NL1026091C2 - New cyclic guanosine monophosphate derivatives are phosphodiesterase 9 inhibitors useful to treat conditions, diseases or symptoms of e.g. hyperglycemia, dyslipidemia, metabolic syndrome, type 1 diabetes and cardiovascular diseases - Google Patents

Abstract

Cyclic guanosine monophosphate derivatives (I) and their stereoisomers and/or prodrugs or salts are new. Cyclic guanosine monophosphate derivatives of formula (I) and their stereoisomers and/or prodrugs or salts are new. [Image] A : 8 heterocyclic derivatives e.g. purine 2-one derivative of formula (b) or 2,4,6-tirazole imidazole derivative of formula (c); ring P+C : 3-8C (hetero)cycloalkyl or (hetero)aryl (all optionally substituted with 1-3 substituents of halo, 1-5C alkyl, 1-5C alkoxy or CF 3); J : O, S, N(R 1> 5>), N(R 1> 5>)CO, CON(R 1> 5>), SO 2N(R 1> 5>) or N(R 1> 5>)SO 2; R 1> 0>CO 2H, CONR 3> 0>R 3> 1>, NR 3> 0>R 3> 1> or N(R 1> 5>)SO 2R 4> 0>; R 1>, R 2>H or 1-3C alkyl; R 3>1-8C alkyl, 3-8C (hetero)cycloalkyl, 3-8C (hetero)cycloalkyl-methyl or (hetero)aryl (all optionally substituted with 1-3 substituents of halo, OH, oxo, 1-5C alkyl or 1-5C alkoxy); R 1> 5>H or 1-5C alkyl; either R 3> 0>, R 3> 1>H, 1-5C alkyl, 3-8C (hetero)cycloalkyl or (hetero)aryl (all optionally substituted with 1-3 substituents of halo, oxo, 1-5C alkyl, CO 2R 4> 0>, COR 4> 0>, OR 4> 0>, CONR 5> 0>R 5> 1>, NR 5> 0>R 5> 1> or SO 2R 4> 0>); or NR 3> 0>R 3> 1>5-8 membered heterocycloalkyl ring (the ring optionally having 1 additional heteroatom of N, O or S) (optionally substituted with 1-3 of halo, oxo, 1-5C alkyl, CO 2R 4> 0>, COR 4> 0>, OR 4> 0>, CONR 5> 0>R 5> 1>, NR 5> 0>R 5> 1> or SO 2R 4> 0>); R 4> 0>H, 1-5C alkyl, 3-8C (hetero)cycloalkyl or (hetero)aryl; either R 5> 0>, R 5> 1>H, 1-5C alkyl, 3-8C (hetero)cycloalkyl or (hetero)aryl; or NR 5> 0>R 5> 1>5- to 8-membered heterocycloalkyl ring (the ring optionally having 1 additional heteroatom of N,O or S); and x : 0-6. [Image] ACTIVITY : Antidiabetic; Antilipemic; Cardiovascular-Gen. MECHANISM OF ACTION : Phosphodiesterase 9 (PDE9) inhibitor. (I) were tested for PDE9 inhibitory activity using biological assays. The results showed that (I) exhibited a median inhibitory concentration value of less than 50 nM.

Description

•; # ι. ·

PDE9 - BRAKES FOR TREATMENT OF TYPE 2 DIABETES, ME-5 TABOLIC SYNDROME AND CARDIOVASCULAR DISEASE

FIELD OF THE INVENTION

The present invention relates to novel cyclic guanosine monophosphate (hereinafter referred to as cGMP) specific phosphodiesterase type 9 inhibitors (PDE9 inhibitors) for the treatment of a variety of diseases, in particular diabetes, including type 1 and type 2 diabetes, hyperglycaemia, dyslipidemia, impaired glucose tolerance, metabolic syndrome and / or cardiovascular disease. In addition, the invention relates to methods and processes relating to these inhibitors and to compositions containing these inhibitors.

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BACKGROUND OF THIS INVENTION

The group of phosphodiesterase enzymes hydrolyses the cyclic nucleotides cGMP and cyclic adenosine monophosphate (cAMP). cGMP and cAMP play a central role in the control and regulation of a range of cellular processes, both physiological and pathophysiological processes.

The PDE9 enzyme has been identified as a new member of the group of phosphodiesterase enzymes (PDE) that selectively hydrolyses cGMP over cAMP. See D.A.

Fisher et al., J. Biol. Chemistry, vol 273, no. 25, 15559-15564 (1998), which publication is incorporated herein by reference. It was discovered that PDE9 is present in a range of human tissues, namely the testes, the brain, the small intestine, the skeletal muscle, the heart, the lung, the thymus and the spleen. We have now discovered that PDE9 is present in smooth muscle cells within the human vein system of a range of tissues.

It is believed that insulin is the sag. Vian blood cells, at least in part by the action 102609j-. ♦ '2 of nitrogen monoxide (NO). Nitrogen monoxide that is generated in the endothelium then stimulates the production of cGMP in blood cells and causes it to relax or widen. This opening of the blood cells allows more blood to flow, which is particularly important when there is a higher blood flow to critical organs, such as the heart. is required. Patients with insulin resistance have been shown to have a reduced release of NO from the endothelium. This reduced release of NO comes not only from insulin but also from other important vasodilators such as acetylcholine. This so-called "endothelial disorder" contributes to the risk factors for cardiovascular disease associated with metabolic syndrome. The vascular effect of insulin contributes to the regulation of metabolism, in particular, but necessarily limited to, the metabolism of glucose.

Nitric oxide also has an effect on the uptake of glucose by the skeletal muscle. That is, treatment with an NO donor compound, such as nitroprusside, or with an analogue of cGMP, increases glucose uptake in vitro (transport through the GLUT4 glucose transporters). This pathway independent of vasodilation is described in G.J. Etgen, D.A. Fryburg and E.M. Gibbs in Diabetes, £ 6, 1997> p. 1915-1919, which publication is incorporated herein by reference. In summary, NO and cGMP have a direct target tissue (the skeletal muscle) and have an effect on the vasculature that influences, mediates or imitates insulin action.

There are two major forms of diabetes mel-litus. Type 1 diabetes, or insulin-dependent diabetes, is the result of an absolute deficiency of insulin, the hormone that controls the use of carbohydrates. Type 2 diabetes, or non-insulin-dependent diabetes, often occurs with normal or even elevated levels of insulin and appears to be the result of the 1 0260 91 - ♦ 3.

K *. . "inability of tissues to properly respond to insulin. Complications of type 1 and 2 diabetes include retinopathy, nephropathy, neuropathy and heart disease due to a coronary artery defect, and are thought to be caused by a coronary artery. combination of factors including excessive glycosylation of proteins and increased flux through the polyol pathway These disorders are thought to be the result of excessive levels of circulating glucose.

10 Metabolic syndrome, as in this description and according to the Adult Treatment Pane III (A. III; National Institutions of Health: Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults 15 (Adult Treatment Panel III), Executive Summary; Bethesda, MD, National Institutes of Health, National Heart, Lung and Blood Institute, 2001 (NIH Publication number 01-3670) is defined, acts as a person to three or more of the following criteria: 1. Obesity of the lower abdomen: circumference of the waist> 102 cm in men and> 88 cm in women 2. Hyper-triglyceridemia: a 150 mg / dl (1.695 mmol / l) 3. Low HDL cholesterol : <40 mg / dl (1.036 mmol / l) in men and <50 mg / dl (1.295 mmol / l) in women 25 4. High blood pressure: a 130/85 mmHg 5. High glucose in fasting: a 110 mg / dl (a 6.1 mmol / l)

The metabolic syndrome can also be associated with and / or sometimes referred to as syndrome X and / or insulin resistance syndrome.

Cardiovascular disorders, diseases and / or disorders, as defined in this specification, include systemic (or essential) high blood pressure, high blood pressure of the lung (e.g. high blood pressure of the veins of the lung, high blood pressure of the lung at 35), heart attack, heart disease due to a rejection.

. coronary artery, atherosclerosis, stroke, thrombosis. bose, disorders with reduced openness of the veins 102 6091 - 4 'κ * (eg angioplasty after percutaneous damage due to surgery on the coronary artery), peripheral vein disease. kidney disease (in particular those associated with diabetes), angina (including stable, unstable and variant (Prinzmetal) angina), and any condition in which an increased blood flow leads to an improved function of the organs. In particular, the vascular disease is systemic high blood pressure.

Accordingly, there is a need for a PDE9-10 inhibitor that leads to clinically relevant improvements in blood pressure, serum glucose, insulin; fats, uric acid and / or procoagulation factors. This treatment can take place on its own or in combination with other therapeutics.

I5. '' _ · '

SUMMARY OF THE INVENTION

The invention provides compounds of Formula (I), 20 N (CH 2) x R 10 (O.

a stereoisomer or prodrug thereof, or a pharmaceutically acceptable salt of that compound, stereoisomer or prodrug thereof; wherein A, P, J, X and R10 are as defined below; and pharmaceutical preparations thereof; and its use in the treatment of a variety of diseases, including diabetes, including type 1 and type 2 diabetes, hyperglycemia, dyslipidemia, impaired glucose tolerance, metabolic syndrome and / or cardiovascular disease.

