 mmol) in dry CHCl 3 (10 mL) were then heated under reflux for 16 h. This gave, after filtration, the urea derivative 123. Treatment of 123 with BBr 3 gave A124. 15 Preparation of 8-Hydroxy-quinazoline-2-thiourea (A125) (Scheme A16) The 2-urea derivative 123 (1 mmol) and Lawesson's reagent (0.7 mmol) in THF (10 mL) was heated under reflux for 16 h, allowed to cool and concentrated. 20 Subsequent column purification on silica gave 124. Treatment of 124 with BBr 3 , according to Example 6, then provided A125.

WO 2004/031161 PCT/AU2003/001303 - 57 Preparation of 2-Acylamido-8-hydroxy-quinazolines (A126 - A127) (Scheme A17)

NINH

2 N NH NH OMe OMe O 81 OMe sAR1 122 125-126 127-128 NH N q N NH q N- NH OH OH SIRI A126 - A127 A128 - A129 Scheme Al 7 5 The 2-amine 122 was acylated under standard conditions: Ac 2 0 gave 125; benzoic anhydride gave 126. Respective treatment of 125 and 126 with BBr 3 (Example 6) provided A126 and A127. Preparation of 2-Thioacylamido-8-hydroxy-quinaxolines (A128 - A129) 10 (Scheme A17) The acylamido compounds 125 and 126 were individually treated with Lawesson's reagent according to conditions previously described (Example 21), which furnished, after standard workup, 127 and 128. Subsequent 0-methyl ether cleavage 15 with BBr 3 then gave A128 and A129, respectively. Preparation of 2-(acylamido)-methyl-8-hydroxy-quinazolines (A130) (Scheme A18) N N OH NH 2 OH NHCOR A10 A130 20 Scheme A18 Standard acylation of the amine A10 using a range of acid anhydrides provided, after column purification, the 2-(acylamido)-methyl derivatives A130.

WO 2004/031161 PCT/AU2003/001303 - 58 Preparation of 2-(Azole)-8-hydroxy-quinazolines (A131 - A134) (Scheme A19) " N N N 'N N CI NIR N R OMe 113 OMe OH R = 5-membered heterocyclic A131 -A134 Scheme A19 5 A mixture of the 2-chloride 113 (0.5 mmol) and pyrazole (2.5 mmol) was heated at 175 "C in a steel autoclave for 48 h. The crude product was then treated with BBr 3 according to the procedure described in Example 6. Subsequent purification by column chromatography gave 2-pyrazol-1-yl-quinazolin-8-ol (A131). 10 The above procedure was repeated using imidazole, 2-methylimidazole and 1H-1,2,3-triazole to give A132 - A134. Preparation of 2-Aryl(or heterocyclic)-8-hydroxy-quinazolines (A135 - A138) (Scheme A20) N I NR NN cl:N- R CN--R OMe OMe OMe OH 113 129 A135 - A138 R = ary, heteroary 15 Scheme A20 According to a literature 2 procedure, the 2-chloride 113 (5 mmol) was treated with acetyl chloride and sodium iodide in AcCN. Standard workup followed by 20 column chromatography on silica furnished the 2-iodo-compound 129. To a stirred solution of 129 (0.1 mmol) and PdCl 2 (PPh 3

)

2 (5 mg) in THF (2.5 mL) under an argon atmosphere at RT was added over 5 min 2-pyridylzinc bromide (0.37 mL of a 0.5 M solution in THF, 0.185 mmol). After 2 h, saturated NH 4 Cl (5 mL) was added and the mixture extracted with dichloromethane (10 mL x 3). The 25 combined extracts were washed with H 2 0 and brine, dried and concentrated. Subsequent column chromatography on silica gave 8-methoxy-2-pyrid-2-yl quinazoline. The 8-0-methyl ether was cleaved according to the procedure of Example 6, to give 2-(pyrid-2-yl)-8-hydroxy-quinazoline (A135).

WO 2004/031161 PCT/AU2003/001303 - 59 The reaction was repeated using: 2-(methylthio)phenylzinc iodide, 2 (ethoxycarbonyl)phenylzinc iodide and 6-methylpyridylzine bromide to give A136 A138. 5 Preparation of 5-Bromo-8-hydroxy-quinazoline (A139) (Scheme A21) Br OH OH Al A139 Scheme A21 Bromination of 8-hydroxy-quinazoline (Al) with N-bromosuccimide, 10 according to the method previously described by Gerson and McNeil, 15 gave 5-bromo 8-hydroxy-quinazoline (A139). Preparation of 8-Hydroxy-quinazoline-2-sulfonic acid (2-pyridin-2-yl-ethyl)-amide (A140 and A141) (Scheme A22) 15 OMe OMe OMe OMe Lawesson's N reagentoN Mel N SOCI, H 2 0 's N N O "' N S N SMe N S Hl 0 OMe H OMe H OMe 131 OMe C32 95 130 11132 I RNH 2 OH OMe " N Example 6 N BBra N N- S N"-S NXS'( OH 0 -0 O 1e NH OH OH NHR OMe H A141 A140 133 Scheme A22

RNH

2 : preferably, ethylamine, histamine, 2-(2-aminoethyl)pyridine, 2 (2-methylaminoethyl)pyridine 20 WO 2004/031161 PCT/AU2003/001303 - 60 Preparation of 2-Alkyl(or aryl or heterocyclic)sulfanyl-8-hydroxy-quinazoline (A142 and A143) (Scheme A23) OMe OH OH BBr 3 N RBr N Example 6 H I '" N S N - S N- S OMe OH OH R OH R 130 134 A142 A143 R = alkyl, aryl, R = alkyl, aryl, heteroaryl heteroaryl Scheme A23 5 RBr: preferably, 2-bromopyridine, 2-bromothiazole, 4-bromo-1H imidazole, 4-bromo-1-methylimidazole Reductive amination of amines (A4 - A7) from Example 3 to give A142 - A153 (Scheme A24) RCHO (reductive amination) N N Example 3 N OH NHR 1 OH NR 1 R2 A4 - A7 A142 -A153 10 Scheme A24 RCHO: preferably, 4-imidazolecarboxaldehyde, 2 thiazolecarboxaldehyde, 2-pyridinecarboxaldehyde 15 WO 2004/031161 PCT/AU2003/001303 - 61 Preparation of 5-fluoro-, 7-fluoro, 5,7-difluoro, 5-chloro-7-fluoro and 7-chloro-5 fluoro-quinazolines (A154 - A158) (Scheme A25) F F Nc OH Al OH OH A154 A156 ci F F ' C F N' F / OH OH OH A157 A158 A155 5 Scheme A25 The synthesis of the fluorinated quinazoline derivatives (A154 - A158) followed literature methods1 8 for 8-hydroxy-quinoline. 10 Preparation of 8-Hydroxy-quinazoline-7-carboxylic acid amides (A160) (Scheme A26) N ~N "N HOOC RHN- 1 ) OH 0 OH A160 A159 15 Scheme A26 8-Hydroxy-quinazoline (Al) was converted into 8-hydroxy-quinazoline-2-carboxylic acid (A159) following the literature method 20 for the carboxylation of 8-hydroxy 20 quinoline. Subsequent conversion of A159 into the amide A160 followed the procedure described in Example 8. PART B: SYNTHESIS OF 8-HYDROXY-QUINOXALINE DERIVATIVES 25 Charts B1 - B4 show the routes to the 8-hydroxy-quinoxaline derivatives. The synthetic procedures were, unless otherwise indicated, analogous to those previously described in Part A.

WO 2004/031161 PCT/AU2003/001303 - 62 CHART B1 F N N) ~DER Cl N OH B7 DER F F N N )#N IDER N F N' OH B6 OH B8 DER

SO

2 OH I N Cl NC OH Bi OH B2 OH B3 OH B4 I Cl CI NND -- DER R OH B9 OH B10 DER : Cl N OH B5 Cl R = aryl, heterocyclic DER N (5- and 6-membered) -- DER R NDE OH B11 R = aryl, heterocyclic (5- and 6-membered) DER: derivatives via substitution of 2- and /or 3-chloro/cyano group(s) to a subject in need thereof e.g. NR 1

R

2

(R

1 = H, alkyl; R2 = alkyl, aryl, heterocyclic), SR, CONHR, alkylamino WO 2004/031161 PCT/AU2003/001303 - 63 CHART B2 NHCOR O NH.C(O).R ,z N R R N Me Me N Me OH B23 OH B19* OH B22* R = alkyl, aryl R = aryl, heterocyclic 1 (5- and 6-membered) N' NH N > H2 ,-z NN CI NN 4 N MeN Me H 2 N N Me OH NHR OH B18* OH OH R= alkyl, substituted alkyl H N 0 N NN NMe Me N CHO NHCe H OHB12* OH B13* OH Me C1 OH B20* Cl N Me NMe OH B24 OH B27* OH NsOR OH B15* NH 2 N NH.C(S).NH 2 R = H, alkyl N Me B17A N MeI C1 OH B21* CN M N CN Cl N Me I" N ' Me CI OH B25 OH B28 N OH B16* NH.C(O).R R = alkyl, aryl, NH 2 OH N, OR I SR = H, alkyl N R "' B17B ' N I R N' Me N C1 NIMe OH B29 OH B16* NH.C(S).R OH B26 R- aryl, heterocyclic R = alkyl, aryl, NH 2 R = aryl, heterocyclic (5- and 6-membered) (5- and 6-membered) DER: derivatives via substitution of 3-chloro/cyano group to a subject in need thereof e.g. NR 1

R

2 (R1 = H, alkyl;

R

2 = alkyl, aryl, heterocyclic), SR, CONHR, alkylamino * - and halogen-substitution, preferably at the 5 and/or 7-position(s) WO 2004/031161 PCT/AU2003/001303 - 64 CHART B3 OH OMe OMe OMe N R1 N R1 D NH 2 N R N R 2 N R 2

NH

2 B N R 2 OH OMe OMe OMe B34* R 1 , R 2 = aryl, heteroaryl R 1 , R 2 = H, alkyl, methylhalo alkyl and derivatives of these thereof; preferably A C CONHR, methylamino, oximes, -CH 2

NR

1

R

2

(R

1 = H, alkyl; R 2 = alkyl, OMe OMe acyl) N N OH N Me N NXR1 OMe OMe N R 2 DER OH -- R 1 , R 2 = H, alkyl, methylhalo OH OMe OH N N NCHO N OH NHR OMe OH B36* B39* OH DER N N OH N A = pyruvic aldehyde (Ref 16) OH B37* N OR B= O (Ref 17) N R2 DER OH B38* NR 1

R

2 R1 R1 = H, alkyl R2 = alkyl, acyl C = glyoxal DER: derivatives via substitution of 3 chloro/cyano group to a subject in need D = R (Ref 16) thereof e.g. NR 1

R

2

(R

1 = H, alkyl; OH 2=alkyl, aryl, heteroaryl, SR, N C R 2 CONHR, alkylamino * - and halogen-substitution, preferably N at the 6- and 7-positions OH NR 1

R

2 DER R1 = H, alkyl R2 = alkyl, acyl WO 2004/031161 PCT/AU2003/001303 - 65 CHART B4 OH OMe OMe OMe N R1 N 1 D NH 2 N R1

R

2

R

2

NH

2 N R 2 OH OMe OMe OMe B34* R 1 , R 2 = aryl, heterocyclic et H alkyl, methylhalo, alkyl and derivatives of these thereof; preferably A C CONHR, methylamino, oximes, -CH 2

NR

1

R

2

(R

1 = H, alkyl; R 2 = alkyl, OMe OMe acyl) N N OH N Me N N R1 OMe OMe N) N R 2 DER OH

R

1 , R 2 = H, alkyl, methylhalo, methylamino OH OMe OH B35* N N N N CHO OH NHR OMe OH B36* B39* OH DER N N OH A = pyruvic aldehyde (Ref 16) OH B37* NsOR B= 0 (Ref17) N F R2 DER OH B38* NR1R2 R1 R1 = H, alkyl R2 = alkyl, acyl C = glyoxal DER: derivatives via substitution of 3 chloro/cyano group to a subject in need D = (Ref 16) thereof e.g. NR 1

R

2

(R

1 = H, alkyl; OH 2 = alkyl, aryl, heterocyclic), SR, R 2 0 CONHR, alkylamino N N - and halogen-substitution, preferably N DER at the 6- and 7-positions OH NR 1

R

2 R1 = H, alkyl R2 = alkyl, acyl WO 2004/031161 PCT/AU2003/001303 - 66 PART C: SYNTHESIS OF 8-HYDROXY-CINNOLINE DERIVATIVES Charts C1 and C2 show the routes to the 8-hydroxy-cinnoline derivatives. The synthetic procedures were, unless otherwise stated, analogous to those 5 previously described in Part A. CHART C1 -R C N NN N N Ci N Cl[ N. CI OH OH OH Ci SOOH NHR N N N OH OH OH I 1 NH, NHCOR N N N N N N N N N N N N OMe OH OH OH OH Ci' C1 CI -N ON N lN N NN N N N N OH OMe OH OH R R N OMe N N OH R = alkyl, aryl WO 2004/031161 PCT/AU2003/001303 - 67 CHART C2 0 NHR COOH N -N OH OH KR2

CH

2 OH CHO NHR

NHR

1 reductive N N NN nomination N N N tN e OH OH OH OH OR N OH

NH

2 NHCOR N N N OH OH NHR NgN OH PART D: SYNTHESIS OF 4,7(4,10)-PHENANTHROLIN-5-OL DERIVATIVES 5 Charts D1 - D4 show the routes to the 4,7(4,10)-phenanthrolin-5-ol derivatives. The synthetic procedures were, unless otherwise stated, analogous to those previously described in Part A. The reactions shown in Chart D1 were also repeated using 4,7-phenanthrolin-5,6-diol (D3) instead of 4,7-phenanthrolin-5-ol (D1) as starting material.

WO 2004/031161 PCT/AU2003/001303 - 68 CHART D1 (0,S) N N (0,S) OH NC HOOC RHN N N N - )m I

-

- 0 N CN N COOH N OH OH OH NHR HNglycerol HN H.O OHC ''' ~(Skraup synthesis) N N ' Ni NH N N N NRHH) OHOH OH OH OH RH NN NHR N NN N N S N- R N C I N N OH OH O H N. O RH NHR R = aryl, heteroaryl R H lyayay (5- or 6-membered) I ,sklayay ROCHN HN HN.CO(S).HN N NHCOR N NH, HCOS.H OH OH OH RHN N N NHR

OH

WO 2004/031161 PCT/AU2003/001303 - 69 CHART D2 NH, glycerol e Me (Skraup synthesis) N Me N M e N Me OH OH OH NN N N N CHO N OH OH NHR "N Nj OH N, R = H alkyl OR N OH NHR R =H, acyl WO 2004/031161 PCT/AU2003/001303 - 70 CHART D3 N N NHCOR OH N N N NH.CO(S).NH 2 N NH 2 OH OH

NH

2 N N glycerolI (Skraup synthesis) N 0 N 0 N 0 N Cl H H H OH OH OH OH N N R OH R = aryl, heteroaryl (5- or 6-membered) WO 2004/031161 PCT/AU2003/001303 - 71 CHART D4 NH, N H2glycerolNN (Skraup synthesis) N N Br N OH OH OH D2 N 0 2 N N R N OH R/ OH N

H

2 N N OH N RHN N OH R = alkyl, aryl, acyl Skraup synthesis of 4,7-Phenanthrolin-5-ol (D1) 5 A stirring mixture of 3-hydroxy-p-phenylenediamine (0.185 mol), glycerol (1.17 mol), arsenic solution (100 mL; prepared from 123 g of arsenic pentoxide in 100 mL H 2 0) and diluted sulphuric acid (400 mL; prepared by adding 240 mL of concentrated sulphuric acid to 200 mL H 2 0) was heated under reflux for 4 h, allowed to cool and then made alkaline with concentrated ammonia. The mixture was 10 extracted with benzene. Removal of the benzene afforded 4,7-Phenanthrolin-5-ol (D1).

WO 2004/031161 PCT/AU2003/001303 - 72 PART E: SYNTHESIS OF 4-HYDROXY-ACRIDINE DERIVATIVES The 4-hydroxy-acridines were prepared via Ullman condensation 4

'

1 4 of a substituted 2-halobenzoic acid and a substituted aniline as shown in Chart El. Hence, 5 condensation of aniline itself with 2-bromo-3-nitro-benzoic acid gave 4-hydroxy acridine (E1). In an analogous fashion, o-anisidine gave 4-amino-5-hydroxy-acridine and 4,5-dihydroxy-acridine. Further derivatives of these acridines (Chart E2) were prepared using analogous reaction conditions previously described for the synthesis of compounds in Parts A - D. 10 CHART E1 HOOC OOc substituted NH, Br NSsu Substitute Substituted H NO 2 NO2 0 0 Susiue:-N C N Susiue H, Substituted H NO Substituted O OH Substituents on aniline include alkyl, methoxy, halogen WO 2004/031161 PCT/AU2003/001303 - 73 CHART E2 O ClR Ullman N ndensation N z NH 2 N N

NO

2

NO

2

NO

2 HR1 = H, alkyl N R2 = alky, aryl, acyl OH El- H Ri =H, alkyl R2 = alkyl, aryl, acyl O OH CN CHO NR1R2 - CCNq N OH = H, alkylOH R alkyl, aryl, acyl 0 NHN COOHO NH OH OH OH OH O WO 2004/031161 PCT/AU2003/001303 - 74 PART F: SYNTHESIS OF 4,5(4,8)-DIHYDROXY-PHENAZINE DERIVATIVES The compounds, 4,5- and 4,8-dihydroxy-phenazines (F1 and F2), were prepared according to the literature 3 procedure. The synthesis of derivatives of these 5 compounds, as summarized in Charts F1 - F3, unless otherwise stated, followed reaction conditions analogous to those previously described in Parts A - E. CHART F1 0 2 N_- N

NH

2 N OMe OMe OMe OMe

NH

2

NH

2

NH

2

NH

2 Br Br N N N N N # N "NH ', N N'N,. OH OH OH OH OHF2 OH OH OH

NR

1

R

2

NR

1

R

2

NR

1

R

2 NR1R 2 S0 2 0H SO 2 OH N ' N: NN

R

2

R

1 N N NR 1

R

2 N N OH OH OH OH OH OH R1 = H, alkyl R 1 = H, alkyl

R

2 = alkyl, acyl R 2 = alkyl, acyl N C " N CI N. OH OH c# N - c OH OH WO 2004/031161 PCT/AU2003/001303 - 75 CHART F2 OMe OMeN

NH

2 + 02NN 'N OMe OMe

NH

2 OH NH 2 OH OH Br OH HN N N N OH NH 2 OH NH 2 OH F3 OH Br NHR OH NHR OH SO 2 OH OH NN4 N4 RHN # N NHRN OH NHR OH NHR OH SO 2 OH R = alkyl, acyl R = alkyl, acyl HC

OH

WO 2004/031161 PCT/AU2003/001303 - 76 CHART F3

SO

2 OH -CI -- C -Cl -CI NN- -CI- N ICiJ N OH OH OH OH AI - NH 2 N, N N CN CN -CN

NH

2 NC N Ci NC OH OH OH OH 4c I N N CCI N OH OH

NH

2 NC, NH2C -CONHR OH CiN OH N N0 CI NRR1 = H, alkyl 0 OH

R

2 = alkyl, acyl B O ON B0 N N N NN N N N 0 7R R N O 2 NR RC =D Ci N c I # N2 OH O H WO 2004/031161 PCT/AU2003/001303 - 77 REFERENCES 1. F.E. King, N.G. Clark and P.M.H. Davis, J. 1949, 3012-3016. 5 2. E.J. Alford, H. Irving, H.S. Marsh and K. Schofield, J. 1952, 3009-3017. 3. A. Sugimoto, S. Kato, H. Inoue and E. Imoto, Bull. Chem. Soc. Jpn., 1976, 49(1), 337-338. 10 4. A. Corsini and E.J. Billo, J Inorg. NucL. Chem., 1970, 32, 1241-1255. 5. R.N. Iyer, N. Anand and M.L. Dhar, J Sci. Ind. Res., 1956, 15C, 1-7. 6. A. Albert and A. Hampton, J. 1952, 4985-4993. 15 7. H. Gerson, M. W. McNeil and S.G. Schulman, J. Org. Chem., 1971, 36, 1616 1618. 8. A. Atsushi, N. Kazuo, H. Kinichi and 0. Masana, Radioisotopes, 1974, 23(1), 6 20 9. 9. H. Gerson, M.W. McNeil and A.T. Grefig, J. Org. Chem., 1969, 34, 3268-3270. 10. H. Gerson and M.W. McNeil, J. Org. Chem., 1971, 8, 821-824. 25 11. S. Wagaw and S.L. Buchwald, J. Org. Chem., 1996, 61, 7240-7241. 12. R.C. Corcoran and S.H. Bang, Tetrahedron Lett., 1990, 31, 6757-6758. 30 13. (a) A. Dondoni, G. Fantin, M. Fogagnolo, A. Medici and P. Pedrini, Synthesis, 1987, 998-1001. (b) A. Dondoni, F.L. Merchan, P. Merino, I. Rojo and T. Tejero, Synthesis, 1996, 641-646. 14. S. Issmaili, G. Boyer and J.-P. Galy, Synlett, 1999, 641-643. 35 15. H. Gerson and M.W. McNeil, J. Org. Chem., 1972, 37, 4078-4082.

