OA13337A - Pyrido[2,3-d]pyrimidine-2,4-diamines as PDE 2 inhibitors. - Google Patents

Description

013337

PYRIDOf2.3-d1PYRIMIDINE-2.4-DIAMINES

AS PPE 2 INHIBITORS

FIELD OF THE INVENTION

The Invention relates to certain pyrido[2,3-d]pyrimidine-2,4-diamines useful asPDE 2 inhibitors; pharmaceutical formulations thereof; combinations thereof; anduses thereof.

BACKGROUND OF THE INVENTION

The phosphodiesterase (Λε) family of enzymes régulâtes intracellularlevels of secondary messenger cAMP or cGMP through hydrolytic control.Phosphodiesterase Type II (PDE 2) possesses a low affinity catalytic domain and anallosteric domain spécifie for cGMP. The low affinity catalytic site can hydrolyze bothcAMP and cGMP with a lower apparent Km for cGMP over cAMP. However, whencGMP binds to the allosteric site, the catalytic site undergoes a conformationalchange showing high affinity for cAMP. PDE 2 shows the highest expression in thebrain, but is also found in many other tissues as well and, therefore, has a broadarray of function and potential therapeutic utiiity (J. A. Beavo, et al., Rev. Physio.Biochem. Pharm., 135, 67 (1999)). Examples of PDE 2 function and therapeuticpotential are in neuronal development, leaming, and memory (W. C. G. vanStaveren, et. al., Brain Res., 888.275 (2001) and J. O’Donneil, et. al., J. Pharm. Exp.Ther., 302, 249 (2002)); prolactin and aldostérone sécrétion (M. O. Velardez, et. al.,Eur. J. Endo., 143, 279 (2000) and N. Gallo-Payet, et al., Endo., 140, 3594 (1999));bone cell différentiation, growth, and bone résorption (C. Allardt-Lamberg, et. al.,Biochem. Pharm., 59.1133 (2000) and S. Wakabayashi, et. al., J. Bone. Miner. Res.,17, 249 (2002)); immunological response (M. D. Houslay, et. al., Cell. Signal., 8, 97(1996); vascular angiogenesis (T. Keravis, et. al., J. Vase. Res., 37, 235 (2000);infiammatory cell transit (S. L. Wolda, et ai., J. Histochem. Cytochem., 47, 895(1999); cardiac contraction (R. Fischmeister, et al., J. Clin. Invest., 99, 2710 (1997),P. Donzeau-Gouge, et. al., J. Physiol., 533, 329 (2001), and D. J. Paterson, et. al.,Card. Res., 52, 446 (2001)); platelet aggregation (R. J. Haslam, et al., Biochem. J.,323. 371 (1997); female sexual arousal disorder (FSAD) (C.P. Wayman, et al.,European Patent Application Publication Nos. EP 1 097 7707 and 1 0977 06); andhypoxie pulmonary vasoconstriction (J. Haynes, et al., J. Pharm. Exp. Ther., 276,752 (1996)). It has been shown that EHNA (erythro-9-(2-hydroxy-3-nonyl)adenine), a -2- 013337 potent adenosine deaminase inhibiteur, selectively inhibits PDE 2, however, the use ofEHNA as a PDE 2 based therapeutic agent is limited due to low potency in inhibitingPDE 2, and high potency in inhibiting adenosine deaminase (R. Fischmeister, et. al.,Mol. Pharm.,48,121 (1995)). 5 It has now been found that certain pyrido[2,3-d]pyrimidine-2,4-diamine dérivatives of formula (I) hereinbelow inhibit PDE 2 and, therefore, are useful in thetreatment of physioiogical disorders mediated though the cAMP or cGMP cellular-signaling pathway. U.S. Pat. Nos. 5,547,954 and 5,710,157 disclose certain 2,4-diamino-5,6- 10 disubstituted- and 5,6,7-trisubstituted-5-deazapteridines, compositions thereof, anduses thereof in controiling insects in agricultural crops. « SUMMARY OF THE INVENTIONThe invention provides compounds of formula (I)

prodrugs thereof, and the pharmaceuticatly acceptable salts of the compounds orprodrugs, wherein η, X, and Y are as defined hereinbelow; pharmaceuticalcompositions thereof; combinations thereof; and uses thereof.

DETAILED DESCRIPTION OFTHE INVENTION

The présent invention provides compounds of formula (I) -3-

the prodrugs thereof, and the pharmaceuticaliy acceptable salts of the compounds orprodrugs, wherein: R1 and R2 are hydrogen or methoxy, provîded R1 and R2 are not bothhydrogen or both methoxy; n is 1, 2,3, or 4; X is a bond; O; S; C=O; -N(R)-, wherein R is hydrogen or -(Cr C3)alkyl; -C(OH)-; or -SO2; and Y is benzoxazolyl; benzothiazolyl; benzofurazanyl; benzofuranyl;benzothiadiazolyl; benzisoxazolyl; benzisothiazolyl; benzimidazolyl; pyridyl; isatinyl;oxindolyl; indazolyl; indolyl; phenyl; thienyl; or furanyl; wherein Y is optionallysubstituted independently with from one to three halogen; trifluoromethyl; methoxy; -C(=O)CH3; cyano; -C(CH3)2OH; -CH(CH3)OH; -CH(CF3)OH; -C(C=O)CF3; -SO2NH2; - C(=O)OCH3; -CHzCOOH; pO ; thiazolyl; or oxadiazolyl. A generally preferred subgroup of the compounds of formula (I) comprisesthose compounds wherein X is a bond and Y is benzofurazanyl; thienyi; pyridyi; orphenyl, wherein phenyl is optionally substituted independently with one or twohalogen; trifluoromethyl; methoxy; -C(=0)CH3; cyano; -C(CH3)2OH; -CH(CH3)OH; -CH(CF3)OH; -C(C=O)CF3; -SO2NH2; -C(=O)OCH3; -CH2COOH; thiazolyl; oroxadiazolyl.

An espedally preferred subgroup of the compounds of formula (I) comprisesthose compounds wherein X is a bond, n is 2 or 3, and Y is thienyl; pyridyl; or phenyl,wherein phenyl is optionally substituted independently with one or Iwo methoxy;halogen; -C(CH3)2OH; CH(CF3)OH; or-C(C=O)CF3. A cyclic group may be bonded to another group in more than one way. If noparticular bonding arrangement is specified, then ail possible arrangements areintended. For example, the term ''pyridyl" indudes 2-, 3-, or 4-pyridyl, and the term"thienyl" indudes 2- or 3-thienyl. 013337 -4-

The compounds and intermediates of the présent invention may be namedaccording to either the IUPAC (International Union for Pure and Applied Chemistry)or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature Systems.

The carbon atom content of the various hydrocarbon-containing moieties maybe indicated by a prefix designating the minimum and maximum number of carbonatome in the moiety, i.e., the prefix “-(Ca-Cb)alkyr indicates an alkyl moiety of theinteger "a" to "b" carbon atoms, inclusive.

The term "alkyl" dénotés straight or branched, monovalent chains of carbonatoms. Examples of alkyl groupe include methyl, ethyl, propyl, isopropyl, butyl,isobutyl, and the like.

The term "halogen" représente chloro, fluoro, bromo, and iodo.

The term "prodrug" refers to a compound that is a drug precursor which,following administration, releases the drug in vivo via a Chemical or physiologicalprocess (e.g., upon being brought to physiological pH or through enzyme activity). Adiscussion of the préparation and use of prodrugs is provided by T. Higuchi and W.Stella, "Prodrugs as Novel Delivery Systems", Vol. 14 of the ACS Symposium Sériés,and in "Bioreversible Carriers in Drug Design", ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press (1987).

The term "mammàl" means animais including, for example, dogs, cats, cows,sheep, horses, and humans. Preferred mammals include humans of either gender.

The term "salts" refera to organic and inorganic salts of a compound offormula (I), or a prodrug thereof. These salts can be prepared in situ during the finalisolation and purification of a compound, or by separately reacting a compound offormula (I), or a prodrug thereof, with a suitable organic or inorganic acid or base andisolating the sait thus formed. Représentative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, besylate, palmitate,stéarate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate,fumarate, succinate, tartrate, naphthylate, mesyiate, gfucoheptonate, lactobionate,and laurylsulphonate salts, and the like. These may also include cations based on thealkali and alkaline earth metals, such as sodium, lithium, potassium, calcium,magnésium, and the like, as well as non-toxic ammonium, quatemary ammonium,and amine cations induding, but not limited to, ammonium, tétraméthylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, 013337 -5- ethylamine, and the like. For additional examples see, for example, Berge, et al., J.Pharm. Sci., 66,1-19 (1977).

The term "substituted" means that a hydrogen atom on a molécule has beenreplaced with a different atom or molécule. The atom or molécule replacing thehydrogen atom is denoted as a "substituent"

As used herein, the term “therapeutically effective amount” means an amountof a compound that is capable of treating a described pathological condition.

The terms “treat”, “treatmenf, and “treating" include preventative (e.g.,prophylactic) and palliative (e.g., healing or curative) treatment, or the act of providingpreventative or palliative treatment.

The compounds of formula (I) may contain asymmetric or chiral centers and,therefore, exist in difFerent stereoisomeric forms. It is intended that àll stereoisomericforms of the compounds and prodrugs of formula (I) as well as mixtures thereof,including racemic mixtures, form part of the présent invention. In addition, the présentinvention embraces ail géométrie and positlonal isomers. For example, if a compoundor prodrug of formula (I) incorporâtes a double bond(s), both the c/s- and trans-forms, as well as mixtures thereof, are embraced within the scope of the invention.

Diastereomeric mixtures can be separated into their individual diastereomerson the basis of their physical Chemical différences by methods well-known to those ofordinary skill in the art, such as by chromatography and/or fractional crystallization.Enantiomers can be separated by converting the enantiomeric mixture into adiasteriomeric mixture by reaction with an appropriate optically active compound(e.g., alcohol), separating the diastereomers and converting (e.g., hydrolyzing) theindividual diastereomers to the corresponding pure enantiomers.

The compounds and prodrugs of the compounds of formula (i) may exist inunsolvated as well as solvated forms with pharmaceuticaliy acceptable solvents, suchas water, éthanol, and the like, and it is intended that the invention embrace bothsolvated and unsolvated forms.

It is also possible that the compounds and prodrugs of formula (I) may existas tautomeric isomers in equilibrium, and ail such forms are embraced within thescope of the invention.

The présent invention also embraces isotopically-labeled compounds offormula (I), which are identical to those recited herein, but for the fact that one ormore atoms are replaced by an atom having an atomic mass or mass number 013337 -6- different from the atomic mass or mass number usually found in nature. Examples ofisotopes that can be incorporated into compounds of formula (I) include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2H,3H, 13C, 14C, 15N, 18O,170,31P, 32P, 35S, 1eF, and ^Cl, respectively. The compounds offormula (I), the prodrugs thereof, and the pharmaceutically acceptable salts of thecompounds and prodrugs, that contain the aforementioned isotopes and/or otherisotopes of the other atoms are intended to be within the scope of the instantinvention.

Certain isotopically-labeled compounds of formula (I), for example thosecompounds into which radioactive isotopes such as 3H and 14C are incorporated, areuseful in compound and/or substrate tissue distribution assays. Tritiated, i.e., aH, andcarbon-14, i.e., 14C, isotopes are particularly preferred for their relative ease ofpréparation and facile détection. Furthermore, substitution with heavier isotopes suchas deuterium, i.e., ZH, may afford certain therapeutic advantages resuiting fromgreater metabolic stability, for example, increased In vivo half-life, or reduced dosagerequirements and, hence, may be preferred in some circumstances. The isotopically-labeled compounds of formula (I) can generally be prepared by carrying outprocedures analogous to those disdosed in the Schemes and/or Examples set forthhereinbelow, by substituting an isotopically-labeled reagent for a non-isotopically-labeled reagent.

In another aspect, the invention provides methods of treating PDE 2-mediatedconditions, diseases, or symptoms in a mammal in need of such treatment whichmethods comprise administering to the mammal a therapeutically effective amount ofa compound of formula (I), a prodrug thereof, or a pharmaceutically acceptable sait ofthe compound or prodrug; or a pharmaceutical composition comprising a compoundof formula (I), a prodrug thereof, or a pharmaceutically acceptable sait of thecompound or prodrug, and a pharmaceutically acceptable vehicle, carrier, or diluent.

Preferred conditions, diseases, or symptoms treatable according to theprésent methods include osteoporosis, pulmonary hypertension, female sexualarousal disorder, diminished memory or cognition, platelet aggregation, vascularangiogenesis, dementia, cancer, arrhythmia, thrombosis, bone fracture and/or defect,delayed or non-union fracture, spinal fusion, bone in-growth, cranial facialreconstruction, or hypoxia. An especiaily preferred condition is bone fracture and/ordefect. 013337 -7- ln another aspect, the invention provides methods for inhibiting PDE 2 activityin a mammal in need of such inhibition which methods comprise administering to themammal a PDE 2-inhibiting amount of a compound of formula (I), a prodrug thereof,or a pharmaceutically acceptable sait of the compound or prodrug; or apharmaceutical composition comprising a compound of formula (I), a prodrug thereof,or a pharmaceutically acceptable sait of the compound or prodrug, and apharmaceutically acceptable vehicle, carrier, or diluent.

The compounds of formula (I), the prodrugs thereof, and the pharmaceuticallyacceptable salts of the compounds and prodrugs, may be administered to a mammalat dosage levels in the range of from about 0.001 mg to about 200 mg per day. For anormal adult human having a body mass of about 70 kg, a dosage in the range offrom about 0.01 mg to about 100 mg per kg body mass is typically preferred,however, some variability in the general dosage range may be required dependingupon the âge and mass of the subject being treated, the intended route ofadministration, the particular compound being administered, and the like. Thedétermination of dosage ranges and optimal dosages for a particular mammaliansubject is within the abilrty of one of ordinary skill in the art having benetit of theinstant disclosure.

