US008329697B2

(12) United States Patent (10) Patent No.: US 8,329,697 B2 Garbaccio et al. (45) Date of Patent: Dec. 11, 2012

(54) IMIDIZO 1.2-APYRAZINES USEFUL AS (56) References Cited AHCY HYDROLASE INHIBITORS U.S. PATENT DOCUMENTS (75) Inventors: Robert M. Garbaccio, Lansdale, PA 4,507,294 A 3, 1985 Bristol et al. (US); Antonella Converso, Elkins Park, 56.6 f 3.3: Ret , al PA (US); Mark E. Fraley, North Wales, 4. W w Wa (a. PA (US); Timothy J. Hartingh, Blue 2004/0204429 A1 10, 2004 Yuan Bell, PA (US) FOREIGN PATENT DOCUMENTS WO WO 2009/108546 * 9/2009 (73) Assignee: Merck Sharp & Dohme Corp., OTHER PUBLICATIONS Rahway, NJ (US) Jordan, V. C. Nature Reviews: Drug Discovery, 2, 2003, 205.* (*) Notice: Subject to any disclaimer, the term of this Hackam, et al. JAMA, 296(14), 2006, 1731-1732.* patent is extended or adjusted under 35 * cited b U.S.C. 154(b) by 163 days. c1ted by examiner Primary Examiner — Douglas MWillis (21) Appl. No.: 12/811,911 (74) Attorney, Agent, or Firm — Kenrick L. Vidale; John C. Todaro (22) PCT Filed: Feb. 18, 2009 (57) ABSTRACT (86). PCT No.: PCT/US2O09/034344 The present invention is directed to AHCY inhibitors of for mula (I) which are useful in the treatment of diseases charac S371 (c)(1), terized by high homocysteine levels, such as Alzheimer's (2), (4) Date: Jul. 7, 2010 disease. The invention is also directed to pharmaceutical compositions comprising the compounds, and to the use of (87) PCT Pub. No.: WO2009/108546 the compounds and compositions in the treatment of diseases

PCT Pub. Date: Sep.e 3,a 9 2009 characterized by high homocysteine levels.

(65) Prior Publication Data (I) US 2010/0305135 A1 Dec. 2, 2010 NH2 Related U.S. Application Data N21 N2N (60) ProvisionalrOV1S1Onal apol1Cat1Onapplication No.NO. 61/067,210,Z 1U, Tilledfiled onOn Feb.Feb N-N / 26, 2008. OH (51) Int. Cl. X A6 IK3I/495 (2006.01) Y (52) U.S. Cl...... 514/249; 544/350 6H (58) Field of Classification Search ...... 514/249; 544/350 See application file for complete search history. 7 Claims, No Drawings US 8,329,697 B2 1. 2 IMIDIZO 1.2-APYRAZINES USEFUL AS 2003 207):425-8. Further, the known AHCY inhibitor AHCY HYDROLASE INHIBITORS 3-deaza-adenosine (DZA) has been shown to prevent oxida tive damage and cognitive impairment in mice. Shea et al. CROSS-REFERENCE TO RELATED Neuromolecular Medicine 2004, 5:173-182. In clinical stud APPLICATIONS ies, folate deficiency was associated with neurological disor derS Such as Alzheimer's disease. Ho et al., Neurobiology of This application claims the benefit under 35 U.S.C. 119(e) Disease, 2003 14:1, 32-42. of U.S. Provisional Application No. 61/067,210, filed 26 Feb. Seshadri et al, N Engl. J. Med, 2002, 346:476-483, in a 2008. study of data from the Framingham Heart Study, found that 10 increased homocysteine levels in plasma was an independent BACKGROUND OF THE INVENTION risk factor for dementia and Alzheimer's Disease. See also 1. Field of the Invention Morris, Lancet Neurology 2003, 2:425-428. The invention is directed to a class of S-adenosylhomocys Hyperhomocysteinemia is a known risk factor for arterial teine hydrolase (AHCY) inhibitors, their salts, pharmaceuti 15 vascular disease and venous thrombosis. Gellekink et al., Eur cal compositions comprising them and their use in therapy of J Hum Genet. 2004 12(11):942-8; cardiovascular disease, the human body. In particular, the invention is directed to a Levine etal, Prog Neuropsych Biol Psych 2005, 29(7): 1181 class of AHCY inhibitors, which are useful in the treatment of 91; Schizophrenia, Haidemenos et al. Prog Neuropsychop diseases characterized by high homocysteine levels. harmacol Biol Psychiatry. 2007 15:31(6):1289-96; and bipo 2. Description of Related Art lar disorder, Levine et al. Elevated homoscysteine levels have S-adenosylhomocysteine, known as AdoHcy, is an inter also been shown to be risk factors for stroke and Parkinson's mediate in the metabolism of the Sulfur-containing amino Disease. Herrmann et al. Fortschr Neurol Psychiatr. 2007 acids methionine and cysteine. AdoHcy is formed by the 75(9):515-27. Further, animal studies suggest that increased donation of a methyl group from S-adenosylmethionine homocycsteine levels may be a factor in osteoporosis. Her (SAM) to biomolecules undergoing methylation reactions. 25 rmann etal, Clin. Chem. 2007, 53(8): 1455-61. AdoHcy is then metabolized by the enzyme S-adenosylho One possible method for treating diseases via the AHCY mocysteine hydrolase, known as AHCY, SAHH or SAH pathway is to develop mechanisms for clearing homocysteine hydrolase, which reversibly hydrolyses AdoHcy to adenosine from the body. For example, compounds or Substances that and homocysteine. Homocysteine can be remethylated back increase the rate of vitamin B clearance of homocysteine in to methionine or undergo a series of metabolic steps leading 30 Vivo may find utility as agents for treating disease associated to the biosynthesis of glutathione or cysteine. Following the with high levels of homocysteine. A second method of inter hydrolysis of AdoHcy, homocysteine can also be secreted fering with the AHCY pathway is to decrease production of from the body or converted into the anti-oxidant, glutathione, homocytseine, i.e. to develop compounds that can inhibit by a series of transulfuration pathway reactions. Glutathione production of homocysteine in vivo. For example, com is a major anti-oxidant in the body. The relative ratio between 35 pounds or substances which inhibit AHCY may decrease the oxidized and reduced forms of glutathione is thought to be an extent of hydrolysis of S-adenosyl homocysteine into important indicator of oxidative state. homocysteine and adenosine. Furthermore, the ratio between SAM and AdoHcy is criti The inventors have identified a novel group of compounds cal for many biological processes, as AdoHcy can inhibit which act as inhibitors of S-adenosyl homocysteine hydro many methyltransferases that use SAM as a methyl donor. 40 lase, thereby inhibiting the hydrolysis of S-adenosyl Thus, the rate of conversion of AdoHcy to Hey is a critical homocysteine into homocysteine and adenosine. regulator of many biological reactions involving phospholip ids, proteins, and nucleic acids. Chiang, Pharmacol Ther BRIEF SUMMARY OF THE INVENTION 1998, 77, 2, 115-134. Various nucleosides and nucleoside derivatives act as inhibitors of AHCY. Chiang. 45 The present invention is directed to novel compounds of Homocysteine metabolism is also dependent on the nutri generic formula (I) ents folate, vitamin B12 and vitamin B6. Obeid etal, FEBS Letters 2006, 580:2994-3005. These nutrients are cofactors for the enzymes that remethylate Hey back to methionine NH2 (folate, B12) or convert it to glutathione (B6). 50 AHCY is a 432 amino acid protein, which is a thioether N 21 2N. hydrolase. AHCY is a cytosolic enzyme that has been found in a wide variety of cells. Walker, et al. Can. J. Biochem. 1975 N- N / 53: 312-319. The sequence of AHCY is disclosed in Interna tional Patent Application Publication No. WO 2005/015221. 55 wOH X AHCY catalyzes the conversion of S-adenosyl-homocys V teine to homocysteine and adenosine. Because of the key role Y of AdoHcy in the synthesis of cysteine, and the role of S-ad enosylmethionine as a universal methyl donor, misregulation OH of AHCY can affect methylation of phosphlipids, proteins, 60 DNA and RNA. orpharmaceutically acceptable salts thereof, which are useful Epidemiological evidence demonstrates that increased lev as AHCY inhibitors. els of homocysteine are associated with many diseases, The invention is further directed to methods of treating a including cardiovascular disease, stroke, and neurodegenera patient (preferably a human) for diseases or disorders which tive diseases such as Alzheimer's Disease. Hyperhomocys 65 are characterized by high homocysteine levels, such as Alzhe teinemia, which may be caused by folic acid deficiency, can imer's disease, by administering to the patient a therapeuti contribute to Alzheimer's Disease. Morris, Lancet Neurol. cally effective amount of a compound of general formula (I), US 8,329,697 B2 3 4 or a pharmaceutically acceptable salt thereof. The invention is The invention is also directed to medicaments or pharma also directed to pharmaceutical compositions which include ceutical compositions for treating diseases or disorders char an effective amount