WO 2004/031161 PCT/AU2003/001303 - 78 16. X.-H. Bu, H. Liu, M. Du, K.M.C. Wong, V. W.W. Yam and M. Shionoya, Inorg. Chem., 2001, 40, 4143-4149. 17. S. Antoniotti and E. Dunach, Tetrahedron Lett., 2002, 43, 3971-3973. 5 18. H. Gerson, M.W. McNeil, R. Parmegiani and P.K. Godfrey, J. Med. Chem., 1972, 15, 987-989, and references cited therein. 19. I.Y. Postovskii and N.G. Koshel, Khim. Geterotsikl. Soedin., 1970, 7, 981-985. 10 20. V.V. Ragulin, I.R. Ragulina and L.G. Shakirov, Zhurnal Prikladnoi Khimii, 1994, 67(7), 1227-1229. 15 (3 PREPARATION OF 8-HYDROXY-[1,6]NAPHTHYRIDINES A range of 7-substituted-8-hydroxy-[1,6]naphthyridines can be prepared using the route shown in Scheme 9 (see for example: Anthony and coworkers, Patent WO 02/30931 A2). Hence, 2,3-pyridinedicarboxylic acid is readily transformed through a 20 series of reactions into 8-hydroxy-[1,6]naphthyridine-2-carboxylic acid methyl ester. The 7-methyl ester can then be converted, e.g. under Curtius conditions, into the corresponding 7-amino compound. The amino functionality, in turn, can be further elaborated using known methods. For example, diazotisation in the presence of HCl provides the corresponding chloride. Further elaboration of the chloride can readily be 25 achieved using known methods. Scheme 9 N COOH N N 30 N COOH MeOOC N MeOOC N OH 0 N N N --------- V- 7-(5- or 6-membered optionally substituted HN2 N CIN aryl or heterocyclyl), 7-XR (X = 0, S), 7-NRIR etc 0 0 35 7-amides etc *prepared according to Anthony et al, Patent WO 02130931 A2 WO 2004/031161 PCT/AU2003/001303 - 79 5,7-Disubstituted-8-hydroxy-[ 1,6]naphthyridines can be accessed from an advanced intermediate such as 5,8-dihydroxy-[1,6]naphthyridine-7-carboxylic acid methyl ester (prepared according to the procedure of Albert and Hampton, J. Chem. 5 Soc., 1952, 4985; Blanco and coworkers, J. Heterocyclic Chein., 1996, 33, 361). Chlorination of 5,8-dihydroxy-[1,6]naphthyridine-7-carboxylic acid methyl ester with POCl 3 or SOC1 2 provides 5-chloro-8-hydroxy-[1,6]naphthyridine-7-carboxylic acid methyl ester. Elaboration of the chloride into a range of 5-substituted-8-hydroxy [1,6]naphthyridines can be achieved using known methods. Further elaboration using 10 literature methods provides a range of 5,7-disubstituted-8-hydroxy-[1,6]naphthyridines. Derivatives of 8-hydroxy-[1,6]naphthyridine substituted at 2-,3- and/or 4 positions can be synthesised using an appropriately substituted 2,3-pyridinedicarboxylic acid. For example, the commercially available 6-methyl-2,3-pyridinedicarboxylic acid 15 can be transformed into 8-hydroxy-2-methyl-[1,6]naphthyridine-7-carboxylic acid methyl ester (Scheme 10). Other substituted 2,3-pyridinedicarboxylic acids can readily be prepared using known methods (see for example: Wepplo, US Patent 4,460,776). Further elaboration of the 8-hydroxy-2-methyl-[1,6]naphthyridine provides a range of analogs such as 2-(methylamino)methyl and 2-(alkylamino)methyl derivatives. Various 20 7-substituted derivatives of these compounds can be prepared from the corresponding 7-methoxycarbonyl, 7-chloro or 7-amino compounds using known methods. Scheme 10 N 25 MeOOC- R OH HOOH ON O 3 0 N COOH 0eOOC N-- MeQOC' N HN N' OH 0 0 35 N ' RN' N N'N' Ri Nc H,N31 N OH 0 OH WO 2004/031161 PCT/AU2003/001303 - 80 (4) PREPARATION OF PYRIDO[3,2-dPYRIMIDIN-4-OL, [1,7]NAPHTHYRIDIN-8-OL, PYRIDO[2,3-d]PYRIDAZIN-8-OL, [1,6]NAPHTHYRIDIN-8-OL, PYRIDO[3,4-b]PYRAZIN-5-OL, 5 PYRIDO[3,4-b]PYRAZIN-8-OL, [1,5]NAPHTHYRIDIN-4,8-DIOL, [1,5]NAPHTHYRIDIN-8-OL AND PYRIDO[4,3-d]PYRIMIDIN-8-OL General 10 The following reagents were sourced commercially: - amines: ethylamine, histamine, 2-(2-aminoethyl)pyridine, 2-(2-methylaminoethyl)pyridine; aldehydes: 4-imidazolecarboxaldehyde, 2-thiazolecarboxaldehyde and 2 pyridinecarboxaldehyde, azoles: pyrazole, imidazole, methylimidazole and 1H-1,2,3 triazole, boronic acids: phenylboronic acid, 2-(trifluoromethyl)phenylboronic acid, 2 15 methoxyphenylboronic acid, o-tolylboronic acid, 2-fluorophenylboronic acid, 3 methoxyphenylboronic acid, 4-methoxyphenylboronic acid, rn-tolylboronic acid, 3,5 difluorophenylboronic acid, 2,4-difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid and 4-fluorophenylboronic acid; and organozinc reagents: 2-pyridylzinc bromide, 2-(methylthio)phenylzinc iodide, 2-(ethoxycarbonyl)phenylzinc 20 iodide and 6-methylpyridylzinc bromide (0.5 M solution in THF) (Aldrich). 3 Pyridylboronic acid was purchased from Frontier Scientific and pyrido[3,2 d]pyrimidin-4-ol from Ambinter, France. 2-Aminomethylthiazole was prepared according to the literature.

1 Solvents were analytical grade and used as supplied. THF was distilled from sodium and benzophenone under argon. 1H NMR spectra (6, relative 25 to TMS) were recorded on a Varian Unity 300 spectrometer unless otherwise indicated; J-Values are given in hertz. Mass spectral data were recorded on a Micromass Quattro II mass spectrometer. The synthesis of derivatives of 8 classes of compounds: pyrido[3,2 d]pyrimidin-4-ol (A), [1,7]naphthyridin-8-ol (B), pyrido[2,3-d]pyridazin-8-ol (C), 30 [1,6]naphthyridin-8-ol (D), pyrido[3,4-b]pyrazin-5-ol (E), pyrido[3,4-b]pyrazin-8-ol (F), and [1,5]naphthyridin-4,8-diol, [1,5]naphthyridin-8-ol (G) and pyrido[4,3 d]pyrimidin-8-ol (H), is described.

WO 2004/031161 PCT/AU2003/001303 - 81 NN N NN NNN N N N NNN OH OH OH OH A B C D 5 OH N N N N OH OH OH OH OH 10 E F G H Charts Al - A3 summarise the routes used in the synthesis of a series of pyrido[3,2-d]pyrimidin-4-ol derivatives. The preparation of the derivatives of [1,7]naphthyridin-8-ol is described in Charts BI - B2. Compound BI was prepared 15 following the literature method 2 for the synthesis of a similar compound. The synthesis of the pyrido[2,3-d]pyridazin-8-ol itself and derivatives C1 and C2 followed the method previously described 3 by Brzezinski and coworkers. Further derivatives in this series were prepared following the routes summarised in Charts Cl - C4. Charts Dl D2 show the routes to a series of [1,6]naphthyridin-8-ol derivatives. The synthesis of 20 compound D1 used the method 4 of Blanco and coworkers. Other members in this series have been prepared following the routes shown in Charts D1 - D2. Chart El describes the route to a series of pyrido[3,4-b]pyrazin-5-ol derivatives. Condensation of 1,2,3 triaminopyridine with 2,3-dihydroxy-1,4-dioxan gave pyrido[3,4-b]pyrazin-5-ol (El), the parent compound in this series; the same reaction employing 2,3,4-triaminopyridine 25 as starting material (Chart Fl)gave pyrido[3,4-b]pyrazin-8-ol (F1). Further derivatives in the pyrido[3,4-b]pyrazin-5-ol and pyrido[3,4-b]pyrazin-8-ol classes were prepared using the routes shown in Charts El and Fl. Charts G1 - G2 show the routes employed in the synthesis of a range of [1,5]naphthyridin-4,8-diol and[ 1,5]naphthyridin-8-ol derivatives. The synthesis of compound GI, the precursor to the parent compound, 30 [1,5]naphthyridin-4,8-diol, followed the route described by Brown and Dewar. Pyrido[4,3-d]pyrimidin-8-ol (Hi) was prepared 4 following the route shown in Chart Hl, employing 4,5-pyrimidinedicarboxylic acid as starting material. Further derivatives in this class were prepared using routes as shown in Charts Hl and H2.

WO 2004/031161 PCT/AU2003/001303 - 82 CHART Al

RNH

2 N CHO N eQ (reductive N N ~ N- ' Nyr aminiN)

NH

2 N Me N2CHO N OH OH OH OH N NH2OR R =H, alkyl; H2SO4 I R = alkyl, aryl, acyl N COOH N OH OH N'OR

RNH

2 , R = H, alkyl, aryl DCC N DH 2 , Pd/C N - N C(O).NHR N OH N, N OH NH 2 5 WO 2004/031161 PCT/AU2003/001303 - 83 CHART A2 HO YN MeO N CHO MeO N MeO N N N NR 1

R

2 N NN OH N N R = H, alkyl; On OBn OBn R2 = alkyl, heterocyclic; A2 acyl HO N MeO H MeO H MeO R NR NyZ - N -f- HZ; N N- RR N RN N R H CI NR OH OH OBn OH R = aryl, heterocyclic R = aryl, heterocyclic R1 = H, alkyl; (5- and 6-membered) (5- and 6-membered) R2 = alkyl, heterocyclic; acyl MeO N KN "N N -f-H 5NH 2 (N HR OBn NNH NR R 2 OH OH R = aryl, heterocyclic R = H, alkyl; (5- and 6-membered) MeO N R2 = alkyl, heterocyclic; Y N N acyl N NH.C(O).R OBn R = alkyl, aryl, NH 2 NMeO N HO N MeO H HO N N H N HC ). NN N( N.NNH -Z -f-HZ -' HCS. N NH.C(S).R H NH.C(S).R N NH.C(O).ROOH OBn OBn OBn R = alkyl, aryl, NH 2 NIR =alkyl aryl, NH- 2 N N,_N, N NMeO H NH HO N N NH.C(S).R N NH.C(S).R N / N NH.C(O).R N NH.C(O).R OH OH OH OH R = alkyl, aryl, NH 2 R = alkyl, aryl, NH 2 HO N N, / N NH.C(S).R

OH

WO 2004/031161 PCT/AU2003/001303 - 84 CHART A3 (N -( N R N R R OH OBn R = CONHR', -CH 2

N(R

2

)R

3 R = SR (R 1 = alkyl, aryl, - from CHART Al heterocyclic), aryl, heterocyclic,

NR

2

R

3

(R

2 = H, alkyl; R 3 =alkyl, heterocyclic, acyl), NH.CO(S).NH 2 , SO2NHR4 N- from CHART A2 N R OMeI R = C(O)NHR 1 , -CH 2

N(R

2

)R

3 N R OH R = SRI (R' = alkyl, aryl, heterocyclic), aryl, heterocyclic, NH 2 NR2R3 (R2 = H, alkyl; R3 =alkyl, heterocyclic, acyl), NH.CO(S).NH 2 ,

SO

2 NHR4, C(O).NHR6, -CH 2 N(R2)R3; R5 = group from R WO 2004/031161 PCT/AU2003/001303 - 85 CHART BI II 11 - HO "' N- - NZ N C(0).NHR HO C(O).NHR OH OH MeO NMeO NN C(O).NHR N C(O).NHR OBn OH N R MeO MeO N MeO N MeO C N R INZ NH 2 H -N, 'H I~ NN NIN COON OH0nnOBn OBn R aryl, heterocyclic BI n (5- and 6-membered) M e O M e O R M e O C M e O N NIN N R N R N C I N CN OH OBn OBn OBn R = aryl, heterocyclic R = aryl, heterocyclic (5- and 6-membered) (5- and 6-membered HO R MeO N MeO N N N N N N N" NH O N R O N NR 1

R

2 O N NH 2 OH 0I OH OBn OBnO R R =aryl, heterocyclic R 1 = H, aikyl; 1 H (S) (5- and 6-membered) = alkyl, aryl, R NH2, alkyl, aryl heterocyclic MO N N HO N N OH OH R = NH 2 , alkyl, aryl R = NH 2 , alkyl, aryl HO N OH N : NH

OHHO

WO 2004/031161 PCT/AU2003/001303 - 86 CHART B2 N R N O N R OH OMe OMe 2-R-substituted compound from CHART B31 N ~~N :' N NC R CN R OMe OMe RiHOOC N' 1N 'N H N'- '- NX I NY R R NOR N R OH OH OH R = aryl, heterocyclic

NH

2 , NR2R3 (R2 = H, alkyl;

R

3 = alkyl, aryl, heterocyclic, acyl), SR 4 i 2 2 N.7 N R OH R = H, alkyl;

R

2 = alkyl, aryl, acyl WO 2004/031161 PCT/AU2003/001303 - 87 CHART C1 """N 0 b'( N N II NHR OH R OH R = alkyl, aryl, heterocyclic - H~ ' -N - N K N MeO NHPh MeO N MeO N HO N 0 OH OBn OBn C' I Rf N N R N RN C OH OH OBn R = aryl, heterocyclic, R = aryl, heterocyclic,

NH

2 , NRIR2 (R' = H, NH 2 , NR 1

R

2 (R'= H, alkyl, R 2 = alkyl, alkyl, R 2 alkyl, heterocyclic, acyl), heterocyclic, acyl),

NHC(O).NH

2 , NH.C(S)NH 2

NH.C(O)NH

2 , NH.C(S).NH 2 WO 2004/031161 PCT/AU2003/001303 - 88 CHART C2 ~N"N _______ ."N NHPh - Me Nh Me N OHCN 0 OH OH C2 - N N N HOOC OH RO N OBn NRR 2 OH R = H, alkyl; R2 = alkyl, aryl, alkylamino, acyl 0 N Z N NN "N NHR OH NH 2 OH N N NR R 2 OH R = H, alkyl; R2 = alkyl, aryl, alkylamino, acyl WO 2004/031161 PCT/AU2003/001303 - 89 CHART C3 """N R"N OH R = aryl, heterocyclic, NH 2 , NR 1

R

2 (R' = H, alkyl; R2 = alkyl, heterocyclic, acyl), SR 3

(R

3 = alkyl, heterocyclic),

SO

2

NHR

4 , NH.CO(S).NH 2 , CONHRs, -CH 2

NH

2 , -CH 2

N(R')R

2 -from CHARTS C1 AND C2 K: "N R N OMe CI CN N N R N -N R N OMe OH

NR

1

R

2 CONHR' """N """N I I lK R N R N OH OH R1 = H, alkyl; R2 = alkyl, aryl, heterocyclic, acyl WO 2004/031161 PCT/AU2003/001303 - 90 CHART C4 n-BuLi; Me DMF; H+; NHPh MeNH.NH 2 _ R 1 NHh R' N N 0 OH

R

1 OMe or Me Me N

R

1 N OMe R' = aryl, heterocyclic, NH 2 , NR2R3 (R 2 = H, alkyl;

R

3 = alkyl, heterocyclic, acyl), SR 4

(R

4 = alkyl, heterocyclic),

SO

2 NHR5, NH.CO(S).NH 2 , CONHR6, -CH 2

NH

2 , -CH 2 N(R2)R3 - from CHARTS C1 AND C2

OR

2 CHO N-R 'N N 'N NN I I

R

1 N R N N

R

1 N OH OMe OH

R

2 = H or alkyl

R

2 = H, alkyl; R3 = alkyl, aryl,

NH

2 alkylamino, acyl N

R

1 N N

OH

WO 2004/031161 PCT/AU2003/001303 - 91 CHART D1 OH OMe OMe N N "':N EtOOC N EtOOC N EtOOC N C1 OH OMe OMe D1 CI OH N 6 N EtOOC N EtOOC N R R N R OMe OH OH R = SRI (R' = alkyi, aryl, R = SR 2

(R

2 alkyl, aryl, heterocyclic), aryl, heterocyclic, heterocyclic), aryl, heterocyclic,

NR

2

R

3 H alkyl; R 3 alkyl, NR 3

R

4

(R

3 = H, alkyl; R 4 = alkyl, R N heterocyclic, acyl), NH.CO(S).NH 2 , aryl, heterocyclic, acyl),

ISO

2

NHR

5

NH.CO(S).NH

2 ,

NHS

2 NHR5 EtOOC'' N0 OH R 6

=CONHR

7 , -CH 2

N(R

3 e)R 4 OH C R = SR (R = alkyl, aryl, heterocyclic), aryl, heterocyclic,

NR

2

R

3

(R

2 = H, alkyl; R 3 = alkyl, EtOOC N R OH heterocyclic, acyl), NH.CO(S).NH 2 , 1 OH

SO

2

NHR

5 IR = SRI (R1 = alkyl, aryl, heterocyclic), aryl, heterocyclic,

NR

2

R

3

(R

2 = H, alkyl; R 3 = alkyl, heterocyclic, acyl), NH.CO(S).NH 2 , N, - "N SO 2

NHR

5 OHO RR = SRR (R a = alkyl, aryl, h caheterocyclic), aryl, heterocyclic,N NR R (R = H, alkyl; R=alkyl, OH heterocyclic, acyi), NH.CO(S).NH 2 , R =SR 1 I (R 1 =alkyl, aryl,

SO

2

NHR

5 heterocyclic), aryl, heterocyclic,

NR

2

R

3

(R

2 = H, alkyl; R 3 = alkyl, heterocyclic, acyl), NH.CO(S).NH 2 ,

SO

2

NHR

5 WO 2004/031161 PCT/AU2003/001303 - 92 CHART D2 OMe OMe N EtOOC N EtOOC N CN OMe OMe prepared according to route shown in CHART Dl N CN OH N CHO N N COOH N C(O)NHR OH OH OH N N OH N,OR OH N R =H, alkyl R = H, alkyl;

R

2 = alkyl, aryl, acyl N OH

NH

2 WO 2004/031161 PCT/AU2003/001303 - 93 CHART El

NH

2 0 0~O N N - ~ N N

NH

2 00.

NH

2 OH El

NNH

2 0 N C hfMN N ,- + N Me N, NH 2 0 MeN

NH

2 OH NN N CH=NOR -CHO NCOOH OH R =H or alkyl OH OH N CH2 NH N 2 N CONHR N N :l R N;(N) OH OH OH R = H, alkyl; R2 = alkyl, aryi, acyl NHN R 1 N2 I~ NNy: H 2 NH 2I 1)cd ~~O NH NX N2H R N R NH2 OH R 1, R2 = alkyl, substituted alkyl, aryl, heterocyclic, OH example: Me H22 N NH 2 M Me N N R(R N rNH 2 NOf;R2 N R(R)ONH

NH

2 OH O Ar N Py NO: Ar N M N N H Me RAr N N- N Ar OHN N ~ MeC N CONHR

NH

2 N Me 'N N CONHR NOH OH O~Py N N H=NOR O Py N CH=NOR OH N Ar N '~ X KN CHNRR 2 OH N N CH 2

NR

1

R

2 XR OH R', R2 = H or substituted R' = H, alkyl; R2 = H, alkyl, aryl, acyl WO 2004/031161 PCT/AU2003/001303 - 94 CHART F1 N NH 2 +U -OUo N

NH

2 N

NH

2 OH Fi N zYNH 2 0 <H N N N -Me

N

2 0 MeN

NH

2 OH N CH=NOR N CHO N -COOH N N"N OH R = H or alkyl OH OH N! CHNH -) NCONHR OH OH OH R = H, alkyl; R2 = alkyl, aryl, acyl N NN N- N11 N R' N" N R'NR + -C- I '-R2NRR

N-NH

2 /X N2 H 2 2 NN NH2 OH R1, R2 = alkyl, substituted alkyl, aryl, heterocyclic, OH NH22 R(R N M e 0 CONHR N' NH 2 O -XNR N R 2

(R

1 )N Y NH 0 N Me N N-) CONHR

NH

2 OH OH 0 Me' SPy O N C H = NOR O Py " N CH=NOR OH N~ N Ar NrN 'N N CHNRR 2 N N OHN ArN' OH N. -N: CH 2

NRR

2 OH

R

1 , R 2 = H or substituted R1 = H, alkyl;

R

2 = H, alkyl, aryl, acyl WO 2004/031161 PCT/AU2003/001303 - 95 CHART GI OMe H OH OMe N N N N COOEt N N O OH OMe GI OH OMe NN CN N NC 1:N'V N OH OMe OH 0 OH OMe H N NHR N COOH N O RHN I __I N HOOC N 0 N 0 OH OH H OMe OH N'OR OH OMe N N CHO N Cl N"N N OHC N C N N OH OH OMe RO R= H, alkyl I OH NH 2 OH NR 1

R

2 OH NN R N N R N

NH

2 OH NRR 2 OH OH R1 = H, alkyl; R = NH 2 , SR' (R' = alkyl, aryl, R2 = alkyl, aryl, heterocyclic), aryl, heterocyclic, acyl

NR

2

R

3

(R

2 = H, alkyl; R 3 = alkyl, heterocyclic, acyl), NH.CO(S).NH 2 ,

SO

2

NHR

5 WO 2004/031161 PCT/AU2003/001303 - 96 CHART G2 O Cl N COOEt N COOEt N COOEt N N N OMe H OMe OMe GI N CH 2 OH N COOH N N DER OMe OMe N CH=NOR N CHO N N N H NN N OH OH OH R= H or alkyl I 4 DER

CH

2

NH

2 N N RE - 'N~ R 2 ER N N OH OH R1 = H or alkyl; R2 = alkyl, substituted alkyl, acyl N CH 2 NH.C(O)R 'N 7 N OH DER: derivatives with substitutions at 2- and 4 6-positions as in CHART GI

DER

WO 2004/031161 PCT/AU2003/001303 - 97 CHART HI OH CI N COOH NNN"N N N N "N I N COOH E0C N EtOOC N OH OMe OH D1 I H1 R R N N N'N R N EtOOC OH OH R SRI (R' = alkyl, aryl, R SRI (R1 = alkyl, aryl, heterocyclic), aryl, heterocyclic, heterocyclic), aryl, heterocyclic,

NR

2

R

3 (R' =H, alkyl; R 3 =alkyl, NR2R3 (R2 = H, alkyl; R3 = alkyl, heterocyclic, acyl), NHCO(S).NH 2 , heterocyclic, acyl), NH.CO(S).NH 2 , S0 2

NHR

5

SO

2

NHR

5 ,R6 = CONHR 7 , -CH 2 N(R )R9

(R

8 = H, alkyl; R 9 = H, alkyl, aryl, acyl) N OHH ( = SR (R R alkyl, aryl, heterocyclic), aryl, heterocyclic,

NR

2

R

3

(R

2 = H, alkyl; R 3 = alkyl, N heterocyclic, acyl), NH.CO(S).NH 2 , OH SO 2

NHR

5 - 89N

R

6 = CONHR, -CH 2 N(R=)Ra

(R

8 N = H, alkyl; R H, alkyl, aryl, acyl) WO 2004/031161 PCT/AU2003/001303 - 98 CHART H2 OH Cl yCOOH N COOHN 'N "N N N N N "N Me N COOH EtOOC N Me EtOOC N Me Me OH OMe OH DER R R N 'N "N N 'N ''N DER DER N Me EtOOC N Me OH OH R = SR' (R' = alkyl, aryl, R = SR 1 (R' = alkyl, aryl, heterocyclic), aryl, heterocyclic, heterocyclic), aryl, heterocyclic,