102 60 9I

». »5

DETAILED DESCRIPTION OF THE INVENTION

The invention provides compounds of Formula 5 (I), 10 10 N (CH 2) x R 10. 0) a stereoisomer or prodrug thereof, or a pharmaceutically acceptable salt of that compound, stereoisomer or prodrug, wherein: | · ·

a

20 R 10, HN ^ S ^. 1 IO> j I y-r 2 ·? V> N% N-N-f (a) R 11 (b) R 3; (c) R 3;

25 O R1 O

ην '^ ιΓ ^ ° \ Λ ^ ν ^ VW'. W r3 I (e) R3 (f) R3; 30 A 9 R1 HN ^ Sr ^ N Τ'ιΤ'Λ ΰΧ / (g) · R3; or, (h) R 3; 35, 1 02 6091 6 P, including the carbon atoms to which it is attached, (C 3 -C 8) cycloalkyl, (C 3 -C 8) heterocycloalkyl, aryl or heteroaryl; optionally and independently substituted with 1 to 3 substituents independently selected from halogen, (C 1 -C 5) alkyl, (C 1 -C 5) alkoxy and trifluoromethyl; is; J is O, S, -N (R 15) -, -N (R 15) CO -, -CON (R 15) -, -SO 2 N (R x 5) -, -N (R 15) - SO 2 -; x is 0, 1, 2, 3, 4, 5 or 6; 10. R10 is -C02 H, -CONR30 R3x, -NR30R3x or -N (RX5) SO2R4 °; R 1 and R 2 are independently H or (C 1 -C 3) alkyl; R 3 is (C 1 -C 8) alkyl, (C 3 -C 8) cycloalkyl, (C 3 -C 6) cycloalkyl-methyl, (C 3 -C 8) heterocycloalkyl, (C 3 -C 6) heterocycloalkyl-methyl, aryl or heteroaryl; optionally and independently substituted with 1 to 3 substituents independently selected from halogen, hydroxy, oxo, (C 1 -C 5) alkyl and (C 3 -C 5) alkoxy; is; R15 is H or (Cx -C8) alkyl; R30 and R31 are separately and independently of one another H, (Cx-C8) alkyl, (C3-C8) cycloalkyl, (C3-C8) heterocycloalkyl, aryl or heteroaryl, wherein R30 and R31 are optionally and independently substituted. are with 1 to 3 substituents independently selected from halogen, oxo, (Cx-C5) alkyl, -C02R4, -COR40, -OR40, -CONR50RS1, -NR50R51 and -S02R40; or R30 and R31 together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocycloalkyl ring, which ring optionally has 1 additional heteroatom independently selected from N, O and S, the 5- to 8-membered heterocycloalkyl ring is optionally and independently substituted with 1 to 3 substituents independently selected from halogen, oxo, (C 1 -C 5) alkyl, -CO 2 R 40, -COR 40, -OR 40, -CONR 50 R 51, -NR 50 R 5, -SO 2 R 40; R 40 is H, (C 1 -C 5) alkyl, (C 3 -C 8) cycloalkyl, (C 3 -C 8) heterocycloalkyl, aryl or heteroaryl; R50 and R51 are each independently and independently H, (C 1 -C 5) alkyl, (C 3 -C 8) cycloalkyl, (C 3 -C 8) heterocycloalkyl, aryl or heteroaryl; or R50 and RS1 together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocycloalkyl ring, which ring optionally has 1 additional heteroatom independently selected from N, 0 and S.

One skilled chemist will appreciate that the compounds represented by the general formula (I) (a) include the two compounds (I) (a) (i) and (I) (a) (ii): O yR1 Μ HN'xWsV'N \ HNxMvVssN \ 15 JU> NJ ^ / N-R '<iKa) (i) r3; mm k5

A general preferred subgroup of compounds of Formula (I) includes the compounds wherein: A is (a), (b), (c) or (h); R1 and R2 are H; R 3 is (C 3 -C 6) alkyl or (C 3 -C 5) cycloalkyl; P is (C 3 -C 8) cycloalkyl or aryl; J is 0 or S; and x is 1, 2 or 3.

A more preferred subgroup of the compounds of Formula (I) includes the compounds wherein: A is (a) or (b).

The naming of the compounds and intermediates of the present invention can be according to the nomenclature systems of the IUPAC (International Union for Pure and Applied Chemistry) or the CAS (Chemical Abstracts Service, Columbus, OH).

The number of carbon atoms of the various hydrocarbon-containing radicals in this specification may be indicated by a prefix indicating the minimum or maximum number of carbon atoms of the radical, for example. the prefix (Ca-Cb) alkyl means an alkyl group with the integer, "a" to the integer "b", 5 carbon atoms. Thus, for example, (C 1 -C 6) alkyl refers to an alkyl group with one to six carbon atoms.

The term "alkoxy" refers to unbranched or branched, monovalent, saturated aliphatic chains of carbon atoms bonded to an oxygen atom. Examples of alkoxy groups include methoxy, ethoxy, propoxy, butoxy, isobutoxy, tert-butoxy and the like.

The term "alkyl" refers to unbranched or branched, monovalent, saturated aliphatic chains of carbon atoms and includes, for example, methyl, ethyl, propyl, is-15. opropyl, butyl, isobutyl, pentyl, isopentyl, hexyl and the like. equal.

The term "alkenyl" refers to a hydrocarbon with a branched or branched chain with one or more double bonds and includes, for example, vinyl, allyl and the like.

The term "aryl" refers to a cyclic aromatic hydrocarbon. Examples of aryl groups include phenyl, naphthyl, anthracenyl, phenanthrenyl and the like.

The term "cycloalkyl" denotes a saturated monocyclic or bicyclic cycloalkyl group optionally condensed with an aromatic hydrocarbon group. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, indanyl, tetrahydronaphthalinyl and the like.

The term "halogen" denotes chlorine, bromine, fluorine and iodine atoms and / or substituents.

The term "heteroaryl" denotes a monocyclic or polycyclic aromatic hydrocarbon group in which one or more carbon atoms are replaced by heteroatoms such as nitrogen, oxygen and sulfur. If the heteroaryl group contains more than one heteroatom, the heteroatoms may be the same or different. Examples of hotter roaryl groups include benzofuranyl, benzothienyl, benzyl

T02 609H

9,.

•% midazolyl, benzoxazolyl, benzothiazolyl, chromenyl, furyl, imidazolyl, indazolyl, indolizinyl, indolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazalazyl, oxadiazalazyl, oxadiazalazyl, oxadiazalazyl, oxadiazalazyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrido [3,4-b] indolyl, pyridyl, pyrimidyl, pyrrolyl, quinolizinyl, quinolyl, quinoxalinyl, thiadiazolyl, thiatriazolyl, thiazolyl, thienyl, triazyl, triazyl, triazinyl, triazyl, triazinyl of such.

The term "heterocycloalkyl" refers to a saturated monocyclic or polycyclic cycloalkyl group optionally condensed with an aromatic hydrocarbon group, wherein at least one of the carbon atoms has been replaced by a heteroatom such as nitrogen, oxygen and sulfur. If the heterocycloalkyl group contains more than one heteroatom, the heteroatoms may be the same or different. Examples of such heterocycloalkyl groups include azabicycloheptanyl, azetidinyl, indolinyl, morpholinyl, piperazinyl, piperidyl, pyrrolidinyl, tetrahydrofuryl, tetrahydroquinolinyl, tetrahydroindazolyl, tetrahydroindolyl, tetrahydroisochinolinyl, tetrahydropyloxy, tetrahydropyloxy, tetrahydropyloxy, tetrahydropyloxy, tetrahydropyloxy, tetrahydropyloxy, tetrahydropyloxy, and tetrahydrofuran. thioxanyl and the like.

A cyclic group can be bound to another group in more than one way. If no particular way of binding is specified, all possible ways are meant. For example, the term "pyridyl" includes 2-, 3-or 4-pyridyl and the term "thienyl" includes 2- or 3-thienyl.

The term "mammal" means animals including, for example, dogs, cats, cows, sheep, horses, and humans. Preferred mammals include humans.

The term "oxo" means a carbonyl group formed by the combination of one or more carbon atoms and one or more oxygen atoms.

The term "pharmaceutically acceptable" means that the carrier, vehicle, diluent or adjuvant in question is generally chemically and / or physically compatible with the other components that it contains. preparation contains and. is physiologically compatible with its recipient.

The term "prodrug" refers to a compound that is a precursor drug that, after administration, releases the drug in vivo via a chemical or physiological process (e.g., because it is brought to physiological pH or by enzymatic activity). A discussion of the synthesis and use of prodrugs is provided by T. Higuchi and W. Stella in "Prodrugs as No-Sheet Delivery Systems" vol. 14 of the ACS Symposium Series and in Bioreverible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon 15 Press, 1987, both of which are incorporated herein by reference.

The term "salts" and "pharmaceutically acceptable salts" refers to organic and inorganic salts of a compound of Formula (I), or a steroisomer or a prodrug thereof. These salts can be prepared in situ during the final isolation and purification of a compound or by independently reacting a compound of Formula (I), or a steroisomer or a prodrug thereof, with a suitable organic or inorganic acid or base and isolating the salt formed in that way. Representative salts include the hydrobromide, hydrochloride, sulfate, nitrate, bisulfate, acetate, oxalate, besylate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate , tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate and lauryl sulfonate salts and the like. They can also be cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium and the like, as well as non-toxic ammonium, quaternary ammonium and amine cations, including, but not limited to, ammonium, tetramethylammonium, te 1 02 60 91 -% 9 11 l. . ".

traethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine and the like.

For further examples, see, for example, Berge, et al., J. Pharm. Sci., 66, 1-19 (1977), which publication is incorporated herein by reference.

A salt of a compound of Formula (I) can be easily prepared by suitably mixing solutions of a compound of Formula (I) and the desired acid or base. The salt can precipitate from the solution and can be collected by filtration or can be obtained by evaporation of the solvent.

The term "substituted" means that a hydrogen atom of a molecule has been replaced by another atom or molecule. The atom or molecule that replaces the hydrogen atom is referred to as a "substituent".

The "-" symbol indicates a covalent bond.

The term "solvent that is inert under the reaction conditions" or "inert solvent" refers to a solvent or mixture of solvents that does not interact with starting materials, reagents, intermediates, or products in a manner that adversely affects their desired properties. affects.

The terms "treat," "treated," or "treated," as used herein, include preventative (e.g., prophylactic), mitigating or healing use or outcome.

The compounds of Formula (I) may contain asymmetric or chiral centers and may therefore exist in various stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of Formula (I), as well as mixtures thereof, including racemic mixtures, form part of the present invention. In addition, the present invention includes all geometric and positional isomers. For example, if a compound of Formula (I) comprises a double bond, both the cis and trans forms, as well as mixtures thereof, are covered by the invention. .

1 026091 - t 12

Diastereoisomeric mixtures can be separated into their individual diastereomers based on their physicochemical properties by methods known to those skilled in the art, such as by chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with a suitable optically active compound. (e.g. an alcohol), separating the diastereomers and converting (e.g. hydrolyzing) the individual diastereomers into the corresponding pure enantiomers. Also, some of the compounds of Formula (I) may be atropisomers (e.g., substituted biaryls), and these are also considered part of the invention.

The compounds of Formula (I) may exist in both pharmaceutically acceptable solvents such as water, ethanol and the like in both unsolvated and solvated form, and the invention includes both solvated and unsolvated form.

It is also possible that the compounds of Formula (I) may exist as tautomeric isomers, and all such forms are covered by the invention.

The present invention also encompasses isotope-labeled compounds of Formula (I) which are identical to those mentioned herein, with the difference that one or more of the atoms are replaced by an atom having an atomic mass or atomic number that is different from the atomic mass or number that is normally found in nature. Examples of isotopes that can be incorporated into compounds of Formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as 2 H, 3 H, 13 C, 14 C, 15 N, 180, 170, 31P, 32P, 35S, 18F and 36C1. Compounds of Formula (I), the stereoisomers and prodrugs thereof and the pharmaceutically acceptable salts of the compounds, stereoisomers and prodrugs containing the isotopes and / or other isotopes of the aforementioned isotopes and / or other isotopes I ».