of a compound of formula (I), or a phar acterized by high homocysteine levels, such as Alzheimer's maceutically acceptable salt thereof, and a pharmaceutically disease, hypertension, cardiovascular disease, stroke and acceptable carrier, and the use of the compounds and phar 5 Schizophrenia, which comprise a compound of formula (I), or maceutical compositions of the invention in the treatment of a pharmaceutically acceptable salt thereof, and a pharmaceu Such diseases. tically acceptable carrier. The invention is further directed to a method for the manu DETAILED DESCRIPTION OF THE INVENTION facture of a medicament or a composition for treating diseases 10 or disorders characterized by high homocysteine levels, such as Alzheimer's disease, hypertension, cardiovascular disease, In one embodiment, the invention is directed to compounds stroke or Schizophrenia, comprising combining a compound of general formula (I) of formula (I) with one or more pharmaceutically acceptable carriers. NH2 15 Specific embodiments of formula (I) are described herein as Examples 1 ((1S,2R,5S)-5-(8-aminoimidazo[1,2-a pyrazin-3-yl)-3-(hydroxymethyl)cyclopent-3-ene-1,2-diol) N 21 2N and 2 (1R,2S,3S)-3-(8-aminoimidazo[1,2-alpyrazin-3-yl)cy clopentane-1,2-diol, and pharmaceutically acceptable salts N- N / thereof. Where a variable occurs more than once in Formula (I) or X WOH in a substituent thereof, the individual occurrences of that V variable are independent of each other, unless otherwise Y specified. OH 25 As used herein, in particular in the definitions of R', the term “alkyl by itself or as part of another substituent, means a saturated Straight or branched chain hydrocarbon radical and pharmaceutically acceptable salts thereof, wherein having the number of carbon atoms designated (e.g., Co X-Y is selected from the group consisting of alkyl means an alkyl group having from one to ten carbon (1) CH, CRR, 30 atoms). Preferred alkyl groups for use in the invention are (2) CH-CR'; C. alkyl groups, having from one to six atoms. Exemplary R" is selected from the group consisting of alkyl groups include methyl, ethyl, n-propyl, isopropyl. n-bu (1) hydrogen, tyl, isobutyl, tent-butyl, pentyl, hexyl, and the like. Co alkyl (2) hydroxyl, means a bond. (3) —C alkyl, wherein said alkyl is optionally substi 35 As used herein, the term “halo' or “halogen includes tuted with fluoro, chloro, bromo and iodo. (a) halogen, or The compounds of the invention may have one or more (b) hydroxyl; and asymmetric centers. Compounds with asymmetric centers R is selected from the group consisting of give rise to enantiomers (optical isomers), diastereomers (1) hydrogen, 40 (configurational isomers) or both, and it is intended that all of (2) hydroxyl, the possible enantiomers and diastereomers in mixtures and (3) —C alkyl, wherein said alkyl is optionally substi as pure or partially purified compounds are included within tuted with the scope of this invention. The present invention is meant to (a) halogen, or encompass all such isomeric forms of the compounds of (b) hydroxyl. 45 formula (I). In one embodiment, R' is selected from the group consist Formula (I) is shown above without a definite stereochem ing of istry at certain positions. The present invention includes all (1) hydrogen, and Stereoisomers of formula (I) and pharmaceutically acceptable (2) —C alkyl, wherein said alkyl is optionally substi salts thereof. tuted with 50 The independent syntheses of the enantiomerically or dias (a) halogen, or tereomerically enriched compounds, or their chromato (b) hydroxyl. graphic separations, may be achieved as known in the art by In another embodiment, R' is selected from the group appropriate modification of the methodology disclosed consisting of herein. Their absolute stereochemistry may be determined by (1) hydrogen, and 55 the X-ray crystallography of crystalline products or crystal (2) —CHOH. line intermediates that are derivatized, if necessary, with a In one embodiment, the invention is directed to methods of reagent containing an asymmetric center of known absolute treating a patient (preferably a human) for diseases charac configuration. terized by high homocysteine levels, such as Alzheimer's If desired, racemic mixtures of the compounds may be Disease, hypertension, cardiovascular disease, stroke and 60 separated so that the individual enantiomers or diastereomers Schizophrenia, by administering to the patient a therapeuti are isolated. The separation can be carried out by methods cally effective amount of a compound of general formula (I). well known in the art, Such as the coupling of a racemic The invention is also directed to the use of a compound of mixture of compounds to an enantiomerically pure com formula (I) for treating diseases or disorders characterized by pound to form a diastereomeric mixture, followed by separa high homocysteine levels. Such as Alzheimer's disease, 65 tion of the individual diastereomers by standard methods, hypertension, cardiovascular disease, stroke and Schizophre Such as fractional crystallization or chromatography. The 18. coupling reaction is often the formation of salts using an US 8,329,697 B2 5 6 enantiomerically pure acid or base. The diastereomeric Alzheimer's disease. Other conditions that may be treated by derivatives may then be converted to the pure enantiomers by the compounds of the invention include Parkinson's Disease, cleavage of the added chiral residue. The racemic mixture of hypertension, Schizophrenia, including the positive and nega the compounds can also be separated directly by chromato tive symptoms of Schizophrenia and treatment of cognitive graphic methods using chiral stationary phases, which meth impairment due to schizophrenia, bipolar disorder, cancer, ods are well known in the art. cardiovascular disease, viral infections and osteoporosis. Alternatively, any enantiomer or diastereomer of a com In certain embodiments, the compounds of the invention pound may be obtained by Stereoselective synthesis using are useful in treating Alzheimer's Disease. For example, the optically pure starting materials or reagents of known con compounds may be useful for the prevention of dementia of figuration by methods well known in the art. 10 During any of the above synthetic sequences it may be the Alzheimer's type, as well as for the treatment of early necessary or desirable to protect sensitive or reactive groups stage, intermediate stage or late stage dementia of the Alzhe on any of the molecules concerned. This may be achieved by imer's type. means of conventional protecting groups, such as those Potential schizophrenia conditions or disorders for which described in Protective Groups in Organic Chemistry, ed. J. F. 15 the compounds of the invention may be useful include one or W. McOmie, Plenum Press, 1973, and T. W. Greene & Wuts, more of the following conditions or diseases: Schizophrenia Protective Groups in Organic Synthesis, John Wiley & Sons, or psychosis including Schizophrenia (paranoid, disorga 1991. The protecting groups may be removed at a convenient nized, catatonic or undifferentiated), Schizophreniform dis sequent stage using methods known from the art. order, schizoaffective disorder, delusional disorder, brief psy The term “substantially pure” means that the isolated mate chotic disorder, shared psychotic disorder, psychotic disorder rial is at least 90% pure, and preferably 95% pure, and even due to a general medical condition and Substance-induced or more preferably 99% pure as assayed by analytical tech drug-induced (phencyclidine, and other dissocia niques known in the art. tive anaesthetics, amphetamine and other psychoStimulants The term “pharmaceutically acceptable salts' refers to and ), psychosispsychotic disorder, psychosis associ salts prepared from pharmaceutically acceptable non-toxic 25 ated with affective disorders, brief reactive psychosis, bases or acids including inorganic or organic bases and inor Schizoaffective psychosis, 'schizophrenia-spectrum' disor ganic or organic acids. The compounds of the invention may derS Such as Schizoid or schizotypal personality disorders, or be mono, di or tris Salts, depending on the number of acid illness associated with psychosis (such as major depression, functionalities present in the free base form of the compound. manic depressive (bipolar) disorder, Alzheimer's