In yet another aspect, the invention provides pharmaceutical compositionscomprising a combination of a PDE 2 inhibitor, an EP2 sélective agonist; and apharmaceutically acceptable vehicle, carrier, or diluent; and methods of treatingosteoporosis, pulmonary hypertension, female sexual arousal disorder, diminishedmemory or cognition, platelet aggregation, vascuiar angiogenesis, dementia, cancer,arrhythmia, thrombosis, bone fracture and/or defect, deiayed or non-union fracture,spinal fusion, bone in-growth, cranial facial reconstruction, or hypoxia using suchcompositions. An especially preferred condition is bone fracture and/or defect.

In yet another aspect, the invention provides methods of treating bonefracture and/or defect in a mammal in need of such treatment which methodscomprise administering to the mammal a therapeutically effective amount of a PDE 2inhibitor, a prodrug thereof, or a pharmaceutically acceptable sait of said inhibitor orprodrug.

Any PDE 2 inhibitor, including the compounds of formula (I) herein, can beemployed in the methods and combinations of the invention. Examples of knownPDE 2 inhibitors comprise EHNA, 6-(3,4-dïmethoxy-benzyl)-1-[(1-hydroxy-ethyl)^{- 013337 -8- phenyl-butyl]-3-methyl-1,5-dihydro-pyrazolo[3,4-d]pyrimidine-4-one (BAY-60-7550;U.S. Pat No. 6,174,884), and 9-(1-acetyl-4-phenyl-butyl)-2-(3,4-dimethoxy-benzyl)~1,9-dihydropurin-6-one (U.S. Pat No. 5,861,396). Additional examples of PDE 2inhibitors are disclosed in U.S. Pat Nos. 5,861,396; 5,401,774; 6,458,796; and6,555,547; and in PCT International Application Publication No. 98/32755.

Any EP2 sélective receptor agonist can be employed in the combinationaspects of the présent invention, however, a generally preferred class of EP2sélective receptor agonists, disclosed in commonly-assigned U.S. Patent Number6,498,172, comprises compounds of Formula AA

Formula AA prodrugs thereof, and the pharmaceutically acceptable salts thereof, wherein G, A,B, K, M, Q, and Z are as defined therein.

Generally preferred compounds of Formula AA are (3-(((pyridine-3-sulfonyl)-(4-pyrimidin-5-yl-benzyl)-amino)-methyl)-phenyl)-acetic acid; (3-(((5-phenyl-furan-2-ylmethyl)-(pyridine-3-sulfonyl)-amino)-methyl)-phenyl)-acetic acid; (3-(((pyridine-3-sulfonyl)-(4-pyrimidin-2-yl-benzyl)-amino)-metliyl)-phenyl)-acetic acid; (3-(((pyridine-3-sulfonyl)-(4-thiazol-2-yl-benzyl)-amino)-methyl)-phenyl)-acetic acid; (3-(((4-pyrazin-2-yl-benzyl)-(pyridine-3-suHonyl)-amino)-methyl)-phenyl)-acetic acid; (3-(((4-cyclohexyl-benzyl)-(pyridine-3-sutfonyl)-amino)-methyl)-phenoxy)-acetic acid; (3-(((pyridine-3-sulfonyl)-(4-pyridin-2-yl-benzyl)-amino)-methyl)-phenoxy)-acetic acid; (3-(((pyridine-3-sulfonyl)-(4- pyridin-3-yl-benzyl>amino)-methyi)-phenoxy)-acetic acid; (3-(((pyridine-3-sulfonyl)-(4-pyridfn-4-yl-benzyl)-amino)-methyl)-phenoxy)-acetic acid; (3-(((pyridine-3-sulfonyl)-(4-thiazol-2-yl-benzyl)-amino)-methyl)-phenoxy)-acetic acid; 5-(3-((pyridine- 3-sulfonyl)-(4-thiazol-2-yl-benzyl)-amino)-propyl)-thiophene-2-cartxîxylic acid; (3-(((2,3-dihydra-benzo[1,4]dioxin-6-yimethyl)-(pyridine-3-sulfonyl)-amino)-methyl)-phenyl)-acetic acid; and (3-((benzofuran-2-ylmethyl-(pyridine-3-suiftMiyl)-amino)-methyl)-phenyl)-acetic acid; (3-(((4-butyl-benzyl)-(pyridine-3-sulfonyl)-amino)-methyl)-phenyl)-acetic acid; (3-((berLænesulfonyl-(4-butyl-benzyl)-amino)-methyl)-phenyl)-acetic acid; (3-(((4-butyl-benzyl)-(1-methyl-1 H-imidazole-4-sulfonyl)-amino)-methyl)-phenyl)-acetic acid; and (3-(((4-dimethylamino-benzyl)-(pyridine-3-sulfonyl)-amÎno)- 013337 -9- methyl)-phenyl)-acetic acid; (3-(((4-dimethylamino-benzyl)-(pyridine-3-sulfonyl)-amino)-methyl)-phenoxy)-acetic acid and (3-(((4-fert-butyl-benzyl)-(pyridine-3-suifonyl)-amino)-methyl)-phenoxy)-acetic add; frans-(3-(((3-(3,5-dichloro-phenyl)-allylHpyddine-3-sulfonyl)-amino)-methyl)-phenyl)-aoetic acid; (3-(((2-(3,5-dichloro-phenoxy)-ethyl)-(pyridine-3-sulfonyl)-amino)-methyl)-phenoxy)-acetic acid; prodrugsthereof, and pharmaceutically acceptable salts of the compounds and the prodrugs.

An especially preferred compound of Formula AA is (3-(((4-ferf-butyl-benzyl)-(pyridine-3-sulfonyl)-amino)-methyl)-phenoxy)-acetic acid, a prodrug thereof, or apharmaceutically acceptable sait of the compound or prodrug. A particularly preferredsait is the sodium sait

Another generally preferred class of EP2 sélective receptor agonists useful inthe combination aspects of the Invention comprises the compounds, prodrugs, andpharmaceuflcally-acceptable salts of Formula BB below, which are disciosed incommonly-assigned U.S. Pat. No. 6,288,120

K-M

Formula BB wherein A, B, K, M,Q, and Z are as defined therein.

Generally preferred compounds of Formula BB are 7-[(2'-hydroxymethyl- biphenyl-4-ylmethyl)-methanesulfonyl-aminol-heptanoic acid; 7-{[4-(3-hydroxymethyl-thiophen-2-yl)-benzyl]-methanesulfonyl-amino}-heptanoic acid; 7-[(2'-chloro-blphenyl- 4-ylmethyl)-methanesulfbnyl-amino]-heptanoie acid; 7-{[4-(1-hydroxy-hexyl)-benzyl]-methanesulfonyl-aminoj-heptanoic acid; 7-[(4-butyl-benzyl)-methanesulfonyl-amÎno]-heptanoic acid; 7-{[5-(1~hydroxy-hexyl)-thiophen-2-ylmethyl]-methanesulfonyl-amino}-heptanoic acid; (3-{[(4-butyl-benzyl)-methanesulfonyl-amino]-methyl}-phenyl)-aceticacid; 7-{[3-(3-Chloro-phenyl)-propyi]-methanesuifonyl-amino}-heptanoic acid; 7-{[3-(3,5-Dichloro-phenyl)-propyO-methanesulfonyl-amino}-heptanoic acid; 5-(3-{[3-(3-chloro-phenyl)-propyl]-methanesulfonyl-amino}-propyl)-thÎophene-2-carboxylic acid;7-((2-(3,5-dichloro-phenoxy)-ethyl]-methanesuifonyl-amino}-heptanoic acid; 5-(3-{[2-(3,5-dichloro-phenoxy)-ethyl]-methanesu,fonyl-amino}-propyl)-thiophene-2-carboxylicacid; N-[2-(3,5-dichloro-phenoxy)-ethyl]-N-[6-(1 H-tetrazol-5-yl)-hexyl]- methanesulfonamide; frans-(4-{[3-(3,5-dichloro-phenyl)-allyl]-methanesulfonyl- 013337 -10- amino)-butoxy)-acetic acid; frans-N-[3-(3,5-dichloro-phenyl)-allyl]-N-[6-(1 H-tetrazolyi- 5-yl)-hexyl]-methanesulfonamide; frans-5-(3-{[3-(3,5-dichloro-phenyl)-allyl]- methanesulfonyi-amino)-propyl)-thiophene-2-carboxylic acid; frans-[3-({[3-(3,5-dichloro-phenyl)-allyO-methanesulfonyl-amino}-methyl)-phenyl]-acetic acid; theprodrugs thereof, and the pharmaceutically acceptable salts of the compounds andprodrugs.

An especially preferred compound of Formula BB is 7-[(4-butyl-benzyl)-methanesulfonyl-aminoj-heptanoic acid, a prodrug thereof, or a pharmaceuticallyacceptable sait of the compound or prodrug. A preferred sait Is the monosodium sait.

Other EP2 sélective receptor agonists useful in the combination aspects of theprésent invention comprise the prostaglandln receptor agonists disclosed in U.S. Pat.Nos. 6,531,485; 6,376,533; 6,124,314; 5,877,211; 5,716,835; 5,698,598; 'and5,462,968; U.S. Published Pat. Appl’n. No. 2002/187961; N. Duckworth, et al.,Journal of Endocrinology, 172 (2), 263-269 (2002); K. Tani, et al., Synlett, 2,239-242(2002); K. Tani, et al., Bioorganic & Médicinal Chemistry, J0 (4), 1107-1114 (2002);K. Tani, et ai., Bioorganic & Médicinal Chemistry, 10 (4), 1093-1106 (2002); J.Michelet, et al., EP 1 175 891 A1 ; K. Tani, et al., Bioorganic & Médicinal ChemistryLetters, H (15), 2025-2028 (2001); J. Y. Crider, et al., International Journal ofEnvironmental Studies, 58 (1), 35-46 (2000); J. Y. Crider, et al., Journal of OcularPharmacology and Therapeutics, 17 (1), 35-46 (2001); D. F. Woodward, et al.,Journal of Ocular Pharmacology and Therapeutics, IJ (3), 447-54 (1995); A. T. Niais,et al., Cardiovascular Drug Reviews, IJ (2), 165-79 (1993); and D. F. Woodward, etal., Prostaglandine, 46 (4), 371-83 (1993).

In the combination aspects of the invention, the EP2 sélective receptoragonists may be administered to mammals at dosage levels ranging from about0.001 mg/kg to about 100 mg/kg body mass per day. For a normal adult humanhaving a body mass of about 70 kg, a dosage in the range of from about 0.01 mg/kgto about 50 mg/kg body mass is typicaliy preferred, however, some variability in thegeneral dosage range may be required depending upon the âge and mass of thesubject belng treated, the intended route of administration, the particular compoundbeing administered, and the like. The détermination of combination dosage rangesand optimal dosages for a particular mammalian subject is within the ability of one ofordinary skilf in the art having benefit of the instant disciosure. 013337 -11-

Pharmaceutical compositions suitable for parentéral injection may comprisepharmaceutically acceptable stérile aqueous or nonaqueous solutions, dispersions,suspensions, or émulsions, and stérile powders for extemporaneous reconstitutioninto stérile injectable solutions or dispersions. Examples of suitable aqueous andnonaqueous carriers, vehicles, and diluents include water, éthanol, polyols (such aspropylene glycol, polyethylene glycol, glyceral, and the like), suitable mixtures thereof,vegetabie oils (such as olive oil), and injectable organic esters such as ethyl oleate.Proper fluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the case of dispersions,and by the use of surfactants.

The pharmaceutical compositions of the invention may further compriseadjuvants, such as preserving, wetting, emulsifying, and · dispersing agents.Prévention of microorganism contamination of the instant compositions can beaccomplished with various antibacterial and antifungal agents, for example,parabens, chlorobutanol, phénol, sorbic acid, and the like. It may also be désirable toinciude isotonie agents, for example, sugars, sodium chloride, and the like. Prolongedabsorption of injectable pharmaceutical compositions may be effected by the use ofagents capable of delaying absorption, for example, aiuminum monostearate andgelatin.

Solid dosage forms for oral administration include capsules, tablets, powders,and granules. In such solid dosage forms, the active compound is admixed with atleast one inert conventional pharmaceutical excipient (or carrier) such as sodiumcitrate or dicalcium phosphate, or (a) filière or extenders, as for example, starches,lactose, sucrose, mannitol, and silicic acid; (b) binders, as for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrroiidone, sucrose, and acacia;(c) humectants, as for example, glyceroi; (d) disintegrating agents, as for example,agar-agar, calcium carbonate, potato or tapioca starch, alginic acid certain complexsilicates, and sodium carbonate; (e) solution retardera, as for example, paraffin; (f)absorption acceleratora, as for example, quatemary ammonium compounds; (g)wetting agents, as for example, cetyl alcohol and glyceroi monostearate; (h)adsorbents, as for example, kaolin and bentonite; and/or (i) lubricants, as forexample, talc, calcium stéarate, magnésium stéarate, solid polyethylene glycols,sodium lauryl sulfate, or mixtures thereof. In the case of capsules and tablets, thedosage forms may further comprise buffering agents. 013337 -12-

Solid compositions of a similar type may also be employed as fillers in soft orhard filied gelatin capsules using such excipients as lactose or milk sugar, as well ashigh molecular weight polyethylene giycols, and the like.