NR

2

R

3

(R

2 = H, alkyl; R 3 = alkyl, NR 2

R

3

(R

2 = H, alkyl; R 3 = alkyl, heterocyclic, acyl), NH.CO(S).NH 2 , heterocyclic, acyl), NH.CO(S).NH 2 ,

SO

2

NHR,,R

6 = CONHR', -CH 2

N(R

8 )R' SO2NHR5 (Ra = H, alkyl; R 5 = H, alkyl, aryl, acyl) R N 'N 'NN N Me DER OH R = SR 1 (R' = alkyl, aryl, heterocyclic), aryl, heterocyclic, DER :' N NR 2

R

3 (R2 = H, alkyl; R3 = alkyl, R6 N Me heterocyclic, acyl), NH.CO(S).NH 2 , OH

SO

2

NHR

5

R

6 = CONHRI, -CH 2 N(R")R9 (R8 = H, alkyl; R 9 = H, alkyl, aryl, acyl) DER: derivatives thereof , prepared via substitution of the 2-methyl group to a subject in need thereof e.g. COOH, CONHR, alkylamino, alkylamido, oxime, alkyloxime WO 2004/031161 PCT/AU2003/001303 - 99 REFERENCES 1. (a) A. Dondoni, G. Fantin, M. Fogagnolo, A. Medici and P. Pedrini, Synthesis, 1987, 998-1001. (b) A. Dondoni, F.L. Merchan, P. Merino, I. Rojo and T. 5 Tejero, Synthesis, 1996, 641-646. 2. A Albert and A. Hampton, J., 1952, 4985-4993. 3. J.Z. Brzezinski, H.B. Bzowski and J. Epsztajn, Tetrahedron, 1996, 52, 3261 10 3272. 4. M.Blanco, M.G. Lorenzo, I. Perillo and C.B. Schapira, J Heterocyclic Chem., 1996, 33, 363-366. 15 5. S.B. Brown and M.J.S. Dewar, J. Org. Chem., 1978, 43, 1331-1337. (5) PREPARATION OF 8-HYDROXY-6H-[1,61NAPHTHYRIDIN-5-ONE, 4 HYDROXY-4a,8a-DIHYDRO-PYRANO[3,2-bPYRIDIN-2-ONE, 8 20 HYDROXY-6H-[1,61NAPHTHYRIN-5-ONE, DIBENZO[a,glQUINOLIZIN-8-ONE AND 4-HYDROXY-1H PYRIDO[3,2-d1PYRIMIDIN-2-ONE General 25 The following reagents were sourced commercially: - amines: ethylamine, histamine, 2 (2-aminoethyl)pyridine, 2-(2-methylaminoethyl)pyridine; aldehydes: 4 imidazolecarboxaldehyde, 2-thiazolecarboxaldehyde and 2-pyridinecarboxaldehyde, azoles: pyrazole, imidazole, methylimidazole and 1H-1,2,3-triazole, boronic acids: 30 phenylboronic acid, 2-(trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o-tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, m-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid and 4-fluorophenylboronic acid; and organozinc reagents: 2-pyridylzinc bromide, 2 35 (methylthio)phenylzinc iodide, 2-(ethoxycarbonyl)phenylzinc iodide and 6 methylpyridylzinc bromide (0.5 M solution in THF) (Aldrich). 3-Pyridylboronic acid was purchased from Frontier Scientific. 2-Aminomethylthiazole was prepared according to the literature.

4 Solvents were analytical grade and used as supplied. THF WO 2004/031161 PCT/AU2003/001303 - 100 was distilled from sodium and benzophenone under argon. 1H NMR spectra (8, relative to TMS) were recorded on a Varian Unity 300 spectrometer unless otherwise indicated; J-Values are given in hertz. Mass spectral data were recorded on a Micromass Quattro II mass spectrometer. 5 The synthesis of derivatives of 5 classes of compounds: 8-hydroxy-6H [1,6]naphthyridin-5-one (A), 4-hydroxy-4a,8a-dihydro-pyrano[3,2-b]pyridin-2-one (B), 8-hydroxy-6H-[1,6]naphthyrin-5-one (C), dibenzo[a,g]quinolizin-8-one (D) and 4 hydroxy-lH-pyrido[3,2-d]pyrimidin-2-one (E), is described. H O 0 NIN 0 0O 'N HN 'N 'N~~ NI N OH OH OH A B C 0 H N 0 Y N N N 0 OH OH D E 10 Condensation of ethyl 3-amino-picolin-2-ate with acetic anhydride according to the method of Zhou and coworkers' gave 8-hydroxy-6H-[1,6]naphthyridin-5-one (Al). By substituting ethyl 3-amino-picolin-2-ate respectively with ethyl 5-chloro-3-amino picolin-2-ate and ethyl 6-methyl-3-amino-picolin-2-ate, the corresponding 2-methyl 15 and 3-chloro-8-hydroxy-6H-[1,6]naphthyridin-5-one derivatives were obtained. Further derivatisation of these systems as shown in Charts Al - A4 gave a series of 8-hydroxy 6H-[1,6]naphthyridin-5-ones. The preparation of a series of 4-hydroxy-4a,8a-dihydro pyrano[3,2-b]pyridin-2-ones is described in Charts B1 - B2. The method of Blanco and coworkers2 was employed for the synthesis of C2 which upon subsequent acid 20 hydrolysis provided 8-hydroxy-6H-[1,6]naphthyrin-5-one (C1). Further derivatives in this series were prepared following routes shown in Charts Cl - C2. Charts D1 - D2 show the preparation of a series of dibenzo[a,g]quinolizin-8-ones. The key step, involving the construction of the ring system, followed the method previously described 3 by Colye and coworkers. Chart El shows the routes employed in the 25 preparation of a series of 4-hydroxy- 1 (alkylated)-pyrido[3,2-d]pyrimidin-2-one derivatives.

WO 2004/031161 PCT/AU2003/001303 - 101 CHART A1 1

-~NH

2 N COOEt H O N MeO N MeO N MeO N N N N N Cl )-RI OH OMe OMe OMe Al H H 0 N0 O N MeO N N N N H NN N CN N -N R HN N R OH CN OH R OH OHH OO R' NH 2 , SIR 2

(FR

2 = alkyl, aryl, heterocyclic), aryl, heterocyclic, 41NR 3 R 4 (R 3 =H, alkyl; R 4 = alkyl, RMo N heterocyclic, acyl), NH.CO(S).NH 2 , I S0 2

NHR

5 COO R N CH O N I N CON O M e R HR=aNH R = alkyl0 a O N N N e0R=SR 2

(R

2 =alkylaryl, R = alkyl heteroaryl), aryl, heterocyclic, OH R OH NHRNR 3

R

4

(R

3 =H, alkyl; R 4 = alkyl, heterocyclic, acyl), NH.CO(S).NH 2 , Me

SO

2

NHR

5 R 0 N N O nHN:J N N COOH R 6 O)H NHR R 6 = alkyl, alkylamino, acyl OH R = alkyl The above reactions were repeated by substituting

NH

2 C N NH 2 with COOEt N COOEt WO 2004/031161 PCT/AU2003/001303 - 102 CHART A2

NH

2 Me N COOEt HR R H NH.CO.CHR.COOrt 0 N -- N meN N' Me COOEt R N MeR N 2 Me aky OH OH R OH R alkyl R =alkyl R alkyl RR= alkyl; 2 = RCH 2

N(R

3

)R

4

(R

3 = H, alkyl; R 4 = alkyl, acyl), R H H H-CH 2

NH

2 , -CH=NOR5 (R 5 =H, N 0 N 0 NO alkyl), CONHR CH=NH N N N -NR-N COO lHR = alky R= H, alkyl, aryl H 10 N 0 N H N0 N NR N' OH N OH NHR 2 RRN R2 OH NH 2 lkR = alkyl; R = alkyl ak;= HR alkyl= S= alkyl, aryl, acyl The above reactions were repeated by substituting

NH

2 Cl NH 2 with Me N COOt N COOEt 5 WO 2004/031161 PCT/AU2003/001303 - 103 CHARTA3 H 0 N N N CH=NOR OH R H, aikyi

NH

2 R Me N COOEt H M H HR HN ' N Me HN N me HN N R R6 OH R OH R OH i) malonic ester li) NaOEt, EtOH H R

NH.CO.CH

2 .COOEt 0 N 0 N 0 N Me N COOEt Me Me N R' OH OH OH R alkyl R -CH 2

N(R

2 )R3

(R

2 = H, alkyl; R 3 = alkyl, acyl),

-CH

2

NH

2 , -CH=NOR 4

(R

4 = H, H H alkyl), CONHR 5 O N 0 N N CH=NOR N CHO OH OH R =H, alkyl H O N N R N OH N OHH NHN OH NH-12

R

1 = H, alkyl;R R2 = alkyl, aryl, acyl H IO N I " HN N R N OH NH 2 HN N R' OH /N

R

1 R2 R6 = alkyl, alkylamino, acyl The above reactions were repeated by substituting NH2 Cl NH 2 N CO with N COOEt N COORt WO 2004/031161 PCT/AU2003/001303 - 104 CHART A4

-NH

2 N COOEt H 0 N M eO R N M eO R N M eO N X N R R RN'R N OH1- R; N ' CI OH OMe OMe OMe I H' MeO 0 N R N CN R N R' OMe O R alkyl R' NH,, SR 2

(R

2 = alkyl, aryi, heterocyclic), aryl, heterocyclic, MeO N NR R (R = H, alkyl; R = alkyl, - N heterocyclic, acyl), NH.CO(S).NH 2 , R N COOH SONHR 5 OMe Me OI 0 N R N R1 OH NR1 = NH 2 , SR 2

(R

2 = alkyl, aryl, OH NHR heterocyclic), aryl, heterocyclic, NR3R4 (R3 = H, alkyl; R4 = alkyl, heterocyclic, acyl), NH.CO(S).NH2, Me S0 2

NHR

5 0 N N RR R OH NHR The above reactions were repeated by substituting

NH

2 w C NH 2 OH NHRN with N COOEt N COOEt WO 2004/031161 PCT/AU2003/001303 - 105 CHART B1 C OC(O)Me N COOEt 0 0 0 0 0 0 0 0 ON N Cl NR OH OMe OMe OMe BI I I O 0 N0 0 0 0 N CN N R H R OMe OH R6 OH R = NH 2 , SR 2

(R

2 = alkyl, aryl, heterocyclic, aryl, heterocyclic, NR3R (R = H, alkyi; R =alkyl, N OOH heterocyclic, acyl), NH.CO(S).NH 2 , OMe SO 2

NHR

5 R = alkyl, alkylamino, acyl O 0 N VNY OH NHR O 0 NH N

R

6 OH NHR

R

6 = alkyl, alkylamino, acyl The above reactions were repeated by substituting OAc CI OAc NY with N COOEt N COOEt WO 2004/031161 PCT/AU2003/001303 - 106 CHART B2 OC(O)Me O0 0 Me N COOEt Me HN Me HN CH=NOR OH R OH1 R3 OH R H or alkyl

R

3 = alkyl, alkylamino, acyl O 00 0 N CH=NOR N CHO OH R = H or alkyl OH 00 O 0 N N OH N OH

NH

2 = H, alkyl; alkyl, aryl, acyl O 00 0 HN HN 1I 13V R OH NH 2 R OH N R R2

R

1 = H, alkyl;

R

2 = alkyl, aryl, acyl

R

3 = alkyl, alkylamino, acyl The above reactions were repeated by substituting OAc Ci OAc with Me N COOEt N COOEt WO 2004/031161 PCT/AU2003/001303 - 107 CHART Cl 0 0 0 HN NHN NHN O MeOOC* N N NHR OH OH OH C2 HN HN N N RON OH N 1 N,'2 OH OH Cl R = H, alkyl; R2 = alkyl, aryl, acyl 0 2NHN N Me N O NH 2 N OH N OH C3 WO 2004/031161 PCT/AU2003/001303 - 108 CHART C2 0 R~ R R N R 1 R* N R OH OH 2 alky R' = NH2, SR 2

(R

2 = alkyl, aryl, heterocyclic), aryl, heterocyclic,

NR

3

R

4

(R

3 = H, alkyl; R 4 = alkyl, heterocyclic, acyl), NH.CO(S).NH 2 ,

SO

2

NHR

5 O OMe OMe OMe R*N R* R R N R R' N C OH OMe me OMe 7-R-substituted derivatives from CHART CI 2 R 0 Me, N OH H OHO 0 0 0 MN NHN NM N R CHO O RRN CHO R COO R! N CONFIR 1 OH OH OH O 0 0 RN RN N 2 N COON CONHR 1 OH N R OH N, OR OH R 2 = alkyi R = H, alkyl; = H, alkyl alkyl R2 = aikyl, ary, acyl MN N RN N RN N O R N R N R N MeN OH NH 2 OH OH NH 2 N N 2O2 R alkyl R a lkyl R NN ONO O R<RR R2 WO 2004/031161 PCT/AU2003/001303 - 109 CHART D1 OH 0 S H 2 N Me ONHN M OH OHC OH0 N + rD _~N R:0 H ,N H alkyl R Me'N Me N OH OH N O 0 0 N -~ N N N. - N' "N 'X HOOC N -~ OHC N NHR OH N H NOHN I 0 0 N -~ N NN

NR

1 R' OH NO CC OHN Rl= H, alkyl; R =H, alkyl R2 = alkyl, aryl, acyl 0 -~ N N

NH

2

OH

WO 2004/031161 PCT/AU2003/001303 - 110 CHART D2 OH 0 +N

-

0 H 2 N .I-I N MeO HMeO N OR OH R 0 N N -' NN H2'N N CIN* HO N OH N.OH "OH N 0 0 10 N N -~ N HN N - R'N N N~N 2 I O H-N -1 RN O H O H" IS N (S) NH 2 R' = H-, aikyl;ROH N R= alkyl, aryl, R alkyl, aryl, heterocyclic heterocyclic 0 0 -~ N -~ N N ~ 0 R N 0~ Is" N' OH 'N HR OH N R= ary, heterocyclic WO 2004/031161 PCT/AU2003/001303 - 111 CHART D3 OH 0 + N - o H 2 NN O Z' X H 2N NR 1 N OH R' = halogen, alkyl, CN, NH 2 OH OH 0 N 0 + 'N ~N Me N H 2 N N N N OH RI = halogen, alkyl, CN, NO 2 OH = halogen, alkyl, CN, NO 2 N N N47 N / HOOC N NHR OH OH RI = halogen, amide, amino, R halogen, alkyl, CN, NO 2 substituted amino OH 0 "N 0 + "N 7 N MeO N R MeO N OH R= halogen, alkyl, CN, NO 2 OH N R halogen, alkyl, ON, NO 2 ON NN 0

R

2 N OH Q l N R' R= SR 3

(R

3 alkyl, aryl, OH N heterocyclic), aryl, heterocyclic, R 1 = halogen, alkyl, ON, NO 2

NR

4

R

5

(R

4 = H, alkyl; R 5 =alkyl, heteroaryl, acyl), NH.CO(S).NH 2 ,

SO

2

NHR

6 R' = halogen, amide, amino, substituted amino WO 2004/031161 PCT/AU2003/001303 - 112 CHART El R O R N' R' OH R R= NH 2 , SR 2

(R

2 = alkyl, aryl, heterocyclic, O NR 3

R

4

(R

3 = H, alkyl; R 4 = alkyl, acyl, N CI heterocyclic), NH.CO(S).NH 2 , SONHR 5 N N- Cl R R OBn O N CHO O

NH

2

N

OBn OBn R R = alkyl, substituted 0 N alkyl OY N CN OBn R O N S N R 1 OH R'= COOH, CONHR 3 R R 0 N N Me N R 1 OBn OH R = -CH=NOR, alkylamino, alkylamido WO 2004/031161 PCT/AU2003/001303 - 113 REFERENCES 1. Z.-L. Zhou, J.M. Navratil, S.X. Cai, E.R. Whittemore, S.A. Espitia, J.E. Hawkinson, M. Tran, R.M. Woodward, E. Weber and J.F.W. Keana, Bioorg. 5 Med. Chem., 2001, 9, 2061-2071. 2. M. Blanco, M.G. Lorenzo, I. Perillo and C.B. Schapira, J. Heterocyclic Chem., 1996, 33, 363-366. 10 3. J.D. Colye, J.F. Challiner, E.J. Haws and G.L. Newport, J Chem. Res., M, 1985, 3748-3747. 4. (a) A. Dondoni, G. Fantin, M. Fogagnolo, A. Medici and P. Pedrini, Synthesis, 1987, 998-1001. (b) A. Dondoni, F.L. Merchan, P. Merino, I. Rojo and T. 15 Tejero, Synthesis, 1996, 641-646. (6 PREPARATION OF 9-HYDROXY-PYRIDO[1,2-a]PYRIMIDIN-4-ONE 20 9-Hydroxypyrido[1,2-a]pyrimidin-4-one itself was synthesised according to a literature procedure (Dennin and coworkers, J. Heterocyclic Chem., 1991, 28, 1287). Derivatives of 9-hydroxypyrido[1,2-a]pyrimidin-4-one can be prepared using known methods (see for example: Yale and coworkers, US Patent 4,022,897). For example, condensation of commercially available 2-amino-3-hydroxypyridine with an 25 appropriate 2-substituted-3-oxo-propionic acid ethyl ester (for methods of preparation, see for example: Marabout and coworkers, Patent FR 2,765,582) produces an intermediate enamine which under dehydrating conditions yields the desired 3 substituted-9-hydroxypyrido[1,2-a]pyrimidin-4-one (Scheme 11). Further elaboration at the 8-position can be achieved using known reported methods (see for example: 30 Smirnov and coworkers, Chemistry of Heterocyclic Compounds, 1992, 12, 1425).

WO 2004/031161 PCT/AU2003/001303 - 114 Scheme 11 OH EtOOC R 1 tONOC R' R N HHO NH NY 5 OH OH 0 0 N R R R OH OH 10 Scheme 12 shows an alternate route to some functionalised 9 hydroxypyrido[1,2-a]pyrimidin-4-ones where an appropriately substituted 2-amino-3 hydroxypyridine is condensed with a 2-substituted-3-oxo-propionic acid ethyl ester. For example, 2-amino-3-chloro-3-hydroxypyridine (see Gudmundsson and coworkers, 15 Synthetic Communications, 1997, 27(5), 861, for preparation) and 2-(4-chlorophenyl) 3-oxo-propionic acid ethyl ester gives an 8-chloro derivative; the latter can be further elaborated using known methods. Scheme 12 20 C1 OH EtOOC EtOOC R + I N N NH2 HO C, NH OH 25 0 0 C N R N R C O N C H N OH OH WO 2004/031161 PCT/AU2003/001303 - 115 Q1 PREPARATION OF 8-HYDROXY-4(3H)-OUINAZOLINONE, 9 HYDROXYPYRIMIDO[1,6-alPYRIMIDIN-4-ONE AND 9 HYDROXYPYRIDO[1,2-a]PYRIMIDIN-4-ONE 5 General The following compounds/reagents were sourced commercially: amines: ethylamine, histamine, 2-(2-aminoethyl)pyridine, 2-(2-methylaminoethyl)pyridine; aldehydes: 4 imidazolecarboxaldehyde, 2-thiazolecarboxaldehyde and 2-pyridinecarboxaldehyde, 10 azoles: pyrazole, imidazole, methylimidazole and 1H-1,2,3-triazole, boronic acids: phenylboronic acid, 2-(trifluoromethyl)phenylboronic acid, 2-methoxyphenylboronic acid, o-tolylboronic acid, 2-fluorophenylboronic acid, 3-methoxyphenylboronic acid, 4 methoxyphenylboronic acid, n-tolylboronic acid, 3,5-difluorophenylboronic acid, 2,4 difluorophenylboronic acid, 3-thiopheneboronic acid, 3-fluorophenylboronic acid and 15 4-fluorophenylboronic acid; and organozinc reagents: 2-pyridylzinc bromide, 2 (methylthio)phenylzinc iodide, 2-(ethoxycarbonyl)phenylzinc iodide and 6 methylpyridylzinc bromide (0.5 M solution in THF) (Aldrich). 3-Pyridylboronic acid was purchased from Frontier Scientific. 2-Aminomethylthiazole was prepared according to the literature.' Solvents were analytical grade and used as supplied. THF 20 was distilled from sodium and benzophenone under argon. 1H NMR spectra (8, relative to TMS) were recorded on a Varian Unity 300 spectrometer unless otherwise indicated; J-Values are given in hertz. Mass spectral data were recorded on a Micromass Quattro II mass spectrometer. 25 The synthesis of derivatives of 3 classes of compounds: 8-hydroxy-4(3H) quinazolinone (A), 9-hydroxypyrimido[1,6-a]pyrimidin-4-one (B) and 9 hydroxypyrido[1,2-a]pyrimidin-4-one (C), is described. 0 0 0 4 4 N H / N N 2 8 N 2 N 2 OH OH OH A B C 30 The method of Iyer and coworkers was employed in the synthesis of compounds Al and A2. Further derivatisation of these compounds to provide a series of 8-hydroxy- WO 2004/031161 PCT/AU2003/001303 - 116 4

(

3 H)-quinazolinones employed routes shown in Charts Al - A2. Halogenation of the 5- and/or 7-positions in the 8-hydroxy-4(3H)-quinazolinone system followed methods previously described 3 in the literature for the reactions on 8-hydroxyquinolines. The synthesis of the parent system in classes B and C, compounds B1 and C1, employed the 5 procedure previously reported 4 by Dennin and coworkers. Further derivatisation of BI and C1 followed routes shown in Charts BI- B2 and Cl - C2, respectively. CHART A1 0 Me - DER N '_Me OH R=alkyl O 0

CONH

2

CH

3 CoCI, CON )MD pyridine DEHR N

H

2 NHC(O)Me O N Me * * N- Me OMe OMe OMe OH Al 0 0 0 DER CH=NOR N CHO N COOH OH OH OH 0 2 R - H, alkyl S NR2 N CH=NOR 0 0O OH N HN O H, alkyl

R

2 alkyl N" N N N OH NH 2 OH N(R 1

)R

2 OH NHR R' = H, alkyl;

R

2 =alkyl, acyl 0 UNDER I DER N DER N OH NH 2 * R = DER : -and derivatives with N'R3 R = alkyl halogen-substitution, preferably at 5- and/or 7-position(s) N indicated by asterisk OH N(Ri)R2 R1 = H, alkyl; R2 = alkyl, acyl;

R

3 = alkyl WO 2004/031161 PCT/AU2003/001303 - 117 CHARTA2 O 0 O OOH HCONH 2 NH - NH' NH

NH

2 N N * N OMe OMe OH OH A2 N RI N OMe OMe OH N NI R =alkylN C OMe OMe O 0 0 0 N NH / NH N R N R 1 N R N CN N CN OH OH OH OH R = alkyl R' = SR 2

(R

2 = alkyl, aryl, R alkyl heterocyclic), aryl, heterocyclic,

NR

3

R

4

(R

3 = H, alkyl; R 4 alkyll, O heterocyclic, acyl), NH.CO(S).NH 2 , R

SO

2

NHR

5 NH N N COOH N COOH OH OH R = alky O O NH NR * N5 O' N' _' O OH NHR4 OH NH4 * - and derivatives with halogen-substitution, R = alkyl preferably at the 5- and/or 7 position(s) indicated by asterisk 5 WO 2004/031161 PCT/AU2003/001303 - 118 CHARTBI O 0 NN N_______N N NO 2 NN NH 2 N N N N NN OH OH OH B1 O RO H 0 NN N R 2 N N N,R N N R N N N O OH OH OH Rl = alkyl; R = alkyl R2 = alkyl, alkylamino R = NH 2 , alkyl, aryl 5 CHART B2 0 0 0 NNN COOEt N"N CHO N CH=NOR NN N N OH OH OH B24 00 N NRIR N NH 2 N N OH OH Rl = H, alkyl; R2 = alkyl, aryl, alkylamino, acyl WO 2004/031161 PCT/AU2003/001303 - 119 CHART CI 0 00 N NO 2 N N N N OH OH , OH C1 O Ri 0 I H 0 7N N.