disease and Free bases and salts derived from inorganic bases include 30 post-traumatic stress syndrome), including both the positive aluminum, ammonium, calcium, copper, ferric, ferrous, and the negative symptoms of Schizophrenia and other psy lithium, , manganic salts, manganous, potassium, choses; cognitive disorders including dementia (associated Sodium, Zinc, and the like. with Alzheimer's disease, ischemia, multi-infarct dementia, Salts in the Solid form may exist in more than one crystal trauma, vascular problems or stroke, HIV disease, Parkin structure, and may also be in the form of hydrates. Salts 35 son's disease, Huntington's disease, Pick's disease, derived from pharmaceutically acceptable organic non-toxic Creutzfeldt-Jacob disease, perinatal hypoxia, other general bases include salts of primary, secondary, and tertiary amines, medical conditions or Substance abuse); delirium, amnesiac Substituted amines including naturally occurring Substituted disorders or age related cognitive decline. amines, cyclic amines, and basic ion exchange resins, such as In one embodiment, the present invention provides a arginine, betaine, caffeine, choline, N,N'-dibenzylethylene 40 method for treating cognitive disorders, comprising: admin diamine, diethylamine, 2-diethylaminoethanol, 2-dimethy istering to a patient in need thereof an effective amount of a laminoethanol, ethanolamine, ethylenediamine, N-ethyl compound of the present invention. Particular cognitive dis morpholine, N-ethylpiperidine, glucamine, glucosamine, orders are dementia, delirium, aninestic disorders and age histidine, hydrabamine, isopropylamine, lysine, methylglu related cognitive decline. At present, the text revision of the camine, morpholine, piperazine, piperidine, polyamine res 45 fourth edition of the Diagnostic and Statistical Manual of ins, , purines, theobromine, triethylamine, trimethy Mental Disorders (DSM-IV-TR) (2000, American Psychiat lamine, tripropylamine, tromethamine, and the like. ric Association, Washington D.C.) provides a diagnostic tool When the compound of the present invention is basic, salts that includes cognitive disorders including dementia, may be prepared from pharmaceutically acceptable non-toxic delirium, amnestic disorders and age-related cognitive acids, including inorganic and organic acids. Such acids 50 decline. As used herein, the term “cognitive disorders' include acetic, trifluoroacetic, benzenesulfonic, benzoic, includes treatment of those mental disorders as described in camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, DSM-IV-TR. The skilled artisan will recognize that there are glutamic, hydrobromic, hydrochloric, isethionic, lactic, alternative nomenclatures, noSologies and classification sys maleic, malic, mandelic, methanesulfonic, mucic, nitric, tems for mental disorders, and that these systems evolve with pamoic, pantothenic, phosphoric, succinic, Sulfuric, tartaric, 55 medical and Scientific progress. Thus the term “cognitive p-toluenesulfonic acid, and the like. disorders' is intended to include like disorders that are The present invention is directed to the use of the com described in other diagnostic Sources. pounds of formula (I) disclosed herein as AHCY hydrolase Potential cardios vascular conditions or disorders for which inhibitors in a patient or Subject such as a mammal in need of the compounds of the invention may be useful include ath Such activity, comprising the administration of an effective 60 erosclerosis, hypertension, hyperlipidemia, coronary heart amount of the compound. In addition to humans, a variety of disease, peripheral vascular disease, dyslipidemia, hyperbe other mammals can be treated according to the method of the talipoproteinemia, hypoalphalipoproteinemia, hypercholes present invention. terolemia, hypertriglyceridemia, familial-hypercholester The compounds of the present invention have utility in olemia, angina, ischemia, cardiac ischemia, stroke, treating or ameliorating Alzheimer's disease. The com 65 myocardial infarction, reperfusion injury, angioplastic rest pounds may also be useful in treating or ameliorating other enosis, vascular complications of diabetes, obesity (including diseases characterized by high homocysteine levels, such as abdominal obesity) and endotoxemia. US 8,329,697 B2 7 8 The compounds of the invention may also be useful in the alone. Additionally, the compounds of the present invention treatment of the metabolic syndrome. According to one may be used in combination with one or more other drugs that widely used definition, a patient having metabolic syndrome treat, prevent, control, ameliorate, or reduce the risk of side is characterized as having three or more symptoms selected effects or toxicity of the compounds of the present invention. from the following group of five symptoms: (1) abdominal Such other drugs may be administered, by a route and in an obesity; (2) hypertriglyceridemia; (3) low high-density lipo amount commonly used therefor, contemporaneously or protein (HDL); (4) high blood pressure; and (5) sequentially with the compounds of the present invention. elevated fasting glucose, which may be in the range charac Accordingly, the pharmaceutical compositions of the present teristic of Type 2 diabetes if the patient is also diabetic. Each invention include those that contain one or more other active of these symptoms is defined clinically in the recently 10 ingredients, in addition to the compounds of the present released Third Report of the National Cholesterol Education invention. The combinations may be administered as part of a Program Expert Panel on Detection, Evaluation and Treat unit dosage form combination product, or as a kit or treatment ment of High Blood Cholesterol in Adults (Adult Treatment protocol wherein one or more additional drugs are adminis Panel III, or ATP III), National Institutes of Health, 2001, NIH tered in separate dosage forms as part of a treatment regimen. Publication No. 01-3670. Patients with metabolic syndrome 15 Examples of combinations of the compounds of the present have an increased risk of developing the macrovascular and invention include combinations with anti-Alzheimer's Dis microvascular complications that are listed above, including ease agents, for example beta-secretase inhibitors; alpha 7 atherosclerosis and coronary heart disease. nicotinic agonists, such as ABT089, SSR180711 and As stated above, the compounds of the invention may be MEM63908: ADAM10 ligands or activators; gamma-secre used for treating stroke. One class of stroke patients to which tase inhibitors, such as LY450139 and TAK 070; gamma a compound of the invention may be administered is a patient secretase modulators; tau phosphorylation inhibitors; glycine at risk for stroke. As used herein, the term “patient at risk for transport inhibitors; LXR Bagonists; ApoE4 conformational stroke’ means an individual who has had a previous stroke, or modulators; NR2B antagonists; androgen receptor modula has a risk factor for stroke. Known risk factors for stroke tors; blockers of AB oligomer formation; 5-HT4 agonists, include atherosclerosis, arterial hypertension, lipohyalinosis, 25 such as PRX-03140; 5-HT6 antagonists, such as GSK hyperlipidemia, hypercholesterolemia, atrial fibrillation, 742467, SOS-518, FK-962, SL-65.0155, SRA-333 and xalip Smoking, inflammatory markers (including C-reactive pro roden; 5-HT1a antagonists, such as lecozotan; p25/CDK5 tein), infection, homocysteine, sleep-disordered breathing, inhibitors; NK1/NK3 receptor antagonists: COX-2 inhibi cerebral autosomal dominant arteriopathy with subcortial inf tors; HMG-CoA reductase inhibitors; NSAIDs including ibu arcts and leuko-encephalopathy (CADASIL), migraine, 30 profen; vitamin E; anti-amyloid antibodies (including anti sickle-cell anemia, antiphospholipid antibody syndrome, amyloid humanized monoclonal antibodies). Such as arterial dissection, cocaine abuse and obesity. bapineuzumab, ACC001, CAD106, AZD3102, H12A11V1; A second class of patients to which a compound of the anti-inflammatory compounds such as (R)-flurbiprofen, invention may be administered are acute stroke patients, i.e., nitroflurbiprofen, ND-1251, VP-025, HT-0712 and EHT-202; patients who have suffered ischemic stroke within the last 7 35 PPARgamma agonists, such as pioglitaZone and rosiglita days. One class of acute stroke