Solid dosage forms such as tablets, dragees, capsules, and granules can beprepared with coatings and shells, such as enteric coatings and others weli-known toone of ordinary skili in the art. They may also comprise opacifying agents, and canalso be of such composition that they release the active compound(s) in a delayed,sustained, or controlled manner. Examples of embedding compositions that can beemployed are polymeric substances and waxes. The active compound(s) can also bein micro-encapsulated form, if appropriate, with one or more of the above-mentionedexcipients.

Liquid dosage forms for oral administration indude pharmaceuticallyacceptable émulsions, solutions, suspensions, syrups, and élixirs. In addition to theactive compounds, the liquid dosage form may contain inert diluents commoniy usedin the art, such as water or other solvents, solubilizing agents and emulsifiers, as forexample, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide,oils, in particular, cottonseed Oit, groundnut oil, corn germ oïl, olive oil, castor oil, andsesame seed oil, glycerol, tetrahydrofurfuiyl alcohol, polyethylene giycols and fattyacid esters of sorbitan, or mixtures of these substances, and the like.

Besides such inert diluents, the pharmaceutical composition can also inciudeadjuvants, such as wetting agents, emuisifying and suspendlng agents, sweetening,flavoring, and perfuming agents.

Suspensions, in addition to the active compound(s), may further comprisesuspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide,bentonite, agar-agar, and tragacanth, or mixtures of the aforementioned substances,and the like.

Compositions for rectal or vaginal administration preferably comprisesuppositories, which can be prepared by mixing an active compound(s) with suitablenon-irritating excipients or carriers such as cocoa butter, polyethylene glycol or asuppository wax, which are solid at ordinary room température, but liquid at bodytempérature, and therefore, melt in the rectum or vaginal cavity thereby releasing theactive component. 013337 -13-

Dosage forms for topical administration may comprise ointments, powders,sprays and inhalants. The active agent(s) are admixed under stérile condition with apharmaceutically acceptable carrier, vehicle, or diluent, and any preservatives,buffers, or propellants that may be required. A generally preferred composition for administering PDE 2 inhibitors,induding the compounds of formula (I), as weli as the combinations comprising PDE2 inhibitors and EP2 sélective receptor agonists in the treatment of bone fractures,comprises an injectable, flowable composition that provides sustained release at alocal site of injection by forming a biodégradable solid or gel depot, matrix, or implant.An example of such an administration System comprises a slow-release,biodégradable polymer-based delivery System. See, for example, U.S. Published PatAppl’n. No. 2003-0104031 A1. ,

Such a polymer-based delivery System generally comprises atherapeutically useful agent(s), dissolved or dispersed in a flowable, biodégradable,thermoplastic polymer solution or dispersion in an organic solvent. Upon injection ofthe composition, the organic solvent diffuses away from the injection site, causingthe polymer to precipitate or gel, thereby entrapping the agent(s) in a sustained-release depot. The agent(s) is subsequently released by diffusion from, andérosion of, the polymeric matrix. The matrix siowly erodes by hydrolysis andeventually disappears from the site of administration. The molecular weight andconcentration of the polymer can contrai the in vivo release of the agent(s) as weilas the dégradation rate of the matrix.

The polymer-based delivery System provides sustained release of an activeagent(s) in vivo for a sustained period of tirne with minimum or reduced burst in apatient in need thereof. A large burst of agent(s) would resuit in poor local tolerationdue to local effects (e.g., irritation) and would minimize the amount of agent(s)available for efficacy. The advantage this administration method offers is that itminimizes or reduces the initial burst, but still delivers the agent(s) at efficaciousieveis for sustained periods of tirne upon a single local injection.

The polymeric System is prepared by contacting the flowable compositionwith a gélation medium to coagulate or gel the composition into a solid,microporous polymeric matrix, or a gel polymeric matrix. The flowable compositioncontains a thermoplastic polymer or copolymer in combination with a suitabtesolvent. The polymère or copolymere, which form the body of the matrix, are 013337 -14- substantially insoluble, preferably essentially completely insoluble, in water andbodily fluids. The insolubility of the matrix body enables it to fonction as a single sitefor the controlled release of the agent(s). The polymers or copolymers are alsobiocompatible and biodégradable and/or bioerodlble within the body of an animal,e.g., mammal. The biodégradation enables the patient to metabolize and excretethe polymeric matrix such that there is no need for surgical removal. Because thefiowable composition and polymer System are biocompatibie, the insertion processand the presence of the polymer System within the body do not cause substantieltissus irritation or necrosis at the implant site. The composition of the présentinvention is administered as a fiowable composition directly into bodily tissues.

Suitable thermoplastic polymère for incorporation into the solid matrix of thecontrolled-release- System are solids, pharmaceutically compatible andbiodégradable by cellular action and/or by the action of bodily fluids. Examples ofappropriate thermoplastic polymère include polyesters of diols and dicarboxylicacids or of hydroxycarboxylic acids, such as poiylactides, polyglycolides, andcopolymers thereof. More preferably the polymer comprises the copolymer, poly-lactic-co-glycolic acid (abbreviated PLGH) which, upon hydrolysis, produces lacticand glycolic acid. The burst of release of this copolymer can be minimized furtherby the addition of polyethylene glycol (PEG) to form the PEG end-cappedl PLGH.

Preferred materials comprise poiylactides, polyglycolides, and copolymersthereof. These polymère can be used to advantage in the polymer System in partbecause they show excellent biocompatibility. They produce little, if any, tissueirritation, inflammation, necrosis, or toxiclty. In the presence of water, the polymèreproduce lactic and glycolic acid, respectively, which are readily metabolized. Thepoiylactides can also incorporais glycolide monomer to enhance the resuitlngpolymeric dégradation. These polymère are also preferred because they effectivelycontrol the rate of release of agent(s) from the polymeric System, and because theyresuit in the local rétention of the agent(s) at the site of the site of administration.

The solubility or miscibility of a thermoplastic polymer in the organic solventof the composition will vary according to factors such as crystallinity, hyd rophilicity,capacity for hydrogen bonding, and the molecular weight of the polymer.Consequently, the molecular weight and the concentration of the polymer in thesolvent are adjusted to achieve desired miscibility, as well as a desired release ratefor the incorporated agent(s). 013337 -15-

The flowable composition of thermoplastic polymer, solvent, and theagent(s) comprises a stable flowable substance. A homogenous solution of theagent(s) in an organic solvent preferabiy results. The thermoplastic polymer issubstantially soluble in the organic solvent. Upon placement of the flowablecomposition into the body, the solvent dissipâtes and the polymer solidifies or gelsto form the polymeric System having the agents) within a solid or gel polymericmatrix.

For certain preferred polymère, the molecular weight of the polymer orcopolymer is adjusted to be within a range of about 0.2 to about 0.4 inhérent viscosity(I.V. in dedliters/g) for effective sustained release of the bone growth promotlngcompound. The typical rate of release of the incorporated agent(s) occure at an I.V.of about 0.2 (about 8,000 to about 16,000 molecular weight) or about 0.3 (about23,000 to about 45,000 molecular weight), but can vary depending on the particularcomponents of the composition. For most système, it is preferred to adjust themolecular weight of the polymer to about 0.2 I.V. for an effective sustained release ofthe agent(s).

For a poly(DL-lactide) or a lactide-co-glycolide polymer System, the desiredmolecular weight range is about 0.2 to about 0.4 I.V., with an I.V. of about 0.2being preferred. The molecular weight of a polymer can be modified byconventional methods.

Especially preferred, commercially available thermoplastic polymèrecomprise the following: PLGH copolymer with 1:1 ratio of lactic and glycolic acidwith an inhérent viscosity of about 0.2 dl/g (commercially available from BoehringerIngelheim as Copolymer RESOMER® RG 502 H) (about 12,000 molecular weight);PLGH copolymer with 1:1 ratio of lactic and glycolic acid with an inhérent viscosityof about 0.3 dl/g (commercially available frorn Boehringer Ingelheim as CopolymerRESOMER® RG 503 H)(about 37,000 molecular weight); PLGH copolymer with 1:1ratio of lactic and glycolic acid with an inhérent viscosity of about 0.4 dl/g(commercially available from Boehringer Ingelheim as Copolymer RESOMER® RG504 H) (about 47,000 molecular weight); and polyethylene glycol (PEG) end-capped PLGH copolymer with 1:1 ratio of lactic and glycolic acid with an inhérentviscosity of about 0.79 dl/g (commercially available from Boehringer Ingelheim asPLG-PEG) (about 52,000 molecular weight). 013337 -16-

The solvents employées in the thermoplastic compositions are preferablypharmaceutically acceptable, biocompatible, and will dissipate into bodily fluid insitu such that they may be classed as having a solubility in water ranging fromhighly soluble to insoluble. Preferably, they cause relatively little, if any, tissueirritation or necrosis at the site of the injection and implantation. Preferably, thesolvent will hâve at least a minimal degree of water solubility. When the organicsolvent is water-insoluble or is minimally soluble in water, the solvent will slowlydisperse from the flowable polymeric composition. The resuit will be an implantthat, during the course of its life, may contain varying amounts of residual solvent.Preferably, the organic solvent has a moderate to high degree of water solubility sothat it will faciiely disperse from the polymeric composition into bodily fluids. Mostpreferably, the solvent disperses rapidly from the polymeric composition so as toquickly form a solid implant. Concomitant with the dispersion of solvent, thethermoplastic polymer coagulâtes or gels into the solid polymeric System.Preferably, as the thermoplastic polymer coagulâtes, the solvent dispersion causespore formation within the polymer System. As a resuit, the flowable compositioncontaining thermoplastic polymer, solvent, and agent(s) will form a porous solidpolymer system. Also, when the solvent is slightly water-soluble, or Is water-insoluble, the solvent dispersion may resuit in the formation of a solid porousimplant, or if some solvent remains with the implant, the resuit may be formation ofa gel implant having few or no pores.

Suitable solvents indude those liquid organic compounds meeting theforegoing criteria. A generally preferred solvent comprises N-methyl-2-pyrrolidone(NMP).

The solvents for the thermoplastic polymer flowable compositions arechosen for compatibility and appropriate solubility of the polymer and solvent.Lower molecular weight thermoplastic polymers will normally dissolve more readilyin the solvents than high molecular weight polymère. As a resuit, the concentrationof a thermoplastic polymer dissolved in the various solvents diffère depending upontype of polymer and its molecular weight. Convereely, the hïgher molecular weightthermoplastic polymère will tend to coagulate, gel or solidifÿ faster than the very lowmolecular weight thermoplastic polymers. Moreover, the higher molecular weightpolymers tend to give higher solution viscosities than the low molecular weightmateriels. Thus, for advantageous injection efflciency, in addition to advantageous 013337 -17- release rate, the molecular weight and the concentration of the polymer in thesolvent are controiled.

Upon formation of the polymer System from the flowable composition, theagent(s) becomes incorporated into the poiymeric matrix. After insertion of theflowable composition to form the poiymeric System, the agent(s) is released fromthe matrix into the adjacent tissues or fluids by diffusion and dégradationmechanisms. Manipulation of these mechanisms also can influence release of theagent(s) into the surroundings at a controiled rate. For example, the poiymericmatrix can be fôrmulated to dégradé after an effective and/or substantiel amount ofthe agent(s) is released from the matrix. Thus, release of the agent(s) from thematrix can be varied by, for example, the solubility of the agent(s) in water, thedistribution of the bone growth-promoting compound within the matrix, or the size,shape, porosity, solubility, and biodegradability of the polymer matrix, among otherfactors. The release of the agent(s) from the matrix is controiled relative to itsinhérent rate by varying the polymer molecular weight to provide a desired durationand rate of release.

For example, a preferred dosage form of the agent(s) comprises a lyophileto be reconstituted with a solution of PLGH in NMP before administration. Thedosage form, consisting of the lyophilized compound in one syringe (syringe A) anda solution of PLGH in NMP in a second syringe (syringe B), is known as the A/Breconstitution System. The contents of both syringes are mixed togetherimmediately prior to dose delivery at or near site. After reconstitution, the contentsare transferred into a graduated dosing syringe for delivery. The administereddosage forms will be a solution and will resuit in the dispersion of the compoundwith PLGH in NMP at desired strengths of, for example, 5 and 50 mgA/ml (mgA/mlrefera to the free acid équivalent of the sodium sait form of the agent(s)). Thedosage form is a parentéral (e.g., subcutaneous, intramuscular, or intramedullary)sustained-release injection for local administration. This compound in a slow-release polymer matrix (depot injection) is designed for administration at or near asite, and is not intended for intravenous administration. To provide adéquate shelf-life stability for the dosage form, a two-syringe System (A/B), as described above,may be used, preferably with the sodium sait form of the compound. A uniphaseformulation, preferably with the free acid form of the compound, is a preferredalternative formulation. Based on the agent(s) and polymer stability, stérile filtration 013337 -18- 10 15 of the agent(s) and irradiation of the polymer solution may be prefemed formanufacturlng a stable stérile product. In one embodiment, the dosage form can bemanufactured and shipped as separate aluminum pouches containing syringesfilled with the lyophile form of the agent(s) in one pouch and the polymer solution inthe other pouch. Delivery containers, Systems, and methods for the lyophilization ofbone growth promoting compounds are described in published PCT InternationalPatent Application Publication No. WO 01/73363. Other methods of administrationinclude local administration by injection to a particular site or delivery by a cathéterto a site. Additional examples can be found in U.S. Provisional Application No.60/335,156, filed November 30, 2001.

The teachings of ali U.S. Pat. Nos. disclosed herein are incorporated byréférencé in their entirety.