2 )7N N, R N N>yR N N NR OH OH OH

R

1 = alkyl; R alkyl, aryl R2 = alkyl, aryl, alkylamino R = NH 2 , alkyl, aryl 5 CHART C2 0 0 0 COOEt CHO CH=NOR OH OH OH C2I 0 0

NR

1

R

2 N NH 2 O OH OH COOH R = H, alkyl; N R 2 = alkyl, alkylamino, N acyl OH 0 / :N C(O).NHR

OH

WO 2004/031161 PCT/AU2003/001303 - 120 REFERENCES 1. (a) A. Dondoni, G. Fantin, M. Fogagnolo, A. Medici and P. Pedrini, Synthesis, 1987, 998-1001. (b) A. Dondoni, F.L. Merchan, P. Merino, I. Rojo and T. 5 Tejero, Synthesis, 1996, 641-646. 2. R.N. Iyer, N. Anand and M.L. Dhar, J Sci. Ind. Res. A, General, 1956, 15C, 1 7. 10 3. H. Gerson, M.W. McNeil, R. Parmegiani and R.K. Godfrey, J. Med. Chem., 1972, 15, 987-989, and references cited therein. 4. F. Dennin, D. Blondeau and H. Sliwa, J. Heterocyclic Chem., 1991, 28, 1287 1291. 15 General The following compounds/reagents were sourced commercially: 2,4 dichloroanisole, 2,4-dichlorobenzoic acid, 2,3-pyridine dicarboxylic acid, 6-methyl-2,3 20 pyridinedicarboxylic acid and 2-amino-3-hydroxypyridine (Aldrich). These compounds were prepared according to literature procedures: 5-chloro-8-hydroxy [1,6]naphthyridine-7-carboxylic acid methyl ester (PB 1045) from 5,8-dihydroxy [1,6]naphthyridine-7-carboxylic acid methyl ester (Albert and Hampton, J Chem. Soc., 1952, 4985; Blanco and coworkers, J. Heterocyclic Chein., 1996, 33, 361); 9 25 hydroxypyrido[1,2-a]pyrimidin-4-one (PB 1048) (Dennin and coworkers, J. Heterocyclic Chem., 1991, 28, 1287); 8-hydroxy-3H-quinazolin-4-one (PB 1049) (Iyer and Dhar, J. Sci. Ind. Res., 1956, 15C, 1). Solvents were analytical grade and used as supplied. THF was distilled from sodium and benzophenone under argon. 1H NMR spectra (6, relative to TMS) were recorded on a Varian Inova 400 spectrometer unless 30 otherwise indicated; J-Values are given in hertz; multiplicity: s = singlet, d = doublet, m = multiplet, sep = septet, t = triplet, q = quartet. Electrospray mass spectral data were recorded on a Micromass Quattro II mass spectrometer.

WO 2004/031161 PCT/AU2003/001303 - 121 Example 1 (A) 6,8-Dichloro-3-methylquinoxalin-5-o1 (1064) and 6,8-Dichloro-2 methylquinoxalin-5-ol (1065) 5 Step A: Preparation of 6,8-Dichloro-3-methylquinoxalin-5-ol and 6,8-Dichloro-2 methylquinoxalin-5-ol CI cl C1

NH

2 NN 10 C N 2 C N C N OMe OMe OMe CI ci C4N N N' N Nr 15 OH OH To a stirred suspension of 2,3-diamino-4,6-dichloroanisole (prepared from 2,4 dichloroanisole according to the conditions reported by Philips' Gloeilampenfabrieken, N.V. (1973), Patent GB 1303711) (1.48 g, 7.15 mmol), 2 M AcOH (14.2 mL) and 4 M 20 NaOAc solution (8.8 mL) at 60 0 C was added 40% aqueous pyruvic aldehyde solution (1.15 mL). The mixture was then stirred at the same temperature for a further 40 min. The precipitate was isolated via filtration and washed with H 2 0 (5 mL x 2). This solid (1.21 g) was then dissolved in ethyl acetate and the solution was filtered through a short plug of SiO 2 -gel (ethyl acetate/hexanes, 1:3). This provided a 1:1 mixture of 6,8 25 dichloro-5-methoxy-2-methylquinoxaline and 6,8-dichloro-5-methoxy-3 methylquinoxaline as a pale pink solid (1.13 g). To an ice-cooled solution of the mixture of the 2-methyl- and 3-methyl compounds (2.48 g, 0.01 mol) in dichloromethane (30 mL) was added BBr 3 (20.1 mL of a 1 M solution in dichloromethane). The cooling bath was removed after 1 h and the 30 mixture was left to stir at 30 "C for 16 h. MeOH was added and the solution was concentrated. The latter process was repeated four more times. Dichloromethane (20 mL) was added to the remaining residue and the pH of the mixture was adjusted to 7 (conc NH 4 0H). The organic layer was dried, concentrated, and filtered through a short plug of SiO 2 -gel (dichloromethane/MeOH, 19:1) to give a 1:1 mixture of 6,8-dichloro 35 2-methylquinoxalin-5-ol and 6,8-dichloro-3-methylquinoxalin-5-o as a light yellow solid (1.51 g). A sample of this mixture was extracted with diethyl ether (3x). The ethereal extracts were concentrated and the residue recrystallised from isopropanol to WO 2004/031161 PCT/AU2003/001303 - 122 yield 6,8-dichloro-2-methyl-quinoxalin-5-o1 as yellow needles. The remaining solid after extraction with diethyl ether was washed with MeOH to give 6,8-dichloro-3 methyl-quinoxalin-5-ol as a pale pink solid. 6,8-Dichloro-3-methylquinoxalin-5-ol: 'H NMR (CDCl 3 ): 5 8.88 (s, 1 H), 8.01 5 (br, 1 H), 7.79 (s, 1 H), 2.83 (s, 3 H). 6,8-Dichloro-2-methylquinoxalin-5-ol: 1 H NMR (CDCl3): 6 8.68 (s, 1 H), 7.95 (br, 1 H), 7.83 (s, 1 H), 2.87 (s, 3 H). Step B: Preparation of 8-Benzyloxy-5,7-dichloro-2-nethylquinoxaline 10 C1 C1 Cj N N N ' OH OBn 15 A mixture of 6,8-dichloro-3-methylquinoxalin-5-o1 (150 mg, 0.655 mmol), benzyl bromide (0.20 mL, 1.68 mmol), KOH (0.09 g) and EtOH (10 mL) was heated under reflux for 16 h, cooled, and concentrated. Dichloromethane (20 mL) was added and the mixture washed with H20 (5 mL x 2), dried and concentrated. The residue was purified via SiO 2 -gel chromatography (dichloromethane/MeOH, 1:0 - 150:1) to give 8 20 benzyloxy-5,7-dichloro-2-methylquinoxaline (98 mg, 47%). - 1H NMR (CDCl 3 ): 6 8.82 (s, 1 H), 7.81 (s, 1 H), 7.58 (in, 2 H), 7.38 (in, 2 H), 5.45 (s, 2 H), 2.85 (s, 3 H). Step C: Preparation of 8-Benzyloxy-5,7-dichloroquinoxaline-2-carboxaldehyde Cl C) 25 C# N N Ci N Cl N CHO OBn OBn A suspension of the 2-methyl product from Step B (93 mg, 0.291 mmol) and 30 SeO 2 (100 mg) in dioxane (4 mL) and H20 (0.4 mL) was heated under reflux for 3 h. The mixture was cooled, filtered (celite), and concentrated. Dichloromethane was added and the insoluble material was filtered off. The solvent was evaporated and the residue purified by SiO 2 -gel chromatography (dichloromethane) to yield the desired aldehyde as an off-white solid (57 mg, 59%). 35 8-Benzyloxy-5,7-dichloroquinoxaline-2-carboxaldehyde: 'H NMR (CDCl 3 ): 6 10.31 (s, 1 H), 9.48 (s, 1 H), 8.06 (s, 1 H), 7.58 - 7.35 (in, 5 H), 5.55 (s, 2 H).

WO 2004/031161 PCT/AU2003/001303 - 123 (B) 6,8-Dichloro-3-(dimethylaminomethyl)quinoxalin-5-ol hydrochloride (1066) N N N C CHO Cl C HCI 5 OBn NMe2 OH NMe2 Sodium triacetoxyborohydride (50 mg, 0.236 mmol) was added to a stirred solution of the aldehyde from Step C (57 mg, 0.171 mmol), dimethylamine hydrochloride (15.1 mg, 0.185 mmol) and Et 3 N (0.023 mL) in 1,2-dichloroethane (2 10 mL). After 1 h, dichloromethane (20 mL) was added and the resulting solution was washed with sat'd NaHCO 3 solution (5 mL x 2), dried, and concentrated. The residue was purified via Si0 2 -gel chromatography (dichloromethane/MeOH, 1:0 - 75:1) to yield the desired (8-benzyloxy-5,7-dichloro-quinoxalin-2-ylmethyl)dimethylamine as a pale yellow solid (38 mg, 61%). - 'H NMR (CDC1 3 ): 6 9.11 (s, 1 H), 7.84 (s, 1 H), 7.58 15 -7.35 (in, 5 H), 5.47 (s, 2 H). A solution of (8-benzyloxy-5,7-dichloro-quinoxalin-2-ylmethyl)dimethylamine (38 mg, 0.105 mmol) in conc HCl (2 mL) was stirred at room temperature for 16 h. The orange solution was concentrated to dryness and the remaining residue was washed with diethyl ether (2 ml x 3) to give the desired product as a light yellow solid (27 mg, 20 83%). 6,8-Dichloro-3-(dimethylaminomethyl)quinoxalin-5-ol hydrochloride: 'H NMR (DMSO-d 6 ): a 8.75 (s, 1 H), 7.78 (s, 1 H), 4.40 (br, 2 H), 2.13 (s, 6 H); mass spectrum: m/z 272, 274, 276 (M + 1, 100%, 66%, 11%). 25 Example 2 (A) 5,7-Dichloro-3-(4-fluorophenyl)-8-hydroxy-3H-quinazolin-4-one (1055) Step A: Preparation of 4,6-Dichloro-3-hydroxy-2-nitrobenzoic acid 30 cl Ci ci Ci ci COOH COOH 3 COOH COOH COOH Ci CI CI Cl NO, Cl NO, NO, NH, NHAc NHAc OH 35 Tin (II) chloride hydrate (50 g, 0.29 mol) was added to a solution of 2,4 dichloro-5-nitrobenzoic acid (prepared according to a reported procedure (Golstein, H. and Schaaf, E., Helv. Chim. Acta, 1957, 5 7(23), 132)) (10.0 g, 0.045 mol) in EtOH WO 2004/031161 PCT/AU2003/001303 - 124 (200 mL). The mixture was stirred at 70 "C for 0.5 h, cooled and poured onto ice. The pH of the mixture was adjusted to 8 (sat'd NaHCO 3 ). The suspension was left to stir at room temperature for 5 h and re-acidified to pH 5 (glacial HOAc). The resulting white suspension was continuously extracted with ethyl acetate, the extracts combined, 5 washed with brine, dried and concentrated to yield the desired amine as an off-white solid (8.8 g, 96%). 5-Amino-2,4-dichlorobenzoic acid:'H NMR (CD 3 0D): 6 7.30 (s, 1 H), 7.27 (s, 1 H). Acetic anhydride (27 mL) was added to 5-amino-2,4-dichlorobenzoic acid (8.0 10 g, 0.041 mol) in glacial HOAc (150 mL). The solution was stirred at room temperature for 0.5 h and concentrated to yield the desired acetamide as a white solid (9.6 g, 96%). 5-Acetamido-2,4-dichlorobenzoic acid: 'H NMR (CD 3 0D): a 8.32 (s, 1 H), 7.62 (s , 1 H), 2.19 (s, 3 H). 5-Acetamido-2,4-dichlorobenzoic acid (9.6 g, 0.039 mol) was added in small 15 portions over 30 min to a stirred ice-cooled solution of fuming nitric acid (1.8 mL, 0.043 mol) and conc sulfuric acid (120 mL). After the addition was complete, more fuming nitric acid (17 mL) and conc sulfuric acid (80 mL) were added at 30 min and 60 min intervals. The reaction mixture was then left to stir for an additional 2.5 h at 0 *C, allowed to warm to 12 -16 0 C and left to stir at this temperature until all starting . 20 material was consumed (about 3 h). The solution was poured onto ice and extracted with ethyl acetate (3x). The organic extracts were combined, washed with brine, dried, and concentrated to give 3-acetamido-4,6-dichloro-2-nitrobenzoic acid as an orange solid (9.8 g, 86%). 3-Acetamido-4,6-dichloro-2-nitrobenzoic acid: 'H NMR (CD 3 0D): 3 8.01 (s, 1 25 H), 2.13 (s, 3 H). 3-Acetamido-4,6-dichloro-2-nitrobenzoic acid (9.7 g, 0.033 mol) was added to a solution of KOH (18.7 g, 0.034 mol) in H 2 0 (85 mL). The solution was heated under reflux for 18 h and cool to room temperature. Conc HCl was added to adjust the pH to 0. The mixture was diluted with ethyl acetate and H 2 0 and left to stir at room 30 temperature for 30 min. The layers were separated; the aqueous layer was extracted with ethyl acetate (3x), the extracts combined with the original organic layer, washed with brine, dried and concentrated to yield 4,6-dichloro-3-hydroxy-2-nitrobenzoic acid as a dark red solid (7.4 g, 89%); m.p. 188-189 "C (lit. m.p. (Linderberg, M., Hellberg, S., Bjork, S. et al, Eur. J Med. Chem., 1999, 34, 729-744): 186 C (dec)). 35 4,6-Dichloro-3-hydroxy-2-nitrobenzoic acid: 'H NMR (CD 3 0D): 8 7.79 (s, 1 H); mass spectrum: m/z 250, 252, 254 (M+ - 1, 100%, 66%, 11%).

WO 2004/031161 PCT/AU2003/001303 - 125 Step B: Preparation of 2-Anino-4,6-dichloro-N-(4-fluorophenyl)-3-hydroxybenzamide c5 cc 0 c 0 F I' COOH N N IN- HH OH OH OH CDI (2.2 g, 0.0 13 mol) was added to a stirred solution of 4,6-dichloro-3 hydroxy-2-nitrobenzoic acid (3.0 g, 0.012 mol) in anhydrous THF (30 mL) and DMF (2 mL). After 40 min, 4-fluoroaniline (4 mL, 0.042 mol) was added. The solution was 10 heated under reflux overnight, cooled, and concentrated to a dark brown viscous oil. The oil was purified via SiO 2 -gel chromatography (dichloromethane/methanol, 19:1 9:1) to yield 4,6-dichloro-N-(4-fluorophenyl)-3-hydroxy-2-nitrobenzamide as a light brown oil (2.7 g, 65%). 4,6-Dichloro-N-(4-fluorophenyl)-3-hydroxy-2-nitrobenzamide: 'H NMR 15 (CD 3 0D): 6 7.58 (m, 2 H), 7.44 (s, 1 H), 7.07 (m, 2 H). A suspension of tin (II) chloride hydrate (8.8 g, 0.039 mol), 4,6-dichloro-N-(4 fluorophenyl)-3-hydroxy-2-nitrobenzamide (2.7 g, 0.008 mol) and EtOH (70 mL) was heated at 70 C for 1 h and left to cool to room temperature. The mixture was poured onto ice, basified to pH 8 (sat'd NaHCO 3 ), and stirred at room temperature for 1.5 h 20 before re-acidifying to pH 6.5 with glacial HOAc. The suspension was filtered; the filtrate was concentrated to give a beige solid. The beige solid was combined with the filter cake and extracted with hot ethyl acetate (5x). The extracts were combined to provide 2-amino-4,6-dichloro-N-(4-fluorophenyl)-3-hydroxybenzamide as a solid (2.0 g, 79%). 25 2-Amino-4,6-dichloro-N-(4-fluorophenyl)-3-hydroxybenzanide: 1H NMR

(CD

3 0D): 6 7.69 (in, 2 H), 7.09 (in, 2 H), 6.73 (s, 1H). Step C: Preparation of 5,7-Dichloro-3-(4-fluorophenyl)-8-hydroxy-3H-quinazolin-4 one (1055) 30 o N F Cl N F H CI NH 2 ci- N OH OH A solution of CDI (0.65 g, 4.0 mmol) and formic acid (0.17 mL, 3.6 mmol ) in 35 anhydrous THF (18 mL) and DMF (2 mL) was stirred at room temperature for 4 h. A solution of the amine from Step B (1.01 g, 3.2 mmol) in THF (15 mL) was then added, the solution heated under reflux for 15 h, and concentrated. The resulting brown solid WO 2004/031161 PCT/AU2003/001303 - 126 was washed with ethyl acetate to yield the title compound as white needles (0.49 g, 47%). 5,7-Dichloro-3-(4-fluorophenyl)-8-hydroxy-3H-quinazolin-4-one: 1H NMR (CDCl 3

/CD

3 0D, 9:1): 5 8.10 (br, 1 H), 7.50 (s, 1 H), 7.36 (m, 2 H), 7.21 (m, 2 H); mass 5 spectrum: n/z 325, 327, 329 (Mi + 1, 100%, 66%, 11%). (B) 5,7-Dichloro-3-cyclopropyl-8-hydroxy-3H-quinazolin-4-one (1061) ci CI 0 cI 0 ci 0 10 H c Hc CI NO, CI_ No" Ci- NH , CI N3 OH OH OH OH The following compounds were prepared using the methods described in Example 3: thus, replacing 4-fluoroaniline with cyclopropylamine in Step B (reaction 15 was performed in a sealed tube at 70 "C) gave, after SiO2-gel chromatography (dichloromethane/MeOH, 9:1), 4,6-dichloro-N-cyclopropyl-3-hydroxy-2 nitrobenzamide (43%).- 1 H NMR (CD 3 0D): 6 7.37 (s, 1 H), 2.73 (m, 1 H), 0.74 (m, 2 H), 0.59 (m, 2 H); mass spectrum: n/z 289, 291, 293 (M+ - 1, 100%, 66%, 11%). Reduction of 4,6-dichloro-N-cyclopropyl-3-hydroxy-2-nitrobenzamide with tin 20 (II) chloride hydrate using the conditions as described in Step B yielded 2-amino-4,6 dichloro-N-cyclopropyl-3-hydroxybenzamide (93%). - 'H NMR (CD 3 0D): 8 6.67 (s, 1 H), 2.86 (m, 1 H), 0.79 (m, 2 H), 0.62 (m, 2 H). CDI-mediated coupling of formic acid with 2-amino-4,6-dichloro-N cyclopropyl-3-hydroxybenzainide (as described in Step C) gave, after washing with 25 ethyl acetate and MeOH, 5,7-dichloro-3-cyclopropyl-8-hydroxy-3H-quinazolin-4-one as a white solid (40%). - 1 H NMR (CDCl 3

/CD

3 0D, 9:1): 6 8.30 (s, 1 H), 7.46 (s, 1 H), 3.22 (m, 1 H), 1.21 (m, 2 H), 0.94 (m, 2 H); mass spectrum: m/z 271, 273, 275 (M* + 1, 100%, 66%, 11%). 30 (C) 5,7-Dichloro-8-hydroxy-3H-quinazolin-4-one (1067) cI CI 0 CI 0 cI 0 cl COOH C NH2 : cl NH2 3 C NH IN O, cI N O, CI q N H, C1 N3 OH OH OH OH 35 A solution of 2,4-dichloro-5-hydroxy-6-nitrobenzoic acid (Step A) (4.40 g, 17.6 mmol) and thionyl chloride (2 mL, 27 mmol) in toluene (60 mL) was heated under WO 2004/031161 PCT/AU2003/001303 - 127 reflux for 1 h and allowed to cool to room temperature. The solution was added to conc NH4 0 H (40 mL) at 0 0 C. The cooling bath was removed and the mixture stirred at room temperature for 2 h and then concentrated to give crude 4,6-dichloro-3-hydroxy-2 nitrobenzamide as a brown solid. - 'H NMR (CD 3 0D): 5 7.33 (s, 1 H); mass spectrum: 5 in/z 249, 251, 253 (M+ - 1, 100%, 66%, 11%). Reduction of 4,6-dichloro-3-hydroxy-2-nitrobenzamide with tin (II) chloride hydrate according to the conditions as described in Step B gave 2-amino-4,6-dichloro 3-hydroxybenzamide as a beige solid (2.5 g, 62% (2 steps)). - 'H NMR (CD 3 0D): 5 6.69 (s, 1 H). 10 According to the procedure described in Step C, 2-amino-4,6-dichloro-3 hydroxybenzamide and formic acid in the presence of CDI gave, after washing the crude product with MeOH, 5,7-dichloro-8-hydroxy-3H-quinazolin-4-one as a white solid (36%). - 'H NMR (DMSO-d 6 ): 6 8.12 (s, 1 H), 7.55 (s, 1 H); mass spectrum: m/z 229, 231, 233 (M* - 1, 100%, 66%, 11%). 15 Example 3 (A) 7-Morpholin-4-yl-[1,6]naphthyridin-8-ol (1053) 20 Step A: Preparation of 7-Anino-8-isopropoxy-[1, 6]naphthyridine N N NN' N N ' N MeOOC MeOOc N H2N N OH O 0 25 2-Bromopropane (0.9 mL) was added to a stirred mixture of 8-hydroxy [1,6]naphthyridine-7-carboxylic acid methyl ester (prepared according to Anthony and coworkers, Patent WO 02/30931 A2) (1.00 g, 4.90 mmol), K 2 C0 3 (2.79 g) and DMSO (15 mL). After 40 h at 50 "C, sat'd NH 4 C1 (20 mL) was added and the mixture extracted 30 with dichloromethane (10 mL x 3). The extracts were combined and concentrated. Diethyl ether (40 mL) was added to the residue and the resulting mixture washed successively with H 2 0 (5 mL x 2) and brine (10 mL), and dried. Solvent removal gave 8-isopropoxy-[1,6]naphthyridine-7-carboxylic acid methyl ester (1.06 g, 88%). - 'H NMR (CDCl 3 ): 6 9.16 (dd, J=1.9 and 4.3, 1 H), 9.03 (s, 1 H), 8.34 (dd, J=1.9 and 8.5, 1 35 H), 7.63 (dd, J=4.3 and 8.5, 1 H), 5.33 (sep, J=6.2, 1 H), 4.04 (s, 3 H), 1.42 (d, J=6.2, 6

H).