13 other atoms contain are included in the present invention.

Certain isotope-labeled compounds of Formula (I), for example those compounds in which radioactive isotopes such as 3 H and 14 C are incorporated, are useful in determining the distribution of tissues of compounds and / or substrates. Tritiated, i.e., 3 H, and carbon-14, i.e., 14 C, isotopes are particularly preferred because of the relative simplicity of preparation and their easy detection. Furthermore, substitution with heavier isotopes, such as deuterium, i.e., 2H, may provide certain therapeutic benefits that result from greater metabolic stability, for example, an increased half-life in vivo or a lower necessary dosage, and therefore they may, in some circumstances, preferred.

The isotope-labeled compounds of Formula (I) can generally be prepared by carrying out methods analogous to those described in the Schemas and / or Examples described below, such as by replacing a isotope-labeled reagent by an isotope-labeled reagent. .

In another aspect, the invention provides methods for treating conditions including diabetes, including type 1 and type 2 diabetes, hyperglycemia, dyslipidemia, impaired glucose tolerance, metabolic syndrome and / or cardiovascular disease, which involve administering a therapeutically effective amount of a compound of Formula (I), a stereoisomer or pro.

A drug thereof, or a pharmaceutically acceptable salt of the compound, stereoisomer or prodrug; or a pharmaceutical composition comprising a compound of Formula (I) or a stereoisomer or prodrug thereof) or a pharmaceutically acceptable salt of the compound, stereoisomer or prodrug and a pharmaceutically acceptable carrier, vehicle or diluent, to a mammal in need of such treatment. A preferred

1 02609H

♦ r 14 disorder 'includes diabetes, metabolic syndrome and / or cardiovascular disease.

In another aspect, the invention provides methods for inhibiting the activity of PDE9 in a mammal in need of such inhibition, which involves the administration of a PDE9 inhibitory amount of a compound of Formula (I), a prodrug thereof or a pharmaceutically acceptable salt of the compound or prodrug; or a pharmaceutical composition comprising a compound of Formula (I) or a prodrug thereof, or a pharmaceutically acceptable salt of the compound, or the prodrug and a pharmaceutically acceptable carrier, vehicle or diluent.

The compounds of Formula (I) can be administered to a mammal in dosages in the range of about 0.1 mg to about 3,000 mg per day. For a normal adult human with a body weight of about 70 kg, a dosage in the range of about 0.01 mg to about 100 mg per kg of body weight is usually sufficient, and preferably of about 0.1 mg up to about 10 mg per kg. Some variability in the general range of dosage may be required depending on the age and weight of the patient being treated, the intended mode of administration, the particular compound being administered, and the like. The determination of dosage ranges and the optimum doses for a particular mammal to be treated is within the competence of a person skilled in the art who can benefit from the present description.

According to the methods of the present invention, a compound of Formula (I), a stereoisomer or prodrug thereof or a pharmaceutically acceptable salt of the compound, stereoisomer or prodrug may be administered in the form of a pharmaceutical composition comprising a pharmaceutically acceptable carrier, vehicle or diluent. Accordingly, a compound of Formula (I), a stereoisomer or prodrug thereof or a pharmaceutically acceptable salt may be separated from the compound, stereoisomer or prodrug or in combination in any of the usual forms. of dosage, including oral, buccal, sublingual, ocular, topical (e.g., 5. transdermal), parenteral (e.g., intravenous, in-tramuscular, or subcutaneous), rectal, intracistemal, in-travaginal, intraperitoneal, intravesical, local (e.g., powder, ointment) or drops), nasal and / or inhalable forms of dosage are administered to a patient.

Pharmaceutical compositions suitable for parenteral injection may include pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstituting sterile injectable solutions or dispersions in situ. Examples of suitable aqueous and non-aqueous vehicles, vehicles and thinners include water, ethanol, polyols (such as propylene glycol, polyethylene glycol, glycerol and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. The appropriate fluidity can be maintained, for example, by the use of an envelope such as lecithin, 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 comprise additives such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of microorganism contamination of the present compositions can be achieved with various antibacterial agents and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid and the like. It may also be desirable to add isotonic agents, for example sugars, sodium chloride and the like. Prolonged absorption of injectable pharmaceutical preparations can be obtained by using 1026007 ♦ 16.

Of agents capable of delaying absorption, for example aluminum monostearate and gelatin.

Solid dosage forms for oral administration include capsules, tablets, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one normal inert pharmaceutical excipient (or carrier) such as sodium citrate or dicalcium phosphate, or (a) fillers, such as for example starches, lactose, sucrose, manitol and silica; (b) binders such as, for example, carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) moisture-retaining agents such as, for example, glycerol; (d) disintegrating agents such as, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicate complexes and sodium carbonate; (e) solution retarders such as, for example, paraffin; (f) absorption accelerators such as, for example, quaternary ammonium compounds; (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate; (h) absorbents such as, for example, kaolin and bentonite; and / or (i) lubricants such as, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate or mixtures thereof. In the case of capsules and tablets, the dosage forms may further comprise buffering agents.

Solid dosage forms may be formulated as altered-release and pulsed-release dosage forms containing adjuvants such as those described above for immediate-release dosage forms, together with other adjuvants that change the rate of release and that are coated and / or are included in the material of the dosage form. Substances that change the rate of release include, but are not limited to, hydroxypropyl methylcellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, polyethylene oxide, xanthan gum, copolymer of ammonium methacrylate, hydrogenated. 1. 02.60 B.] - 17. %. I ·.

castor oil, carnauba wax, paraffin wax, cellulose acetate phthalate, hydroxypropyl methyl cellulose phthalate, copolymer of methacrylic acid and mixtures thereof. Dosage forms with an altered release and a pulsed release. 5. to be able to contain one or a combination of additives that change the rate of release.

The pharmaceutical compositions of the invention may further comprise rapid dispersion or dissolution dosage formulations (FDDFs) containing the following components: aspartame, potassium acesulfame, citric acid, sodium croscarmellose, crospovidone, diascorbic acid, ethyl acrylate, ethyl cellulose, gelatin, hydroxypropyl methyl cell -lulose, magnesium stearate, mannitol, methyl methacrylate, mint flavor, polyethylene glycol, fumed silica, silicon dioxide, sodium starch glycolate, sodium stearyl fumarate, sorbitol and xylitol. The terms disperse or dissolve as used in this specification to describe FDDFs depend on the solubility of the drug being used, that is, a rapid dispersing dosage form can be prepared if the drug is insoluble, and that a dosage form for rapid dissolution can be prepared if the drug is soluble.

Solid preparations of a similar type can also be used as filler in soft or hard filled gelatin capsules, with additives such as lactose, milk sugar and high-polyethylene glycols. molecular weight and the like.

Dosage forms such as tablets, dragees, capsules and granules can be prepared with coatings and casings, such as enteric coatings and other coatings known to those skilled in the art. They may also comprise opacifying agents and may also have such a composition that they release the active compound (s) in a delayed, stable or controlled manner. Examples of coating compositions that can be used are polymeric compounds and waxes. The active compound (s) may also be in a microencapsulated form, if appropriate with one or more of the additives described above.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may include inert diluents normally used in the art, such as water or other solvents, solubility enhancing agents, and emulsifiers, such as, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, benzyl benzoate, propylene glycol, 1 , 3-butylene glycol, oils, in particular cotton seed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, or mixtures of these substances and the like.

In addition to such inert diluents, the pharmaceutical compositions may also contain additives such as wetting agents, emulsifiers and suspending agents, sweeteners, flavors and fragrances.

In addition to the active compound (s), the pharmaceutical compositions may further comprise suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene-sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and dragant gum or mixtures of these substances and the like.

Pharmaceutical compositions of the present invention can also be made suitable for veterinary use treatments wherein a compound of the present invention or a veterinary acceptable salt thereof, or veterinary acceptable solvates or prodrugs thereof, is administered in the form of a suitable acceptable formulation. in accordance with normal veterinary practice, and the veterinarian will determine the dosage schedule and method of administration that are most suitable for a particular animal.

102608 "*" * 2.9

The present invention further comprises the combination of a PDE9 inhibitory compound of Formula (I) and one or more other antidiabetic and / or cardiovascular agents.

The present invention further comprises the combination of a PDE9 inhibitor, as described in Formula (I), and one or more other active compounds selected from: a) a PGI2 prostaglandin such as prostacyclin or iloprost; b) an antagonistic compound for the α-adrenergic receptor, also known as α-adrenoceptor antagonists, α-receptor antagonists or α-blockers; suitable compounds for use herein include: the α-adrenergic receptor antagonists as described in PCT application WO99 / 30697 published June 14, 1998, the content of which relates to α-adrenergic receptor antagonists is incorporated herein by reference, and which include selective αι-adrenoceptor antagonists, α2-adrenoceptor antagonists and non-selective adrenoceptor antagonists. Suitable αι-adrenoceptor antagonists include: phentolamine, phentolamine mesylate, trazodone, alfuzosin, indoramine, naphthopidil, tamsulosin, dapiprazole, phenoxylbenzamine, idzoxane, eferaxane, yohimbine, rauwolfa-alkaloine, doxa-azozine, doxa-azozine, doxa-azozine, doxa-azozine, doxa-azozine α 2 blockers from US 6,037,346, dibenarmine, tolazoline, trimazosin and dibenarnine, α-adrenergic receptors as described in U.S. Pat. Nos. 4,188,390; 4,026,894; 3,511,836; 4,315,007; 3,527,761; 3,997,666; 2,503,059; 4,703,063; 3,381,009; 4,252,721 and 2,599,000, each of which is incorporated herein by reference; or α-adrenoceptor antagonists include: clonidine, papaverine, papaverine hydrochloride, optionally in the presence of a carotonic agent such as pirxamine; c) an NO dorior compound (NO agonist); suitable NO. donor compounds for use herein include organics

• A

chemical nitrates such as mono-, di- or tri-nitrates or organic nitrate esters including glyceryl trinitrate (also known as nitroglycerin), isosorbide-5-mononitrate, isosorbide dinitrate, pentaerythritol tetranitrate, erythritylte-5-tranitrate, sodium nitroprinoid-nitroprussidone-nitroprussidone-nitroprussidone-nitrate-proline molsidomine, S-nitroso-N-acetylpenicillamine (SNAP), S-nitroso-N-glutathione (SNO-GLU), N-hydroxy-L-arginine, amyl nitrate, linsidomine, linsidomine hydrochloride, (SIN-1) S-nitroso -N-cysteine, diazenium di-10 let, (NONOaten), 1,5-pentanedinitrate, L-arginine, ginseng, zizfi fructus, molsidomine and nitrosylated maximalysyl derivatives such as NMI-678-11 and NMI-937 as described in the published PCT application WO 0.012.075; d) a potassium channel opener; suitable potassium channel openers for use herein include nicorandil, cromokalim, levcromakalim, lemakalim, pinacidil, cliazoxide, minoxidil, charybdotoxin, glyburide, 4-aminopyridine and barium chloride; e) a compound that influences the action of the atrial natural factor (also known as atrial natural peptide), such as neutral endopeptidase (NEP) inhibitors; f) a compound that inhibits the angiotensin-converting enzyme (ACE), such as alacepril, alindapril, altiopril, benazepril, betaazeprilat, captopril, ceronapril, cilazapril, cilazaprilat, delapril, enalapril, enalaprilat, fosinopril, indolapril, imidril , libenzapril, lisinopril, mo-exepril, moveltipril, pentopril, perindopril, quinapril, quinaprilat, ramipril> rentiapril, spirapril, temocapril, teprotide, trandolapril and zofenopril or a double ACE / NEP inhibitor that is both a ACE and a compound that is both ACE and ACE. NEP inhibits, such as, for example, omapatrilat, fasidotril and mixanpril; g) an angiotensin II receptor (ARB) blocker such as candesartan, eprosartan, irbesartan, losartan, olmesartan, olmessartan, medoxomil, saralasin, telmisartan and valsartan; 1 02 60 91 4 λ.