The compounds of formula (I), the prodrugs thereof, and the pharmaceuticallyacceptable salts of the compounds and prodrugs, may be prepared according to theexemplary synthetic routes disclosed in the Schemes and Examples hereinbelow, aswell as by other conventional organfc préparative methods known, or apparent in lightof the instant disdosure, to one of ordinary skill in the relevant art. The methodsdisclosed in the instant Schemes are intended for purposes of exemplifying theinstant invention and are not to be construed in any manner as limitations thereon. 20

STEP2

H2N-(CH2)n-X-Y (VI) -► (i)

DIPEA; DMSO; 120° C

In Scheme 1, Step 1, 2,4-dichloro-pyrido[2,3-d]pyrimidine (IV) is reacted withan appropriately-substituted benzylamine (V) in the presence of a tri-substitutedamine base, such as triethylamine (TEA) or diisopropylethylamine (DIPEA), or an 013337 -19- aromatic base, such as pyridine. The réaction is typically effected in a polar alcoholicsolvent, such as methanol (MeOH), éthanol (EtOH), or isopropanol (1PA), at atempérature ranging from about 0° C to about 100° C. Preferably, the reaction iseffected in the presence of D1PEA in éthanol at about room température (RT). InStep 2, the resulting condensation product (VI) is then reacted with an appropriately-substituted amine (VII) in the presence of a tri-substituted amine base, such as TEAor DIPEA, or an aromatic base, such as pyridine, to afford compound (I). Thereaction is typically effected in a polar aprotic solvent, such as N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), N-methylpyrrolidinone, orsulfoiane, at an eievated température ranging from about 60* C to about 250* C.Preferably, the reaction is effected in the presence of DIPEA in DMSO at betweenabout 90* C to about 120° C.

Although Scheme 1 has been depicted as a discreet, two-step process inwhich intermediate (VI) is isolated and then reacted with amine (VII), it has also beenfound convenient to prépare and react (IV) in situ with amine (VII) in a single-step. Insuch process, an aprotic solvent, preferably DMSO, is employed. This reaction is alsoeffected in the presence of DIPEA in DMSO at a température of between about 90* Cto about 120° C.

PREPARATIVE EXPERIMENTAL

Unless noted otherwise, ali reagents employed were obtained commercially.Uniess noted otherwise, the following experimental abbreviations hâve the meaningsindicated:

AcOH - acetic acid dec - décomposition DMAP - 4-dimethylaminopyridine

EtOAc - ethyl acetate hr- hour(s) LAH - lithium aluminum hydridemin - minute(s) MS - mass spectrometryNMR - nuclear magnetic résonanceTHF-tetrahydrofuranp-TsOH - p-toluenesulfonic acid 013337 -20-

Preparation 1 (2-Chloro-pvridof2.3-dlPvrimidin-4-vlH3.5-dimethoxy-benzyl)-amine Το a stired solution of 2,4 dichloro-pyrido[2,3-d]pyrirnidine (1.3 g, 6.7 mmol)and 3,5-dimethoxybenzylamine (1.1 g, 6.7 mmol) in 30 mL EtOH at RT was addedTEA (4 mL, 28.7 mmol). A precipitate formed that was filtered off and washed withcold EtOH followed by hexanes to give 1.8 g of the title compound (82%) as a solid.mp 185°C (dec). 1H-NMR (DMSO-de) 5: 9.5 (t, 1H), 8.9 (dd, 1H), 8.7 (dd, 1H), 7.5 (m,1H), 6.5 (d, 2H), 6.4 (t, 1H), 4.6 (d, 2H), 3.7 (s, 6H). MS (m/z, %): 331 (100).Préparation 2 (2-Chloro-pvridof2.3-d1pvrimidin-4-vl)-(3.4-dimethoxv-benzvl)-amine

This compound was prepàred in a manner analogous to that described inPréparation 1 using appropriate starflng matériels. 1H-NMR (DMSO-de) δ: 9.5 (t, 1H),8.9 (dd, 1K), 8.7 (dd, 1H), 7.0 (s,TH), 6.9 (d, 2H), 4.6 (d, 2H), 3.7 (s, 3H) 3.7 (s, 3K).MS (m/z, %): 331 (100).

Préparation 3 2-(4-Aminomethvl-phenvl)-propan-2-ol

To a stirred solution of 4-(1-hydroxy-1-methyl-ethyl)-benzonitrile (2.0 g, 12.4mmol) in 30 mL THF at 0°C was added dropwise 1.0 N LAH in THF (26 mL, 26.1mmol). The mixture was allowed to warm to RT, and then refluxed for 20 min. Themixture was then cooled to 0°C and quenched with 5 mL MeOH added dropwise. Themixture was diluted with 300 mL chloroform and washed with water (1 X 80mL), driedover magnésium sulfate, and concentrated to give 1.9 g (95%) of the title compoundas a solid. 1H-NMR (CDCI3) δ: 7.45 (d, 2H), 7.26 (d, 2H), 3.83 (s, 2H), 1.57 (s, 6H).GC-MS (m/e, %): 164 (M+, 15), 150 (80), 132 (75), 106 (100).

Préparation 4 3-(4-Acetvl-phenvh-propionitrile A mixture of 4-(2-bromoethyl)acetophenone (2.0 g, 8.8 mmol) and KCN (0.6g, 8.8 mmol) in 30 mL DMSO was heated at 75°C for 4 hr. The mixture was dilutedwith water and extracted with EtOAc. The organic extract was washed successivelywith water and brine, dried, and concentrated to give an oïl. Chromatography on silicagel eluting with 40% EtOAc/hexanes gave 0.8 g of an oil. 1H-NMR (CD3OD) δ: 7.9 (d,2H), 7.4 (d, 2H), 3.3 (t, 2H), 2.8 (t. 2H), 2.6 (s, 3H). GC-MS (m/e, %): 173 (M+, 20),158 (100). 013337 -21-

Préparation 5 3-Γ4-(1 -Hvdroxv-1 -methyl-ethyD-phenvil-propionitrileTo a stirred solution of 3 M methyl magnésium chloride in THF (3.3 mL, 9.8 mmol), further dïluted with 10 mL THF, was added dropwise a solution of 3-(4-acetyl-phenyl)-propionitrile (0.7 g, 3.9 mmol) in 10 mL THF at -40°C. The reaction mixturewas aliowed to slowly warm to RT overnight, cooled to 0°C, then quenched withaqueous AcOH added dropwise. The reaction mixture was diluted with water andextracted with EtOAc. The organic extract was washed successively with water and

I brine, dried, and concentrated to give 0.8 g of an oil. 1H-NMR (CDgOD) δ: 7.4 (d, 2H),7.2 (d, 2H), 2.9 (t, 2H), 2.7 (t, 2H), 1.5 (s, 6H). GC-MS (m/e, %): 189 (M+, 5), 174(100).

Préparation 6

2-l4-(3-Amino-propvl)-phenvn-propan-2-olTo a stirred solution of 1M LAH in THF, further diluted with 20 mL THF, at 0°C was added dropwise a solution of 3-[4-(1-hydroxy-1-methyl-ethyl)-phenyl]propionitrile(0.8 g, 4.0 mmol) in 10 mL THF. The reaction was aliowed to slowly warm to RT thenrefluxed for 4 hr. The reaction mixture was then cooled to 0°C and quenched withMeOH added slowly dropwise. The mixture was diluted with chloroform and washedwith water. The organic extract was filtered through diatomaceous earth, the filtrateconcentrated then diluted with ethyl acetate, dried, and concentrated to give 0.5 g ofan oil. 1H-NMR (CD3OD) δ: 7.4 (d, 2H), 7.1 (d, 2H), 2.6 (m, 4H), 1.7 (m, 2H), 1.5 (s,6H). MS (m/e, %): 194 (M++1,100), 176 (90).

Préparation 7 3-f4-(2-Methvl-ri .31dioxolan-2-vl)-phenvn-propionitrileA mixture of 3-(4-acetyl-phenyl)propionitrile (2.2 g, 13 mmol), ethylene glycol (2.8 mL, 51 mmol), and a catalytic amount of p-TsOH (~200 mg) in 100 mL toluenewas refluxed over a Dean-Stark trap for 18 hr. The mixture was diluted with EtOAcand washed successively with 5% sodium bicarbonate solution, water and brine,dried (MgSO4), and concentrated to give an oil. Chromatography on silica gel eiutingwith EtOAc/hexane solution gave 2.5 g of an oil. 1H-NMR (CDCIg) δ: 7.4 (d, 2H), 7.2(d, 2H), 4.0 (m, 2H), 3.8 (m, 2H), 2.9 (t, 2H), 2.6 (t, 2H), 1.6 (s, 3H). MS (m/e, %): 216(M+-1,1), 202 (100).

Préparation 8 3-f4-(2-Methylf1,31dioxolan-2-vl)-phenvn-propvlamine 013337 -22-

To a stirred solution of 3-[4-(2-methyl-[1,3]dioxolan-2-yl)-phenyl]-propionitrile(2.4 g, 11 mmol) in 30 mL THF was added dropwise a solution of 1M LAH in THF.The mixture was allowed to warm to RT then refluxed for 1 hr. The mixture wascooled to 0°C then quenched with MeOH added dropwise. The mixture was dilutedwith chloroform and washed with water. The resulting suspension was filteredthrough diatomaceous earth and the filtrats layers separated. The organic extractwas dried over MgSO4 and concentrated to glve 2.4 g of an oil. 1H-NMR (CDCI3) δ:7.4 (m, 2H), 7.1 (d, 2H), 4.0 (m, 2H), 3.8 (m, 2H), 2.7 (m, 2H), 2.6 (m, 2H), 1.7 (m,2H), 1.6 (s, 3H). MS (m/e, %): 221 (M+, 10), 206 (60), 189 (100).

Préparation 9 2.2.2-T rifluoiO-1 -(4-iodo-phenvl)-ethanol

To a stirred solution of 4-iodobenzaldehyde (2.0 g, 8.6 mmol) in 20 mL THF atRT was added 0.5M solution of trimethyl(trifluoromethyl)silane in THF (19 mL, 9.5mmol) followed by tetrabutylammonium fluoride (112 mg, 0.4 mmol). The mixture wasstirred at RT ovemight, poured Info 0.1 N hydrochloric acid, and extracted withEtOAc. The organic extract was in tum washed with water and brine, dried (MgSO4),and concentrated to give 2.6 g of an oil. 1H-NMR (CDCI3) 5: 7.7 (d, 2H), 7.2 (d, 2H),5.0 (m, 1H), 2.7 (d, 1H). MS (m/e, %): 302 (M+, 100), 233 (100).

Préparation 10 2-{3-r4-(2.2.2-TrifluoiO-1-hvdroxy-ethvl)-phenvll-prop-2-vnvf}-isoindole-1.3-dione

To a stirred suspension of 2,2,2-trifluoro-1-(4-iodo-phenyl)-ethanol (2.6 g, 8.5mmol.), N-propargylphthalimide (1.6 g, 8.5 mmol.),dichlorobis(triphenylphosphine)palladium (298 mg, 0.43 mmol.) and copper (I) iodide(82 mg, 0.43 mmol) in 20 mL THF at RT was added 5 ml TEA. The mixture wasdeaerated briefly under a stream of nitrogen and then refluxed for 6 hr. The mixturewas diluted with chloroform, washed with water, dried over MgSO4, and concentratedto give a solid. The soiid was tiiturated with EtOAc to afford 2.2 g of a solid. 1H-NMR(CDCI3) δ: 7.9 (m, 2H), 7.7 (m, 2H), 7.5 (d, 2H), 7.4 (d, 2H), 5.0 (m, 1H), 4.7 (s, 2H),MS (m/e, %): 359 (M*, 100).

Préparation 11 243-r4-(2.2.2-trilfuoro-1-hvdrox¥-ethv0-phenvll-propvl)-lsùindole-1.3.dione A mixture of 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-prop-2-ynyl}-isoindole-1,3-dione (2.2 g, 6.1 mmol) and 10% Pd/C (220 mg) in 150 mL EtOH and150 mL THF was shaken under 50 psi hydrogen at RT in a Parr apparatus for 2 hr. 013337 -23-

An additional 220 mg of 10% Pd/C was added and the mixture shaken under 50 psihydrogen at RT for an additional 2 hr. An additional 220 mg of 10% Pd/C was thenadded and the mixture shaken under 50 psi hydrogen at RT overnight. The mixturewas filtered through diatomaceous earth and the filtrate concentrated to give an oii.Chromatography on silica gel eluting with EtOAc/hexanes afforded 1.8 g of an oii. 1H-NMR (CDCIs) δ: 7.8 (m, 2H), 7.7 (m, 2H), 7.3 (d, 2H), 7.2 (d, 2H), 4.9 (m, 1H), 3.7 (t.2H), 2.7 (t, 2H), 2.0 (m, 2H). MS (m/e, %): 363 (M+, 15), 345 (35), 325 (50), 161(100).

Préparation 12 1-r4-(3-Amino-propvl)-phenvn-2,2,2-trifluorc>-ethanolTo a stirred suspension of 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenylJ- propyi}-isoindole-1,3-dlone (1.8 g, 5.0 mmol.) in 50 mL MeOK at'RT was addedhydrazine hydrate (0.46 ml, 15.0 mmol). The mixture was stirred at RT for 18 hr. Thereaction mixture was filtered and the filtrate concentrated to give an oii. The oii wastriturated with chloroform, filtered, and the filtrate concentrated to give 1.0 g of an oii.1H-NMR (CDCIa) δ: 7.3 (d, 2H), 7.2 (d, 2H), 5.0 (m, 1H), 2.6 (m, 2H), 2.5 (m, 2H), 1.6(m,2H). MS (m/e, %): 363 (M+, 10),216(100).