WO 2004/031161 PCT/AU2003/001303 - 128 A solution of 8-isopropoxy-[1,6]naphthyridine-7-carboxylic acid methyl ester (1.06 g, 4.30 mmol) in MeOH (10 mL) was added dropwise over 5 min into a stirred solution of hydrazine hydrate (2.5 mL), left to stir for a further hour, and concentrated. This provided the desired crude 7-acyl hydrazide as a golden syrup (1.06 g). 5 8-Isopropoxy-[ 1,6]naphthyridine-7-carboxylic acid hydrazide: 'H NMR (CDCl 3 ): 6 9.16 (dd, J=1.8 and 4.0, 1 H), 9.08 (br, 1 H), 9.07 (s, 1 H), 8.35 (dd, J=1.8 and 8.2, 1 H), 7.64 (dd, J=4.0 and 8.2, 1 H), 6.02 (br, 1 H), 5.39 (sep, J=6.2, 1 H), 4.10 (br, 2 H), 1.45 (d, J=6.2, 6 H). To a stirring ice-cooled solution of the 7-acyl hydrazide (1.06 g, 4.30 mmol) in 10 1 M HCl (6 mL) was added a solution of sodium nitrite (334 rug, 4.84 mmol) in H 2 0 (1.5 mL) dropwise at such a rate that the temperature did not rise above 5 0 C. The mixture was left to stir at 0 0 C for a further 20 min and then concentrated in vacuo. HOAc (5 mL) and H20 (5 mL) were added and the resulting orange solution heated at 95 C for 24 h, cooled, and concentrated. H 2 0 (10 mL) was added, the pH of the 15 mixture was adjusted to 8 (conc NH 4 0H), extracted with dichloromethane (10 mL x 3), and dried. Solvent removal gave the 7-amine as an orange solid (467 mg, 53%). 7-Amino-8-isopropoxy-[1,6]naphthyridine: 1H NMR (CDC1 3 ): 3 8.88 (dd, J=1.5 and 4.0, 1 H), 8.62 (s, 1 H), 8.07 (dd, J=1.5 and 8.2, 1 H), 7.15 (dd, J=4.0 and 8.2, 1 H), 5.16 (sep, J=6.2, 1 H), 4.83 (br, 2 H), 1.39 (d, J==6.2, 6 H); mass spectrum: n/z 204 (M* 20 +1,100%). Step B: Preparation of 7-Chloro-8-isopropoxy-{1, 6]naphthyridine 25 HN NC 0 0 Sodium nitrite (1.8 g, 0.026 mol) was added portionwise into a stirred solution of the amine (1.00 g, 4.92 mmol) in conc HCl (20 mL) at 0 - 5 "C. The mixture was left 30 to stir at this temperature for a further h and then allowed to warm to room temperature over 1 h. Ice was added and the pH of the mixture was adjusted to 8 (conc NH 4 0H). The mixture was then extracted with dichloromethane, the extracts washed with brine, dried and concentrated. Purification on Si0 2 -gel chromatography (dichloromethane/MeOH, 200:3) provided the desired 7-chloride as a colourless solid 35 (400 mg, 36%). 7-Chloro-8-isopropoxy-[1,6]naphthyridine: 1 H NMR (CDC1 3 ): 8 9.04 (dd, J=1.8 and 4.2, 1 H), 8.76 (s, 1 H), 8.23 (dd, J=1.8 and 8.4, 1 H), 7.46 (dd, J=4.2 and 8.4, 1 H), WO 2004/031161 PCT/AU2003/001303 - 129 5.14 (sep, J=6.2, 1 H), 1.38 (d, J=6.2, 6 H); mass spectrum: m/z 223, 225 (M* + 1, 100%, 33%). Step C: Preparation of 7-Morpholin-4-yl-[1,6]naplithyridin-8-ol (1053) 5 cN N N 0 0 0 01-1j 10 A mixture of the 7-chloride from Step B (100 mg, 0.448 mmol) and morpholine (2 mL) was heated at 180 C for 20 h and then left to cool to room temperature. The reaction mixture was extracted with MeOH and the extracts concentrated. The residue was purified using Si02-gel chromatography (dichloromethane/MeOH, 20:1) to yield the desired product as a yellow solid (78 mg, 64%). 15 To a stirred solution of 8-isopropoxy-7-morpholin-4-yl-[1,6]naphthyridine (149 mg, 0.545 mmol) in dichloromethane (10 mL) at 0 0 C was added BC1 3 (1.5 mL of a 1 M solution in dichloromethane). The dark orange solution was left to stir at room temperature for 24 h. MeOH (10 mL) was added and the mixture concentrated. This process was repeated four times. Further washing of the remaining residue with diethyl 20 ether (5 mL x 2) provided 7-morpholin-4-yl-[1,6]naphthyridin-8-ol hydrochloride as a dark red solid (quantitative yield). - 1 H NMR (CD 3 OD): 6 9.09 (dd, J=1.5 and 5.1, 1 H), 9.02 (s, 1 H), 8.99 (dd, J=1.5 and 8.2, 1 H), 7.79 (dd, J=5.1 and 8.2, 1 H), 3.95 (m, 4 H), 3.80 (m, 4 H). A solution of the 7-morpholin-4-yl-[1,6]naphthyridin-8-ol hydrochloride in 25 dichloromethane (20 mL) was washed with sat'd NaHCO 3 solution (2 x), dried, and concentrated. Solvent removal gave, after SiO 2 -gel chromatography (dichloromethane/MeOH, 9:1), 7-morpholin-4-yl-[1,6]naphthyridin-8-ol as an orange solid (97 mg, 77%). - 1H NMR (CD 3 0D): 8 8.91 (dd, J=1.4 and 4.0, 1 H), 8.60 (s, 1 H), 8.32 (dd, J=1.4 and 8.2, 1 H), 7.41 (dd, J=4.0 and 8.2, 1 H), 3.89 (m, 4 H), 3.47 (m, 4 30 H).

WO 2004/031161 PCT/AU2003/001303 - 130 (B) 8-Hydroxy-2-methylaminomethyl-[1,6]naphthyridine-7-carboxylic acid (4 fluorophenyl)amide (1070) Step A: Preparation of 6-Methylpyridine-2,3-dicarboxylic acid 2-isopropyl ester 5 COOH COOH N COOH N 0 6-Methyl-2,3-pyridinedicarboxylic acid (10.0 g, 0.055 mol) and Ac 2 0 (50 mL) 10 were heated at 120 C for 4 h, cooled, and concentrated to a brown oil. Isopropanol was added to the brown oil and the solution heated at 80 "C overnight. The volatiles were removed in vacuo and the residue gave, after washing with diethyl ether, 6 methylpyridine-2,3-dicarboxylic acid 2-isopropyl ester as a straw-coloured solid (8.8 g, 71%). - 1H NMR (CDCl 3 ): 6 8.24 (d, J=8.2, 1 H), 7.34 (d, J=8.2, 1 H), 5.34 (sep, J=6.2, 15 1 H), 2.68 (s, 3 H), 1.39 (d, J=6.2, 6 H). Step B: Preparation of 3-Hydroxynethyl-6-methylpyridine-2-carboxylic acid isopropyl ester COH 20 K KN OH 20N 0 OT _ N 0 0 r 2 0 A mixture of 6-methylpyridine-2,3-dicarboxylic acid 2-isopropyl ester (7.31 g, 0.33 mol) and thionyl chloride (65 mL) was heated under reflux for 1 h. The 25 resulting solution was concentrated in vacuo. The residue was washed with anhydrous THF (2x) to give the crude acid chloride as a brown oil. To a stirred ice-cooled solution of the acid chloride in THF (50 mL) was added sodium borohydride (1.26 g, 0.33 mol) in THF (15 mL) dropwise over 20 min. After 1 h at 0 C, the reaction mixture was poured onto ice and extracted with dichloromethane (3x). The extracts were combined, 30 washed with brine, dried and concentrated. The residue gave, after purification via SiO 2 -gel chromatography (dichloromethane/MeOH, 19:1), 3-hydroxymethyl-6 methylpyridine-2-carboxylic acid isopropyl ester as a light brown solid (3.42 g, 48%). 'H NMR (CDCl 3 ): 6 7.76 (d, J=7.9, 1 H), 7.31 (d, J=7.9, 1 H), 5.32 (sep, J=6.2, 1 H), 4.74 (s, 2 H), 1.45 (d, J=6.2, 6 H). 35 WO 2004/031161 PCT/AU2003/001303 - 131 Step C: Preparation of3-[(Benzenesulfonyl(methoxycarbonylmethyl)amino)nethyl]-6 inethylpyridine-2-carboxylic acid isopropyl ester

SO

2 Ph OH N COOMe 5 N-O N/ O 0 01r0 According to the conditions reported by Anthony et al (WO 02/30931 A2) for the preparation of 3-[(benzenesulfonyl(methoxycarbonylmethyl)amino)methyl] 10 pyridine-2-carboxylic acid isopropyl ester, a solution of diisopropyl azodicarboxylate (1.07 g, 5.30 mmol) in THF (5 mL) was added dropwise over 40 min to a stirred solution of 3-hydroxymethyl-6-methyl-pyridine-2-carboxylic acid isopropyl ester (739 mg, 3.53 mmol), N-(phenylsulphonyl)glycine methyl ester (810 mg, 3.53 mmol) and triphenylphosphine (1.39 g, 5.30 mol) in THF (15 mL) The resulting solution was then 15 left to stir at room temperature overnight. The reaction mixture was concentrated to give crude 3-[(benzenesulfonyl(methoxycarbonylmethyl)amino)methyl]-6 methylpyridine-2-carboxylic acid isopropyl ester as a golden syrup. This was used in the subsequent step without further purification. - Selected 1 H NMR (CDCl 3 ): 6 4.73 (s, 2 H), 4.00 (s, 2 H), 3.54 (s, 3 H), 2.70 (s, 3 H). 20 Step D: Preparation of 8-Hydroxy-2-methyl-[1, 6]naphthyridine- 7-carboxylic acid methyl ester SOPh N COOMe N 25 N MeOOC N O OH To a stirred solution of crude 3 [(benzenesulfonyl(methoxycarbonylmethyl)amino)methyl]-6-methylpyridine-2 carboxylic acid isopropyl ester (from Step C) in MeOH (20 mL) at 0 C was added a 30 solution of NaOMe in MeOH (prepared from Na (200 mg) in MeOH (4 mL)) dropwise over 20 min. The resulting solution was left to stir at 0 C for a further hour and concentrated. Ethyl acetate (50 mL) and ice-H 2 0 (50 mL) were added. The layers were separated; the aqueous layer was extracted with ethyl acetate (2x), the pH adjusted to 6 (5 M HCl) and extracted with dichloromethane (20 mL x 3). The extracts were 35 combined, dried, and concentrated to give pure 8-hydroxy-2-methyl-[1,6]naphthyridine 7-carboxylic acid methyl ester as a cream solid (708 mg, 98% (2 steps)). - 1H NMR WO 2004/031161 PCT/AU2003/001303 - 132 (CDC1 3 ): 6 8.87 (s, I H), 8.26 (d, J=8.5, 1 H), 7.62 (d, J=8.5, I H), 5.40 (br, 1 H), 4.13 (s, 3 H), 2.91 (s, 3 H). Step E: Preparation of 8-Hydroxy-2-nethyl-[, 6]naphthyridine-7-carboxylic acid (4 5 fluorophenyl)amide hydrochloride MeOOC N N OH 0 OH 10 A solution of 8-hydroxy-2-methyl-[1,6]naphthyridine-7-carboxylic acid methyl ester (0.34 g, 1.56 nmol) and 4-fluorobenzylamine (0.5 mL) in toluene (4 mL) was heated under reflux for 16 h and allowed to cool to room temperature. Subsequent solvent removal gave a cream solid which, after washing with diethyl ether, gave 8 hydroxy-2-methyl-[1,6]naphthyridine-7-carboxylic acid (4-fluorophenyl) amide, as an 15 off-white solid (0.40 g, 87%). - 1H NMR (CDCl 3 ): 6 8.57 (s, 1 H), 8.44 (br, 1 H), 8.15 (d, J=8.4, 1 H), 7.52 (d, J=8.4, 1 H), 7.37 (in, 2 H), 7.07 (in, 2 H), 4.67 (d, J=6.2, 2 H), 2.87 (s, 3 H). Step F: Preparation of 8-Hydroxy-2-(mnethylamino)methyl-[l, 6]naphthyridine- 7 20 carboxylic acid (4-fluorophenyl)amide hydrochloride (1070) H- -H I- - H N F CHO N 0 OH 0 OH 0 OH HN, .HCI 25 Se0 2 (100 mg, 0.901 mmol) was added to a solution of 8-hydroxy-2-methyl [1,6]naphthyridine-7-carboxylic acid (4-fluorophenyl)amide (150 mg, 0.482 mnol) in 1,4-dioxane (8 mL) and H 2 0 (0.1 mL) and the mixture was heated at 55 C for 3 h. The mixture was cooled to room temperature and filtered (celite). The filtrate was 30 concentrated to give crude 2-aldehyde as a pale orange solid. - 1H NMR (CDCl 3 ): 5 10.37 (s, 1 H), 8.75 (s, 1 H), 8.46 (d, J=8.4, 1 H), 8.45 (br, 1 H), 8.26 (d, J=8.6, 1 H), 7.40 (in, 2 H), 7.30 (br, 1 H), 7.08 (in, 2 H), 4.70 (d, J=6.0, 2 H). To a stirred solution of the crude 2-aldehyde and methylamine hydrochloride (50 mg, 0.741 mmol) in 1,2-dichloroethane(5 mL) was added Et 3 N (0.07 mL). Sodium 35 triacetoxyborohydride (102 mg, 0.482 mmol) was then added and the mixture left to stir at room temperature overnight. MeOH was added and the resulting solution was concentrated. Saturated NaHCO 3 solution was added and the resulting pale orange solid WO 2004/031161 PCT/AU2003/001303 - 133 was isolated via filtration, washed with H 2 0 and dried under vacuum. MeOH (10 mL) was added to the solid and the remaining insoluble material was filtered off. The filtrate was concentrated; conc HC1 was added and the solution evaporated to dryness which afforded 8-hydroxy-2-(methylamino)methyl-[1,6]naphthyridine-7-carboxylic acid (4 5 fluorophenyl)amide hydrochloride (PB 1070) as a pale yellow solid (45 mg, 30%). - IH NMR (CD 3 0D): 5 8.92 (s, 1 H), 8.64 (d, J=8.4, 1 H), 7.82 (d, J=8.4, 1 H), 7.43 (m, 2 H), 7.06 (in, 2 H), 4.69 (s, 2 H), 4.68 (br, 1 H), 4.65 (s, 2 H), 2.91 (s, 3 H). Example 4 10 (A) 3-(4-chlorophenyl)-9-hydroxypyrido[1,2-alpyrimidin-4-one (1069) Step A: Preparation of 2-(4-Chlorophenyl)-3-oxo-propanoic acid ethyl ester 15 CI c)I 5C COOH c COOEt C COOEt OH A solution of (4-chlorophenyl)acetic acid (20.0 g, 0.12 mol) and conc sulfuric 20 acid (2 g) in EtOH (150 mL) was heated under reflux for 19 h, cooled, and then concentrated. Diethyl ether (100 mL) was added and the solution was washed with sat'd NaHCO 3 solution (40 nL x 2), dried, and the volatiles removed. This provided (4 chlorophenyl)acetic acid ethyl ester as an oil (22.3 g, 96%). Ethyl formate (7.46 g, 0.101 mol) was added dropwise to a stirred ice-cooled 25 suspension of (4-chlorophenyl)acetic acid ethyl ester (20.0 g, 0.101 mol) and NaH (0.131 mol) in diethyl ether (100 mL). The mixture was then allowed to warm to room temperature overnight. The mixture neutralised (1 M HCl); the organic layer was isolated, dried, and the solvent removed to give 2-(4-chlorophenyl)-3-oxo-propanoic acid ethyl ester as a colourless solid (19.8 g, 87%). 1 H NMR (CDCl 3 ): 8 12.14 (d, 30 J=12.8, 1 H), 7.31 - 7.18 (m, 4 H), 4.29 (q, J=7.2, 2 H), 1.57 (br, 1 H), 1.29 (t, J=7.2, 3

H).

WO 2004/031161 PCT/AU2003/001303 - 134 Step B: Preparation of 3-(4-Chlorophenyl)-9-hydroxypyrido[1,2-a]pyrimidin-4-one (1069) OH CIEtOOC / l N NH 2 5 2 c COOEt c cl" OH OH OH 2- Amino-3-hydroxypyridine (5.51 g, 0.05 mol) and 2-(4-chlorophenyl)-3-oxo propanoic acid ethyl ester (13.6 g, 0.06 mol) in EtOH (100 mL) were heated under 10 reflux for 15 h and then allowed to cool. The precipitate was isolated via filtration to yield a 9:1 mixture of the enamine and the cyclized product (11.1 g). Enamine: 'H NMR (DMSO-d 6 ): 5 10.67 (d, J=12.4, 1 H), 8.10 (d, J=12.4, I H), 7.72 (d, J=4.8, 1 H), 7.37 (m, 4 H), 7.16 (in, 1 H), 6.86 (dd, J=4.8 and 8.0, 1 H), 4.16 (q, J=7.2, 2 H), 1.18 (t, J=7.2, 3 H). 15 A solution of the mixture of enamine and the cyclized product (11.1 g) in diethylbenzenes (100 mL) was heated at 160 0 C for 8 h and cooled. The precipitate was isolated via filtration, washed successively with hexanes and EtOH, and dried. This provided 3-(4-chlorophenyl)-9-hydroxypyrido[1,2-a]pyrimidin-4-one as a straw coloured solid (7.03 g, 74%). 20 3-(4-Chlorophenyl)-9-hydroxypyrido[1,2-a]pyrimidin-4-one: 'H NMR (DMSO d6): 6 8.64 (dd, J=2.0 and 6.0, 1 H), 8.61 (s, 1 H), 7.89 (in, 2 H), 7.51 (in, 2 H), 7.30 (in, 2 H); mass spectrum: m/z 273, 275 (lX +1, 100%, 33%). (B) 3-(4-Chlorophenyl)-9-hydroxy-8-iodopyrido[1,2-alpyrimidin-4-one (1063) 25 0 c ci N IN OH OH 30 To a stirred suspension of 3-(4-chlorophenyl)-9-hydroxypyrido[1,2-a]pyrimidin 4-one (2.73 g, 0.01 mol) and iodine (2.54 g, 0.01 mol) in EtOH (100 mL) was added hydrogen peroxide (1.94 g of 30% w/v aqueous solution). The mixture was left to stir at room temperature for 6 days. The solid was isolated via filtration, washed with EtOH (20 mL x 3), and dried to give a dark green powder (3.56 g). Subsequent 35 recrystallisation from DMF provided 3-(4-chlorophenyl)-9-hydroxy-8-iodopyrido[1,2 a]pyrimidin-4-one as an orange-brown solid (1.80 g).