patients are those who have Zone; CB-1 receptor antagonists or CB-1 receptor inverse suffered stroke within the last 3 days. Another class of acute agonists, such as AVE 1625; antibiotics such as doxycycline stroke patients are those who have suffered stroke within the and rifampin: N-methyl-D-aspartate (NMDA) receptor last 48 hours, for example within the last 24 hours. As com antagonists, such as memantine, neramexane and EVT 101; mon in the art of treating stroke, patients may be classified 40 cholinesterase inhibitors such as galantamine, rivastigmine, according to the period of time when stroke occurred. So, for donepezil, tacrine, phenserine, ladostigil and ABT-089: example, one class of acute stroke patients are those who have growth hormone secretagogues such as ibutamoren, ibuta suffered stroke within the last 18 hours. Another class of acute moren mesylate, and capromorelin; histamine H receptor stroke patients are those who have suffered stroke within the antagonists such as ABT-834, ABT 829, GSK 1892.54 and last 12 hours. Another class of acute stroke patients are those 45 CEP16795; AMPA agonists or AMPA modulators, such as who have suffered stroke within the last 8 hours. Another CX-717, LY 451395, LY404187 and S-18986; PDE IV class of acute stroke patients are those who have suffered inhibitors, including MEM1414, HT0712 and AVE8112: stroke within the last 6 hours. Another class of acute stroke GABA inverse agonists; GSK3? inhibitors, including patients are those who have suffered stroke within the last 4 AZD1080, SAR502250 and CEP16805; neuronal nicotin ic hours. Another class of acute stroke patients are those who 50 agonists; selective M1 agonists; HDAC inhibitors; and micro have suffered stroke within the last 3 hours. tubule affinity regulating kinase (MARK) ligands; or other A third class of patients to which a compound of the present drugs that affect receptors or enzymes that either increase the invention may be administered are patients who have suffered efficacy, safety, convenience, or reduce unwanted side effects stroke more than 7 days previously, who are typically in need or toxicity of the compounds of the present invention. of restorative treatment. 55 Examples of other active ingredients that may be adminis The subject or patient to whom the compounds of the tered in combination with a composition of the present inven present invention is administered is generally a human being, tion, and either administered separately or in the same phar male or female, characterized by high homocysteine, but may maceutical composition, include agents for the treatment of also encompass other mammals, such as dogs, cats, mice, diabetic conditions, such as rats, cattle, horses, sheep, rabbits, monkeys, chimpanzees or 60 (a) insulin sensitizers including (i) PPARYagonists, such as otherapes or primates, for which treatment of the above noted the glitaZones (e.g. troglitaZone, pioglitaZone, englitaZone, disorders is desired. MCC-555, rosiglitazone, balaglitazone, and the like) and The compounds of the present invention may be used in other PPAR ligands, including PPARO/Y dual, agonists, such combination with one or more other drugs in the treatment of as KRP-297, muraglitazar, naveglitazar, tesaglitazar, and diseases or conditions for which the compounds of the 65 TAK-559; PPARC. agonists, such as fenofibric acid deriva present invention have utility, where the combination of the tives (gemfibrozil, clofibrate, fenofibrate and bezafibrate); drugs together are safer or more effective than either drug and selective PPARY modulators (SPPARyMs), such as dis US 8,329,697 B2 9 10 closed in WO 02/060388, WO 02/08188, WO 2004/019869, nolol, atenolol, bisoprolol, carvedilol, metoprolol, or meto WO 2004/020409, WO 2004/020408, and WO 2004/066963: prolol tartate), alpha adrenergic blocking drugs (e.g., (ii) biguanides, such as metformin and phenformin, and (iii) doxazocin, prazocin or alpha methyldopa) central alpha adr protein tyrosine phosphatase-1B (PTP-1B) inhibitors; energic agonists, and peripheral vasodilators (e.g. hydrala (b) insulin or insulin mimetics; Zine); (c) sulfonylureas and other insulin secretagogues, such as (o) glucokinase activators (GKAS). Such as those disclosed , glyburide, , , and megli in WO 03/015774; WO 04/076420; and WO 04/081001; tinides, such as and ; (p) inhibitors of 11 B-hydroxysteroid dehydrogenase Type (d) C-glucosidase inhibitors (such as acarbose and migli 1, such as those disclosed in U.S. Pat. No. 6,730,690; WO tol); 10 03/104207; and WO 04/058741; (e) glucagon receptor antagonists, such as those disclosed (q) inhibitors of cholesteryl ester transfer protein (CETP), in WO 97/16442: WO 98/04528, WO 98/21957; WO Such as torcetrapib; and 98/22108; WO 98/22109; WO 99/01423, WO 00/39088, and (r) inhibitors of fructose 1,6-bisphosphatase, such as those WO 00/69810; WO 2004/050039; and WO 2004/069158: disclosed in U.S. Pat. Nos. 6,054,587; 6,110,903; 6,284,748; (f) GLP-1, GLP-1 analogues or mimetics, and GLP-1 15 6,399,782; and 6,489,476; receptor agonists, such as exendin-4 (exenatide), liraglutide (s) dipeptidyl peptidase IV (DP-IV) inhibitors; (N,N-2211), CJC-1131, LY-307161, and those disclosed in (t) PPARö agonists such as those disclosed in WO97/ WO 00/42026 and WO 00/59887; 28149; and (g) GIP and GIP mimetics, such as those disclosed in WO (u) agents intended for use in inflammatory conditions 00/58360, and GIP receptor agonists: Such as , non-steroidal anti-inflammatory drugs, glu (h) PACAP. PACAP mimetics, and PACAP receptor ago cocorticoids, aZulfidine, and cyclo-oxygenase 2 selective nists such as those disclosed in WO 01/23420; inhibitors, including etoricoxib and rofecoxib. (i) cholesterol lowering agents such as (i) HMG-CoA Examples of anti-hypertensive agents that can be used in reductase inhibitors (lovastatin, simvastatin, pravastatin, combination with the compounds of the invention include cerivastatin, fluvastatin, atorvastatin, itavastatin, and rosuv 25 angiotensin II receptor antagonists (for example losartan, astatin, and other statins), (ii) sequestrants (cholestyramine, candesartan, irbesartan, Valsartan, telmisartan, and eprosar colestipol, and dialkylaminoalkyl derivatives of a cross tan, including all stereoisomers, pharmaceutically acceptable linked dextran), (iii) nicotinyl , nicotinic acid or a salt salts, hydrates, and crystalline forms thereof). ACE inhibitors thereof, (iv) PPARC. agonists, such as fenofibric acid deriva (for example, alacepril, benazepril, captopril, ceronapril, tives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (v) 30 cilaZapril, delapril, enalapril, enalaprilat, fosinopril, imi PPARO/Y dual agonists, such as naveglitazar and muragli dapril, lisinopril, moveltipril, moexipril, perindopril, tazar, (vi) inhibitors of cholesterol absorption, such as beta quinapril, ramipril, spirapril, temocapril, and trandolapril), sitosterolandezetimibe, (vii) acyl CoA:cholesterol acyltrans neutral endopeptidase inhibitors (e.g., thiorphan and phos ferase inhibitors. Such as avasimibe, and (viii) antioxidants, phoramidon), aldosterone antagonists, renin inhibitors (e.g. Such as probucol; 35 urea derivatives of di- and tri-peptides), endothelin receptors (i) PPARgamma agonists and partial agonists, including antagonists, vasodilators, blockers (e.g., glitaZones and non-glitaZones (e.g. pioglitaZone, englitaZone, , , Veraparmil, , , MCC-555, rosiglitaZone, balaglitaZone, netoglitaZone, , nimodipins, ), acti T-131, LY-300512, and LY-818; vators (e.g., , , cromakalim, , (k) anti-obesity compounds, such as fenfluramine, dexfen 40 aprilkalim, loprazolam), diuretics (e.g., hydrochlorothiaz fluramine, phentermine, sibutramine, orlistat, neuropeptide ide), sympatholitics, beta-adrenergic blocking drugs (e.g., Y1 or Y5 antagonists, B3 adrenergic receptor agonists, mel , atenolol, bisoprolol, carvedilol, metoprolol, or anocortin-receptor agonists, in particular melanocortin-4 metoprolol tartate), alpha adrenergic blocking drugs (e.g., receptor agonists, ghrelin antagonists, bombesin receptor doxazocin, prazocin or alpha methyldopa) central alpha adr agonists (such as bombesin receptor Subtype-3 agonists), a 45 energic agonists, and peripheral vasodilators (e.g. hydrala cannabinoid CB1 receptor antagonist/inverse agonist, 5-FIT Zine). (serotonin) inhibitors, and melanin-concentrating hormone