21 h) a substrate for NO synthase such as L-arginine; i) a calcium channel blocker such as amlodipine, verapamil, pranidipine, azelnidipine and vatanidipine;

* I

j) an antagonist of endothelin receptors or an inhibitor of the endothelin-converting enzyme; k) a cholesterol-lowering agent, such as statins, such as, for example, calciumatorvastatin (Lipitor), sodium cerivastatin (Baycól), sodium fluvastatin (Lescol), lovastvastatin (Mevacor), sodium pragvastatin (Pravachol) and simvastatin (Zocor); l) one. antibiotic platelet or antithrombotic agent, for example, tPA, uPA, warfarin, hirudin and other thrombin inhibitors, aspirin, plavix, cilastozol, heparin and inhibitors. thromboplastin activating factor; 15 m) a PDE5 inhibitor (such as 5- [2-ethoxy-5- (4-methyl-1-piperzazinylsulfonyl) phenyl] -1-methyl-3-n-propyl-1,6-dihydro-7H " Pyrazolo [4,3-d] pyrimidin-7-one (sildenafil); (6, 12, 12?) - 2,3,6,7,12,12a-hexahydro-2-methyl-6- (3,4- methylenedioxyphenyl) pyrazino [2 ', 1': 6.1] pyrido [3,4-b] indole-1,4-20 dione (tadalafil, IC-351); 2- [2-ethoxy-5- ( 4-ethyl-piperazi-ne-1-yl-1-sulfonyl) -phenyl] -5-methyl-7-propyl-3-i-imidazo [5,1-f] [1,2,4] triazine-4- one (vardenafil); 5- [2-ethoxy-5- (4-ethyl-piperazin-1-yl-sulfonyl) -pyridin-3-yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro- 7'-pyrazolo [4,3-d] pyrimidin-7-one-one and 5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2,6-dihydro-7β-pyrazolo 4,3-d] pyri-midihe-7-one); n) a beta-blocker, diuretic antagonist or aldostrone antagonist.

30

When a combination of active agents is administered, the agents can be administered simultaneously, separately or sequentially.

Compounds of Formula (I) can be prepared according to the following reactions from the exemplary synthesis processes ("schemes") and according to other normal organic preparative methods. These methods form further aspects of: The general formulas are represented by Roman numerals I, II, III, IV, etc. Subgroups of these general formulas are represented by Ia,

Ib, Ic, etc. ...... IVa, IVb, IVc, etc. It should be clear, for example, that reference to Formula * Ia "refers to the compound represented by Formula (I) together with a ring structure that is selected from the ring structures that are part of Group A. In this example, structure "(a)." It should also be understood that reference to Formula Hm or Illm refers to compounds represented by Formulas II or III, together with a ring structure selected from the ring structures that are part of Group Q, in this example structure "(m)." It should further be understood that these methods are only intended to clarify the present invention and that they should not be construed as limiting this in any way.

Target compounds of Formula (I) may contain primary or secondary amine groups or carboxylic acid groups in protected form that require deprotection as a final step.

The use of such protecting groups is known to those skilled in the art. For a detailed reference to the use of protecting groups see: T.A. Greene, P.G.M. Wuts "Protecting Groups in Organic Synthesis *, Vol.

2 Wiley and Sons, 1991, which publication is incorporated herein by reference.

Compounds of the general Formula (I) wherein A is (a), (b), (d), (f), (g) or (h) can be prepared from compounds of the general Formula (II) according to Scheme 1. Suitable conditions are known to those skilled in the art, and include a base-catalyzed cycle, wherein reagents such as potassium tert-butoxide, sodium hydroxide and potassium carbonate in an alcohol solvent such as ethanol or isopropanol or a mixture of water and alcohol become alcoholic. used. The reaction can be carried out at a temperature between room temperature and 1026091. . . · K J.

23 boiling point of the solvent and optionally in the presence of hydrogen peroxide.

Also compounds of the general Formula (I) in which A is (a), (b), (d), (f), (g) or (h) can be made by condensing compounds of Formula (III) with esters IVb under basic catalysis. Typically, the compounds of Formula III and IVb are treated with a base such as potassium tert-butoxide in a protic solvent such as 1-butanol at elevated temperatures of from about 80 ° C to about 120 ° C for about 2 hours to about 24 hours. The reaction mixture can also be heated in a microwave. Usually, the reaction mixture is heated to a temperature of about 150 ° C to about 200 ° C, preferably at about 180 ° C for about. 15 five minutes to about 30 minutes.

Scheme 1 '· O · _. 1 20. J &), w.

I π 25 '. OR '+ Xzs

X (CH 2) x -R 10 - (CH 3 J x -R 10 H 2 N)

rva jvb m

Compounds of Formula II can be prepared by reacting compounds of Formula IVa with compounds of Formula III. Such reactions where an amide bond becomes. can be formed under a 1 02 6091

t I

A wide range of circumstances are known to those skilled in the art. Compounds of Formula IVa can be activated, for example, by treatment with a reagent such as 1,1-carbonyldiimidazole (CDI) or fluoro-Ν, Ν, Ν ', N'-tetramethylformamidinium hexafluorophosphate (TFFH), or with a combination of reagents such as azaben- zotriazole-1-yloxytris (pyrrolidino) -phosphonium hexafluoro-phosphate (PyAOP) and 1-hydroxy-7-azabenzotriazole (HOAt), followed by the addition of the compound of Formula 10 (III).

Q is selected from ""; £ · "X> m *: (n) R"; (o) V; w **; (Q) R *; (r) rS; (s) RS; or, (t) rS.

Also, compounds of Formula II can be prepared by the addition of a peptide coupling agent such as O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyliironium hexafluorophosphate (HATU) or 1-propane phosphoric anhydride (T3P ) to a mixture of the compounds of Formula III and IVa. This reaction is carried out in a suitable solvent such as dichloromethane, pyridine, N, N-dimethylformamide (DMF), N, N-dimethylacetamide (DMA) or 1-methyl-2-pyrrolidone at a temperature between 0 ° C and the temperature boiling point of the solvent. The reaction is preferably carried out by activating the compound of Formula IVa with T3P and triethylamine at room temperature.

Compounds of Formula III (n) wherein R 2 and R 3 are as defined above can be prepared from a suitable amine and a compound of Formula 1026091 ~ V 25 (Scheme 2). The compound of Formula V is first treated with a trialkyl orthoester, preferably triethyl orthoformate, in a non-protic solvent such as acetonitrile at a temperature of about 50 ° C to about 80 ° C for about 0.5 hour to about three hours. The R3-NH2 compound of Formula VII is then added and the reaction is cooled and stirred at room temperature for about 12 hours to about 24 hours.

10

Scheme 2 I o

HjN'I-'V-k * ^ CN EM \' f / ™ *

15 / R R 2 H 2 N + and R 7 R 7 R + R

H 2 N 2 NH 2 EtO

R3 V VI 'vn ΠΙ (η) V V1 20

Compounds of Formula III (m) wherein R 1 and R 3 are as defined above can be prepared from the corresponding nitro compounds of Formula VIII by reduction (Scheme 3). The reduction is usually carried out by catalytic hydrogenation. The compound of Formula VIII is treated with a noble metal catalyst, preferably Pd / C, in a reaction inert solvent such as ethanol under a hydrogen atmosphere of 15-45 psi at room temperature for about one hour to about six hours o'clock.

1 02 6091 - bm

Schedule 3 "·" · 4 26. _ HoA ^ R ’5 H2N’ X ^ (Ο, Ν '^ Ν' R3 * 8 r3

m (m) Vin. IX

.-.---. / "... o o / 10. Etc k / roA ^ R1.

R9 R3

ΧΠ X R = H

X 1 R = Et 15 Compounds of Formula VIII wherein R 1 and R 3 are as defined above are prepared from the appropriate compound of Formula IX via a reaction in which an amide bond is formed. In general, the compound of Formula IX is first converted to the acid chloride by reaction with oxalyl chloride in a reaction inert solvent such as methylene chloride containing a catalytic amount of DMF at room temperature for about four hours to about 24 hours. The intermediate acid chloride is then treated with an excess of ammonia in a suitable solvent, such as tetrahydrofuran, dioxane or mixtures thereof, at about ° C to about room temperature for about one hour to about 24 hours.

Compounds of Formula IX wherein R1 and R3 are as defined above are prepared by nitrating a compound of Formula X. Usually, the compound of Formula X is dissolved in sulfuric acid and then treated with nitric acid at elevated temperatures of about 40 ° C to about 80 ° C, preferably at about 60 ° C for about one hour to about six hours.

1 02 60 91 -, 27 4 Μ I.

. Compounds of Formula X wherein R 3 and R 3 are as defined in the above can be prepared by saponifying a compound of Formula XI. Usually the compound of Formula XI is treated with an alkali base, such as sodium hydroxide, in a protic solvent, such as methanol or ethanol, at a temperature of about 30 ° C to about 80 ° C for one hour to about six hours.

Compounds of Formula XI wherein R1 and R3 are as defined above can be prepared by reacting a compound of Formula XII with a hydrazine. Typically, the compound of Formula XII is treated with hydrazine in a protic solvent, such as ethanol, at room temperature for about 10 hours to about 24 hours.