WO 2004/031161 PCT/AU2003/001303 - 135 3-(4-Chlorophenyl)-9-hydroxy-8-iodopyrido[1,2-a]pyrimidin-4-one: 1H NMR (DMSO-d 6 ): 8 8.57 (s, 1 H), 8.35 (d, J=7.2, 1 H), 7.88 (in, 2 H), 7.64 (d, J=7.2, I H), 7.51 (in, 2 H); mass spectrum: m/z 399, 401 (M* + 1 , 100%, 33%). 5 The following assays are used in the assessment of new compounds according to the invention for suitability for use in the methods of the invention. Example 5 - Assessment of Compounds of Formula I or II 10 The following Assays were used in the assessment of the compounds of formula I or II for suitability for use in the methods of the invention. Assay 1. Fluorometric H202 Assay A fluorometric assay was used to test the ability of a test compound to 15 inhibit hydrogen peroxide generation by AB in the presence of copper based on dichlorofluoroscein diacetate (DCF; Molecular Probes, Eugene OR). The DCF solution (5mM) in 100% dimethyl sulphoxide (previously purged with argon for lhr at 20 0 C) was deacetylated in the presence of 0.025M NaOH for 30min and neutralised at pH 7.4 to a final concentration of 1mM. Horseradish peroxidase(HRP) stock solution was 20 prepared to 1gM at pH 7.4. The reactions were carried out in PBS, pH 7.4 in a 96 well plate (total volume =250pl/well). The reaction solutions contained AB 1-42 at concentrations in the range of 50nM to 1 M, copper-glycine chelate (Cu-Gly), was prepared by adding CuCl 2 to glycine in the ratio of 1:6 and added to the AB in the proportion 2Cu-Gly: 1AB ), reducing agents including dopamine (5pM) or ascorbic 25 acid, deacetylated DCF 100gM, and HRP, 0.1 M. 1-10pM EDTA or another chelator may also be present as a control for free copper, but was not required for the assay to function. The reaction mixture was incubated at 37C for 60 min. Catalase (4000 units/ml) and H 2 0 2 (1-2.5gM) standards in PBS pH 7.4 may be included as positive controls. Fluorescence was recorded using a plate reader with excitation and emission 30 filters at 485nM and 530nM respectively. H 2 0 2 concentration may be established by comparing fluorescence with the H 2 0 2 standards. Inhibition of AB H 2 0 2 production was assayed by including a given concentration of test compound(s) in the test wells. Assay 2. Neurotoxicity Assays 35 Primary cortical neuronal cultures Cortical cultures were prepared as previously described (White et al., 1998). Embryonic day 14 BL6Jxl29sv mouse cortices were removed, dissected free of WO 2004/031161 PCT/AU2003/001303 - 136 meninges and dissociated in 0.025% (wt/vol) trypsin. Dissociated cells were plated in 48 well culture plates at a density of 2 x 106 cells/mL in MEM with 25% (vol/vol) FCS and 5% (vol/vol) HS and incubated at 37'C, 2hrs. Media was then replaced with Neurobasal media (Invitrogen Life Technologies) and B27 supplements (Invitrogen 5 Life Technologies). Cultures were maintained at 37"C in 5% CO 2 . Prior to experimentation, the culture medium was replaced with Neurobasal media and B27 minus antioxidants (Invitrogen Life Technologies). Primary cerebellar granule neuronal cultures 10 Cerebella from post-natal day 5-6 (P5-6) mice were removed and dissected free of meninges and dissociated in 0.025% trypsin. Cerebellar granule neurons (CGN) were plated in 24 well culture plates at 350 000 cells/cm2 in BME (Invitrogen Life Technologies) supplemented with 10% Fetal Calf Serum (FCS), 2 mM glutamine and 25 mM KCl. Gentamycin sulphate (100 tg/mL) was added to all plating 15 media and cultures were maintained at 37 0 C in 5% CO 2 . Assay 3. Assays for Cell Viability (a) MTS Assay for Cell Viability 20 Cell viability is determined using the MTS assay. Culture medium is replaced with fresh neurobasal medium plus B27 supplements minus antioxidants. 1/10th volume MTS solution (Cell Titre 96 Aqueous One, Promega Corporation) and incubated at at 37 0 C, 2hrs. 200 microlitre aliquots are measured with a spectrophotometer at 560 nm. 25 (b) LDH Assay for Cell Viability Cell death is determined from culture supernatants free of serum and cell debris using the lactate dehydrogenase (LDH) Cytotoxicity Detection Kit (Boehringer Ingelheim) according to the manufacturer's instructions. 30 (c) Assay for A/i Neurotoxicity and AfiNeuroprotection Neuronal cortical cells were cultured for five days as per Assay 2. On day six the neurobasal (NB) media (Invitrogen Life Technologies) and B27 supplement (Invitrogen Life Technologies) were replaced with NB media and B27 supplement (no 35 antioxidants). On day six, test compounds were individually added to the neuronal cell cultures: WO 2004/031161 PCT/AU2003/001303 - 137 The test compounds were dissolved in 100% DMSO to a concentration of 2.5 mM (10mM if excess compound was weighed out per vial - then diluted to 2.5mM). 2.5mM stock solution was serially diluted 1 in 10 to give working solutions of 250uM, 25uM, 2.5uM. 5 Af6 preparation: As was initially dissolved in 20mM NaOH to a concentration of 1mM and sonicated for 5 minutes. The peptide was then diluted in H20 and 10 X PBS to a final concentration of 200uM As in 1X PBS. The peptide was again sonicated for 5 10 minutes and then spun at 14000 rpm for 5 min and transferred to a fresh tube. The test compounds were dissolved in 100% DMSO to a concentration of 2.5 mM (10mM if excess compound was weighed out per vial - then diluted to 2.5mM). 2.5mM stock solution was serially diluted 1 in 10 [in NB media and B27 (no antioxidants)] to give working solutions of 250uM, 25uM, 2.5uM. Test compounds 15 were not added directly to cells, instead they were added to a 48 well 'Drug Plate' as comprised below: Preparation of "Drug Plate": To a 48 well plate add: Well 1: 515 ul NB+B27(no antioxidant)* + 24 ul 25uM test compound + 60ul AP 20 diluent** Well 2 : 515 ul NB+B27(no antioxidant) + 24 ul 250uM test compound + 60ul As diluent Well 3 : 515 ul NB+B27(no antioxidant) + 24 ul test compound diluent*** + 60ul As1 42 25 Well 4: 515 ul NB+B27(no antioxidant)+ 24 ul 2.5uM test compound + 60ul AP 1-42 Well 5 : 515 ul NB+B27(no antioxidant) + 24 ul 25uM test compound + 60ul AP 1-42 Well 6: 515 ul NB+B27(no antioxidant)+ 24 ul 250 uM test compound + 60ul Ap1-42 diluent Well 7: 515 ul NB+B27(no antioxidant) + 24 ul test compound diluent + 60ul AP 1-42 30 diluent Well 8 : 600 ul NB+B27(no antioxidant) N.B. 60ul As 1-42 equals 20ul As 1-42 per well equals 20 uM AP 1-42 The Drug Plate was incubated at 370 C for 15 mins. 200 ul of each well 35 was added in triplicate to the corresponding cell plate. The cell plate was incubated at 37 C, for 4 days.

WO 2004/031161 PCT/AU2003/001303 - 138 * NB media + B27 (no antioxidants) , ** AP diluent 2mM NaOH, 1 X PBS *** PBT diluent 10% DMSO in NB+B27(no antioxidant) 5 Completion of the assay: On the 4 th day after treating the cells the assay is completed by adding MTS to the cells. (d) Assay for Test Compound Cytoxicity 10 Neuronal cortical cells were cultured for five days as per Assay 2 in NB media and B27 supplement. On day six the test compounds were added to the neuronal cell cultures in NB media and B27 supplement minus antioxidants. Test compounds were dissolved in 100% DMSO to a concentration of 15 2.5 mM (10mM if excess compound was weighed out per vial - then diluted to 2.5mM). 2.5mM stock solution was serially diluted 1 in 10 to give working solutions of 250uM, 25uM, 2.5uM. Test compounds were not added directly to cells, instead they were added to a 48 well 'Drug Plate' as comprised below: Preparation of "Drug Plate": 20 To a 48 well plate add: Well 1: 576 ul NB+B27(no antioxidant)* + 24 ul 2.5uM test compound Well 2 : 576 ul NB+B27(no antioxidant) + 24 ul 25uM test compound Well 3 : 576 ul NB+B27(no antioxidant) + 24 ul 250uM test compound Well 4: 576 ul NB+B27(no antioxidant)+ 24 ul 2.5uM test compound 25 Well 5 : 576 ul NB+B27(no antioxidant)+ 24 ul 25uM test compound Well 6 : 576 ul NB+B27(no antioxidant) + 24 ul 250uM test compound Well 7 : 576 ul NB+B27(no antioxidant) + 24 ul test compound diluent** Well 8 : 600 ul NB+B27(no antioxidant) The Drug Plate was incubated at 370 C for 15 mins. 200 ul of each well 30 was added in triplicate to the corresponding cell plate. The cell plate was incubated at 37 C, for 4 days, (2 compounds are tested on each plate of cells). * NB media and B27 (no antioxidants) , ** PBT diluent 10% DMSO in NB+B27 (no antioxidants) 35 WO 2004/031161 PCT/AU2003/001303 - 139 On completion of the assay, 1/10 volume MTS was added per well of plate (ie 25ul/ 250 ul). The plates were incubated at 37C for 2hrs, and then absorbance was read at 560nm. 5 Assay 4. Caspase Assay To measure caspase activity in neuronal cultures, growth medium is removed, cells are washed twice with control salt solution (pH 7.4) and ice-cold cell extraction buffer is added directly to the cultures. The extraction buffer consists of 20 mM Tris (pH 7.4), 1 mM sucrose, 0.25 mM EDTA, 1 mM dithiothreitol (DTT), 0.5 10 mM PMSF, 1% Triton X-100 (Tx-100) and 1 ptg/mL of pepstatin and aprotinin. After incubation for 15 min on ice, the extraction buffer is removed, centrifuged for 5 min at 4 0 C in a microcentrifuge and 100 .L of supernatant is added to each well of a 96 well plate. 100 [L of 200 1 M substrate (either DEVD-pNA, VEID-pNA or IETD-pNA for caspases 3, 6 and 8 respectively) is added to each well to give a final concentration of 15 100 pM substrate. Plates are incubated at 37'C for 2, 4, 6 or 24 hr and the absorbance is determined at a wavelength of 415 nm (Abs415). The absorbance reading is compared to a known standard of pNA alone. Assay 5. Annexin V Assay 20 To determine the level of annexin V binding to cells, cultures are washed twice with control salt solution (pH 7.4) followed by the addition of annexin V-FITC at a concentration of approximately 0.5 pig/mL in control salt solution (pH 7.4). Propidium iodide (10 tg/mL) is also added to the cultures at the same time. Cells are incubated in the dark for 30 min at ambient temperature and subsequently washed three times with 25 fresh control salt solution. Analysis of FITC fluorescence (ex. 488 nm, em. 510 nm) is determined using a Leica DMIRB microscope. Photographs are taken with a Leica MPS 60 camera attachment using ASA400 colour film, and negatives are scanned into Adobe Photoshop v2.0.1. 30 Assay 6. Lipoprotein Oxidation Assay Two different assays of metal-mediated lipid peroxidation can be utilized. The first assay involves measuring the oxidative activity of metallated proteins. This is determined by mixing dialyzed metallated or native protein (at designated concentrations) with 0.5 mg/mL LDL for 24 hr (37'C). Lipid peroxidation 35 (LPO) is measured using a lipid peroxidation assay kit (LPO 486, Oxis International Inc. Portland, OR) as per kit instructions. The level of LPO is determined by comparing absorbance (486 nm) with LDL alone (100% LPO). The second assay is WO 2004/031161 PCT/AU2003/001303 - 140 used to measure the LPO activity of native proteins in the presence of free, non-protein bound Cu. This involves adding non-metallated peptides (140 pM) to 0.5 mg/mL LDL together with 20 ptM Cu-gly and assaying for LPO as for the metallated proteins. The level of LPO is determined by comparing the absorbance (486 im) with LDL + Cu-gly 5 (100% LPO). As a negative control, LDL is also exposed to dialysed Cu-gly solutions comparable to those used to Cu-metallate the proteins. Assay 7. Cytotoxicity Induced by Cu-Metallated Proteins Proteins or synthetic peptides are mixed with metal-glycine solutions at 10 equimolar or two-fold metal to protein concentration. Metal-protein mixtures are incubated overnight at 37 0 C and then extensively dialysed (24 hr against two changes of dH 2 0 (3 L/change) at room temperature) using mini-dialysis cups with a 3,500 kilodalton cut-off (Pierce, Rockford, IL). Dialysis of proteins against PBS pH 7.4 resulted in metallated proteins with identical activity to dH20 dialysis. 15 To determine their neurotoxic effects, metallated proteins, native proteins or peptides are added to two day-old primary cortical neuronal cultures. The cultures are also exposed to Cu-gly (5 or 10 tM) or LDL. Positive control cultures are treated with Cu-gly + LDL or the LPO product, 4-hydroxy-nonenol (HNE, Sigma Chemicals). Cultures are assayed for cell death using the lactate dehydrogenase (LDH) 20 assay kit (Roche Molecular Biochemicals, Nunawading, Australia) according to the manufacturer's instructions. Assay 8. Acridine Orange Assay for AP-Mediated Loss of Lysosomal Acidification Cultured mouse cortical neurons are treated with AP 1-42 (20 M) for 16 25 h and then stained with 5 mg/ml acridine orange (AO) for 5 min at 37 0 C. 15 min at 37*C. The AO-induced fluorescence is measured with a red filter on a fluorescence microscope. AO is a lysosomotropic weak base which accumulates in the endosomal/lysosomal compartments and displays orange fluorescence during incubation. AO is sequestered inside the lysosomes as long as there is a substantial 30 proton gradient over the lysosomal membranes. Treatment of cells with As 1-42 disrupts the lysosomal membrane proton gradient and relocalises AO into the cytosol, as indicated by the loss of orange fluorescence within 16-24 hr. Assay 9. Human Brain Amyloid Solubilisation Assay 35 This assay was performed in order to assess the ability of a test compound to mobilise As from the insoluble to the soluble phase of an extract of tissue from post mortem human AD brain.

WO 2004/031161 PCT/AU2003/001303 - 141 Up to 0.5 g of plaque-bearing cortex without meninges was homogenized using a DIAX 900 homogenizer (Heudolph and Co, Kelheim, Germany) or other suitable device for three 30-second periods at full speed in up to 2 ml of ice cold phosphate-buffered saline, pH 7.4. To obtain the phosphate-buffered saline 5 extractable fraction, the homogenate was centrifuged at 100,000 x g for 30 min and the supernatant removed. Alternatively, the tissue was freeze dried then pulverised to form a powder which was then weighed out into aliquots for extraction as above. A 1Opl aliquot of supernatant was removed after centrifugation and mixed with an equal volume of 2XTris-Ticene SDS sample buffer, pH 8.3, containing 8% SDS, 10% 2 10 mercaptoethanol. Samples were then heated for 10 mins at 90*C and separated by gel electrophoresis. The insoluble fraction of the cortical samples was obtained by resuspending the initial pelleted sample in 1 ml of phosphate-buffered saline. A 50-[d aliquot of this suspension was then boiled in 200 ml of sample buffer as above. Tris-Tricine polyacrylamide gel electrophoresis was performed by 15 loading appropriately diluted samples on to 10% to 20% gradient gels (Novex, San Diego, CA) followed by transfer on to 0.2-pm nitrocellulose membrane (Bio Rad,Hercules, CA). As was detected by using monoclonal antibody W02, which detects residues 5 through 8, 17 (or another suitable antibody) in conjunction with horseradish peroxidase-conjugated rabbit anti-mouse IgG (Dako, Denmark), and 20 visualized by using enhanced chemiluminescence (eg ECL; Amersham Life Science, Buckinghamshire, UK). Each gel included three lanes containing 0.5, 1, and 2 ng of synthetic Ap 40 (Keck Laboratory, Yale University, New Haven, CT) as reference standards. Blot films were scanned by using a suitable imaging system such as the 25 UVP gel documentation system, and densitometry performed using suitable software, eg UVP Labworks. The dynamic range of the film/scanner was determined by using a step tablet (No. 911ST600, Kodak, Rochester NY), a calibrated film exposed by the manufacturer to provided steps of known increasing intensity. The quantifiable range of signal intensity for densitometric analysis of the mono- and dimeric As bands was 30 based on the comparison with a curve obtained by scanning and densitometry of the step tablet. Samples in which the signal intensity is low after preliminary assay may be re-assayed by using synthetic standards of lower or higher concentration. All samples were analysed several times, and gel loadings and dilutions were adjusted to fit within the quantifiable region of the standard curve. The insoluble AP 35 being comprised of the pelletable fraction derived from the insoluble amyloid plaque from the above cortical samples and the soluble fraction comprising monomeric and/or oligomeric soluble AP.

WO 2004/031161 PCT/AU2003/001303 - 142 Several gels were run per test compound with a PBS control included on each gel. Each gel containing varying concentrations of the test compound. A student's 't test' was used to compare the mean of the highest value obtained by the test compound for each gel at any concentration, to the mean of the PBS values taken from the multiple 5 gels. Accordingly a determination can be made of whether the average increase in solubilisation obtained by any test compound is significant compared with PBS alone. Test compounds with a (+) score are compounds which achieved a statistically significant increase in plaque solubilisation over that of PBS alone. A test compound with a (-) score is a compound which does not achieve a statistically significant increase 10 in plaque solubilisation over that of PBS alone Assay 10. Metal Partitioning To assay effects upon the partitioning of various metals, including zinc and copper, following extraction of brain tissue in the presence of a test compound, 15 soluble and insoluble fractions from an extract of human brain tissue are prepared as for the amyloid solubilisation assay. Metals in the two fractions are analysed by inductively-coupled plasma mass spectrometry, following appropriate pretreatment with nitric acid and/or hydrogen peroxide where necessary. 20 Assay 11. Effect of Administration of Test Compounds on AP deposits in Transgenic Animals Transgenic mouse models are available for a number of neurological disorders, including Alzheimer's disease (Games et al., 1995; Hsiao et al., 1996); Parkinson's disease (Masliah et al., 2000); familial amyotrophic lateral sclerosis (ALS) 25 (Gurney et al., 1994); Huntington's disease (Reddy et al., 1998); and Creutzfeld-Jakob disease (CJD) (Telling et al., 1994). We have found that one of the transgenic models for Alzheimer's disease, the APP2576 transgenic mouse (Hsiao et al., 1996) also has a high incidence of cataract. These animal models are suitable for testing the methods of the invention. 30 Transgenic mice of the strain APP2576 (Hsiao et al 1996) are used. Eight to nine month old female mice are selected and divided into groups for treatment. Mice are sacrificed at intervals, and their brains examined to determine whether the treatment with test compounds decreased brain amyloid formation, and the identification of the most effective administration protocol. The levels of soluble and 35 insoluble AP in the brain and serum are determined using calibrated Western blots as per the methodology described for Assay 9. Brain Amyloid Solubilisation Assay. Other mice in each group are tested over a period of up to eight months WO 2004/031161 PCT/AU2003/001303 - 143 for cognitive performance, using a Morris water maze according to standard methods. The general health and well-being of the animals is also measured every day by a blinded operator, using a five point integer scale which subjectively rates a combination of features, including motor activity, alertness and general health signs. 5 Assay 12. Physiochemical Properties Polar Surface Area Calculations (PSA) Polar surface area values were calculated using the web-based program available through "Molinspiration", a package for calculation of molecular properties. 10 Turbidimetric Solubility Measurements The solubility estimate was measured at both pH 2.0 and pH 6.5. This is within the pH range that can be anticipated along the proximal gastrointestinal tract in humans. 15 The compounds were dissolved in DMSO to appropriate concentrations and then spiked into either 0.01M HCl (approx. pH = 2.0) or pH 6.5 isotonic phosphate buffer, the final DMSO concentration being 1%. Samples were then analysed via Nephelometry to determine a solubility range. [as per D. Bevan and R. S. Lloyd, Anal. Chem. 2000, 72, 1781-17871. 20 cLog P values Theoretical Log P values were determined using the ACD Log P software. The values quoted have been calculated from an untrained database and refer to the unionised species. 25 Assay 13. Blood Brain Barrier Penetration The test compounds were dissolved in DMSO and phosphate buffered saline (PBS) was added to obtain solutions at a concentration of 50 JIM in PBS containing 1.25-2.5% DMSO. A trace amount of 14 C-sucrose was added to each stock 30 infusion solution (approx 0.01 pCi/mL) to act as Blood-Brain Barrier (BBB) impermeable marker in order to assess the integrity of the BBB during each perfusion and to estimate the volume of the residual vascular space (RVS) in samples of brain tissue (ie: the volume of fluid remaining inside the lumen of blood vessels at the end of each perfusion). 35 Adult male Spague Dawley rats (180-190g) were anaesthetized with intraperitoneal injections of Urethane (25% w/v) at a dose of 1.0 mL/lOOg body weight. The right common carotid artery was surgically exposed and cannulated for perfusion WO 2004/031161 PCT/AU2003/001303 - 144 of the cerebral circulation. The right external carotid artery (which supplies tissues outside the skull) was then ligated distal to its bifurcation from the right common carotid artery so that all of the infusion solution would pass into the brain via the remaining right internal carotid artery. The heart was then exposed and transected 5 immediately prior to the commencement of the infusion. The rate of the infusion was controlled by a pump set to deliver at 3.2mL/min (approx. 85% of the normal blood supply to the brain for this size of rat). The infusion cannula initially contained a 0.5 mL pre-wash of heparinised PBS (10 IU/ml) that acts to flush blood vessels and to prevent blood from clotting and blocking small vessels. 10 After 1.5 minutes, the infusion pump automatically stopped, the cannula was withdrawn from the carotid artery and a sample of the infusion solution (1-1.5 mL) was then collected from the tip of the infusion cannula. The brain was then dissected free and divided into 3 parts; the right hemisphere together with the right midbrain, the left hemisphere together with the left midbrain and the hindbrain (cerebellum, pons and 15 brainstem). Only the right part of the brain was used for subsequent measurements because perfusion via the right internal carotid artery preferentially supplies the right hemisphere and right midbrain (the left hemisphere and hindbrain receive a variable collateral perfusion). The brain tissue samples from each animal were frozen at -30*C, homogenized and weighed aliquots analysed by LC-MS to give total brain 20 concentration. The analysis was carried out using the Micromass Triple Quad instrument. The mobile phase consisted of an acetonitrile / water gradient (containing 0.05% Formic acid) and the column was a Phenomenex Luna CN. Small aliquots from each brain tissue sample and the corresponding infusion solution were analysed by liquid scintillation counting to determine the level of 25 1C-sucrose. The residual vascular space (RVS) in each brain tissue sample was calculated by dividing the measured concentration of sucrose in brain tissue (dpm/mg) by its concentration in the corresponding infusion solution (dpm/pL). This is the volume of fluid that remains inside blood vessels at the end of each perfusion. Multiplying this RVS by the concentration of the test compound in the infusion solution 30 gives the total residual amount of the test compound that is present inside blood vessels in each brain tissue sample (ie: that which has not crossed the BBB). Subtracting this from the total brain concentration gives the amount of drug in each brain tissue sample that is outside the blood vessels (ie: which has crossed the BBB). Dividing this RVS corrected brain concentration gives the brain uptake ratio (Equation. 1). 35 WO 2004/031161 PCT/AU2003/001303 - 145 Equation 1. [brain ng.mg 3- [RVS ng. 1-1] Brain Uptake Ratio 5 [infusion solution ng.gL-] A total of 5-6 brain perfusion experiments were performed for each of the test compounds and mean brain uptake ratios were calculated. Ratios of greater than 50% indicate compounds that enter the brain 10 extremely rapidly; ratios between 10 and 50% indicate compounds that enter the brain well; ratios less than 10% (not observed) would indicate compounds that enter the brain very slowly and would not be suitable for therapeutic administration; ratios less than 1% (not observed) would indicate compounds that are effectively excluded from the brain. 15 Assay 14. Transgenic Mouse Brain Immunohistochemistry The APP2576 transgenic mouse (Hsiao et al., 1996) as referred to in Assay 11 is utilized in this assay. The contralateral formalin-fixed mouse brain tissue is coronally cut. Sections (10 Im) are taken from the corresponding sites and treated with 20 80% formic acid for antigen retrieval. The primary antibody used is monoclonal antibody 1E8, which recognizes epitopes between residues 18 and 22 of A# (SmithKline Beecham, UK). Immunoreactivity is developed with secondary antibody linked to horseradish peroxidase (using a 3,39-diaminobenzidinechromagen) (Dako) and alkaline phosphatase (using 5-bromo-4-chloro 3-indoxyl phosphate and nitroblue 25 tetrazolium chloride chromagen) (Dako). Plaque abundance per section is assessed by two operators blinded to treatment according to the following scale: 0 = no plaques apparent 1 = plaques present but very sparse 2 = several plaques present 30 3 = numerous plaques visible in restricted areas 4 = plaques abundant and not restricted to any particular area. Intermediate values eg 2.5 are assigned where applicable. Students' t ' test is used for comparisons between groups.