Examples of agents for treating schizophrenia that can be (MCH) receptor antagonists: used in combination with the compounds of the invention (1) ileal bile acid transporter inhibitors; include sedatives, hypnotics, anxiolytics, antipsychotics, (m) agents intended for use in inflammatory conditions 50 antianxiety agents, cyclopyrrolones, imidazopyridines, pyra Such as aspirin, non-steroidal anti-inflammatory drugs Zolopyrimidines, minor tranquilizers, melatonin agonists and (NSAIDs), glucocorticoids, aZulfidine, and selective antagonists, melationergic agents, benzodiazepines, barbitu cyclooxygenase-2 (COX-2) inhibitors; rates, 5HT-2 antagonists, and the like. Such as: adinazolam, (n) antihypertensive agents, such as neutral endopeptidase allobarbital, alonimid, aiprazolam, amisulpride, amitrip inhibitors (e.g., thiorphan and phosphoramidon), aldosterone 55 tyline, amobarbital, amoxapine, aripiprazole, bentazepam, antagonists, renin inhibitors (e.g. enalkrein, RO 42-5892. A benzoctamine, brotizolam, bupropion, buSprione, butabar 65317, CP80794, ES 1005, ES 8891, SQ 34017, aliskiren bital, butalbital, capuride, carbocloral, chloral betaine, chlo (2OS),4(S).5(S).7(S)-N-(2-carbamoyl-2-methylpropyl)-5- ral hydrate, , clonazepam, cloperidone, clo amino-4-hydroxy-2,7-diisopropyl-8-4-methoxy-3-(3-meth razepate, chlordiazepoxide, clorethate, , oxypropoxy)phenyl-octanamid hemifumarate) SPP600, 60 , cyprazepam, , dexclamol, diazepam, SPP630 and SPP635), endothelin receptors antagonists, dichloralphenaZone, divalproex, , , vasodilators, calcium channel blockers (e.g., amlodipine, estazolam, ethchlorVynol, etomidate, fenobam, fluni nifedipine, Veraparmil, diltiazem, gallopamil, niludipine, trazepam, flupentiXol, fluiphenazine, flurazepam, fluvoxam nimodipins, nicardipine), potassium channel activators (e.g., ine, , fosazepam, glutethimide, halazepam, halo nicorandil, pinacidil, cromakalim, minoxidil, aprilkalim, 65 peridol, hydroxy Zine, , lithium, , loprazolam), diuretics (e.g., hydrochlorothiazide), sym lormetazepam, maprotiline, mecloqualone, melatonin, patholitics, beta-adrenergic blocking drugs (e.g., propra mephobarbital, meprobamate, methaqualone, midaflur, US 8,329,697 B2 11 12 midazolam, nefazodone, nisobamate, nitrazepam, nortrip azid, phenelzine, tranylcypromine and selegiline; moclobe tyline, olanzapine, oxazepam, paraldehyde, paroxetine, pen mide: Venlafaxine; dulloxetine; aprepitant; bupropion, tobarbital, perlapine, perphenazine, phenelzine, phenobar lithium, nefazodone, traZodone and Viloxazine; alprazolam, bital, prazepam, , propofol, protriptyline, chlordiazepoxide, clonazepam, chiorazepate, diazepam, quazepam, quetiapine, reclazepam, risperidone, roletamide, halazepam, lorazepam, oxazepam and prazepam, buspirone, secobarbital, Sertraline, Suproelone, temazepam, thior flesinoxan, gepirone and ipsapirone, and pharmaceutically idazine, thiothixene, tracazolate, tranylcypromaine, traZ acceptable salts thereof. odone, triazolam, trepipam, tricetamide, triclofos, trifluop The term “composition” as used herein is intended to erazine, trimetozine, trimipramine, uldazepam, Venlafaxine, encompass a product comprising specified ingredients in pre Zaleplon, Ziprasidone, Zolazepam, Zolpidem, and salts 10 determined amounts or proportions, as well as any product thereof, and combinations thereof, and the like, or the subject which results, directly or indirectly, from combination of the compound may be administered in conjunction with the use specified ingredients in the specified amounts. This term in of physical methods such as with light therapy or electrical relation to pharmaceutical compositions is intended to stimulation. encompass a product comprising one or more active ingredi Alternatively, the compounds of the invention may be used 15 ents, and an optional carrier comprising inert ingredients, as in combination with levodopa (with or without a selective well as any product which results, directly or indirectly, from extracerebral decarboxylase inhibitor such as carbidopa or combination, complexation or aggregation of any two or benserazide), anticholinergics such as biperiden (optionally more of the ingredients, or from dissociation of one or more as its hydrochloride or lactate salt) and trihexyphenidyl(ben of the ingredients, or from other types of reactions or inter Zhexol)hydrochloride, COMT inhibitors such as entacapone, actions of one or more of the ingredients. MOA-B inhibitors, antioxidants, A2a adenosine receptor In general, pharmaceutical compositions are prepared by antagonists, cholinergic agonists, NMDA receptor antago uniformly and intimately bringing the active ingredient into nists, serotonin receptor antagonists and dopamine receptor association with a liquid carrier or a finely divided solid agonists such as alentemol, bromocriptine, fenoldopam, carrier or both, and then, if necessary, shaping the product lisuride, naxagolide, pergolide and pramipexole. It will be 25 into the desired formulation. In the pharmaceutical composi appreciated that the dopamine agonist may be in the form of tion the active compound, which is a compound of formula a pharmaceutically acceptable salt, for example, alentemol (I), is included in an amount Sufficient to produce the desired hydrobromide, bromocriptine mesylate, fenoldopam mesy effect upon the process or condition of diseases. Accordingly, late, naxagolide hydrochloride and pergolide mesylate. the pharmaceutical compositions of the present invention The compounds of the invention may also be used in com 30 encompass any composition made by admixing a compound bination with phenothiazine, thioxanthene, heterocyclic of the present invention and a pharmaceutically acceptable dibenzazepine, butyrophenone, diphenylbutylpiperidine and carrier. indolone classes of neuroleptic agent. Suitable examples of The carrier may take a wide variety of forms depending on phenothiazines include chlorpromazine, mesoridazine, thior the form of preparation desired for administration, e.g., oral idazine, acetophenazine, fluphenazine, perphenazine and tri 35 or parenteral (including intravenous). Thus, the pharmaceu fluoperazine. Suitable examples of thioxanthenes include tical compositions of the present invention can be presented chlorprothixene and thiothixene. An example of a dibenza as discrete units Suitable for oral administration Such as cap Zepine is clozapine. An example of abutyrophenone is halo Sules, cachets or tablets each containing a predetermined peridol. An example of a diphenylbutylpiperidine is amount of the active ingredient. Further, the compositions can . An example of an indolone is molindolone. Other 40 be presented as a powder, as granules, as a solution, as a neuroleptic agents include loxapine, Sulpiride and risperi Suspension in an aqueous liquid, as a non-aqueous liquid, as done. The Subject compound may be used in combination an oil-in-water emulsion or as a water-in-oil liquid emulsion. with acetophenazine, alentemol, aripiprazole, amisulpride, In addition to the common dosage forms set out above, the benzhexol, bromocriptine, biperiden, chiorpromazine, chlo compounds of the invention, or pharmaceutically acceptable rprothixene, clozapine, diazepam, fenoldopam, fluphenazine, 45 salts thereof, may also be administered by controlled release , levodopa, levodopa with benserazide, levodopa means and/or delivery devices. with carbidopa, lisuride, loxapine, mesoridazine, molin Pharmaceutical compositions intended for oral use may be dolone, naxagolide, olanzapine, pergolide, perphenazine, prepared according to any method known to the art for the pimozide, pramipexole, quetiapine, risperidone, Sulpiride, manufacture of pharmaceutical compositions and Such com tetrabenazine, frihexyphenidyl, , thiothixene, tri 50 positions may contain one or more agents selected from the fluoperazine or ziprasidone. group consisting of Sweetening agents, flavoring agents, col Further, the compounds of the invention may be used in oring agents and preserving agents in order to provide phar combination with an anti-depressant or anti-anxiety agent, maceutically elegant and palatable preparations. Tablets may including norepinephrine reuptake inhibitors (including ter contain the active ingredient in admixture with non-toxic tiary amine tricyclics and secondary amine tricyclics), selec 55 pharmaceutically acceptable