Compounds of Formula IIIp wherein R3 is as defined above can be prepared by reducing a compound of Formula XIII (Scheme 4). Typically, the reduction is carried out by treating the nitro compound of Formula XIII with a powder of a transition metal, preferably zinc, and a proton source such as acetic acid or ammonium chloride, in a protic solvent such as water at room temperature for about 30 minutes to about three hours.

25 102 60 81 ·: · t

Schedule 4 28 ·· * ·

O o o

s. ^ yy 80 χ> 02Ν ^ Λ 02N ^^ (R3 R3. R3 m (p) xni xrvr, 10 / HO ^ Λα * o fv

XV

15.

Compounds of Formula XIII wherein R 3 is as defined in the above, theirs are prepared from the corresponding ester compound of Formula XIV by an amidation reaction. Typically, the ester is treated with an excess of ammonia in a reaction inert solvent, such as ethanol or water, at room temperature for about two hours to about 24 hours.

Compounds of Formula XIV, wherein R 3 is as defined above, can be prepared by reaction of the appropriate compound. of Formula XV with ethyloxalyl chloride. Usually the reaction is carried out in an ether solvent, such as diethyl ether or tetrahydrofuran, and catalyzed by a base, preferably an amine base, most preferably triethylamine, at a temperature of about 0 ° C to about room temperature for about one hour until about four o'clock.

Compounds of Formula III (r) wherein R 3 is as defined above can be prepared from the cyanamide XVI and azide XVII (Scheme 5). Typically, the compounds of Formula XVI and Formula XVII are added to a solution of sodium ethoxide in ethanol at an elevated temperature of from about 40 ° C to about 80 ° C for about 30 minutes to about four hours .

5 Schedule 5

O

ηΥΥλ f 10 2: ^ 3 · ni {r) XVI xvil

Compounds of Formula III (t) wherein R1 and R3 are as defined above can be prepared from the corresponding compounds of Formula XVIII by a hydrolysis reaction (Scheme 6). Usually the compound is Formula. XVIII treated with an acid, preferably sulfuric acid, at a temperature of about 10 ° C to about 20 ° C for about two hours to about four hours.

Diagram 6 25 JΓ f1 R1 OEt ·. X> * - Y> -y +

HZN n3 Η2Ν ^ Υ CN

R. R3 ni (t) xvra xix xx 30

Compounds of Formula XVIII are prepared from the. corresponding compounds of Formula XIX and Formula XX. In general, the compound of Formula XIX 35 is slowly added to a solution of the compound of Formula XX in an alcohol solvent, preferably ethanol, at an elevated temperature, preferably at 102 b.p. of the solvent for about one hour to about three hours.

Compounds of Formula II (s) wherein R3 ice cream is as defined above can be prepared from the corresponding compounds of Formula XXI and Formula XXII (Scheme 7). In general, a mixture of the compound of Formula XXI and the compound of Formula XXII in an alcohol solvent, such as methanol, is treated with an amine base, preferably triethylamine, at a temperature of about room temperature to about 50 ° C for about one hour up to about four hours.

Schedule 7 "

15 X

H’N -.- .. 0.

"" Η Λ - + H ^ - 20 ffl (S) XM ΧΧΠ

Compounds of formula Ic wherein R1, R3, P, J, x and R10 are as defined above can be prepared from the corresponding compounds of Formula XXIII and Formula XXIV (Scheme 8). Typically, the compounds are dissolved in a protic solvent, preferably methanol, at elevated temperature, preferably at the boiling point of the solvent, for about two hours to about eight hours. The crude product is then cyclized by reaction with a chlorinating reagent, such as phosphorus oxychloride, in a halogenated solvent, such as dichloroethane, at a temperature of from about 50 ° C to about 90 ° C for about one hour to about four hours.

35 1026091 - 31., "*", "

Scheme 8 - (1) H + 8 (CH1 / J1 -R10 (CH1 / J1 -R10 icx10) - Compounds of formula Ie wherein R1, R3, P, J, x and R10 are as defined above can be prepared from the corresponding compounds of Formula XXV (Scheme 9.) In general, the compound of Formula XXV is treated with trimethylsilyl chloride in an amine base that is used as a solvent, preferably pyridine, at room temperature for about 15 minutes to about 30 minutes. hexamethyldisilazane is added and the mixture is heated to a temperature of about 80 ° C to about 120 ° C, preferably at the boiling point of the solvent, for about six hours to about 24 hours.

1 026091

Scheme 9 32 '1 · O3 or R3 N (CH2) x -R10. ^

Ie j 10 /

NH

; (CHA-R10. R

15 xxvn

The desired compounds of Formula XXV can be prepared by condensing the corresponding compounds of Formula XXVI and Formula XXVII. The reaction is catalyzed by a base, such as sodium ethoxide, in an alcohol solvent, such as ethanol, at elevated temperature, preferably at the boiling point of the solvent for about one hour to about eight hours.

The various carboxylic acid and ester compounds of Formula IV can be easily prepared from commercially available starting materials according to methods known to those skilled in the art. The following general methods are given as an example of some of these methods (Scheme 10).

· 30 1026091-

Schedule 10 33 * ·

Cm η ο j χΓ ^ ΌΒη --- 5 NcHak-R10 Ο. | ^ ° Βη.

X (CH 2 VR10. HO

IVaR = H XXDC

IVbR «Me XXVIII xm TVcReBn 10 Q ^ Aa - CXa0E —- ΓΧλ g '· .- ·' Y OEt OEt

Nchj ^ r10 hS. O

XXXI xxxn 15

Compounds of Formula IVa wherein J, x and R10 are as defined above can be prepared from the corresponding ester compounds of Formula I Vb by a saponification reaction. The compound of For-20 formula IVb can be treated with an alkali base, such as sodium hydroxide, in a protic solvent, such as methanol or ethanol, at a temperature of about 30 ° C to about 80 ° C for about one hour to about six hours.

The compounds of Formula IVa can also be prepared by deprotection of the corresponding benzyl esters (compounds of Formula IVc). This is generally achieved by catalytic hydrogenolysis. The compound of Formula IVc is treated with a noble metal catalyst, preferably Pd / C, in a reaction inert solvent, such as ethanol, under an atmosphere of hydrogen at 15-45 psi at room temperature for about one hour to about six hours.

Compounds of Formula XXVIII can be prepared 35. alkylation of a compound of Formula XXIX. The compound of Formula XXVIII is thus treated with R 10 (CH 2) x L wherein L is a leaving group, under a catalyst with a base, such as potassium carbonate, in a polar aprotic solvent, such as dimethylformamide, at a temperature from room temperature to about 50 ° C for about four hours to about 24 hours. The alkylation can also be carried out under Mitsunobu conditions. In general, the compound of Formula XXVIII is treated with an alcohol such as R 10 (CH 2) x OH, triphenylphosphine and diethyl azodicarboxylate in a reaction-inert solvent, such as tetrahydrofuran, at room temperature for about six hours to about 24 hours.

Similarly, compounds of Formula XXX wherein R10 and x are as defined above can be prepared from the corresponding alcohol compounds of Formula XXXI by an alkylation reaction. In general, the compound of Formula XXXI is treated with R10 (CH2) XL wherein L is a leaving group, under catalysis with a base such as sodium hydride, in a polar aprotic solvent, such as dimethylformamide, at a temperature of about 0 ° C to about room temperature for about four hours to about 24 hours. The compound of Formula XXXI can be prepared from the corresponding ketone compound of Formula XXXVII by a reduction. Typically, the reduction is carried out by treating a ketone with a reducing agent, such as sodium borohydride, in an alcohol solvent, preferably ethanol, at room temperature for about 30 minutes to about four hours.

The compounds described in Examples 1-30 were subjected to liquid chromatography mass spectrometry (LC-MS) under conditions described below.

Conditions for LC-MS

The molecular weight and retention time determinations for the examples were performed on a reverse-phase LC-MS system. The column was a Polaris C18-A, 5 Mm, 20 x 2.0 mm, which was operated at room temperature. The compounds were eluted with a gradient

1 0-2 6Q9H

»35 release system. Eluens A was 0.05% formic acid; 98% water (HPLC quality); 2% acetonitrile and eluent B was 0.005% formic acid in acetonitrile. The initial pump conditions were 95% A, 5% B with a flow rate of 1 ml / min 5 at a pressure of 270 bar. The solvent gradient ran from 5% to 95% eluent B in 3.74 minutes. The compounds were detected by both UV and MS APCI methods. The UV data was obtained with a Hewlett Packard 1100 DAD (Hewlett Packard, Palo Alto, CA). The MS ^ data was obtained with a Micromass ZMD (Micro-mass, Ine., Manchester, UK). The NMR data was obtained with a Varian Unity 400 (Varian, Ine., Palo Alto, CA).

• 15

EXAMPLES

The examples described below are for illustrative purposes only. The compositions, methods, and the various parameters described herein are only intended to clarify various aspects and embodiments of the invention, and are not intended to limit the scope of the invention for which exclusive rights are sought in any way. .

Unless otherwise specified, all reactants were obtained commercially. Unless otherwise indicated, the following experimental abbreviations have the indicated meanings: BOC - t-butyloxycarbonyl. DMAP - dimethylaminopyridine DMF - dimethylformamide DMSO - dimethylsulfoxide 35 ES / MS - electron spray mass spectrometry EtOAc - ethyl acetate.

EtOH - ethanol 6091 3 'φ 36 Η (for example 1Η, 2Η) - hydrogenato (s) LC - liquid chromatography (M-BOC) - mass - BOC MeOH - methanol 5 min - minute MS - niassas spectrometry NMR - nuclear magnetic resonance RT - room temperature THF - tetrahydrofuran 10

Example 1 1- ([2- (3-Isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl) -pyrrolidine-2 -carboxylic acid Sodium hydroxide (0.5 ml of a 15% solution) was added to a solution of 1- {[2- (3-isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo) at room temperature [4,3-d) -pyrimidin-5-ylmethyl) -phenoxy] -acetyl} -pyrrolidine-2-carboxylic acid ethyl ester (81 mg, 0.18 mmol) in methanol (10 ml). The mixture was heated at 58 ° C for 2 hours, after which it was cooled, acidified with 1 N hydrochloric acid and extracted with EtOAc (2x). The combined extracts were dried (MgSO 4), filtered and concentrated to give 40 mg of the title compound. form of a colorless solid was obtained.

MS 440 (M + H) +; X H NMR (400 MHz, CD 3 OD) 6 7.2 (m, 2H); 6.95 (m, 2 H); 4.6 (m, 2H); 4.04 (m, 2H); 3.6 (m, 4H); 3.4 (m, 1 H); 2.3-1.9 (m, 3H); 1.38 (d, 6H, J = 7.1 Hz). C22 H2 SN5 O5. Retention time on LC: 1.7 minutes.