WO 2004/031161 PCT/AU2003/001303 - 146 Assay 15 Pharmacokinetic Profile e Intravenous infusion of test compound; 2 mg/Kg in a suitable vehicle is administered to 2 rats and arterial blood is sampled up to 24 hours. 5 e Oral administration of test compound; 30 mg/Kg in a suitable vehicle is administered via oral gavage to 2 rats and arterial blood is sampled up to 24 hours. * Plasma concentrations of test compound are determined by suitable analytical method. 10 Calculations: CLtot = Dose v CLtotai BA(%)= AUCora * Dose, AUCff d/ A UCl * Doseora CLtota1= total plasma clearance after IV administration Vd = volume of distribution during the elimination phase after IV 15 administration BA = oral bioavailability AUCiv = area under the plasma concentration versus time profile from time zero to infinity after IV administration AUCorai= area under the plasma concentration versus time profile from time 20 zero to infinity after oral administration # = terminal elimination rate constant after IV administration Assay 16 Determination of mouse plasma levels of test compounds 25 PBT 1061 Oral administration of PBT 1061 at 30mg/kg, as a suspension in Na Carboxymethyl Cellulose (CMC) was administered by oral gavage to four mice. Two mice were sacrificed 30 minutes after administration and two mice were sacrificed 60 minutes after administration. Blood was obtained by cardiac puncture and plasma 30 separated by centrifugation. The concentration of PBT1061 was determined by LC/MS using the ZQ instrument. The mobile phase consisted of an acetonitrile (ACN)/water gradient (containing 0.05% Formic acid) and the column was a Phenomenex Polar-RP 4piM 80A (50 x 2mm) column. 35 WO 2004/031161 PCT/AU2003/001303 - 147 The mouse plasma samples were directly injected following a protein precipitation with ACN. The analytical method in plasma was linear in the range of 125 to 10,000 ng/ml (R = 0.9992). Diazepam was used as the internal standard. Recovery of PBT1061 from plasma was ~ 100 %. 5 The concentration of PBT1061 in mouse plasma after dosing orally at 30 mg/kg is given in Table 1. Table 1. Concentrations of PBT1061 in Mouse Plasma after Oral Dosing at 30 mg/kg Mouse Dose Time Conc. ID (mg/Kg) (min) (ng/ml) 2703 30 30 1826.61 2743 30 30 5475.88 2740 30 60 1115.24 2801 30 60 2417.55 10 PBT 1063 Oral administration of PBT 1063 at 30mg/kg, as a suspension in Na Carboxymethyl Cellulose (CMC) was administered by oral gavage to four mice. Two mice were sacrificed 30 minutes after administration and two mice were sacrificed 60 15 minutes after administration. Blood was obtained by cardiac puncture and plasma separated by centrifugation. The concentration of PBT1063 was determined by LC/MS using the single quadrupole instrument. The mobile phase consisted of an acetonitrile (ACN)/water gradient (containing 0.05% Formic acid) and the column was a Phenomenex C8 4ttm 20 80A (50 x 2mm) column. The supplied acute toxicity mouse plasma samples (delivered 02.09.03) were directly injected following a protein precipitation with ACN. Concentrations were determined by comparison to a calibration curve prepared in rat plasma. The analytical method in plasma was linear in the range of 312 to 10,000 ng/ml (R 2 = 0.9976). 25 Diazepam was used as the internal standard. Recovery of PBT1063 from plasma was ~ 42.9 %. The concentrations of PBT1063 in the mouse plasma samples are given in Table 2. 30 Table 2. Concentrations of PBT1063 in Mouse Plasma after Oral Dosing at 30 mg/kg WO 2004/031161 PCT/AU2003/001303 148 Exp. No. Time Cone. (min) (ng/ml) 2835 30 412.81 2843 30 279.39 2814 60 439.49 2677 60 330.66 WO 2004/031161 PCT/AU2003/001303 -149 > C) C ce 1)o in 0 ) 1.1 2io C) t8 0 0 0 u ~ cqr' rio rb -b = ~~~C 00 0C A 6 lb 'Sb 8 =L A c) 6 :L " cq C '6bkn - kn 6b U) A = LvA = 0 40 0 00 00 ON 0 00 Co 00 - n c r 4-* 0-0 l N C C 0 C. 0 0 11 0 rio 20 rio 0 ~0 ' ~ 004 0 -42 >i -~o i 0 CO n kn CA ) b "R r ,: II C i C > 004 1-4) \0 ~ 2 2 44 0a 0 ' ~ 0 t~0 1 ~ WO 2004/031161 PCT/AU2003/001303 - 150 Example 6 - Clinical trial of compound of formula I or II for the treatment of Alzheimer's disease. A Phase II clinical trial of the compound of formula I or II for the treatment of AD was undertaken to study the effects of oral PBT-1 treatment in a 5 randomised, double-blind, placebo-controlled pilot phase 2 clinical trial of moderately severe AD patients. Thirty-six subjects were randomized [18 placebo and 18 PBT-1, with 32 completions], and stratified into more- and less- severely affected groups. The effect of treatment was statistically significant in preventing cognitive deterioration over 36 weeks in the more-severely affected patients (baseline ADAS-cog > 25). The 10 performance of the less-severely affected group (ADAS-cog < 25) deteriorated negligibly over this interval, so cognitive changes could not be discriminated in this stratum. Plasma A#42 declined in the PBT-1 group but increased in the placebo group (p<0.001). Plasma Zn levels rose significantly (=30 %) in the PBT-1 group. 15 Dosage Several considerations drove the choice of dose. In previous studies on transgenic mice, doses of 20-30 mg/kg of PBT-1 orally daily for five days per week were markedly effective at inhibiting AP accumulation after 2-3 months of treatment. The human equivalent dose of 1500-2250 mg/day is close to the prescribed antibiotic 20 dose of PBT-1 (600 mg po qid). However, this magnitude of dose, administered for months, would raise concerns about SMON toxicity. The starting dose of 3.3 mg/kg/day, assuming 75 kg average weight, is within the same order of magnitude of the effective dose in the transgenic mouse model, but only about one tenth of the antibiotic dose. 25 Since there is no data from the transgenic mouse study of the effectiveness of doses less than 20 mg/kg/day, we reasoned that a beneficial effect might require a longer period of treatment than the 9-12 week duration of the mouse study (Chemy et al., 2001). Therefore a trial length of 36 weeks at an average dose which is approximately one-third of what is effective in the transgenic mice is chosen. 30 The final dose of 10 mg/kg/day is half of an effective dose in mice. The starting dose of 3.3 mg/kg/day was within the same order of magnitude of the effective dose in the transgenic mouse model, but only about one tenth of the anti-infective dose. The study was powered to detect biochemical effects on metal and Ag levels that would be in the same magnitude as those seen in the transgenic 35 study.

WO 2004/031161 PCT/AU2003/001303 - 151 EXPERIMENTAL PROCEDURES Ethical issues: In compliance with Australian laws concerning consent from individuals whose cognitive function may be impaired to the extent of being unable to make informed judgements or decisions, "Consent to Special Procedures" 5 administered by the Victorian Civil and Administrative Tribunal was obtained for each participant not able to consent on their own behalf. In addition, third party consent was obtained from all carers. All subjects were stabilized on donepezil prior to commencement of the study. The study was approved by the Royal Melbourne Hospital Research Foundation's Clinical Research and Ethics Committee. 10 Study population: The study took place at the AD clinical trials unit, Mental Health Research Institute of Victoria and at the Royal Melbourne Hospital. Criteria for inclusion in the study were: informed consent; a diagnosis of probable AD by NINCDS-ADRDA criteria (McKhann et al., 1984); AD Assessment Scale-cognitive (ADAS-cog) (Rosen et al., 1984) score of 18-45; Mini Mental State Examination 15 (MMSE) (Folstein et al., 1975) score of 10-24; on donepezil 5mg or 10mg for at least 6 months; relative or carer willing and able to support the trial; able to complete trial examinations; primary sensorial functions intact. Patients were excluded if they had a history or clinical evidence of peripheral or optic neuropathy or had co-existing illnesses or past history that may have 20 affected cognitive function, nerve conduction or illnesses that may have confounded the adverse event profile. The following factors were obtained at baseline to determine if they correlated with outcome measures: age, sex, premorbid IQ [estimated from the National Adult Reading Test (NART)], years of education, and apolipoprotein E (ApoE) allotype. 25 Study design: The study was a double blind, placebo-controlled, parallel group randomized design. Thirty-six patients and their carers were recruited to participate, with patients randomized at a 1:1 ratio to receive either PBT- 1 or placebo. The duration of the study was 36 weeks. PBT-1 oral dosage was 125mg bid from weeks 0-12, increased to 250mg bid from weeks 13-24, and finally, 375mg bid from weeks 30 25-36. Study procedures: Screening procedures consisted of a complete medical history, physical, neurological and ophthalmic examination, blood and urine tests and psychometric tests (ADAS-cog, MMSE). Nerve conduction tests and visual evoked responses were conducted between the screening and baseline visits to provide a 35 baseline measurement. Blood was collected for ApoE allotyping, baseline plasma levels of metals and A#l prior to randomization. All patients continued their study entry dose of donepezil and all patients received 100 mg vitamin B12 intramuscularly every four WO 2004/031161 PCT/AU2003/001303 - 152 weeks. Blood samples were collected by antecubital venepuncture except on weeks 12, 24 and 36 when they were collected by an indwelling catheter. The procedural change did not affect biochemical readouts except for Zn levels which were found to be 5 consistently -10% depressed (probably as a result of differences in platelet activation). Zn data from these intervals were therefore omitted from analysis. Outcome measures: The primary clinical efficacy variable was a change from baseline score on the ADAS-cog conducted at baseline and at weeks 4, 12, 24 and 36. This measure was chosen to allow comparability of treatment effects with current 10 therapeutics such as donepezil, where efficacy trials also used ADAS-cog as their primary outcome measure (Rogers et al., 1998). Although numerous neuropsychological tests could be considered as secondary measures, it was necessary to avoid fatiguing the subjects at review. Therefore the only other cognitive test was the Mini-Mental State Exam (MMSE). The CIBIC+ (clinician interview based impression 15 of change incorporating caregiver information), a subjective observational index was also conducted. Plasma Afl, and plasma zinc and copper were all taken every four weeks. Double antibody capture enzyme-linked immunosorbent assay (ELISA)for A# detection: Polystyrene plates were coated with mAb G210 (for Afl40) 20 or mAb G211 (for A042). Plates were washed and biotinylated mAb W02 was added. Bound antibody was detected with streptavidin-labelled Europium (Perkin Elmer, Vic Australia). The values obtained from triplicated wells were calculated based on standard curves generated on each plate. Plasma samples supplemented with synthetic Afll-40 and Afll -42 were also assayed to confirm measurement reliability across the 25 concentration range of interest. Metal levels: Metals were measured by inductively coupled plasma mass spectrometry as previously described (Chemy et al., 2001). Therapeutic drug monitoring: At weeks 12, 24 and 36, PBT-1 blood levels were assayed by HPLC with appropriate validation studies (Centre for 3 0 Pharmaceutical Research, University of South Australia). Safety measures: Standard adverse event reporting was conducted and biochemical tests, renal and liver function, complete blood examination, serum vitamin B12 and folate levels were documented at each visit. To assess for peripheral and optic neuropathy a neurological examination was conducted at each visit, and visual evoked 35 responses, nerve conduction studies and ophthalmic examination were conducted at screening, week 16 and prior to the final trial visit. An ECG was done at screening and weeks 12, 24 and 36.

WO 2004/031161 PCT/AU2003/001303 - 153 Data preparation and statistical analysis: Data monitoring and management were undertaken by independent contractors (Kendle International and Health Research Solutions, Melbourne). Evidence for efficacy was indicated by a significant difference in change from baseline between treatment arms. Analysis of 5 variance was the principal method of evaluating statistical significance with the treatment arm illness severity at baseline being the primary design factor. Potentially significant covariates were introduced as necessary. Differences between groups on categorical measures were analysed using exact statistical methods in order to maximise power. Based on the assumption of a correlation of 0.60 between measurement 10 occasions, power to detect an effect of one standard deviation difference in change between groups from baseline to week 36 would have been approximately 80% if 15 subjects were recruited per group. Since an attrition rate of 15% has been observed in similar populations, 18 patients were recruited into each arm. 15 RESULTS Subject recruitment and demographics: Thirty-six subjects were recruited over a 12 month period commencing April 2000 (Fig 1). Of these, 32 had sufficient data for per protocol analysis. Two subjects were lost from each arm. The baseline illness severity factor was created, as planned, by division 20 of the sample into two groups at the median ADAS-cog score at baseline (values <25, >25), yielding less-severely and more-severely affected groups (n=8 and 8 in the treatment arm and n=7 and 9 in the placebo arm, respectively). The groups did not differ across demographic, biological and clinical parameters at baseline (Table 4), other than the treatment arm having a higher mean 25 premorbid IQ than the placebo group as estimated using the NART (111.4 compared to 104.9; t(30)=2.27, p=0.031) and a lower level of thyroid stimulating hormone (TSH) (1.14 compared to 2.00 mU/L; t(30)=4.400, p<0.001). The NART and TSH were subsequently provisionally entered into analyses as co-variates but were found to be not significant in any analysis. 30 Clinical effects: Changes in the ADAS-cog score at weeks 4, 12, 24 and 36 from baseline were subject to two-way analysis of variance with factors of treatment arm and baseline illness severity. The means of the changes in ADAS-cog score showed greater deterioration in the placebo treated group at each examination interval, compared to the PBT-1-treated group (Fig. 2A). This trend came close to statistical 35 significance at week 4 [F(1,28)=3.55, p=0.070] and week 24 [(F(1,28)=3.31, p=0.0 8 0] (Fig 2A). As planned in the protocol, the effect of severity of illness was examined by stratification of the sample into subjects less- or more- severely affected (baseline WO 2004/031161 PCT/AU2003/001303 - 154 ADAS-cog values <25, >25). Simple effects tests within level of severity showed the trend in the pooled groups to be separable into non-significant results for the less-severe stratum on all weeks and significant differences in the more-severe stratum at weeks 4 [F(1,28)=7.73, p=0.010] and week 24 [F(1,28)=6.63, p=0.016] (Fig 2B). This trend was 5 maintained at week 36 but narrowly escaped statistical significance [F(1,28)=3.62, p=0.068]. In the more-severely affected groups, the difference in mean change from baseline ADAS-cog score of PBT-1 over placebo at weeks 24 and 36 was a difference of 7.37 (95% CI: 1.51 - 13.24) and 6.36 (95% CI: -0.50 - 13.23) respectively (Fig 2B). Effects on plasma Afl, Zn and Cu: At baseline, there were no significant 10 differences in plasma A#42 levels between treatment arms or severity strata. The variance in individual levels at baseline in plasma A#40/42 was large and led to reduced power of the study to detect any significant differences in mean changes between groups. However, reference of individual AO levels to baseline reference levels markedly decreased variance, and revealed significant treatment effects. Plasma A142 15 showed a significant decline from baseline in the PBT- 1-treated group from week 20 onwards; over the same time, plasma Af42 in the placebo group increased (Fig 3A). Stratification by illness severity as above demonstrated that changes were evident only in the less-severely affected (Fig 3B). Administration of PBT-1 was associated with a significant elevation 20 (40%) of total plasma Zn (Fig 4A) but with no effect on plasma Cu (Fig 4B). Mean baseline levels of Zn (9.4 gM) in the pooled AD groups were below age-related normative values (Wood and Zheng, 1997). The increase in plasma Zn induced by PBT- 1 treatment therefore represented a normalization of levels. In contrast, mean baseline levels of Cu (13.1 pM) were within the age-related normative range (Rahil 25 Khazen et al., 2000). Correlation of plasma A#42/40 levels with Zn/Cu levels assayed on the same or subsequent occasions showed no significant associations. An important result of treatment of AD subjects with PBT-1 is the paradoxical elevation in plasma Zn (Fig. 4A), which is consistent with a restoration in the ZnT3-mediated communication of synaptic zinc with the blood. This also indicates 30 that, in contrast to a typical metal chelator such as desferrioxamine, the mechanism of action of PBT-1 at this dose is not that of a gross tissue chelator. The relatively weak affinity of PBT-1 for the metals appears to be insufficient to cause marked systemic metal depletion in the presence of a re-established equilibrium of metal homeostasis. Blood levels ofPBT-1: Steady state pre-dose levels of PBT-1 at total 35 daily dosages of 250, 500 and 750 mg were 4.03±2.10, 6.74±3.70, 7.60±2.15 pg/ml, respectively, and did not show significant correlations with ADAS-cog, metal or Afl levels assayed on the same or subsequent occasions.

WO 2004/031161 PCT/AU2003/001303 - 155 Table 4 - Baseline demographics and key clinical variables Group Variable Total Sample Clioquinol Placebo P Value (n=32) (n=16) (n=16) Age mean 72.50 73.19 71.81 P=0.65t (SD; min-max) (8.37; 56-87) (8.61; 58-87) (8.35; 56-87) Sex 17 8 9 P=1.00$ (n; % male) (53.1%) (47.1%) (52.9%) ApoE status ApoE4 heterozygote n (%) 15 7 8 P=1.00$ (46.9%) (43.8%) (50.0%) ApoE4 homozygote n (%) 3 2 1 (9.4%) (12.5%) (6.3%) Estimated premorbid IQ NART 108.1 111.4 104.9 P=0.031 mean, (SD; min-max) (8.86; 91-124) (8.04; 94-121) (8.26; 91-124) ADAS-Cog 26.31 25.56 27.06 P=0.57t (7.27; 15-46) (7.67; 15-46) (7.01; 19-41) Age of first diagnosis 70.09 70.88 69.31 P=0.

59 t mean, (SD; min-max) (7.98; 54-83) (8.50; 57-83) (7.61; 54-83) Duration of illness (years) 2.41 2.31 2.56 P=0.

66 t mean (SD; min-max) (1.19; 1-5) (1.08; 1-4) (1.32; 1-5) 5 t Independent sample t-test (all tests 30 df) Exact, two-tailed test. References cited in the description and examples are listed on the following pages, and are incorporated herein by this reference. 10 REFERENCES Abe, Y., Kayakiri, H., Satoh, S. et al, J. Med Chem., 1998, 41, 4062-4079. 15 Albert, A. and Hampton, A., J. Chem. Soc., 1952, 4985-4993. Anthony, N.J., Gomez, R.P., Young, S.D. et al, (2002) Patent WO 02/30931 A2. Antoniotti, S. and Dunach, E., Tetrahedron Letters, 2002, 4, 3971-3973.

WO 2004/031161 PCT/AU2003/001303 - 156 Ariga, T., Kobayashi, K., Hasegawa, A., Kiso, M., Ishida, H., and Miyatake, T. (2001) Characterization of high-affinity binding between gangliosides and amyloid p-protein. Arch. Biochem. Biophys. 388, 225-230. 5 Atwood et al., J. Biol. Chem., 1998, 273(21), 12817-12826. Beyreuther K, Christen Y, Masters CL (eds) Neurodegenerative Disorders: Loss of Function Through Gain of Function. Springer . Berlin. 2001. 189pp. 10 Blanco, M., Lorenzo, M.G., Perillo, I. and Schapira, C.B., J Heterocyclic Chem., 1996, 33, 361-366. Brower V. Harnessing the immune system to battle Alzheimer's: Some of the most 15 promising approaches to fight Alzheimer's diseases aim to develop vaccines. EMBO Rep 2002;3:207-9. Bush AI, Masters CL. Clioquinol's return. Science 2001; 292:2251-2252. 20 Bush Al. Therapeutic targets in the biology of Alzheimer's disease. Current Opinion in Psychiatry 2001; 14:341-348. Cherny RA, Atwood CS, Xilinas ME et al. Treatment with a copper-zinc chelator markedly and rapidly inhibits p-amyloid accumulation in Alzheimer's disease 25 transgenic mice. Neuron 2001; 30:665-676. Corder, E. H., Saunders, A. M., Strittmatter, W. J., Schmechel, D. E., Gaskell, P. C., Small, G. W., Haines, J. L., and Pericak-Vance, M. A. (1993) Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer's disease in the late onset 30 familial disease. Science 261, 921-923. Curtain, C.C., Ali, F., Volitakis, I., Cherny, R.A., Norton, R.S., Beyreuther, K., Barrow, C.J., Masters, C.L., Bush, A.I., and Barnham, K.J. (2001) Alzheimer's disease amyloid P binds copper and zinc to generate an allosterically ordered membrane-penetrating 35 structure containing superoxide dismutase-like subunits. J. Biol. Chem. 276, 20466 20473.