Préparation 13

Trifluoro-methanesulfonic acid benzoi1.2.5loxadiazol-5-vl esterTo a stirred solution of 5-hydroxybenzofurazan (1.8 g, 13.0 mmol), TEA (2.4 mL, 33.0 mmol) and DMAP (79 mg, 7.0 mmol) in 40 mL dichloromethane at -78°Cwas added dropwlse a solution of trifluoromethanesulfonic anhydride (2.8 mL, 17.0mmol) in 10 mL dichloromethane. The mixture was allowed to slowly warm to RT over3 hr, diluted with dichloromethane, and washed with water. The organic extract wasdried and concentrated to give an oii. Chromatography on silica gel eluting withdichloromethane afforded 3.1 g of an oii. 1H-NMR (CDCI3) δ: 8.0 (dd, 1H), 7.8 (dd,1H), 7.3 (dd, 1H). MS (m/e, %): 268 (M+, 60), 146 (60), 69(100).

Préparation 14 2-(Benzof1.2.5loxadiazol-5-vl-prop-2-vnvl)-isoindole-1.3-dioneThis compound was prepared in a manner analogous to that in Préparation 10 using appropriate starting materials. 1H-NMR (CDCI3) δ: 8.2 (s, 1H), 8.0 (dd, 1H),7.9 (m, 2H), 7.8 (m, 2H), 7.5 (dd, 2H), 4.7 (s, 2H). MS (m/e, %): 303 (M+, 100).Préparation 15 2-(3-Benzof 1.2.5~loxadiazol-5-vl-propyl)-isoindole-1,3.dione 013337 -24-

This compound was préparée! in a manner analogous to that in Préparation 11 using appropriate starting matériels. 1H-NMR (CDCI3) 5: 7.8 (m, 2H), 7.7 (m, 3H), 7.6 (s, 1H), 7.2 (m, 1H), 3.8 (t, 3H), 2.8 (t, 2K), 2.1 (m, 2H). MS (m/e, %): 307 (M\40), 290(30), 272(20), 160(100).

Préparation 16 3-Benzof1.2.51oxadiazol-5-yl-propylamineThis compound was prepared in a manner analogous to that in Préparation 12 using appropriate starting materials. 1H-NMR (DMSO-de):8 7.9 (m, 2H), 7.7 (m,1H), 7.5 (m, 1H), 2.7 (m, 2H), 2.5 (m, 2H), 1.7 (m, 2H). MS (m/e, %): 177 (M+, 40),160(100),

Préparation 17

Trifluoro-methanesulfonic acid benzothiazol-6-vl esterThis compound was prepared in manner analogous to that in Préparation 13 using appropriate starting materials. 1H-NMR (CDCI3) δ: 9.1 (s, 1H), 8.2 (d, 1H), 7.9(d, 1H), 7.4 (dd, 1H). MS (m/e, %): 283 (M*, 80), 150 (100).

Préparation 18 2-(3-Benzothiazol-6-vl-prop-2-ynvl)-isoindole-1.3-dioneThis compound was prepared in a manner analogous to that in Préparation 10 substituting trifluoro-methanesulfonic acid benzothiazol-6-yl ester (Préparation 17)for 2,2,2-trifluoro-1-(4-iodo-phenyl)-ethanol. 1H-NMR (dfl-DMSO) δ: 9.4 (s, 1H), 8.3 (d,1H), 8.0 (d, 2H), 7.9 (m, 1H), 7.8 (m, 2H), 7.5 (dd, 1H), 4.6 (s, 2H). MS (m/e, %): 318(M+, 100). 2-(3-Benzothiazol-6-yl-propyl)-isoindole-1.3-dioneThis compound was prepared in a manner analogous to that in Préparation 11 substituting 2-(3-benzothiazol-6-yl-prop-2-ynyl)-isoindole-1,3-dïone (Préparation19) for 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-prop-2-ynyl}-Îsoindole-1,3-dione (Préparation 10). 1H-NMR (CDCI3) δ: 8.9 (s, 1H), 8.0 (d, 1 H), 7.8 (m, 2H), 7.7(m, 2H), 7.3 (dd, 1H), 3.8 (t, 2H), 2.8 (t, 2H), 2.1 (m, 2H). MS (m/e, %): 322 (M+, 80),174(40),162(100).

Préparation 20 3-Benzothlazol-6-vl-propvlamine 013337 -25-

This compound was préparée! in a manner analogous to that in Préparation12 substituting 2-(3-benzothiazol-6-yl-propyl)-isoïndole-1,3-dione (Préparation 20) for2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-propyl}-isoindole-1,3-dione(Préparation 11). MS (mie, %): 192 (M*, 20), 175 (100).

Préparation 21 2-(3-lodo-phenvi)-2-methvl-f 1,31dioxolane A mixture of 3-iodoacetophenone (3.0 g, 12 mmol), ethylene glycol (2.7 mL,48 mmol), and p-toluenesulfonic acid (30 mg) in 50 mL toluene was refluxed under aDean-Stark trap for 18 hr. The mixture was diluted with EtOAc and washedsuccessively with 5% sodium bicarbonate solution, water, and brine, dried overMgSO4, and concentrated to give an oil. Chromatography on silica gel eluting withEtOAc/hexanes gave 3.1 g of an oil. 1H-NMR (CDCI3) 5: 7.8 (m, 1K), 7.6 (dd, 1H), 7.4(dd, 1H), 7.0 (m, 1H), 4.0 (m, 2H), 3.7 (m, 2H) 1.6 (s, 3H). MS (m/e, %): 290 (M*, 20),275 (100).

Préparation 22 2-i3-f3-(2-Methvl-i1,31dioxolan-2-vl)-phenvn-prop-2-vnvl}-isoindole-1.3-dione

This compound was prepared in a manner analogous to that in Préparation 10 substituting 2-(3-iodo-phenyl)-2-methyl-[1I3]dioxolane (Préparation 21) for 2,2,2-trifiuoro-1-(4-iodo-phenyl)-ethanol. 1H-NMR (CDCI3) δ: 7.9 (m, 2H), 7.7 (m, 2H), 7.5(s, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.2 (m, 1H), 4.7 (s, 2H), 4.0 (m, 2H), 3.7 (m, 2H), 1.6 (s, 6H). MS (m/e, %): 347 (M+, 15), 332 (100).

Préparation 23 2-(3-r3-(2-Methyl-f1,31dioxolan-2-yl)-phenyn-propyll-isoindole-1.3-dione

This compound was prepared in a manner analogous to that in Préparation 11 substituting 2-{3-[3-(2-methyl-[1,3]dioxolan-2-yl)-phenyl]-prop-2-ynyl}-isoindole- 1,3-dione (Préparation 22) for 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-prop-2-ynyl}-isoindole-1,3-dione (Préparation 10). 1H-NMR (CDCI3) δ: 7.8 (m, 2H), 7.7 (m,2H), 7.2 (m, 4H), 7.1 (d, 1 H), 4.0 (m, 2H), 3.7 (m, 4H), 2.6 (t, 2H), 2.0 (m, 2H), 1.6 (s,3K). MS (m/e, %): 351 (M+, 5), 336 (100).

Préparation 24 3-r3-(2-Methvl-i1,31dioxolan-2-vl)-phenvn-proDvlamine

This compound was prepared in a manner analogous to that in Préparation 12 substituting 2-{3-[3-(2-methyl-[1,3]dioxolan-2-yl)-phenylJ-propyil}-isoindole-1,3-dione (Préparation 23) for 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-propyl}- 013337 -26- isoindole-1,3-dione. 1H-NMR (CDCI3) δ: 7.3 (m, 3H), 7.1 (dd, 1 H), 4.0 (m, 2H), 3.8 (m,2H)„ 2.7 (t, 2H), 2.6 (t, 2H), 1.8 (m, 2H), 1.6 (s, 3H). MS (m/e, %): 221 (M+, 20), 206(60), 189(100).

Préparation 25 2-(3-lodo-DhenviV-propan-2-ol

To a stirred solution of methyl magnésium chloride (65 mmol) in 100 mL THFat 0°C was added dropwise 3-iodoacetophenone (4.0 g, 16.3 mmol). The reactionmixture was allowed to warm to RT then cooled to 0°C and an additional équivalent ofmethyl magnésium chloride was added. The mixture was allowed to warm to RT andstirred for 5 hr. The reaction mixture was quenched with MeOH, diiuted with water,acidified with glacial AcOH, and extracted with dichloromethane. The organic extractwas washed with* 5% sodium bicarbonate solution then concentrated.Chromatography on silica gel eluting with dichloromethane gave an oil whichsolidified upon standing. MS (m/e, %): 262 (M+, 80), 247 (100).

Préparation 26 24343-(1 -Hvdroxv-1 -methvi-ethvh-phenvn-prop-2-vnviy-isoindoie-1.3-dione

This compound was prepared in a manner analogous to that in Préparation 10 substituting 2-(3-iodo-phenyl)-propan-2-ol (Préparation 25) for 2,2,2-trifluoro-1-(4-iodo-phenyl)-ethanol. MS (m/e, %): 319 (M+, 90), 301 (80), 160 (100).

Préparation 27 24343-(1-Hvd roxv-methvi-ethvll-phenvn-propvll-isoindole-l ,3-dione

This compound was prepared in a manner analogous to that in Préparation 11 substituting 243-[3-(1-hydraxy-1-methyt-ethyl)-phenyl]-prop-2-ynyl}-isoindole-1,3-dione (Préparation 26) for 2-{3-[4-(2,2,2-trifluoro-1-hydrüxy-ethyl)-phenyl]-prop-2-ynylJ-isoindole-1,3-dione (Préparation 10). MS (m/e, %): 305 (M*-H2O,80), 145 (100).Préparation 28 243-(3-Amino-propyl)-phenvfl-propan-2-ol

This compound was prepared in a manner analogous to that in Préparation 12 substituting 243-(3-(1-hydroxy-1 -methyi-ethyl)-phenyl]-propyl}-isoindole-1,3-dione(Préparation 27) for 243-[4-(2,2,2-trifiuoro-1-hydroxy-ethyl)-phenyl]-propyl}-isoindole- 1,3-dione. MS (m/e, %): 193 (M+, 30), 162 (60), 145 (100).

Préparation 29 443-(1.3-Dioxo-l .3-dihydro-isoindol-2-vl)-prop-1 -νηνΠ-benzonitrile 013337 -27-

This compound was prepared in a manner analogous to that in Préparation 10 using appropriais starting materials. 1H-NMR (CDCI3) δ: 7.8 (m, 6H), 7.6 (d, 2H), 4.6 (s, 2H). MS (m/e, %): 286 (M+, 100).

Préparation 30 4-f3-( 1.3-Dioxo-1.3-dihvdro-isoindol-2-vl)-propyll-berizonitrile

This compound was prepared in a manner analogous to that in Préparation 11 substituting 4-(3-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-prop-1 -ynyl]-benzonitrile(Préparation 29) for 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-prop-2-ynyl}-isoindole-1,3-dione (Préparation 10). 1H-NMR (CDCI3) δ: 7.8 (dd, 2H), 7.7 (dd, 2H),7.5 (d, 2H), 7.3 (d, 2H), 3.7 (t, 2H), 2.7 (t, 2H), 2.0 (m, 2H). MS (m/e, %): 290 (M+,60), 161 (100).

Préparation 31 4-(3-Amino-propvl1-benzonitrile

This compound was prepared in a manner analogous to that in Préparation 12 substituting 4-[3-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-propyl]-benzonitrile(Préparation 30) for 2-{3-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)-phenyl]-propyl}-isoindole- 1,3-dione. 1H-NMR (CDCI3) δ: 7.5 (d, 2H), 7.3 (d, 2H), 2.7 (m, 4H), 1.8 (m, 2H). MS(m/e, %): 160 (M+, 20), 143 (100).

Préparation 32 2-Chloro-3,4-dimethoxv-benzaldehvdeoxime A mixture of 2-chloro-3,4-dimethoxybenzaldehyde (1.5 g, 7.5 mmol),hydroxylamine hydrochloride (650 mg, 9.4 mmol), and sodium acetate (1.5 g, 18.8mmol) in 30 ml MeOH and 15 ml water was heated at 65°C for 18 hr. The mixturewas diluted with water and extracted with EtOAc. The organic extract was washedsuccessively with water and brine, dried over MgSO4, and concentrated to give 1.7 gof a solid. 1H-NMR (CDCI3) δ: 8.5 (s, 1H) 7.6 (d, 1H), 6.8 (d, 1H), 3.9 (s, 3H), 3.8 (s,3H). MS (m/e, %): 215 (M+, 40), 199 (100).

Préparation 33 2-Chloro-3.4-dimethoxv-benzvlamine

To a stirred solution of the product of Préparation 32 (1.7 g, 7.8 mmol) in 40mL THF at 0°C was was added siowly dropwise a 1 M solution of LAH in THF (17 mL,17 mmol). The mixture was allowed to siowly warm to RT then refluxed for 2 hr. Themixture was cool to 0°C and the réaction quenched with MeOH added siowlydropwise. The mixture was diluted with water and extracted with chloroform. The 013337 -28- resulting émulsion was filtered through diatomaceous earth and the flltrate layersseparated. The organic extract was washed with water, dried (MgSO4) andconcentrated to give 1.1 g of an oiî. MS (mie, %): 202 (M+, 100).

Préparation 34 3-Ethoxv-4-methoxy-benzaldehvde oxime

This compound was prepared in a manner analogous to that in Préparation 32 using appropriate starting materials. 1H-NMR (CDCfo) S: 8.1 (s, 1H) 7.2 (d, 1H),7.0 (m, 1H), 6.8 (d, 1H), 4.1 (q, 2H), 3.9 (s, 3H), 1.5 (t, 3H). MS (mie, %): 195 (M+,100).

Préparation 35 3-Ethoxv-4-methoxv-benzvlamine

This compound was prepared in a manner analogous to that in Préparation 33 using appropriate starting materials. 1H-NMR (CDCI3) δ: 6.9 (s, 1H) 6.8 (s, 2H), 4.1(q, 2H), 3.8 (s, 2H), 1.5 (t, 3H). MS (mie, %): 181 (M\ 100).