The following compound, Example 2, was prepared from the suitable ester according to the general method described above for the preparation of Example 35 1.

1 026091 - *.

37 »

Example 2 1- ([2- (1'-Cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetyl) -pyrrolidine- 2 (S) -. carboxylic acid C 24 H 27 N 5 O 5, MS 466 (M + H) *. Retention time on LC: 5 2.0 minutes

Example 3 3-Isopropyl-5- [2- (2-oxo-2-piperazin-1-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one-one trifluoroacetate Trifluoroacetic acid (3 ml) was added to 4- {[2-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl} - piperazine-1-carboxylic acid tert-butyl ester (80 mg, 0.16 mmol) and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo and then concentrated 3 times from toluene. The residue was crystallized from methanol / ether with which. 62 mg of the title compound in the form of a colorless solid was obtained.

MS 411 (M + H) +; * 1 H NMR (400 MHz, CD 3 OD) δ 7.23 (m, 2H); 6.98 (m, 2H)? 5.0 (s, 2H); 4.06 (s, 2H), 3.90 (bs, 2H); 3.80 (bs, 2H); 3.4 (m, 1 H); 3.25 (m, 4H); 1.39 (d, 6H, J = 7.0 Hz). C 21 H 26 N 6 O 3. C 2 HF 3 O 2. Retention time on LC: 1.3 minutes.

The following compound, Example 4, was prepared from the appropriate BOC-protected amine according to the general method described above for the preparation of Example 3.

Example 4 1-Cyclopentyl-6- [2- (2-oxo-2-piperazin-1-yl-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d] pyrimidine-4-one C23 H28 N6 O3. MS 437 (M + H) +. Retention time on LC: 1.6 minutes.

1 02 60 91 -. ♦ 38

Example 5 3-Isopropyl-5- [2- (2-morpholin-4-yl-2-oxo-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one Er 1-propane phosphonic acid anhydride (50% solution in ethyl acetate, 0.1 ml, 0.175 mmol) was added to a solution of [2- (3-isopropyl-7-oxo-6,7-dihydro-1H) at room temperature. pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -. phenoxy] -acetic acid (50 mg, 0.15 mmol) and morpholine (50 μΐ, 0.58 mmol) in anhydrous DMF (3 ml). After 20 hours, the mixture was concentrated in vacuo, after which the residue was purified by flash column chromatography (1- * 2- * 4- * 5% MeOH / CH 2 Cl 2 with 0.2% NH 4 OH) to give 38 mg of the title compound in the form of a colorless solid.

15 'MS 412 (M + H) +; X H NMR (400 MHz, CD3 OD) δ 7.21 (m, 2H); 6.97 (m, 2H); 4.93 (s, 2H); 4.03 (bs, 2H); 3.62 (m, 6H); 3.5 (m, 2H); 3.4 (m, 1 H); 1.4 (m, 6H). C21 H25 N5 O4. C2 HF3 O2. Retention time on LC: 1.8 minutes.

20

The following compounds, Examples 6-18, were prepared from the appropriate amine according to the general method described above for the preparation of Example 5.

Example 6 3-Isopropyl-5- [2- (2-oxo-2-pyrrolidin-1-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one C21 H25 N5 O3. MS 396 (M + H) +. Retention time on LC: 1.9 minutes.

Example 7 5- (2- [2- (4-Ethyl-piperazin-1-yl) -2-oxo-ethoxy] -benzyl) -3-isopropyl-1,6-dihydro-pyrazolo [4,3-d pyrimidine-7-one C 23 H 30 N 6 O 3. MS 439 (M + H) +. Retention time on LC: 1.3 minutes.

1-02 60 Ö1 -: «« · 39

Example 8 N, N-Diethyl-2- [2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetamide C21 H27 N5 O3 . MS 398 (M + H) +. Retention time on LC: 2.0 min.

5

Example 9 1-f [2- (3-Isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl} -pyrrolidine-2- carboxylic acid methyl ester C23 H27 N5 O5. MS 454 (M + H) +. Retention time on LC: 1.9 minutes

Example 10 4 - ([2- (3-Isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl) -piperazine-1- Carboxylic acid tert-butyl ester C 26 H 34 N 6 OS. MS 411 (M-BOC) *. Retention time on LC: 2.3 minutes.

Example 11 N- (2-Dimethylamino-ethyl) -2- [2- (3-isopropyl-7-oxo-e, 7-20 dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) - phenoxy] -acetamide C2 H28 N6 O3. MS 413 (M + H) +. Retention time on LC: 1-2 min.

Example 12 1-f [2- (1-Cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-25 d] pyrimidin-6-ylmethyl) -phenoxy] -acetyl) -pyrrolidine-2 - carboxylic acid methyl ester C 25 H 29 N 5 O 5. MS 480 (M + H) +. Retention time on LC: 2.3 min.

Example 13 4-f [2- (1-Cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetyl) -piperazine- 1-carboxylic acid tert-butyl ester C 2 H 36 N 6 O 5. MS 537 (M + H) +. Retention time on LC: 2.6 minutes.

1 026091 - 1 °, »4 °

Example 14 1-Cyclopentyl-6- [2- (2-oxo-2-pyrrolidin-1-yl-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d] pyrimtidine-4-one • . C23H27NSO3. MS 422 (M + H) V Retention time on LC: 2.9 min.

5

Example 15 1- Cyclopentyl-6- [2- (2-morpholin-4-yl-2-oxo-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d 3 -pyrimidin-4-one C 23 H 27 NSO 4. MS 438 (M + H) +. Retention time on LCs 2.8 minutes.

10

Example 16 2- [2- (1-Cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -N- (2-dimethylamino) ethyl) -acetamide C28H30NeO3. MS 439 (M + H) +. Retention time on LC: 15 1.8.min.

Example 17 1- Cyclopentyl-6- {[2- [2- (4-ethyl-piperazin-1-yl) -2-oxo-ethoxy] -benzyl) -1,5-dihydro-pyrazolo [3,4-d pyrimidine-4-one C 2 H 32 N 6 O 3. MS 465 (M + H)% Retention time on LC: 1.6 min.

Example 18 2- [2 - (1-Cyclopentyl-4-oxo-4f5-dihydro-1H-pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -Ν-diethyl-acetamide C23 H29 N5 O3 . MS 424 (M + H) +. Retention time on LC: 2.4 minutes.

Example 19 [2- (3-Isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetic acid Potassium t-butoxide (2 , 9 g, 26 mmol) was added to a solution of 5-isopropyl-4- {2- [2- (ethoxy-carbonyl-methoxy) -phenyl] -acetylamino} -1H-pyrazole-3-carboxylic acid amide (950 mg, 2.44 mmol) in dry isopropanol (20 ml). The mixture was heated at 85 ° C for 20 hours. The reaction mixture was acidified with 1 N hydrochloric acid and extracted with 1: 1 EtOAc / THF (3x). The. combined extracts were washed with brine, dried (MgSO 4), filtered and concentrated to give 770 mg of the title compound as a colorless solid.

MS 343 (M + H) +; X H NMR (400 MHz, CD 3 OD) δ 7.22 (m, 2H); 6.95 δ (m, 2H); 4.77 (s, 2H); 4.08 (s, 2H); 3.4 (m, 1H) ƒ 1.4 (d, 6H, J = 7.1 Hz). C17 H18 N4 O4. Retention time on LC: 1.7 minutes.

The following compound, Example 20, was prepared from the appropriate starting material according to the general method 10 described above for the preparation of Example 19.

Example 20 [2- (1-Cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetic acid C 19 H 20 N 4 O 4. MS 369 (M + H) +. Retention time on LC: 2.9 minutes.

Example 21 3-Isopropyl-5- [2- (5-chloro-2-morpholin-4-yl-ethoxy) -20 benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one

At -78 ° C, ozone was washed through a solution of 3-isopropyl-5- [2- (5-chloro-2-allyloxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7 -one (134 mg, 0.373 mmol) in CH 2 Cl 2 (8 mL) and MeOH (2 mL bubbled until the blue color persisted for 1 minute. After purging with nitrogen, dimethyl sulfide (0.4 mL) was added and allowed to warm the reaction mixture to room temperature.The mixture was concentrated in vacuo and then concentrated three times from CH 2 Cl 2 The residue was dissolved in methanol (8 ml) and dioxane (8 ml) and morpholine (65 μΐ, 0.75 mmol) was added. sodium cyanoborohydride (47 mg, 0.75 mmol) and acetic acid (45 μΐ) were added.The reaction mixture was stirred at room temperature for 20 hours and the reaction was then stopped by the addition of 2 N hydrochloric acid. made basic by the addition of a saturated solution of NaHCO 3 and then extracted with 1: 1 EtOAc / THF (2x). 26091-42 organic layers were washed with brine, dried (Mg-SO 4), filtered and concentrated in vacuo. Flash chromatography (2 - 3% MeOH / CH 2 Cl 2) afforded 78 mg of the title compound as a colorless solid.

MS 432 (M + H) *; ^ -NMR (400 MHz, CD3OD) δ 7.22 (m, 2H); 6.96 (d, 1 H, J = 9.7 Hz); 4.1 (m, 2H); 3.98 (bs, 2H); 3.58 (m, 4H); 2.7 (m, 2 H); 2.43 (m, 4H) 7 1.3 (d, 6H, J = 6.6 Hz).

C21 H26 ClN5 O3. Retention time on LC: 1.2 min.

10

The following compounds, Examples 22-25, were prepared from the appropriate olefin and amine according to the general method described above for the preparation of Example 21.

15

Example 22 3-Isopropyl-5- [2- (2-pyrrolidin-1-yl-ethoxy-benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-bn C 21 H 28 N 5 O 3. MS 382 (M + H) + Retention time on LC: 1.2 min.

20

Example 23 3-Isopropyl-5- [2- (2-morpholin-4-yl-ethoxy) -cyclohexyl-methyl] -1,6-dihydro-pyrazoloC,3,3-d3 -pyrimidine-7-one hydrochloride C21 H33 N5 O3.HCl. MS 404 (M + H) +. Retention time 25 on LC: 1.1 min.

Example 24 5- [5-Fluoro-2- (2-morpholin-4-yl-ethoxy) -benzyl] -3-isopropyl-1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7- on-hydrochloric acid C21 H26 FN5 O3.HCl. MS 416 (M + H) +. Retention time on LC: 1.2 min.

Example 25 3-Cyclopentyl-5- [5-Fluoro-2- (2-morpholin-4-yl-ethoxy) -35-benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one hydrochloride C23 H28 FNSO3.HCl. MS 442 (M + H) \ Retention time on LC: 1.4 min.