excipients which are suitable for tive serotonin reuptake inhibitors (SSRIs), monoamine oxi the manufacture of tablets. These excipients may be, for dase inhibitors (MAOIs), reversible inhibitors of monoamine example, inert diluents, such as calcium carbonate, sodium oxidase (RIMAS), serotonin and noradrenaline reuptake carbonate, lactose, calcium phosphate or sodium phosphate: inhibitors (SNRTs), corticotropin releasing factor (CRF) granulating and disintegrating agents, for example, corn antagonists, a-adrenoreceptor antagonists, neurokinin-1 60 starch, or alginic acid; binding agents, for example starch, receptor antagonists, atypical anti-depressants, benzodiaz gelatin or acacia, and lubricating agents, for example magne epines, 5-HT1A agonists or antagonists, especially S-HT1A sium Stearate, Stearic acidortalc. The tablets may be uncoated partial agonists, and corticotropin releasing factor (CRF) or they may be coated by known techniques to delay disinte antagonists. Specific agents include: , clomi gration and absorption in the gastrointestinal tract and pramine, doxepin, imipramine and trimipramine; amoxapine, 65 thereby provide a Sustained action over a longer period. desipramine, maprotiline, and protriptyline; flu A tablet containing the composition of this invention may oxetine, fluvoxamine, paroxetine and Sertraline; isocarbox be prepared by compression or molding, optionally with one US 8,329,697 B2 13 14 or more accessory ingredients or adjuvants. Compressed tab patches; buccal dosage forms; inhalation powders, sprays, lets may be prepared by compressing, in a Suitable machine, Suspensions, and the like; and rectal Suppositories. the active ingredient in a free-flowing form Such as powder or The terms “effective amount’ or “therapeutically effective granules, optionally mixed with a binder, lubricant, inert dilu amount’ means the amount of the Subject compound that will ent, Surface active or dispersing agent. Molded tablets may be elicit the biological or medical response of a tissue, system, made by molding in a suitable machine, a mixture of the animal or human that is being sought by the researcher, Vet powdered compound moistened with an inert liquid diluent. erinarian, medical doctor or other clinician. Each tablet preferably contains from about 0.1 mg to about As used herein, the term “treatment' or “treating means 500 mg of the active ingredient and each cachet or capsule any administration of a compound of the present invention 10 and includes (1) inhibiting the disease in an animal that is preferably containing from about 0.1 mg to about 500 mg of experiencing or displaying the pathology or symptomatology the active ingredient. of the diseased (i.e., arresting further development of the Compositions for oral use may also be presented as hard pathology and/or symptomatology), or (2) ameliorating the gelatin capsules wherein the active ingredient is mixed with disease in an animal that is experiencing or displaying the an inert Solid diluent, for example, calcium carbonate, cal 15 pathology or symptomatology of the diseased (i.e., reversing cium phosphate or kaolin, or as Soft gelatin capsules wherein the pathology and/or symptomatology). the active ingredient is mixed with water oran oil medium, for The compositions containing compounds of the present example peanut oil, liquid paraffin, or olive oil. invention may conveniently be presented in unit dosage form Other pharmaceutical compositions include aqueous Sus and may be prepared by any of the methods well known in the pensions, which contain the active materials in admixture art of pharmacy. The term “unit dosage form' is taken to mean with excipients suitable for the manufacture of aqueous Sus a single dose wherein all active and inactive ingredients are pensions. In addition, oily Suspensions may be formulated by combined in a Suitable system, such that the patient or person Suspending the active ingredient in a vegetable oil, for administering the drug to the patient can open a single con example arachis oil, olive oil, Sesame oil or coconut oil, or in tainer or package with the entire dose contained therein, and a mineral oil such as liquid paraffin. Oily Suspensions may 25 does not have to mix any components together from two or also contain various excipients. The pharmaceutical compo more containers or packages. Typical examples of unit dosage sitions of the invention may also be in the form of oil-in-water forms are tablets or capsules for oral administration, single emulsions, which may also contain excipients such as Sweet dose vials for injection, or Suppositories for rectal adminis ening and flavoring agents. tration. This list of unit dosage forms is not intended to be The pharmaceutical compositions may be in the form of a 30 limiting in any way, but merely to represent typical examples sterile injectable aqueous or oleaginous Suspension, or in the of unit dosage forms. form of sterile powders for the extemporaneous preparation The compositions containing compounds of the present of such sterile injectable solutions or dispersions. In all cases, invention may conveniently be presented as a kit, whereby the final injectable form must be sterile and must be effec two or more components, which may be active or inactive tively fluid for easy Syringability. The pharmaceutical com 35 ingredients, carriers, diluents, and the like, are provided with positions must be stable under the conditions of manufacture instructions for preparation of the actual dosage form by the and storage; thus, preferably should be preserved against the patient or person administering the drug to the patient. Such contaminating action of microorganisms such as bacteria and kits may be provided with all necessary materials and ingre fungi. dients contained therein, or they may contain instructions for Pharmaceutical compositions of the present invention can 40 using or making materials or components that must be be in a form Suitable for topical use such as, for example, an obtained independently by the patient or person administer aerosol, cream, ointment, lotion, dusting powder, or the like. ing the drug to the patient. Further, the compositions can be in a form suitable for use in When treating or ameliorating a disorder or disease for transdermal devices. These formulations may be prepared via which compounds of the present invention are indicated, conventional processing methods. As an example, a cream or 45 generally satisfactory results are obtained when the com ointment is prepared by mixing hydrophilic material and pounds of the present invention are administered at a daily water, together with about 5 wt % to about 10 wt % of the dosage of from about 0.1 mg to about 100 mg per kg of animal compound, to produce a cream or ointment having a desired body weight, preferably given as a single daily dose or in consistency. divided doses two to six times a day, or in Sustained release Pharmaceutical compositions of this invention can also be 50 form. The total daily dosage is from about 1.0 mg to about in a form suitable for rectal administration wherein the carrier 2000 mg, preferably from about 0.1 mg to about 20 mg per kg is a solid. It is preferable that the mixture forms unit dose of body weight. In the case of a 70 kg adult human, the total suppositories. Suitable carriers include cocoa butter and other daily dose will generally be from about 7 mg to about 1,400 materials commonly used in the art. mg. This dosage regimen may be adjusted to provide the By “pharmaceutically acceptable' it is meant the carrier, 55 optimal therapeutic response. The compounds may be admin diluent or excipient must be compatible with the other ingre istered on a regimen of 1 to 4 times per day, preferably once dients of the formulation and not deleterious to the recipient or twice per day. thereof. The amount of active ingredient that may be combined The terms “administration of or “administering a com with the carrier materials to produce a single dosage form will pound should be understood to mean providing a compound 60 vary depending upon the host treated and the particular mode of the invention to the individual in need of treatment in a of administration. For example, a formulation intended for form that can be introduced into that individual’s body in a the oral administration to humans may conveniently