Préparation 36 2-(3-Pvridin-4-vl-propvl)-isoindole-1.3-dione

To a stirred solution of 4-pyridinepropanoi (2.0 g, 14.5 mmol), phthalimide (2.1g, 14.5 mmol), and triphenylphosphine (4.9 g, 15.2 mmol) in 50 mL THF at 0°C wasadded dropwise diethyl azodicarboxylate (2.5 mL, 16.0 mmol). The mixture wasallowed to slowly warm to RT then stirred ovemight. The mixture was diluted with 0.1N hydrochloric acid and washed with diethyl ether. The aqueous extract was basifiedwith 6N sodium hydroxide and extracted with EtOAc. The organic extract was washedwith 1N sodium hydroxide and water, dried over MgSO4, and concentrated to give 1.8g of a solid. 1H-NMR (CDCI3) δ: 8.5 (s, 2H), 7.8 (m, 2H), 7.7 (m, 2H), 7.2 (m, 2H), 3.7(t, 2H), 2.7 (t, 2H), 2.0 (m, 2H).

Préparation 37 3-Pvridin-4-vl-propvlamine

This compound was prepared in a manner analogous to that in Préparation12 using the title compound of Préparation 36.1H-NMR (CDCI3) δ: 8.4 (m, 2H), 7.1(m, 2H), 2.7 (t, 2H), 2.6 (t, 2H), 1.8 (m, 2H). MS (m/e, %): 136 (M*, 30), 119 (35), 107(100).

The compounds of formula (I) were prepared as described in the followingExamples. 013337 -29-

Example 1 /^-O.S-Dimethoxv-benzvD-A^-^-pvridin^-vl-ethvD-PVrido^.S-cnpvrimidine^/- diamine A mixture of the title compound of Préparation 1 (174 mg, 0.5 mmol), 2-(2-aminoethyl)-pyridine (386 mg, 3.2 mmol), and DIPEA (0.2 mL, 1.1 mmol) in 1 mL ofDMSO was heated at 90°C for 18 hr. The mixture was poured into water andextracted with methylene chloride. The organic extract was washed with brine, driedover MgSO4, and concentrated. The resulting residue was triturated with EtOAc togive 83 mg (40%) of the title compound. m.p. 157-8°C. 1H-NMR (DMSO-de):6 8.6 (dd,1H), 8.5 (d, 1 H), 8.4 (dd, 1H), 7.7 (m, 1H), 7.2 (m, 1H), 7.0 (m, 2H), 6.5 (m, 2H), 6.3(m, 1H), 4.6 (m, 2H), 3.7 (s, 6H), 3.6 (m, 2H), 3.0 (m, 2H). MS (m/z, %): 417 (100).Anal. Calcd. for CjaH^NeOz: C, 66.3; H, 5.8; N, 20.2.. Found: C, 65.3; H, 6.3; N, 18.3.

The compounds of Examples 2 to 5 were prepared in a manner anaiogous tothat described in Example 1 using appropriate starting matériels.

Example 2 A/*-f3.5-Dimethoxv-benzvl)-Ala-f2-Pvridin-3-vl-ethvl)-Pvridof2.3-dlPvrimidine-2.4- diamine m.p. 185-6’ C. Anal. Calcd. for C23H24NeO2: C, 66.3; H, 5.8; N, 20.2. Found: C,64.5; H, 5.6; N, 19.6.

Example 3 AZt-f3.5-Dimethoxy-benzvl)-A/a-f2-pyridin-4-vl-ethyl)-pvridor2.3-dlPvrimidine-2.4- diamine m.p. 234-5°C. Anal. Calcd. for C, 66.3; H, 5.8; N, 20.2. Found: C, 66.0; H, 5.6; N, 20.0.

Example 4 A/*-f3.5-Dimethoxy-benzyl)-/\/2-2-pvridin-3-vlmethvl-Pvridof2.3-dlpvrimidine-2.4- diamine m.p. 222-3°C. Anal. Calcd. for C^H^NsC^: C, 65.7; H, 5.5; N, 20.9. Found: C,65.5; H, 5.5; N, 20.8.

Example 5 A/2-A/4-fi/s-(3.5-dimethoxv-benzvh-pvridQf2.3-dlPvrimidine-2.4-diaminem.p. 159-160°C. Anal. Calcd. for CæHzzNsO.,: C, 65.1; H, 5.9; N, 15.2. Found: C, 65.3; H, 5.9; N, 15.1. 013337 -30-

Example 6 N4-(3.5-Dimethoxv-benzvl)-f/-f2-(4-methoxv-phenvl)-ethvn-pvridof2,3-dlPvrimidine- 2.4-diamine A mixture of the title compound of Préparation 1 (100 mg, 0.5 mmoi), p-methoxyphenethylamine (89 μι, 0.61 mmoi), and DIPEA (105 pL, 0.61 mmol) in 0.8mL of DMSO was heated at 90°C for 18 hr. The compound was isolated and puriftedfrom the crude réaction mixture by directly injection on a reversed-phase préparativeHPLC (Shimadzu Corp.; Kyoto, Japan) using a step gradient of acetonitrile/watercontaining 0.1% ammonium hydroxide as an elutant. Fractions containing the desiredproduct were combined, concentrated, and the residue recrystallized from IPA to give80 mg (60%) of the title compound. m.p. 88-9°C. Anal. Calcd. for CæHsrNgOa: C,67.4; H, 6.1; N, 15.7. Found: C, 67.0; H, 6.3; N, 15.2.

The compounds of Examples 7 to 31 were prepared in a manner analogousto that described in Example 6 using appropriate starting materials. Spécifieexceptions to the reaction and/or purification conditions employed are noted.

Example 7 M-(3.5-Dimethoxv-benzvi)-A<z-(3-phenvl-propvi)-pvridof2.3-dlPvrimidine-2,4-diamine 1H-NMR (DMSO-de): 8.6 (dd, 1 K), 8.4 (d, 1H), 7.2 (m, br, 3K), 7.1 (m, br, 2H),7.0 (m, br, 1H), 6.5 (s, br, 2H), 6.3 (s, br, 1 H), 4.6 (m, br, 2H), 3.7 (s, br, 6H), 2.6 (m,br, 2H), 1.8 (m, br, 2H), 1.7 (m, br, 2H). MS (m/z, %): 417 (100). MS (m/z, %): 430(100).

Example 8 A/2-i2-(4-Chloro-phenvh-ethvll-/V4-(3.5-dimethoxv-benzvl)-pvrÎdof2.3-d|pvrimidine-2.4- diamine m.p. 85-90°C. Anal. Calcd. for C^NgOzCI: C, 64.1; H, 5.4; N, 15.6. Found:C, 63.6; H, 5.7; N, 15.3.

Example 9 A/2-Benzvl-A/4-(3.5-dimethoxv-benzvl)-pvridof2.3-dlPvritnidine-2.4-diamine m.p. 84-85°C. Anal. Calcd. for C^H^NgC^: C, 68.8; H, 5.8; N, 17.4. Found: C,68.5; H, 6.0; N, 17.4Example 10 A/4-(3,5-Dimethoxv-benzvl)-f72-(2-thiophen-2-vi-ethvi>pvridor2,3-d]pvrimidine-2.4- diamine 013337 -31- m.p. 120-1°C. Anal. Calcd. for CæHaNeOzS: C, 62.7; H, 5.5; N, 16.6. Found:C, 62.2; H, 6.0; N, 15.6.

ExampleJl 2-(4-ff4-(3.5-Dimethoxy-benzvlamino)-pvridof2.3-d1pyrimidin-2-vlamino1-methvl)- phenvO-propan-2-ol m.p. 105-9°C. Anal. Calcd. for CaeHæNsOs: C, 68.0; H, 6.4; N, 15.2. Found: C,67.7; H, 6.6; N, 14.4.

Example 12 A/4-(3.5-Dimethoxy-benzvl)-A/2-2-phenethyl-pyridof2.3-d1pyrimidine-2.4-dlamine m.p. 112-4°C. Anal. Calcd. for C24H25NgO2: C, 69.4; H, 6.1; N, 16.9. Found: C,68.5; H, 6.9; N, 15.3.

Example 13 N4-(,3.5-Dimethoxv-benzvl1-A/2-f2-i3.5-dinnethoxv-phenvl1-ethvl1-pvridof2,3- cf|pvrimidlne-2.4-diamine m.p. 95-100°C. Anal. Calcd. for CæHæNsOz,: C, 65.7; H, 6.2; N, 14.7. Found:C, 65.5; H, 6.6; N, 14.5.

Example 14 A/4-f3.4-Dlmethoxy-benzvl)-A/g-r2-f3-fluoro-phenvll-ethvn-pyridof2.3-dlpvrimidine-2.4- diamine m.p. 227-8°C. Anal. Calcd. for CjwH^NeOzF: C, 66.5; H, 5.6; N, 16.2. Found:C, 66.6; H, 5.6; N, 16.2.

Example 15 /V4-(3.4-Dimethoxy-benzvl)-A/2-f2-(2-fluoro-pheny0-ethvn-pvridor2.3-dlpyrimidine-2.4- diamine m.p. 220-2°C. Anal. Calcd. for CmH^NsOzF: C, 66.5; H, 5.6; N, 16.2. Found:C.66.4; H, 5.5; N, 16.1.

Example 16 A/4-(3.4-Dimethoxy-benzvl)-/V2-r2-(4-fluoro-phenvl)-ethyn-pyridoi2.3-dlpyrimidine-2.4- diamine m.p. 225-6°C. Anal. Calcd. for QmH^NsOzF: C, 66.5; H. 5.6; N, 16.2. Found:C, 66.4; H, 5.6; N, 16.2.

Example 17 A/t-f3.4-Dimethoxv-benzvn-/^-phenethvl-pvridof2.3-d1pvrimidine-2.4-diamine 013337 -32- m.p. 217-8°C. Anal. Calcd. for Cz^ælW C, 69.4; H, 6.1; N, 16.9. Found: C,69.4; H, 6.0; N, 16.8.

Example 18 M-(3.4-Dimethoxv-benzvl)-Ala-(4-phenvl-butvl)-pvridoi2,3-dlpvrimidine-2.4-diaminem.p. 194-5°C. Anal. Calcd. for CæHæNsOz: C, 70.4; H, 6.6; N, 15.8. Found: C,70.5; H, 6.6; N, 15.9.

Example 19 /^-(3,4-Dimethoxv-benzvD-M-(2-phenoxv-ethvn-pvridor2,3-d1pvrimidine-2,4-diaminem.p. 227-8°C. Anal. Calcd. for C24H25N5O3: C, 66.8; H, 5.8; N, 16.2. Found: C, 66.7; H. 5.7; N, 16.2.

Example 20 A/4-(3.4-Dimethoxv-benzvl)-A^-(2-trifluoromethvl-benzvl)-Pvridor2.3-dlPvrimidine-2.4- diamine m.p. 227-8°C. Anal. Calcd. for CaHzzNsOzFg: C, 61.4; H, 4.7; N, 14.9. Found:C, 60.6; H, 4.9; N, 14.8.

Example 21 2-f443-f4-(3.4-Dimethoxv-benzvlamino)-pvridof2.3-dlDvrimidin-2-vlamino1-propvll·- phenvl)-propan-2-ol

Reversed-phase préparative HPLC fractions containing crude desired productwere concentrated and the residue was recrystallized from EtOAc. m.p. 198-9°C.Anal. Calcd. for CaH^: C, 69.0; H, 6.8; N, 14.4. Found: C, 68.7; H, 6.9; N, 14.3.Example 22 2-('4-{3-f4-(3.5-Dimethoxv-benzvlamino>-pvridoi2.3-dlPvrimidin-2-vlaminol-propvft~ phenvO-propan-2-ol (amorphous solid) m.p. 70-5°C. 1H-NMR (DMSO-d6): 8.6 (dd, 1H), 8.4 (d, 1H),7.3 (m, br, 2K), 7.1 (m, br, 2H), 7.0 (m, br, 2H), 6.5 (s, br, 2H), 6.3 (s, br, 1H), 4.6 (m,br, 2H), 3.7 (s, br, 6H), 2.6 (m, br, 2H), 1.8 (m, br, 2H), 1.7 (m, br, 2H), 1.4 (s 6H). MS(m/z, %): 430 (100). MS (m/z, %): 488 (M*+1, 100), 470 (50). Anal. Calcd. forCajHæNsOa: C, 69.0; H, 6.8; N, 14.4. Found: C, 67.1 ; H, 7.0; N, 12.1.

Example 23 1-(4-fF4-f3.4-Dimethoxv-benzvlamino)-pvridof2.3-d1pyrimidin-2-vlamino1-methvll·- phenvl)-2.2.2-trifluoiO-ethanol

Reversed-phase préparative HPLC fractions containing crude desired productwere concentrated and the residue was recrystallized from acetonitrile/water. m.p. 01333? -33- 121-2°C. Anal Calcd. for WIAFÿ C, 60.1; H, 4.8; N, 14.0. Found: C, 58.0; H,5.0; N, 13.4.

Example 24 1-(4-f3-f4-(3.4-Dimethoxy-benzvlamino)-pvridoi2.3-dlpvrimidin-2-vlarTiino1-propvll·- phenvll-ethanone

The reaction mixture containing crude product was poured into water and theresulting precipitate was filtered off. The residue was dissolved in 30 mL of MeOHand 10 mL of 1 N hydrochloric acid was added. The mixture was stirred at RT for fourhr, concentrated, neutralized with 5% sodium bicarbonate solution, and extracted withdichloromethane. The organic extracts were combined and washed with brine, driedover MgSO4, and concentrated. The crude product was purified by reversed-phaseHPLC as described in Éxample 6. Fractions containing desired product werecombined, concentrated, and the residue recrystallized from acetonitrile/water tofumish a solid. m.p. 184-5°C. Anal. Calcd. for C27H2eNsO3: C, 68.8; H, 6.2; N, 14.9.Found: C, 68.7; H, 6.0; N, 14.6.