102 6081 - 4 β ι 43.

Example 26 3-Isopropyl-5- [2- (2-morpholir-4-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one hydrochloride 5 Potassium t-butoxide (5.9 g, 48 mol) was added to a solution of 5-isopropyl-4- {2- [2- (2-morpholin-4-yl-ethoxy) -phenyl] -acetylamino} -1H- pyrazole-3-carboxylic acid amide (4.0 g, 9.65 mmol) in dry isopropanol (50 ml). The mixture was heated at 85 ° C for 20 hours. The reaction mixture was concentrated, diluted with brine and extracted with 1: 1 EtOAc / THF (3x). The combined organic layers were washed with brine, dried (MgSO 4, filtered and concentrated. Flash chromatography (2- * 3.5% MeOH / CH 2 Cl 2 with 0.2% NH 4 OH) gave the free base. The product was dissolved in ethanol and treated with an excess of HCl / ether The mixture was concentrated to give 4.1 g of the title compound as a colorless solid.

MS 398 (M + H) · NMR (400 MHz, CD3 OD) 6 7.34 (t, 1H, J = 8.3 Hz); 7.22 (d, 1H, J = 7.5 Hz); 7.07 (d, 1 H, J = 8.3

Hz); 7.01 Cd, 1 H, J = 7.5 Hz); 4.41 (m, 2H); 4.2 (s, 2H); 3.9 (bm, 4H); 3.61 (m, 2 H); 3.4 (m, 1 H); 3.29 (m, 4H); 1.35 (d, 6H, J = 7.1 Hz). C21 H27 N5 O3 .HCl. Retention time at 25 LC: 1.2 min.

The following compounds, Examples 27-31, were prepared from the appropriate amide according to the general method described above for the preparation of Example 26.

Example 27 9- (1,2-Dimethyl-propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one C23 H31 N5 O3. MS 426 (M + H) +.

Retention time on LC: 1.4 minutes.

1 026091 - 4 m »44

Example 28 2- [2- (2-Morpholin-4-yl-ethoxy) -benzyl] -9- (tetrahydrofuran-3-yl) -1,9-dihydro-purine-6-one C 22 H 27 N 5 O 4. MS 426 (M + H) +. Retention time on LC: 0.7 min.

5

Example 29 5- [2- (2-Dimethylamino-ethoxy) -benzyl] -3-isopropyl-1 # 6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one C21 H29 N5 O2. MS 384 (M + H) +. Retention time on LC: 1.1 min.

10

Example 30 3- Cyclopentyl-5- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one hydrochloride C23 H29 N5 O3.HCl . MS 424 (M + H) +. Retention time on LC: 1.4 min.

15

Example 31 3-Cyclobutyl-5- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one hydrochloride C22 H27 N5 O3.HCl . MS 410 (M + H) +. Retention time on LC: 2/1 min.

20

Example 32 9- (1 (R), 2-Dimethylpropyl) - [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one hydrochloride Potassium t- butoxide (143 mg, 1.27 mmol) was added to a solution of 5-amino-1- (1 (R), 2-dimethylpropyl) -1 H -imidazole-4-carboxylic acid amide (50 mg, 0.25 mmol) and (2-morpholin-4-yl-ethoxy) -phenylacetic acid methyl ester (285 mg, 1.02 mmol) in dry 1-butanol (2 ml). The mixture was heated in a microwave at 180 ° C for 30 minutes. The mixture was concentrated and purified by flash chromatography (1-4% MeOH / CH 2 Cl 2). The purified product was dissolved in methanol / ether and treated with an excess of HCl / ether. The resulting solid was filtered and dried to give 73 mg of the title compound in the form of a colorless solid.

1026091 - ". 45.

MS 426 (M + H) +; -NMR (400 MHz, CD3 OD) δ 9.25 (s, 1H); 7.32 (t, 1H, J = 8.3 Hz); 7.22 (d, 1H, J = 7.5 Hz); 7.03 (d, 1 H, J = 8.3 Hz); 7.0 (t, 1H, J = 7.5 Hz); 4.4 (m, 3H); 4.18 (S, 2H); 4.0 (m, 2H); 3.81 (m, 2 H); 3.61 (m, 2 H); Δ 3.59 (m, 2H); 2.12 (m, 2H); 1.55 (d, 3H, J = 7.1 Hz); 0.91 (d, 3H, J = 6.6 Hz); 0.72 (d, 3H, J = 6.6 Hz). C23H31N5O3.HCl. Retention time on LC: 1.4 minutes.

The following compounds, Examples 33-44, were prepared from the appropriate araine and ester according to the general method described above for the preparation of Example 32.

Example 33 9- (2-Methyl-butyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] - 1,9-dihydro-purine-6-one C 23 H 31 N 5 O 3 · MS 426 (M + H) +. Retention time on LC: 2.4 minutes. Example 34 9-Cyclopentyl-2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one C23 H29 N5 O3. MS 424 (M + H) +. Retention time on LC: 2.3 minutes. Example 35 5- [2- (2-Morpholin-4-yl-ethoxy) -benzyl] -3-pyridin-3-yl-1,6-dihydro-pyrazolo [4, 3-d] pyrimidine-7-one C 23 H 24 N 6 O 3. MS 433 (M + H) +. Retention time on LC: 2.0 min.

Example 36 9- (1,2-Dimethyl-propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one hydrochloride C23 H31 N5 O3. HCl. MS 426 (M + H) +. Retention time on LC: 1.4 minutes.

Example 37 35 9-Isopropyl-2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purin-6-one hydrochloride C21 H27 N5 O3. HCl. MS 398 (M + H) +. Retention time on LC: 1.0 min.

1 02 6091 -: * 46

Example 38

2- [2- (2-Morpholin-4-yl-ethoxy) -benzyl] -9- (tetrahydrofuran-2-ylmethyl) -1,9-dihydro-purine-6-one hydrochloride C23 H29 N5 O4.HCl. MS 440 (M + H) +. Retention time on LC: 0.9 minutes.

Example 39 9- (1-Isopropyl-2-methyl-propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one hydrochloride C25 H3 SN5 O3. HCl. MS 454 (M + H) +. Retention time on LC: 1.7 minutes.

Example 40

9- (1-Ethyl-propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one hydrochloride C23 H31 N5 O3. HCl. MS

15 426 (M + H) +. Retention time on LC: 1.4 minutes.

Example 41 9-Cyclopentyl-8-methyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one hydrochloride C24 H31 N5 O3.HCl. MS 438 (M + H) +. Retention time on LC: 1.4 minutes.

Example 42 3- Cyclopentyl-5- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -3,6-dihydro- [1,2,3] triazolo [4,5-d] pyrimidine - 7-on-hydrochloride C 22 H 28 NeO 3 .HCl. MS 425 (M + H) \ Retention time on LC: 1.3 minutes.

Example 43 1-Cyclopentyl-6- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1.5-30 dihydro-pyrazolo [3,4-d] pyrimldin-4-one hydrochloride C23 H29 N5 O3. HCl. MS 424 (M + H) +. Retention time on LC: 1.3 minutes.

Example 44 9-Cyclopentyl-2- [2- (3-morpholin-4-yl-propoxy) -benzyl] -1.9-35 dihydro-purine-6-one hydrochloride C 24 H 31 N 5 O 3 · HCl. MS 438. (M + H) +. Retention time on LC: 2.4 minutes.

1026091-4, 47

Example 45 N - [(IR, S) 2- (3-propropyl-7-oxo-6,7-dihydro-1H-pyrazolo- [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-- yl] -2-pyrrolidin-1-yl-acetamide hydrochloride At room temperature, pyrrolidine (70 μΐ, 0.83 mmol) was added to a mixture of N- [(IR, 2S) 2- (3-isopropyl) 7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-yl] -2-chloro-acetamide (76 mg, 0.208 mmol) in anhydrous THF ( 3 ml). The mixture was heated at 65 ° C for 2.5 hours, after which the mixture was allowed to cool to room temperature and the mixture stirred for 16 hours. The reaction mixture was concentrated and purified by flash chromatography (2- * 3- * 4% MeOH / CH 3 Cl 2 with 0.2% NH 4 OH). The product was dissolved in methanol / ether and treated with an excess of HCl / ether. The resulting solid was filtered and dried, yielding 20 mg of the title compound as a colorless solid.

MS 401 (M + H) +; ΧΗ-ΝΜΗ (400 MHz, CD3OD) δ 4.07 (s, 2H) / 3.7 (m, 3H); 3.45 (m, 1 H); 3.14 (m, 3H); 2.59 (dd, 1H, J = 14.2, 9.2 Hz); 2.2-1.7 (m, 9H); 1.4 (d, 6H, J = 7.1 Hz); 1.4-1.2 (m, 4H). C21 H32 N6 O2 .HCl. Retention time on LC: 1.0 min.

25

The following compounds, Examples 46-50, were prepared from the appropriate amine according to the general method described above for the preparation of Example 45.

30

Example 46 N - [(IR, 2S) 2- (3-Isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-yl] -2 -morpholin-4-yl-acetamide hydrochloride C 21 H 32 N 6 O 3. HCl. MS.

35 417 (M + H) V Retention time on LC: 1.1 min.

1 0 2 60 91%. · ♦ · '48

Example 47 2-Diethylamino-N - [(1R, 2S) 2-Q-isopropyl-4-oxo-7,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1- yl] -acetamide hydrochloride C21 H34 N6 O2.HCl. MS 403 (M + H) *. Re-5 retention time on LC: 1.13 min.

Example 48 1- f [(IR, 2S) 2- (3-Isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo- [4> 3-d] pyrimidin-5-ylmethyl) -cyclohex-i- ylcarbamoyl] -1-methyl) -pyrrolidine-2 (S) -carboxylic acid methyl ester hydrochloride C23 H324 N6 O4.HCl. MS 459 (M + H) +. Retention time on LC: 1.4 minutes.

Example 49 2-Cyclobutylamino-N- [(1R, 2S) 2-P-isopropyl-4-oxo-7,7-dihydro-1H-pyrazolo [4,3-d 3 pyrimidin-5-ylmethyl) -cyclohex- 1-yl] -acetamide hydrochloride C21 H32 N6 O2. HCl. MS 401 (M + H) +. Retention time on LC: 1.2 min.

Example 50 2- Cyclopropylamino-N - [(1R, 2S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-yl] -acetamide hydrochloride C20 H30 N6 O2.HCl. MS 387 (M + H) +. Retention time on LC: 1.1 min.

25

The following numbered Preparations describe the preparation of certain intermediates used in the preceding Examples.