contain therapeutically useful form and therapeutically useful from about 0.005 mg to about 2.5 g of active agent, com amount, including, but not limited to: oral dosage forms. Such pounded with an appropriate and convenient amount of car as tablets, capsules, syrups, Suspensions, and the like; inject 65 rier material. Unit dosage forms will generally contain able dosage forms, such as IV, IM, or IP and the like; trans between from about 0.005 mg to about 1000 mg of the active dermal dosage forms, including creams, jellies, powders, or ingredient, typically 0.005, 0.01 mg 0.05 mg, 0.25 mg, 1 mg, US 8,329,697 B2 15 5 mg, 25 mg, 50 mg, 100 mg, 200 mg. 300 mg. 400 mg, 500 -continued mg, 600 mg. 800 mg or 1000 mg, administered once, twice or O three times a day. It will be understood, however, that the specific dose level -Y. wO - O - X and frequency of dosage for any particular patient may be BrMg He varied and will depend upon a variety of factors including the i CCN activity of the specific compound employed, the metabolic E. E. THFO C. 1 stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of adminis 10 X w Y- . tration, rate of excretion, drug combination, the severity of the A-2 particular condition, and the host undergoing therapy. Y. / The compounds of the invention may be prepared accord X DMDO ing to the following reaction schemes, in which variables are Ho as defined before or are derived, using readily available start 15 i i Acetone, O C. ing materials, from reagents and conventional synthetic pro cedures. It is also possible to use variants which are them selves known to those of ordinary skill in organic synthesis y art, but are not mentioned in greater detail. A-3 Certain of the starting materials are described in Gallos, J. O / Y. K. etal, J. Org. Chem. 2005: 70(17); 6884-6890, Choi, W.J., X etal, J. Org. Chem. 2004; 69(7): 2634-2636, and Moon, Won NH/MeOH He Jun Choi, et al. Tetrahedron Asymmetry, 13 (11), 1189-1193. i i 100 C., 30 min The following examples are provided to illustrate the 25 invention and are not to be construed as limiting the scope of the invention in any manner. y General Scheme A A-4 According to general scheme A, appropriately substituted 30 HO C N and/or protected cyclopentyl alcohols A-1 can be acylated with methyl chloroformate to produce carbonates A-2. Here X Y can be either saturated (as in X Y) or unsaturated (as in X=Y) and substituted according to the above generic DMF, EtN claims. Carbonates A-2 are then reacted with vinyl magne 35 60 C., 52 h sium bromide in the presence of copper(I) cyanide to provide the vinyl derivates A-3. Regioselective epoxidation of the terminal vinyl group (for example—over an internal X=Y double bond) is achieved using dimethyl dioxirane at low temperature to produce epoxides A-4. Epoxides A-4 are then 40 regioselectively reacted with ammonia under thermal condi tions to give the amino alcohols A-5. Amino alcohols A-5 are coupled with 2,3-dichloropyrazine to produce amino pyra HO zines A-6. Moffat-Swern oxidation of the secondary alcohol 45 in pyrazines A-6 yields the ketone A-7 that is Subsequently TFAA, DMSO cyclized in the presence of trifluoroacetic acid and trfiluoro Her acetic anhydride buffered by the presence of pyridine. These DIPEA, CH2Cl2 conditions provide the heterocyclic imidazopyrazines A-8. -78 C., 2h Imidazopyrazines A-8 are then reacted with ammonia under 50 thermal conditions to provide imidazopyrazines A-9. Subse quent deprotection may vary depending on the nature of the X Y substitution, but acid hydrolysis is utilized to remove the ketal group to provide final compounds A-10. 55

O

1.N.Y. OH C ls o1 60 DMAP, Py TFA, TFAA, Py --- CHCl2, O C. Drt Toluene i. i OC.--> rt, 16 h -X 16h 65 A-1 A-7

US 8,329,697 B2 27 28 the reaction mixture was heated to reflux overnight. Concen oXol-4-yl)imidazol-2-alpyrazine (2-9) (1.5 g., 5.1 mmol. 1 tration and purification via flash chromatography on a silica equiv) in isopropanol (50 mL). The resulting solution was gel column (ethyl acetate/hexanes) yielded desired (S)-2-(3- placed in a high pressure vessel and heated to 100°C. After 16 chloropyrazin-2-ylamino)-1-((3aS.4S,6aR)-2,2-dimethyltet hours, no more starting material was present, the amber solu rahydro-3aFH-cyclopentad 1.3dioxol-4-yl)ethanol (2-7) as tion was concentrated to afford desired 3-((3aS.4S,6aR)-2.2- a colorless oil. "H NMR (500 MHz, CDC1): 8 7.88 (dd. dimethyltetrahydro-3ah-cyclopentad 1.3dioxol-4-yl)imi J=5.5, 2.7 Hz, 1H); 7.60 (d. J=2.7 Hz, 1H); 5.74 (d. J=31.9Hz, dazol-2-alpyrazin-8-amine (2-10) as a tan Solid that was 1H); 4.68-4.63 (m, 1H); 4.34 (t, J=5.4 Hz, 1H); 3.88 (ddd, J=14.5, 6.7, 2.5 Hz, 1H); 3.81.-3.69 (m, 2H); 3.63-3.46 (m, carried to the next step without further purification. "H NMR 2H); 2.15-2.04 (m, 1H); 2.03-1.88 (m, 1H); 1.83-1.72 (m, (500 MHz, CDC1): 87.50 (d. J=4.7 Hz, 1H); 7.36 (t, J=4.6 1H); 1.69-1.52 (m, 1H); 1.50 (d. J=13.2 Hz, 3H); 1.32 (d. 10 HZ, 1H): 7.23 (s, 1H);5.60 (s.2H); 4.78 (s, 1H); 4.64 (d. J–5.7 J–4.5 Hz, 3H). LRMS m/z (M+H) 313.9 found, 314.1 HZ, 1H); 3.44 (d. J–6.9 HZ, 1H); 2.43-2.33 (m, 1H); 2.04-1.93 required. (m,3H); 1.55 (s.3H); 1.34 (s.3H). LRMS m/z (M+H)275.0 found, 275.1 required. 2-(3-chloropyrazin-2-ylamino)-1-((3aS.4R,6aR)-2.2- dimethyltetrahydro-3aH-cyclopentad 1.3dioxol-4- 15 (1R,2S,3S)-3-(8-aminoimidazo 1,2-alpyrazin-3-yl) yl)ethanone (2-8) cyclopentane-1,2-diol (2-11) Oxalyl chloride (2.75 mL, 31.6 mmol. 3.0 equiv) in DCM Concentrated HCl (4 mL) was added to 3-((3aS.4S,6aR)- (35 mL) was added dropwise to DMSO (3.00 mL, 42.1 mmol. 2,2-dimethyltetrahydro-3aFH-cyclopentad 1.3dioxol-4-yl) 4.0 equiv) in DCM (35 mL) at -78° C. After 30 minutes, imidazo[1,2-alpyrazin-8-amine (2-10) (1.20 g, 4.37 mmol. 1 (S)-2-(3-chloropyrazin-2-ylamino)-1-((3aS.4S,6aR)-2.2- equiv) in methanol (20 mL) and the mixture was heated to 65° dimethyltetrahydro-3ah-cyclo-pentad 1.3dioxol-4-yl) C. After 8 hours, no more starting material was present and ethanol (2-7) (3.30 g, 10.5 mmol. 1 equiv) in DCM (35 mL) the mixture was concentrated to afford desired (1R,2S,3S)- was added dropwise. The resulting mixture was stirred at 3-(8-aminoimidazol-2-alpyrazin-3-yl)cyclopentane-1,2- -78° C. for 1 hour, at which point triethylamine (6.60 mL. 25 diol (2-11) as a tan solid. "H NMR (500 MHz, CD OD): 8 47.3 mmol. 4.5 equiv) was added. The cooling bath was 8.06 (d. J=5.8 Hz, 1H); 7.82 (s, 1H): 7.27 (d. J=5.7 Hz, 1H): removed, and after 30 minutes the reaction mixture was con 4.14 (s, 1H); 4.04 (dd, J=9.3, 4.5 Hz, 1H); 3.55 (q, J=9.1 Hz, centrated. The resulting yellow residue was purified via flash 1H); 2.40-2.29 (m. 1H); 2.22-2.10 (m. 1H); 1.92-1.80 (m, chromatography on a silica gel column (ethyl acetate/hex 2H). LRMS m/z (M+H) 235.1 found, 235.1 required. anes) to yield 2-(3-chloropyrazin-2-ylamino)-1-((3aS.4R, 30 In vitro AHCY Activity Assay 6aR)-2,2-dimethyltetrahydro-3aH-cyclopentad 1,3-di The AHCY activity of the compounds of the invention can oxol-4-yl)ethanone (2-8) as a yellow solid. "H NMR (500 be measured by the following in vitro assay. MHz, CDC1): 8 7.92 (d. J=2.7 Hz, 1H); 7.63 (d. J=2.7 Hz, The AHCY enzyme assay is based on the principle of 1H); 5.93 (s, 1H); 4.80 (d. J=5.7 Hz, 1H); 4.73 (t, J=5.4 Hz, fluorescence emission following incubation of a reactive dye 1H); 4.47 (dd, J=19.7, 5.0 Hz, 1H); 4.31 (dd, J=19.7, 4.7 Hz, 35 with the products of an AHCY catalyzed reaction. The reac 1H); 3.17 (d.J=8.1 Hz, 1H); 2.25-2.17 (m, 1H); 1.93-1.84 (m, tive dye (Thioglo 1) binds homocysteine through its thiol 2H); 1.78-1.68 (m. 1H); 1.47 (s.3H); 1.32 (s.3H). LRMS m/z containing moiety. The products of the AHCY catalyzed reac (M+H)312.0 found, 312.1 required. tion, adenosine and homocysteine, are produced from the hydrolysis of SAH (s-adenosylhomocysteine) through 8-chloro-3-((3aS.4S,6aR)-2,2-dimethyltetrahydro 40 AHCY's hydrolase activity. 