Example 25 1-(4-{3-f4-(3.4-Dimethoxv-benzvlamino)-pvridoi2.3-dlpvrimidin-2-vlaminoi-propvl)- phenvR-2.2.2-trifluoro-ethanol m.p. 222-4°C. 1H-NMR (DMSO-de):5 8.6 (dd, 1H), 8.4 (dd, 1H), 7.4 (m, br,2H), 7.3 (m, 1H), 7.0 (m, br, 2H), 6.8 (s, br, 2H), 6.7 (d, 1H), 5.0 (m, br, 1H), 4.6 (s,br, 2H), 3.7 (m, 6H), 3.3 (m, 2H), 2.6 (m, br, 2H), 1.8 (m, br, 2H). MS (m/z, %): 528(M*, 100).

Example 26 /V2-3-(Benzori.2.51oxadiazol-5-vl-propvi1-/V4-(3.4-dimethoxv-benzvn-Dvrido[2.3- dlpyrimidine-2.4-diamine

The product was isolated by chromatography on silica gel eluting with 2.5%MeOH in 2 N ammonia/dichloromethane to give a crude solid. Trituration with EtOAcafforded a solid. m.p. 191-3°C. 1H-NMR (DMSO-de):5 8.6 (dd, 1H), 8.4 (dd, 1H), 7.9(m, br, 1H), 7.7 (m, 1H), 7.5 (m, br, 1H), 7.0 (m, br, 2H), 6.8 (m, 2H), 4.6 (m, br, 2H), 3.7 (m, 6H), 3.4 (m, 2H), 2.7 (m, br, 2H), 1.9 (m, br, 2H). MS (m/z, %): 472 (M+, 100).Example 27 A^-3-(Benzothiazol-6-vl-propvl)-A/4-(3.4-dimethoxv-benzvl)-pvridof2.3-d1pvrimidine- 2,4-diamine 013337 -34-

The product was isolated by chromatography on silïca gel eluting wfth 2.5%MeOH in 2 N ammonia/dichloromethane to give a solid. mp 191-3°C. MS (m/z, %):487 (IVT, 100). Anal. Calcd. for CæHæNeQzS: C, 64.2; H, 5.4; N, 17.3. Found: C, 64.5;H, 5.5; N, 16.8.

Example 28 M-ra^Dimethoxv-benzvO-Af-ÎS-ra-fë-methvl-ri.SI-dioxolan^-vO-phenvII-propvIl· pvridof2.3-dlpvrimidine-2.4-diamine 1H-NMR (DMSO-de):ô 8.6 (d, 1 H). 8.3 (d, 1 H), 7.2 (m, br. 3H), 7.0 (m, br, 3H),6.9 (m, br, 2H), 4.6 (m, br, 2H), 4.1 (m, 2H), 3.9 (m, 2H), 3.7 (m, br, 8H), 2.6 (m, br,2H), 1.8 (m, br, 2H)1.5 (s, 3H). MS (m/z, %): 516 (M++1,100).

Example 29 2-(3-f3-r4-(3.4-Dimethoxv-benzvlamino)-pvridof2.3-dlPvrimidin-2-vlamlnol-propvn- phenvl)-propan-2-ol m.p. 204-5°C. 1H-NMR (DMSO-de):6 8.6 (d, 1H), 8.3 (d, 1H), 7.3 (m, br, 1K),7.2 (m, br, 1H), 7.1 (m, br, 1H), 7.0 (m, br, 3H), 6.8 (s. br, 2H), 4.6 (s, br, 2H), 3.7 (m,br, 6H), 3.3 (m, br, 2H), 2.6 (m, br, 2H), 1.8 (m, br, 2H), 1.4 (s, 6H). MS (m/z, %): 488(M++1,100).

Example 30 4-(3-f4-(3.4-Dimethoxv-benzvlamino>-pvridof2.3-d1pvrimidin-2-vlamino1-propvll· benzonitrile m.p. 220-2°C. 1H-NMR (DMSO-de):5 8.6 (d, 1H), 8.3 (d, 1H), 7.7 (m, br, 2H),7.4 (m, br, 2H), 7.0 (m, br, 2H), 6.8 (s, br, 2H), 4.6 (s, br, 2H), 3.7 (m, br, 6H), 3.3 (m,br, 2H), 2.7 (m, br, 2H), 1.8 (m, br, 2H). MS (m/z, %): 455 (M++1,100).

Example 31 A/4-(3,5-Dimethoxv-benzvl)-A/2-i3-pvridin-4-vl-propvl)-pvridof2.3-cnpvrimidine-2.4- diamine 1H-NMR (CD2CI2):ô 9.1 (d, 1H), 8.9 (m, br, 1H), 8.4 (m, br, 3H), 7.2 (m, br,2H), 7.1 (m, br, 1H), 7.0 (s, br, 1H), 6.9 (d, 1H), 6.6 (d, 1H), 4.8 (d, 2H), 3.7 (s, 3H), 3.7 (s, 3H), 3.6 (m, 2H), 2.7 (m, 2H), 2.0 (m, 2K). MS (m/z, %): 432 (M++1, 20), 256(15), 237 (30), 216 (100). 013337 -35-

The compounds of Examples 32-35 were prepared according to the followinggeneralized procedure. Spécifie exceptions to the reaction and/or purificationconditions employed are noted.

To a stirred solution of 2,4-dichloro-pyrido[2,3-d]pyrimidine (0.75 mmoi) andDIPEA (1.5 mmoi) in 3 mL DMSO was added 0.75 mmoi of an amine correspondingto formula (V), Scheme 1, STEP 1, at RT. The mixture was stirred at RT for one hr,then 2.25 mmoi of an amine corresponding to formula (VII), Scheme 1, STEP 2, andadditional DIPEA (2.25 mmoi) were added, and the mixture heated at 90°C for two hr.Example 32 A/*-f3.4-Dimethoxv-benzyl)-f/-f3-phenvl-propvl)-pvridof2.3-d1pvrimidine-2.4-diamine

The compound was isolated and purified from the crude reaction mixture bydirect injection onto a reversed-phase préparative HPLC using a step gradient ofacetonitrile/water containing 0.1% ammonium hydroxide as an eiutant. Fractionscontaining the compound were combined and concentrated to give a solid. m.p. 190-2°C. 1H-NMR (de-DMSO): δ 8.6 (dd, 1H), 8.4 (dd, 1H), 7.2 (m, br, 3H), 7.1 (m, br, 2H),7.0 (m, br, 2H), 6.8 (m, br, 2H), 4.6 (m, br, 2H), 3.7 (d, br, 6H), 3.3 (m, br, 2H), 2.6(m, br, 2H), 1.8 (m, 2H). MS (m/e, %): 431 (M*+1,50), 430 (M+, 100).

Examole 33 A/4-(3,5-Dimethoxv-benzvl)-Mg-(3-phenoxv-ethvl)-pvridof2,3-dlPvrimidine-2.4-diamine , The compound was isolated and purified from the crude reaction mixture bydirect injection onto a reversed-phase préparative HPLC using a step gradient ofacetonitrile/water containing 0.1% ammonium hydroxide as an eiutant. Fractions.'containing the compound were combined, concentrated, and the residuerecrystallized from IPA/water. m.p. 165-7°C. 1H-NMR (de-DMSO): 8.6 (dd, 1H), 8.4 (d,1H), 7.2 (m, br, 3H), 7.1 (m, br, 2H), 7.0 (m, 1H), 6.9 (m, br, 2H), 6.8 (m, br, 1H), 6.5(d, 2H), 6.4 (s, 1H), 4.6 (m, br, 2H), 4.0 (m, br, 2H), 3.7 (m, br, 8H). MS (m/e, %): 433(M++1, 50), 432 (M+, 100).

Example 34 M-(3-Ethoxv-4-methoxy-benzvi)-/\/2-(3-phenvl-propyl)-pvridof2,3-dipyrimidine-2,4- diamine

The compound was isolated and purified from the crude reaction mixture bydirect injection onto a reverse phase préparative HPLC using a step gradient ofacetonitrile/water containing 0.1% ammonium hydroxide as an eiutant. Fractionscontaining the compound were combined, concentrated, and recrystallized from 013337 -36- acetonitrile/water. m.p. 181-2°C. Anal. Calcd. for C2eH28N5O2: C, 70.4; H, 6.6; N, 15.8.Found: C, 70.7; H, 6.9; N, 15.9.

Example 35 2-(4-f3-f4-(3-Ethoxv-4-methoxv-benzvlamino)-pvridof2.3-dlpvrimidin-2-vlamino1- propvB-phenvO-propan-2-ol

The compound was isolated and purified from the crude reaction mixture bydirect injection onto a reverse phase préparative HPLC using a step gradient ofacetonitrile/water containing 0.1% ammonium hydroxide as an elutant. Fractionscontaining the desired compound were combined, concentrated, and recrystallizedfrom acetonitrile/water. m.p. 150-2°C. Anal. Calcd. for CæHæNeOs: C, 69.4; H, 7.0; N,13.7. Found; C, 69.1; H, 6.9; N, 13.7.

Example 36 1-(4-f3-r4-(3.4-Dimethoxv-benzvlamino)-Dvridof2.3-dlPvrimidin-2-vlamino1-propyll·- phenvl)-2.2.2-trifluoro-ethanone

To a stirred stirred solution of the title compound of Example 25 (150 mg,0.28 mmol) in 15 mL dichloromethane at RT was added 1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (180 mg, 0.43 mmol). The mixture was stirred at RT forsix hr, then an additional portion of 1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (180 mg, 0.43 mmol) was added and the mixture stirred at RT ovemight.The mixture was diluted with chloroform, washed with water, dried over MgSO4, andconcentrated to give an oil. Chromatography on silica gel eluting with 5% MeOH 2Nin ammonia/dlchloromethane affbrded an oil. Trituration with EtOAc yielded 45 mg ofa solid. m.p. 198-200°C. 1H-NMR (de-DMSO): δ 8.6 (dd, 1H), 8.4 (dd, 1H), 7.9 (m, br,2H), 7.5 (m, br, 2H), 7.0 (m, br, 2H), 6.8 (m, br, 2H), 4.6 (m, 2H), 3.7 (m, 6H), 3.3 (m,2H), 2.7 (m, br, 2H), 1.9 (m, br, 2H). MS (m/e, %); 526 (M+, 100).

Example 37 1-f3-(3-f4-Î'3.4-Dimethoxv-benzylamino)-DVridof2.3-dlpvrimÎdin-2-vlaminol-propvll·- phenvh-ethanone A mixture of the title compound of Préparation 27 (419 mg, 0.81 mmol) in 8mL of THF containing 10 mL of 6 N hydrochloric acid was stirred at RT for six hr. Thereaction mixture was extracted with EtOAc, the organic extract was washed with 5% l sodium bicarbonate solution, and then concentrated to give a solid. m.p. 152-5°C. 1H-NMR (de-DMSO): 8.8 (d, 1H), 8.7 (d, 1H), 7.7 (m, 2H), 7.4 (m, 3H), 7.0 (s, 1H), 6.8 013337 -37- (m, 2H), 4.6 (d, 2H), 3.7 (s, 3H). 3.6 (s, 3H), 3.5 (m, 2H), 2.7 (m, 2H), 2.5 (s, 3H), 1.9(m, 2H). MS (m/e, %): 471 (M*c 30), 470 (100).

BIOLOGICAL METHODOLOGIES PPE 2 Enzyme Isolation PPE 2 enzyme was isolated from human platelets with approximately 1.4 L ofblood from multiple donors used to make the platelet pellets. Platelets wereresuspended in approximately 75 mL of lysis buffer [20 mM Tris pH 7.2,5 mM MgCfe,250 mM Sucrose, 1 mM PTT, 1 μ1?2 mL Sigma Protease inhibitor #8340; Sigma-Aldrich; St Louis, MO] and lysed by sonication at 4°C, with 3 rounds of 1 min burststhen spun at 4°C overnight at 100,000x g. Cleared lysâtes were loaded to AKTAExplorer FPLC (Amersham Biosciences; Piscataway, NJ) in a sériés of threechromatographie séparations, with an average load volume of 25 mL. A 5 mL HITrapQ anion exchange column (Amersham Biosciences) was used. A buffer [20 mM TrispH 7.2, 5 mM MgCfe, 1 μΙ/2 mL Sigma Protease inhibitor #8340] was mixed with agradient of B Buffer [20 mM Tris pH 7.2, 500 mM NaCI, 5 mM MgC|2, 1 μΙ/2 mLSigma Protease inhibitor #8340] over 20 column volumes from an initial B Bufferconcentration of 0% to a final concentration of 100%. cGMP-hydrolyzing peaks werenoted with average resolution at low sait-125 mM NaCI (PDE 5) and high sait- 325mM NaCI (PPE 2). Two major cGMP activity fractions (PPE 5 and PPE 2) wereisolated and pooled separately. The PPE 2-pooled fraction total was approximately40 mL which was dispensed in ciyovials of 200 μΙ/vial and placed in storage at -80°C.PDE 2 Enzyme Binding Assav

The inhlbitory activity of the compounds of formula (I) on recombinant orisolated PPE 2, and other PPE’s, was determined using the ^HJcAMP scintillationproximity assay (SPA) kits from Amersham International (Litüe Chalfont, England).The SPA assays were performed using 96 well plates. The PPE SPA yttrium silicatebeads (Amersham Biosciences) bind preferentially to the linear nucléotide, GMP,compared to the cyclic nucléotide, cGMP. 3H-cGMP was added to the réaction andwhen the product, ^H-GMP, was in close proximity to the beads, the scintillant withinthe bead was excited, which was detected using a Packard scintillation counter(Perkin-Elmer Life Sciences; Boston, MA). The enzyme concentration used was inthe linear range and the Km of the enzyme was determined (15 uM). The finalsubstrate concentration was <1/3 of Km (1 μΜ) so that ICæ values would approximate 013337 -38- the «i values. The assay was validated using literature compounds as contrais beforetesting compounds. Then, PDE catalytic activity measurements obtained in thepresence of the test compound, and those obtained in the absence of the testcompound, were compared and the 10« value is determined. 5 The radioactive substrates and the products of the PDE reaction were determined quantitatively using a RACK-BETA 1219 liquid scintillation counter (LKBWallac; Freiburg, FRG). The ICeo values (concentrations with 50% inhibition) weredetermined with 1μΜ cAMP or cGMP using the peak fractions. The data were fittedwith four parameters with the aid of the sigmoidal logistic function. 10 Using the previousiy described PDE 2 enzyme isolated from human platelets and the method for assaying test compounds for inhibition of the enzyme, an ICæ of 1.7 uM was determined for EHNA. In addition, an ICæ of 3 nM for 9-(1-acetyl-4-phenyl-butyi)-2-(3,4-dimethoxy-benzyl)-1,9-dihydropurin-6-one was also determinedusing the aforementioned method. 15 The compounds of formula (I) generally exhibit inhibitory activity, expressed as ICso's against PDE 2, that are <1,000 nM. Ranges of PDE 2 inhibitory activity forthe compounds of formula (I) in Examples 1-37 are setforth in Table 1.