WO 2004/031161 PCT/AU2003/001303 - 157 Czech, C., Forstl, H., Hentschel, F., Monning, U., Besthorn, C., Geigerkabisch, C., Sattel, H., Masters, C., and Beyruether, K. (1994) Apolipoprotein E-4 gene dose in clinically diagnosed Alzheimer's disease: prevalence, plasma cholesterol levels and cerebrovascular change. Eur. Arch. Psychiatry Clin. Neurosci. 243, 291-292. 5 Dennin, F., Blondeau, D. and Sliwa, H., J. Heterocyclic Chem., 1991, 28, 1287-1291. Fassbender, K., Simons, M., Bergmann, C., Stroick, M., Lutjohann, D., Keller. P., Runz, H., Kuhl, S., Bertsch, T., von Bergmann. K., Hennerici, M., Beyreuther, K., and 10 Hartmann, T. (2001) Simvastatin strongly reduces levels of Alzheimer's disease p amyloid peptides AP 42 and AP 40 in vitro and in vivo. Proc. Natl. Acad. Sci. USA. 98, 5856-5861. Folstein MF, Folstein SE, McHugh PR. Mini-mental state: a practical method for 15 grading the cognitive state of patients for the clinician. J. Psychiatr. Res. 1975; 12:189 198. Frears, E. R., Stephens, D. J., Walters, C. E., Davies, H., and Austen, B. M. (1999) The role of cholesterol in the biosynthesis of b-amyloid. NeuroReport 10, 1699-1705. 20 Friedhoff, L. T., Cullen, E. I., Geoghagen, N. S., and Buxbaum, J. D. (2001) Treatment with controlled-release lovastatin decreases serum concentrations of human p-amyloid (AP) peptide. Int. J. Neuropsychopharmacol. 4, 127-130. 25 Games D., Adams D., Alessandrini R., Barbour R., Berthelette P., Blackwell C., Carr T., Clemens J., Donaldson T., Gillespie F., Guido T., Hagopian S., Johnsonwood K., Khan K., Lee M., Leibowitz P., Lieberburg I., Little S., Masliah E., Mcconlogue L., Montoyazavala M., Mucke L., Paganini L., Penniman E., Power M., Schenk D., Seubert P., Snyder B., Soriano F., Tan H., Vitale J., Wadsworth S., Wolozin B., Zhao J., 30 NATURE, 1995, 373 (6514): 523-527. Gilgun-Sherki Y., Melamed E., Offen D., Neuropharmacology, 2001, 40 (8): 959-975. Golstein, H. and Schaaf, E., Helv. Chim. Acta, 1957, 57(23), 132. 35 Gudmundsson, K.J., Hinldey, J.M., Brieger, M.S., Drach, J.C. and Townsend, L.B., Synthetic Communications, 1997, 27(5), 861-870.

WO 2004/031161 PCT/AU2003/001303 - 158 Gurney M.E., Pu H.F., Chiu A.Y., Dalcanto M.C., Polchow C.Y., Alexander D.D., Caliendo J., Hentati A., Kwon Y.W., Deng H.X., Chen W.J., Zhai P., Sufit R.L., Siddique T., SCIENCE, 1994, 264 (5166): 1772-1775. 5 Hartmann, T. (2001) Cholesterol, As and Alzheimer's disease. Trends Neurosci. 24,S45-S48. Hertel, C., Terzi, E., Hauser, N., Jakob-Rotne, R., Seelig, J., and Kemp, J. A. (1997) 10 Inhibition of the electrostatic interaction between p-amyloid peptide and membranes prevents p-amyloid-induced toxicity. Proc. Natl. Acad. Sci. USA. 94, 9412-9416. Hsiao, K., Chapman, P., Nilsen, S., Eckman, C., Harigaya, Y., Younkin, S., Yang, F., Cole, G. (1996) Correlative memory deficits, AP elevation, and amyloid plaques in 15 transgenic mice SCIENCE; 274(5284):99-102. Huang X, Atwood CS, Hartshorn MA et al. The As peptide of Alzheimer's disease directly produces hydrogen peroxide through metal ion reduction. Biochemistry 1999; 38:7609-7616. 20 Iyer, R.N. and Dhar, M.L., J. Sci. Ind. Res., 1956, 15C, 1-7. Ji, S. R., Wu, Y., and Sui, S. F. (2002) Cholesterol is an important factor affecting the membrane insertion of p-amyloid peptide (AP 1-40), which may potentially inhibit the 25 fibril formation. J. Biol. Chem. 277,6273-6279. Karbownik M., Lewinski A., Reiter R.J., Int. J. Biochemistry & Cell Biology, 2001, 33 (8): 735-753. 30 Lee J-Y, Cole TB, Palmiter RD, Suh SW, Koh J-Y. Contribution by synaptic zinc to the gender-disparate plaque formation in human Swedish mutant APP transgenic mice. Proc Natl Acad Sci U S A 2002: Early edition. Lee, S.J., Konishi, Y., Yu, D.T. et al, J Med. Chem., 1995, 38, 3547-3557. 35 Linderberg, M., Hellberg, S., Bjork, S. et al, Eur. J Med. Chem., 1999, 34, 729-744.

WO 2004/031161 PCT/AU2003/001303 - 159 Manfredini S, Pavan B, Vertuani S, Scaglianti M, Compagnone D, Biondi C, Scatturin A, Tanganelli S, Ferraro L, Prasad P, Dalpiaz A, JOURNAL OF MEDICINAL CHEMISTRY, 45 (3): 559-562 JAN 31 2002 5 Marabout, B., Sevrin, M. and Estenne, B.G., (1999) Patent FR 2,765,582. Masliah E., Rockenstein E., Veinbergs I., Mallory M., Hashimoto M., Takeda A., Sagara Y., Sisk A., Mucke L., SCIENCE, 2000, 287 (5456): 1265-1269. 10 McKhann G, Drachman D, Folstein MF, Katzman R, Price D, Stadlen E. Clinical diagnosis of Alzheimer's disease: Report of the NINCDS-ADRDA work group under the auspices of the Department of Health and Human Services Task Force on Alzheimer's Disease. Neurology 1984; 34:939-944. 15 Nunan, J., and Small, D. H. (2000) Regulation of APP cleavage by aX-, P- and S secretases. FEBS Lett. 483, 6-10. Philips' Gloeilampenfabrieken, N.V., (1973) Patent GB 1303711 20 Petersen, R.C, Stevenas, J.C., Ganguli, M., Tangalos, E.G., Cummings, J.L., and DeKosky, S.T. Practice parameter: Early detection of dementia: Mild cognitive impairment Neurology 2001 56 1133-1142. Reddy P.H., Williams M., Charles V., Garrett L., Pike-Buchanan L., Whetsell W.O., 25 Miller G., Tagle D.A., NATURE GENETICS, 1998, 20 (2): 198-202. Rogers SL, Farlow MR, Doody RS, Mohs R, Friedhoff LT. A 24-week, double-blind, placebo-controlled trial of donepezil in patients with Alzheimer's disease. Donepezil Study Group. Neurology 1998; 50:136-45. 30 Rosen WG, Mohs RC, Davis KL. A new rating scale for Alzheimer's disease. Am J Psychiatry 1984; 141:1356-64. Sakaeda T, Tada Y, Sugawara T, Ryu T, Hirose F, Yoshikawa T, Hirano K, 35 Kupczyk-Subotkowska L, Siahaan TJ, Audus KL, Stella VJ, JOURNAL OF DRUG TARGETING, 9 (1): 23-37 2001.

WO 2004/031161 PCT/AU2003/001303 - 160 Schenk, D., Barbour, R., Dunn, W., Gordon, G., Grajeda, H., Guido, T., Hu, K., Huang, J., Johnson-Wood, K., Khan, K., Kholodenko, D., Lee, M., Liao, Z., Lieberburg, I., Motter, R., Mutter, L., Soriano, F., Shopp, G., Vasquez, N., Vandervert, C., Walker, S., Wogulis, M., Yednock, T., Games, D., and Seubert, P. (1999) Immunization with 5 amyloid-p attenuates Alzheimer's disease like pathology in the PDAPP mouse. Nature 400, 173-177. Selkoe, D.J. Alzheimer's disease: genes, proteins and therapy. Physiol Rev 81 (2): 741-766. 10 Shearman MS, Beher D, Clarke EE et al. L-685,458, an aspartyl protease transition state mimic, is a potent inhibitor of amyloid p-protein precursor p-secretase activity. Biochemistry 2000; 29:8698-704. 15 Shiraki, H. The neuropathology of subacute myelo-optico-neuropathy (SMON) in the humans: With special reference to the quinoform intoxication. Jpn J Med Sci Biol 1975; 28 (suppl): 101-164. Simons M, Schwarzler F, Lutjohann D et al. Treatment with simvastatin in 20 normocholesterolemic patients with Alzheimer's disease: a 26-week randomised, placebo-controlled, double-blind trial. Ann of Neurol In Press. Sinha S, Anderson JP, Barbour R et al. Purification and cloning of amyloid precursor protein p-secretase from human brain. Nature 1999;402:537-40. 25 Smirnov, L.D., Nikitin, S.V., Chernyshev, A.I., Sorokin, A.A., Lezina, V.P., Zabrodnyaya, V.G. and Kaganskii, M.M., Chemistry of Heterocyclic Compounds, 1992, 12, 1425-1431. 30 St George-Hyslop, P.H. (2000) Molecular genetics of Alzheimer's disease. Bio. Psychiatry 47, 183-199. T.C. Wang, Y.L. Chen, K.H. Lee and C.C. Tzeng, Tetrahedron Lett., 1996, 37, 6369 6370. 35 Telling G.C., Scott M., Hsiao K.K., Foster D., Yang S.L., Torchia M., Sidle K.C.L., Collinge J., Dearmond S.J., Prusiner S.B., PROCEEDINGS OF THE NATIONAL WO 2004/031161 PCT/AU2003/001303 - 161 ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 11 Oct 1994, 91 (21): 9936-9940. Tani, J., Yamada, Y., Oine, T. et al, J. Med. Chem., 1979, 22, 95-99. 5 Valdez-Gonzalez, T., Inagawa, J., and Ido, T. (2001) Neuropeptides interact with glycolipid receptors: a surface plasmon resonance study. Peptides 22; 1099-1106. Wepplo, P.J., (1984) US Patent 4,460,776. 10 White et al., J Neuroscience, (1998) 18, 6207-6217. Wright, J.S. Johnson, E.R. and DiLabio, G.A. J.Am.Chem.Soc 2001 123 1173-1183. 15 Yale, H.L. and Sheehan, J.T. (1977) US Patent 4,022,897. Yassin MS, Ekblom J, Xilinas M, Gottfries CG, Oreland L. Changes in uptake of vitamin B(12) and trace metals in brains of mice treated with clioquinol. J Neurol Sci 2000; 173:40-44. 20 It will be apparent to the person skilled in the art that while the invention has been described in some detail for the purposes of clarity and understanding, various modifications and alterations to the embodiments and methods described herein may be made without departing from the scope of the inventive concept disclosed in this 25 specification.

Claims (20)

1. A method for the treatment, amelioration and/or prophylaxis of a neurological condition which comprises the administration of an effective amount of a compound of 5 formula I: (R 1 )m N (R 1 )m 10 RH IA in which R is O or S; R' is independently selected from H, optionally substituted alkyl, optionally is substituted alkenyl; optionally substituted alkynyl; optionally substituted aryl; optionally substituted heterocyclyl; an antioxidant; a targeting moiety; CN; halo; CF 3 ; SO 3 H; and OR 2 , SR 2 , SOR 2 , S0 2 R 2 , NR 2 R 3 , (CH 2 )nNR 2 R 3 , HCNOR 2 , HCNNR 2 R 3 , CONR 2 R 3 , CSNR 2 R 3 , NCOR 2 , NCSR 2 , COR 2 , C0 2 R 2 , CSR 2 or SO 2 NR 2 R 3 in which R 2 and R 3 are independently selected from H, optionally substituted alkyl, optionally 20 substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heterocyclyl, an antioxidant or a targeting moiety and n is an integer of I to 10; W is CH, N or NH; U is CH, CO or N; 25 Y is absent or together with the ring to which it is attached forms a 5- or 6 membered optionally substituted aryl or a 5- or 6-membered optionally substituted heterocyclyl; Y' is absent or together with the ring to which it is attached forms a 6-membered N-containing optionally substituted heterocyclyl; and 30 m is an integer from I to 3; salts, hydrates, solvates, derivatives, pro-drugs, tautomers and/or isomers thereof to a subject in need thereof, in which the prodrug is a Ci. 6 alkyl ester or aryl ester moiety located at position 8; 35 with the provisos that: (i) at least one of W and U is other than CH; 2114997_2 (GHMattero) 22/12/09 - 163 (ii) phanquinone or tautomers thereof are excluded i.e., when R is 0, R at position 7 is OH, W and are CH and Y is absent, then Y' is not N 5 (iii) when R is 0, Y is absent, W is CH other than at position 3 Me 10 where W is N, m is 2 and R' is -NH H N_ 20 at position 3, then R 1 at position 2 is not -s- NHAc or - S / NH-S 2 Me ; and (iv) clioquinol i.e, when R is 0, Y and Y' are absent, 25 W and U are CH and m is 2, then R' at position 5 is not chloro and R' at position 7 is not iodo.

2. A method according to claim 1, in which the compound of formula I is selected from the following: 30 0 N8-hydroxy-4(3 H)-q u inazol inones; R OH (R')m.R) 35 I~V'' 3

8-hydroxy-quinazoline; OH (R').R (RI), 8-hydroxy-quinoxaline; N 40 OH 2114997_2 (GHMattera) 22/12/09 - 164 N4 1- [6lnanhthvridin-8-o[; OH 5 -N~\ 9-hydroxypyrimido[ ,6-a]pyrimidin-4-one; OH (R'), 10 (R( , NN 8-hydroxy-cinnoline; OH 15 (RR)V. (R) 4-hydroxy-acridine; OH 20 () M.' ... 4,7(4, 1O)-phenanthrolin-5-oI; OH OH 25 (R 1 ). OH 0 30 N N R ' N\ 9-hydroxypyrido[ 1, 2-a] pyri m id in -4-one; (R')( ~ (R N OH (R) - 165 (R), OH N ( ') [l,5 ]n a p h th y rid in -4 ,8 -d io l; OH 5 )N N R) pyrido[3,4-b]pyrazin-8-ol; N OH 10 () quinaZolin-8-Ol; OH 15 in which R 1 is as defined in claim 1, p is an integer of I to 4 and q is an integer of I or 2. 3. A method according to claim I or 2 in which the compound of formula IA is selected from the following: 20 (i) Formula la N (R 1 )m (R) M N 25 RH Ia in which R, R 1 , m and q are as defined in claim I or 2; (ii) Formula lb 30 Nz (R 1 )m(R g N 35 RH Ib in which R, R', m and q are as defined in claim I or 2; 21499' 2 (GHMattera) 22/12/09 - 166 (iii) Formula Ic N (R')m (R 1 )q 5 N> RH Ic in which R, R', m and q are as defined in claim I or 2; 10 (iv) Formula Id (R 1 )q N ~ ~ (R) 15 N RH Id 20 in which R, R', m and q are as defined in claim I or 2; (v) Formula le 0 N RI)q 25 RH in which R, R', m and q are as defined in claim I or 2; and (vi) Formula If 30 (R 1 )m N (R 1 )m R1 N 35 OH in which R' and m are as defined in claim I or 2. 2114997_ 1 (GKMattere) 10/11/09 - 167 4. A method according to any one of claims I to 3 in which R in the compound of formula I is 0. 5. A method according to any one of claims I to 4 in which R' in the compound of 5 formula I is halo, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted alkyl, OR 2 , SR 2 , (CH 2 )nNR 2 R 3 , CONR 2 R 3 and NCOR 2 in which n, R 2 and R 3 are as defined in claim 1. 6. A method according to any one of claims I to 5 in which R' in the compound of 10 formula I is fluoro, iodo, chloro, optionally substituted phenyl, an optionally substituted unsaturated 3 to 6-membered heteromonocyclic group containing 1 to 4 nitrogen atoms, an optionally substituted saturated 3 to 6-membered heteromonocyclic group containing I to 4 nitrogen atoms, an optionally substituted saturated 3 to 6-membered heteromonocyclic group containing I to 2 oxygen atoms and I to 3 nitrogen atoms, 15 optionally substituted C 1 . 4 alkyl, optionally substituted C 2 . 6 cycloalkyl, optionally substituted CI. 6 alkoxy, optionally substituted thio, CH 2 NR 4 R' in which R 4 and R 5 are independently selected from H and C 14 alkyl or CONH(CH 2 ) 2 R 6 in which R 6 is optionally substituted heterocyclyl. 20 7. A method according to any one of claims I to 6 in which Y in the compound of formula I is an optionally substituted phenyl, an optionally substituted unsaturated 5- or 6-membered heteromonocyclic group containing I to 4 nitrogen atoms or an optionally substituted saturated 5 or 6-membered heteromonocyclic group containing I to 2 oxygen atoms and I to 3 nitrogen atoms. 25 8. A method according to any one of claims I to 7, in which the compound of formula I is as follows: 3ci o NF /C ON 3 0 N , CI N CI N OH OH 1055 1061 35 2114997_1 (GHMatters) 10/11/09 168 (0 0 cO N N N C1 NC C Ne N N OH ~OH Ne OH OH C1 CI 10 N '1 N 0 CI N CI N OH OH NMe 2 15 ci 1066 ci N N c3 1N cI ON 20 OH OH 1064 1065 c 1 N' 25 N N ~ ) 0MeOOC N 0j OH 1053 1045 30 HN N N, 351070 0 H HN 2114997_ 1 IGHMttO) 10/11/09 - 169 CI N CI 5 1 N OH 1063 N O OH 10 1069 N OH 15 1048 N N N 20 C N' C1 N1 N OH OH NMe 2 OH 1026

9. Use according to any one of claims I to 8, in which the neurological condition is acute intermittent porphyria; adriamycin-induced cardiomyopathy; AIDS dementia and 25 HIV-1 induced neurotoxicity; Alzheimer's disease; amylotrophic lateral sclerosis; atherosclerosis; cataract; cerebral ischaemia; cerebral palsy; cerebral tumour; chemotherapy-induced organ damage; cisplatin-induced nephrotoxicity; coronary artery bypass surgery; Creutzfeldt-Jacob disease and its new variant associated with "mad cow" disease; diabetic neuropathy; Down's syndrome; drowning; epilepsy and post 30 traumatic epilepsy; Friedrich's ataxia; frontotemporal dementia; glaucoma; glomerulopathy; haemochromatosis; haemodialysis; haemolysis; haemolytic uraemic syndrome (Weil's disease); haemorrhagic stroke; Hallerboden-Spatz disease; heart attack and reperfusion injury; Huntington's disease; Lewy body disease; intermittent claudication; ischaem ic stroke; inflammatory bowel disease; macular degeneration; 35 malaria; methanol-induced toxicity; meningitis (aseptic and tuberculous); motor neuron disease; multiple sclerosis; multiple system atrophy; myocardial ischaemia; neoplasia; Parkinson's disease; peri-natal asphyxia; Pick's disease; progressive supra-nuclear 2114997_1 (GHMatters) 10/11/09 - 170 palsy; radiotherapy-induced organ damage; restenosis after angioplasty; retinopathy; senile dementia; schizophrenia; sepsis; septic shock; spongiform encephalopathies; subharrachnoid haemorrage/cerebral vasospasm; subdural haematoma; surgical trauma, including neurosurgery; thalassemia; transient ischaemic attack (TIA); traumatic brain 5 injury (TBI); traumatic spinal injury; transplantation; vascular dementia; viral meningitis; viral encephalitis; spinal cord injury; dementia with Lewy body formation; fatal familial insomnia; Gertsmann Straussler Sheinker disease; hereditary cerebral haemorrhage with amyoidoisis-Dutch type; dementia; or Mild Cognitive Impairment (MCI). 10

10. A method according to any one of claims I to 9, in which the neurological condition is a neurodegenerative disorder. I1. A method according to claim 10, in which the neurodegenerative disorder is 15 neurodegenerative amyloidosis.

12. A method according to claim 10 or claim 11, in which the neurodegenerative disorder is sporadic or familial Alzheimer's disease, amyotrophic lateral sclerosis, cataract, Parkinson's disease, Creutzfeldt-Jacob disease and its new variant associated 20 with "mad cow" disease, Huntington's disease, dementia with Lewy body formation, multiple system atrophy, Hallerboden-Spatz disease, diffuse Lewy body disease, fatal familial insomnia, Gertsmann Straussler Sheinker disease, hereditary cerebral haemorrhage with amyloidosis-Dutch type, multiple sclerosis, tauopathies, motor neuron disease or prion diseases. 25

13. A method according to claim 12, in which the neurodgenerative disorder is Parkinson's disease.

14. A method according to any one of claims 10 to 12, in which the 30 neurodegenerative disorder is an Ap-related condition.

15. A method according to claim 14, in which the Ap-related condition is Alzheimer's disease or dementia associated with Down syndrome or one of several forms of autosomal dominant forms of familial Alzheimer's disease. 35

16. A method according to any one of the preceding claims which slows, reduces or arrests the cognitive decline of the subject. 2114997_1 (GHMattera) 10/11/09 - 171 17. A method according to any one of the preceding claims, which further comprises separate, sequential or simultaneous administration of another medicament. 5 18. A method according to claim 17, in which the other medicament is an inhibitor of the acetylcholinesterase active site, an antioxidant, an anti-inflammatory agent or an oestrogenic agent.

19. A method according to any one of the preceding claims, in which the compound 10 of formula IA is administered orally, topically or parenterally.

20. Use of the compound of formula IA as defined in any one of claims I to 8, in the manufacture of a medicament for the treatment, amelioration and/or prophylaxis of a neurological condition. 15

21. Use of a compound of formula IA as defined in any one of claims I to 8 for the treatment, amelioration and/or prophylaxis of a neurological condition.

22. A compound of formula IA as defined in claims I to 8 for use in the treatment, 20 amelioration and/or prophylaxis of a neurological condition.

23. Use of the compound of formula IA as defined in any one of claims I to 8, as a pharmaceutical. 25 24. Use according to claim 23, in which the pharmaceutical is a neurotherapeutic or neuroprotective agent.

25. Use according to claim 23 or claim 24, in which the pharmaceutical is an antiamyloidogenic agent. 30

26. A pharmaceutical or veterinary composition comprising the compound of formula I as defined above in any one of claims I to 8 and a pharmaceutically or veterinarily acceptable carrier. 35 27. A composition according to claim 26 which further comprises another medicament. 2114997_1 (GHMattere) 10/11/09 - 172 28. A composition according to claim 27, in which the other medicament is an inhibitor of the acetyicholinesterase active site, an antioxidant, an anti-inflammatory agent or an oestrogenic agent. 5 29. A compound of formula IA as defined in any one of claims I to 8, with the further proviso that at least one R' is other than H and 1049 is excluded.

30. Compounds of the formulae la, Ib, Ic, Id, le and If as defined in claim 3. 10 31. A compound as defined in claim 8 excluding 1048, 1026 and 1045.

32. A process for the preparation of the compound of formula IA defined in claim 29 as described herein. 15 33. A compound of the formula: CI COOH CI NO 2 20 NHAc

34. Compounds of formula IA, processes for their preparation, pharmaceutical or veterinary compositions containing them or methods or uses involving them, 25 substantially as herein described with reference to the examples and/or drawings. 30 2114997_2 (GHMattero) 22/12/09