3aH-cyclopentad 1.3dioxol-4-yl)imidazo 1,2-a The reaction is optimized in 96-, 384-, 1536- and 3456 pyrazine (2-9) assay plate formats with recombinant expressed human or mouse AHCY enzyme. When run under initial velocity con 2-(3-chloropyrazin-2-ylamino)-1-((3aS,4R,6aR)-2.2- ditions, the assay is suitable for IC50 and% inhibition calcu dimethyltetrahydro-3ah-cyclopentad 1.3dioxol-4-yl) 45 lations. ethanone (2-8) (1.2g, 3.85 mmol. 1 equiv) was dissolved in Materials: toluene (28 mL) and the mixture was cooled to 0°C. Pyridine 1) ThioGlol (catalog 595501, Calbiochem); 100 mM in (3.74 mL, 46.2 mmol. 12 equiv) was added followed by TFA DMSO (2.08 mL, 26.9 mmol. 7 equiv). After 30 minutes, TFAA (3.81 2) SAH (catalog A-9384, Sigma); 4.69 mM with 0.06 NHCl mL, 26.9 mmol. 7 equiv) was added. The resulting mixture 50 3) Aristeromycin (catalog A-0928, Sigma); 100 mM in was stirred for 1 h, then was concentrated and the yellow DMSO residue was purified via flash chromatography on a silica gel 4) NAD (catalog 43410, Sigma); 14.5 mM in water column (ethyl acetate/hexanes) to yield desired 8-chloro-3- 5) EDTA in 500 mM pH 8 (catalog BP2482-100, Fisher) ((3aS.4S,6aR)-2,2-dimethyltetrahydro-3aH-cyclopentad 6) DTT stock 200 mM in water 1.3dioxol-4-yl)imidazo 1,2-alpyrazine (2-9) as a white 55 7) AHCY enzyme (diluted with 100 mM tris pH7.5) solid. "H NMR (500 MHz, CDC1): 88.19 (d. J=4.6 Hz, 1H): Dilutions immediately prior to use: 7.84 (d. J=4.6 Hz, 1H): 7.67 (s, 1H); 4.81 (d. J=5.2 Hz, 1H): 1. SAH 750 uM (10x) (dilute with 100 mM Iris pH 7.5) 4.57 (dd, J=6.0.2.6 Hz, 1H); 3.47 (s, 1H); 2.52-2.41 (m, 1H): 2. Aristeromycin (10x) (dilute to 1 mM with 100 mM Tris 2.09-2.05 (m, 3H); 1.58 (s, 3H); 1.34 (s, 3H). LRMS m/z pH7.5) (M+H)293.9 found, 294.1 required. 60 3. ThioGlol (10x) (dilute to 100 uM in Tris pH7.5) 4. AHCY assay buffer (3.3x): 3 uM DTT, 150 uM NAD, 3 3-((3aS.4S,6aR)-2,2-dimethyltetrahydro-3aH-cyclo mM EDTA pentad 1.3dioxol-4-yl)imidazol-2-alpyrazin-8- Protocol: As per 96-assay plate well amine (2-10) 1. Add5uLAHCY enzyme 30 ngful in 100 mM Tris pH7.5 65 2. Add 5uL DMSO or inhibitor Liquid ammonia (50 mL) was added to 8-chloro-3-((3aS, 3. Add 15 uLAHCY assay buffer 4S,6aR)-2,2-dimethyltetrahydro-3aH-cyclopentad 1.3di 4. Add 15 uL 100 mM Tris pH 7.5 US 8,329,697 B2 29 30 5, Incubate 37° C. 30 min Measure of Cell Viability/Proliferation: 6. Add 5 uL 750 uMSAH 1. Add mixture of 10ul 5 mg/ml MTT labeling reagent plus 90 7. Incubate 37° C. 10 min ul fresh assay media to cells. 8. Add 5ul 100 uM ThioGlol 2. Incubate for 4 hrs at 37° C. in cell culture incubator 9. Incubate 37° C. 15 mins 10. Read plate (EX 380 Em 510) 3. Add 100 ul of solubilization solution. Example 1 was tested according to this protocol (n=4) and 4. Incubate for 16 hrs at 37°C. in cell culture incubator gave an ICs, 4 nM. Example 2 was tested according to this 5. RealDMEMd absorbance at 550 nM and 690 nM. protocol (n=2) and gave an ICso 692 nM. These values attest 6. Express readout as 550 nM-690 nM. to the properties of these compounds as potent inhibitors of 10 Example 1 was tested according to this protocol (n2) and the enzyme AHCY. gave an ICs, 4 nM. Example 2 was tested according to this Cell-Based Assay of AHCY Inhibition protocol (n=2) and gave an ICso 1.4 nM. These values attest The AHCY activity of the compounds of the invention can to the properties of these compounds as potent inhibitors of be measured by the following cell-based assay. 15 AHCY in a cell-based format and their ability to lower This assay measures secreted homocysteine over a period homocysteine concentration intracellularly. of time from adherent HEK293 and/or SH-SY5Y cells as a The following abbreviations are used throughout the text: measure of the ability of a compound to enter a cell in culture AdoHcy S-Adenosylhomocysteine and effectively inhibit AHCY in the context of the cellular Me: methyl milieu. Post treatment collection of the adherent Hek293 Et: ethyl and/or SH-SY5Y cells is also made for a viability and prolif t-Bu: tent-butyl eration measure. Since the amount of secreted homocysteine is limited by the amount of SAH that is converted to homocys i-Pr: isopropyl teine in the cells and its subsequent secretion, an EC50 mea Ph: phenyl sure and % inhibition relative to DMSO control cells can be 25 DCM; dichloromethane made. Since the amount of Secreted metabolites such as THF: tetrahydrofuran homocysteine can also be related to the absolute and cellular Ac: acetyl viability, an MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphe DMAP: 4-Dimethylaminopyridine nyltetrazolium bromide) measure of proliferation and relative DMDO: dimethyldioxirane cytotoxicty (as compared to control conditions) may also be 30 made. NAD: nicotinamide adenine dinucleotide Protocol: DMEM: Dulbecco's modified Eagle's medium 1. The day before compound treatment, plate 20,000 cells MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo (Hek293 or SH-SY5Y) in 96 well cell culture treated plates lium bromide in 100 ul of cell culture media (DMEM and 10% fetal 35 PCR: polymerase chain reaction bovine serum). DMSO: dimethylsulfoxide 2. Add fresh media plus compound (and DMSO treated con TFA: trifluoroacetic acid trols) for 72 hrs. 3. Collect conditioned media for measurement of secreted TFAA. trifluoroacetic acid anhydride homocysteine and perform viability/proliferation assay on 40 Tr triphenylmethyl adherent cells. rt: room temperature Measure of Secreted Homocysteine: aq: aqueous 1. Add 25ul conditioned media, standard or control in 96-well assay plate HPLC: high performance liquid chromatography 2. Add 25ul internal standard 45 MS: mass spectrometry 3. Add 10 ul reduction solution CDCl: chloroform-d 4. Add 50 ul derivatisation reagent CuCN: copper(I) cyanide 5. Incubate for 10 min in PCR (polymerase chain reaction) DIPEA. diisopropylethylamine machine at 60°C., stop reaction at 4°C. or on ice. 6. Add 50 ul precipitating solution 50 DMF: N,N-dimethylformamide 7. Mix well, incubate for 10 min at 4°C. and centrifuge for 10 EtO: diethylether min at 2,000x LRMS: low resolution mass spectrum 8. Inject 20 ul of the supernatant into HPLC system. a. Chromatographic conditions: MeOH: methanol i. Column material: Imtakt Unisom C18, 3 um 55 MgSO: magnesium sulfate ii. Column dimension: 50 mmx4.6 mm NMR: nuclear magnetic resonance iii. Flow rate: 1.5 ml/min Py: pyridine iv. Detection: Fluorescence: Excitation 385 nm, Emission While the invention has been described and illustrated with 515 nm. 60 reference to certain particular embodiments thereof, those V. Injection volume: 20 ul skilled in the art will appreciate that various adaptations, vi. Running time: 3.5 min changes, modifications, Substitutions, deletions, or additions b. Calculation: of procedures and protocols may be made without departing (peak height patient concentration of the calibrator? from the spirit and scope of the invention. It is intended, peak height internal standard patient) 65 therefore, that the invention be defined by the scope of the *F-concentration patient sample (F=IS peak of claims that follow and that such claims be interpreted as calibratori Hcy peak of calibrator) broadly as is reasonable. US 8,329,697 B2 31 32 What is claimed is: 3. The compound of claim 2, or a pharmaceutically accept 1. A compound of formula (I): able salt thereof, wherein X Y is CH CRR, and R' is selected from the group consisting of (1) hydrogen, and (2) —CH2OH. 4. The A compound of claim 1, or a pharmaceutically acceptable salt thereof, whereinX Y is CH=CR', and R' is selected from the group consisting of (1) hydrogen, and OH (2) —C alkyl, wherein said alkyl is optionally substi 10 tuted with (a) halogen, or W (b) hydroxy. OH 5. The compound of claim 1, or a pharmaceutically accept or a pharmaceutically acceptable salt thereof, wherein able salt thereof, wherein CH=CR', and R' is selected from X-Y is selected from the group consisting of 15 the group consisting of (1) hydrogen, and (1) CH, CRR, (2) —CHOH. (2) CH-CR'; 6. The compound of claim 1, which is selected from the R" is selected from the group consisting of group consisting of (1) hydrogen, (2) hydroxy, (3) —C alkyl, wherein said alkyl is optionally substi tuted with (a) halogen, or (b) hydroxy; and 25 R is selected from the group consisting of (1) hydrogen, (2) hydroxy, (3) —C alkyl, wherein said alkyl is optionally Substi tuted with 30 (a) halogen, or (b) hydroxy. 2. The compound of claim 1, or a pharmaceutically accept able salt thereof, wherein X Y is CH CRR, and R' is selected from the group consisting of 35 (1) hydrogen, and or a pharmaceutically acceptable salt thereof. (2) —C alkyl, wherein said alkyl is optionally Substi 7. The pharmaceutical composition comprising a com tuted with pound of any of claims 1-6 or a pharmaceutically acceptable (a) halogen, or salt thereof, and a pharmaceutically acceptable carrier. (b) hydroxy. k k k k k