Table 1

Example PDE 2 Inhibition Example PDE 2 Inhibition 1 ++ 22 ++ 2 ++ 23 + 3 + 24 + 4 + 25 ++ 5 +++ 26 + 6 + 27 + 7 ++ 28 + 8 + 29 + 9 + 30 + 10 +++ 31 + 11 + 32 + 12 ++ 33 ++ 13 ++ 34 + 14 ++ 35 + 013337 -39- 15 4*4· 36 4 16 +++ 37 4 17 +++ 18 +4 19 ++ 20 21 4 PDE 2 Inhibition: +++ (ICæ <50 nM), ++ (ICæ 50-250 nM), + (ICso 250-1,000 nM)

The ability of PDE 2 inhibitors, including the oompounds of formula (I), to treatbone fracture and/or defect, or promote bone in-growth, may be demonstratedaccording to the following protocole. 5 RatTransvèrse Fémoral Fracture Model

Male Sprague-Dawley rats at 3 to 4 months of âge were used. The animaiswere anesthetized with ketamine and xylazine at doses of 100 and 10 mg/kg,respectively. The right hindlimb of each rat was shaved and cleaned. A 1 cmincision was made just latéral to the patella and the fémoral condyle was exposed. 10 A Kirschner wire (0.045" in diameter) was introduced into the intramedullary canalthrough the intercondyiar portion to serve as an internai stabilization. The muscleincision was closed with vicryl and the skin incision was closed with stainless Steelwound clips. The mid-diaphysis of the pinned fémur was fractured by means of athree-point bending device driven by a dropped weight. The rats were permitted full 15 weight-bearing and unrestricted activity after awakening from anesthésia. The testcompounds were administered on various days after surgery by percutaneousinjection onto the fracture site. The animais were sacrificed after treatment and thefémurs were coilected for analysis. Fracture healing was evaluated by usingradiography, histological, and biomechanical test (F. Bonnarens, et al., Journal of 20 Orthopaedlc Research, 2,97-101 (1984).

Studv Protocol and Results in the Rat Transverse Fémoral Fracture Model

Three month-old male rats were subjected to transverse fracture of their rightfémurs under general anesthésia. A single dose (5 mg) of A/*-(3,4-dimethoxy-benzyl)-/^-(3-phenyl-propyi)-pyrido[2,3-d]pyrimidine-2,4-diamine was percutaneously injected 25 into the fracture sites at the completion of fracture génération. Three weeks after theinjections, the rats were sacrificed and the right fémurs were harvested and analyzed.Force-to-failure and stiffness (indices of bone strength) were increased by 19% and 013337 -40- 02%, respectively, in the fémurs treated with /7^-(3,4-dimethoxy-benzyl)-A/z-(3-phenyl-propyl)-pyrido[2,3-d]pyrimidine-2,4-diamin@, compared to those of the fémurs treatedwith placebo.

Rat Perjosteal Injection Mode! 5 Rats were anesthetized with isoflurane (2-3 min) in a conduction chamber located in a fume hood. The right hindlimb of each rat was shaved and cleaned. A 25gauge needle attached to a syringe was pre-filled with a formulation of the testcompound for local injection. The formulation was injected onto the subperiosteum offémur in a volume of 5-15 μΙ_ for 14 days. The rats were sacrificed after dosing, the 10 fémurs were collected, and then analyzed by radiography and dual-energy X-rayabsorptiometry (DEXA). * Studv Protocol and Results In the Rat Periosteal Injection Model

The right fémur of three-week old male Sprague-Dawley rats received a daily

injection of vehicle or test compound five fîmes per week for two weeks. On Day 15,15 ail rate were sacrificed and the right fémurs were collected for analysis. Periostealbone Induction was assessed using radiography and DEXA. Radiography showednew bone formation located on the injection site of ali fémurs treated with testcompound. The bone minerai content (BMC) of the injected région of the fémur (areabetween lesser trochanter and mid-shaft of the fémur) was assessed by DEXA 20 comparing rats treated with test compound with those treated only with vehicle. in thismodel, /V4-(3,5-dimethoxy-benzyl)-/V2-(2-pyridin-4-yl-ethyl)-pyrido[2.3-d]pyrimidine-2,4··diamine, 2-(3-{3-[4-(3,4-dimethoxy-benzylamÎno)-pyrido[2,3-d]pyrimidin-2-ylamino]-propyi}-phenyi)-prapan-2-ol, and //-(S^-dimethojy-benzylj-A^-fS-phenyl-propyl)-pyrido[2,3-d]pyrimidine-2,4-diamine increased BMC by 34%, 49%, and 33% 25 respectively. Additionally, 9-(1-acetyl-4-phenyl-butyl)-2-(3,4-dimethoxy-benzyl)-1,9-dihydropurin-6-one increased BMC by 20% in the aforementioned model.

Claims (14)

1. 013337 -41- C LAI MS

   1. A compound of formula (I)

   the prodrugs thereof, and the pharmaceutically acceptable salts of said compoundsor prodrugs, wherein: R1 and R2 are hydrogen or methoxy, provided R1 and R2 are not bothhydrogen or both methoxy; nis 1,2,3, or 4; X is a bond; O; S; C=O; -N(R)-, wherein R is hydrogen or -(Cr C3)alkyl; -C(OH)-; or -SO2; and Y is benzoxazolyl; benzothiazolyl; benzofurazanyl; benzofuranyl;benzôthiadiazolyl; benzisoxazolyi; benzisothiazolyl; benzimidazolyl; pyridyl; isatïnyi;oxindolyl; indazolyl; indolyl; phenyl; thienyl; or furanyl; wherein Y is optionallysubstituted independently with from one to three halogen; trifluoromethyl; methoxy; -C(=O)CH3; cyano; -C(CH3)2OH; -CH(CK3)OH; -CH(CF3)OH; -C(C=O)CF3; -S02NH2; -C(=O)OCH3; -CH2COOH; h?0; thiazolyl; or oxadiazolyl.
2. 2. A compound of claim 1, wherein X is a bond, and Y is benzofurazanyl; thienyl;pyridyl; or phenyl, wherein phenyl is optionally substituted independently with one ortwo halogen; trifluoromethyl; methoxy; -C(=O)CH3; cyano; -C(CH3)2OH; -CH(CH3)OH;-CH(CF3)OH; -C(C=O)CF3; -SO2NH2; -C(=O)OCH3; -CH2COOH; thiazolyl; oroxadiazolyl.
3. 3. A compound of claim 1 or 2, wherein X is a bond, n is 2 or 3, and Y is thienyl;pyridyl; or phenyl, wherein phenyl is optionally substituted independently with one ortwo methoxy; halogen; -C(CH3)2OH; CH(CF3)OH; or -C(C=O)CF3. 013337 -42- 4. /V2, rf-bis-fi, 5-Dimethoxy-ben2yl}-pyrido[2,3-d]pyrimidine-2,4-diamine; M-(3,5-dimethoxy-benzyl)-/\/2-(2-pyridin-4-yl-ethyl)-pyrido[2,3-d]pyrimÎdine- 2,4-diamine; //-(3,5-dimethoxy-benzyl)-/V2-(2-thiophen-2-yl-ethyl)-pyrido[2,3-d]pyrimidine- 2,4-diamine; W4-(3,5-dimethoxy-benzyl)-W2-2-phenethyl-pyrido[2,3-d]pyrtmidine-2,4- diamine; 7/-(3,5-dimethoxy-benzyl)-/V2-[2-(3,5-dimethoxy-phenyl)-ethyl]-pyrÎdo[2,3-d]pyrimidine-2,4-diamine; 2-(343-[4-(3,4-dimethoxy-benzylamino)-pyrido[2,3-d]pyrimidin-2-ylamina]- propyl}-phenyl)-propan-2-ol; //-(3,4-dimethoxy-benzyl)-A/2-[2-{4-fluoro-phenyl)-ethyl]-pyrido[2,3- d]pyrimidine-2,4-diamine; W4-(3,4-dimethoxy-benzyl)-//-phenethyl-pyrido[2,3-d]pyiimidine-2,4-diamine; or /V4-(3)4-dÎmethoxy-benzyl)-W2-(3-phenyl-propyl)-pyrido[2,3-d]pyrimidine-2,4-diamine; a prodrug thereof, or a pharmaceutically acceptable sait of said compoundor prodrug.
4. 5. A pharmaceutical composition comprising a compound of formula (I) of any ofdaims 1-4, a prodrug thereof, or a pharmaceutically acceptable sait of saidcompound or prodrug, and a pharmaceutically acceptable vehide, carrier, or diluent.
5. 6. Use of a compound of formula (I) of any of daims 1-4, a prodrug thereof, or apharmaceutical acceptable sait of said compound or prodrug; or a pharmaceuticalcomposition comprising said compound of formula said prodrug thereof, or a acceptablesait of said compound or prodrug, and a pharmaceutical acceptable vehicle, carrier, ordiluent in the manufacture of a médicament for treating a PDE 2-mediated condition,disease, or symptom in a mammal in need of such treatment. 013337 -43-
6. 7. A use of claim 6, wherein said condition, disease, or is osteoporosis, pulmonaryhypertension, female sexual arousal disorder, diminished memory orcognition, plateletaggregation, vascular angiogenesis, dementia, cancer, arrhythmia, thrombosis, bonefracture defect, delayed or non-union fracture, spinal fusion, bone in-growth, cranialfacial reconstruction, or hypoxia which method comprises administering to mammal inneed of such treatment a therapeutically effective amount of a compound of formula ofclaim 1, a prodrug thereof, or a pharmaceutical acceptable sait of said compound orprodrug; or a pharmaceutical composition comprising said compound, said prodrugthereof, or said pharmaceutical acceptable sait of said compound or prodrug.
7. 8. A use of claim 6, wherein said condition is bone fracture and/or defect
8. 9. A pharmaceutical composition comprising a PDE 2 inhibitor, an EP2 sélectivereceptor agonist, and a pharmaceuticaliy acceptable vehicie, carrier, or diluent.
9. 10. A composition of claim 9, wherein said PDE 2 inhibitor is /^-(3,5-dimethoxy-benzyl)-A/2-(2-pyridin-4-yl-ethyl)-pyrido[2,3-d]pyrimidin-2,4-diamine; 2-(3-{3-[4-(3,4-dimethoxy-benzylamino)-pyrido[2,3-d]pyrimidin-2-ylamino]-propyl}-phenyl)-propan-2-ol; A/4-(3,4-dimethoxy-benzyl)-/V2-(3-phenyl-propyl)-pyrido[2,3-d]pyrimidine-2,4-diamine; a prodrug thereof, or a pharmaceuticaliy acceptable sait of said compoundor prodrug.
10. 11. A composition of daim 9 or 10, wherein said said EP2 sélective receptor agonist is(3-(((4-ferf-butyl-benzylHpyridine-3-sulfonyl)-amino)-methyl)-phenoxy)-acetic acid, aprodrug thereof, or a pharmaceuticaliy acceptable sait of said compound or prodrug.
11. 12. A use of any of daims 5-8, wherein a therapeutically effective amount of an EP2sélective receptor agonist, or a pharmaceutical composition comprising a combinationof said compound of formula of claim 1 and said sélective receptor agonist are alsoprovided.
12. 13. A use of claim 12, wherein said PDE 2 inhibitor [2, 3-d] pyrimidin-2,4-diamine ; 4-dimethoxy- [2,3-d] 4- [2,3-d] pyrimidine-2,4-diamine ; a prodrug thereof, or apharmaceutical acceptable sait of said compound or prodrug. -44- 013337
13. 14. A use of claim 12 or 13, wherein said sélective receptor agonis! is acid, a prodrugthereof, or a pharmaceutical acceptable sait of said compound or prodrug.
14. 15. Use of a PDE 2 inhibitor, a prodrug thereof, or a acceptable sait of said inhibitor 5 or prodrug in the manufacture of a médicament for treating bone fracture and/or defect ina mammal in need of such treatment. 10