Preparation A1 N- [(IR, 2S) -2- (3-isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo- [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1] -yl] -2-chloro-acetamide

At room temperature, chloroacetyl chloride (95 µl, 1.2 inmol) was added to a suspension of 5- (2-aminocyclohex-1-ylmethyl) -3-isopropyl-1,6-dihydro-pyrazolo [4, 3-d] pyrimi dine-7-one (200 mg, 0.614 mmol) and pyridine (1.2 ml, 14.8 1026091-1> 49 * mmol) in dry CH 2 Cl 2 (8 ml) and dioxane (3 ml). 1'N hydrochloric acid was added to the mixture after 24 hours, after which it was extracted with EtOAc (2x). The combined extracts were washed with 1 N hydrochloric acid and brine, dried (MgSO 4), filtered and concentrated to give 76 mg of the title compound in the form of a light brown solid.

MS 366 (M + H) +; ^ -NMR (400 MHz, d 6 -acetone) δ 8.6 (bs, 1H); 7.5 (bd, 1H, J ≤ 4.5 Hz); 7.35 (m, 1 H), -4., 02 (s, 2 H); 3.42 (m, 1 H); 3.3 (m, 1 H); 2.95 (dd, 1H, J - 14.6, 5.0 Hz); 2.5 (dd, 1H, J = 14, -6, 7.5 Hz); 1.9 (m, 2H); 1.65 (m, 2 H); 1.4 (d, 6H, J = 7.0 Hz); 1.4-1.05 (m, 5H).

Preparation BI

5 - ((IS, 2R) 2-Amino-cyclohex-1-ylmethyl) -3-isopropyl-1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one 10% Pd / C (70 mg ) was added to a suspension of 5 - ((IR, 2S) 2-azido-cyclohex-1-ylmethyl) -3-isopropyl-1,6-dihydro-20-pyrazolino [4,3-d] pyrimidine-7 -one (408 mg, 12.9 mmol) in absolute ethanol (30 ml) containing 1 N hydrochloric acid (2 ml). The mixture was brought under 45 psi of hydrogen for 2 hours and then the reaction mixture was flushed with nitrogen and filtered. The filtrate was concentrated in vacuo, stirred with ether, filtered and dried, yielding 322 mg of the title compound as a colorless solid.

MS. 290 (M + H) +; ^ -NMR (400 MHz, CD 3 OD) δ 3.39 (m, 1H); 3.12 (m, 1H), 2.95 (m, 1H); 2.69 (m, 1 H); 2.01 (m, 2H); 1.85 (m, 2 H); 1.73 (m, 1 H); 1.5-1.2 (m; 4H); 1.4 (d, 6H, J = 7.0 Hz).

102609H

»*> '50;

Preparation C1-5 - ((IS, 2R) 2-Azido-cyclohex-1-ylmethyl) -3-isopropyl-1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one

Potassium t-butoxide (695 mg, 5.68 mmole) was added to a solution of 5-isopropyl-4- [2 - ((1S, 2R) 2-azido-cyclo-hex-1-yl) -acetylamino] -1H-pyrazole-3-carboxylic acid amide (379 mg, 1.14 mmol) in dry isopropanol (10 ml). The mixture was heated at 85 ° C for 20 hours. The reaction mixture was concentrated in vacuo and the residue was 10 acidified with 1 N hydrochloric acid and extracted with BtOAc, (3x) The combined extracts were washed with brine, dried (MgSO 4), filtered and concentrated to give 437 mg of the title compound as a colorless solid.

MS 316 (M + H) 1 H-NMR (400 MHz, CD 3 OD) δ 3.4 (bm, 1H); 3.15 (dt, 1H, J = 10.4, 4.2 Hz); 2.97 (dd, 1H, J = 14.2, 5.3 Hz); 2.48 (dd, 1H, J = 14.2, 8.3 Hz); 2.15-1.5 (m, 9H); 1.4 (d, 6H, J = 7.0 Hz).

20

Preparation D1 5-Isopropyl-4- (2- [2- (2-morpholin-4-yl-ethoxy) -phenyl] - acetylamino) -1H-pyrazole-3-carboxylic acid amide cyclic 1-propane phosphonic anhydride (50% solution in 25 ethyl acetate, 7.3 ml, 0.012 mol) was added at room temperature to a solution of 4-amino-5-isopropyl-1H-pyrazole-3-carboxylic acid amide (1.7 g, 10 mmol), 2- (morpholine-4 -yi-ethoxy) -phenylacetic acid (3.5 g, 13 mmol) and triethylamine (2.85 ml, 20 mmol) in anhydrous DMF (30 ml). After 20 30. hour, the mixture was concentrated in vacuo and the residue was purified by flash chromatography (2- * 6% MeOH / CH 2 Cl 2 with 0.2% NH 4 OH) to give 4.0 g of the title compound as a colorless solid received.

. 35 MS. 416 (M + H) +; X H NMR (400 MHz, CD 3 OD) δ 7.25 (m, 2H); 6.99 (d, 1H, J = 6.9 Hz); 6.95 (t, 1H, J = 7.5 Hz); 4.19 (m, 1026091 -> 1.> 51 2H); 3.7 (m, 2H); 3.59 (m, 2H); 3.01 (m, 1 H); 2.8 (m, 2H); 2.56 (m, 2 H); 1.2 (d, 6H, J = 6.6 Hz).

The following connections. Preparations D2-D6 were prepared from the appropriate amine and acid according to the general method described above for the preparation of D1.

Preparation D2 10 5-Cyclopentyl-4- (2- [2- (2-morpholin-4-yl-ethoxy) -phenyl] -acetylamino) -1H-pyrazole-3-carboxylic acid amide

Preparation D3 5-Isopropyl-4- (2- [2- (2-diethylamino-ethoxy) -phenyl] -acetyl-15 amino) -1H-pyrazole-3-carboxylic acid amide

Preparation D4 1- (1,2-Dimethyl-propyl) -5- {2- [2- (2-torpholin-4-yl-etho-3-cy) -phenyl] -acetylamino) -1H-imidazole-4-carboxylic acid amide

Preparation D5 5- (2- [2- (2-Morpholin-4-yl-ethoxy) -phenyl] -acetylamino) -1- (tetrahydrofuran-3-yl) -1H-imidazole-4-carboxylic acid amide Preparation D6 4- [2- (2-Amino-cyclohexyl) -acetylamino] -5-isopropyl-2H-pyrazole-3-carboxylic acid amide

Preparation E130 4-Amino-5-isopropyl-β-pyrazole-3-carboxylic acid amide 5- Isopropyl-4-nitro-1H-pyrazole-3-carboxylic acid amide (3 9 # 15.1 mmol) and 10% palladium on carbon (500 mg) in ethanol (30 ml) was stirred under hydrogen (50 psi) for 18 hours at room temperature. The reaction mixture was filtered and the solid was washed with methanol (50 ml), dichloromethane (50 ml), ethanol (50 ml) and ethyl acetate (50 ml). The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography on silica gel, eluting with dichloromethane imethanol (9: 1 volume ratio) and to give the title compound (2 , 6 g) in the form of an off-white solid.

^ -NMR (400 MHz, DMSO-d 6) δ 12.20-12.30 (bs, 1H); 7.02-7.14 (bs, 1H); 6.85-6.95 (bs, 1H); 4.30-4.46 (bs, 1 H); 2.90-3.00 (m, 1 H); 1.15 ^ 1.21 (d, 6H) ppm; LRMS (electros-10 pray): m / z [M-H] + 167, [2M-H] * 335; Analysis, found C, 49.86; H, 7.21; N, 33.07. C7 H12 N4 O requires C, 49.99; H, 7.19; N, 33.31.

The following compounds, Preparations E2-E3, were prepared from the appropriate nitropyrazole according to the general method described above for the preparation of E1.

Preparation E2 4-Amino-5-cyclopentyl-1H-pyrazole-3-carboxylic acid amide Preparation E3

4-Amino-5-cyclobutyl-1H-pyrazole-3-carboxylic acid amide Preparation PI

5- 1-propyl-4-nitro-1H-pyrazole-3-carboxylic acid amide

At "0 ° C, oxalyl chloride (6.8 ml, 77.6 mmol) was added dropwise to a suspension of 5-isopropyl-4-nitro-1H-pyrazole-3-carboxylic acid (5.15 g, 25.9 mmol) ) in .30 dichloromethane (80 ml) containing dimethylformamide (0.1 ml) .The reaction mixture was stirred at 0 ° C for 1 hour after which time it was warmed to room temperature and stirred for a further 2 hours. reduced pressure, the residue was dissolved in toluene (100 ml), and ammonia gas was bubbled into the solution for 2 hours, and the reaction mixture was stirred at room temperature under nitrogen for 18 hours.

• I

Stirred, concentrated under reduced pressure and the residue was dissolved in hot methanol (300 ml). The resulting precipitate was filtered off and the filtrate was concentrated under reduced pressure. The residue was azeotroped with water (300 ml), concentrated to about 80 ml under reduced pressure, and the precipitate isolated by filtration. This was washed with water and dried under vacuum to give the title compound (3.1 g) in the form of an orange solid.

X H NMR (400 MHz, DMSO-d 6) δ 7.94-7.99 (bs, 1H); 7.68 - 7.72 (bs, 1 H); 3.45-3.55 (m, 1 H); 1.24-1.30 (d, 6H) ppm; LRMS

(electrospray): m / z [M-Na] + 221, [M-H] * 197.

15

The following compounds, Preparations F2-F3, were prepared from the appropriate nitropyrazole according to the general method described above for the preparation of F1.

20

Preparation F2 5 - Cyc 1 openty 1 -4-nitro-1H-pyrazole -3-carboxylic acid amide Preparation F3 25 5-Cyclobutyl-4-nitro-1H-pyrazole-3-carboxylic acid amide Preparation 61 5-1 sopropyl -4-nitro-1H -pyrazole-3-carboxylic acid 5-Isopropyl-1H-pyrazole-3-carboxylic acid (5 g, 32.5 mmol) was 30. added in portions with concentrated sulfuric acid (25 ml) at room temperature with stirring. The reaction mixture was then heated to 60 ° C and concentrated nitric acid (70%, 6 ml, 90 mmol) was added dropwise while maintaining the temperature at 60 ° C. The reaction mixture was subsequently stirred at 60 ° C. for 3 hours, cooled to room temperature and poured onto 50 ml of ice with stirring. After 15 minutes, the white precipitate was isolated by filtration, after which it was washed with water and dried under reduced pressure to give the title compound (5.2 g) in the form of a white solid was obtained.

5 1 H NMR (400 MHz, DMSO-d 6) δ 13.86-13.93 (bs, 1H); 13.50-13.80 (bs, 1 H); 3.39-3.52 (m, 1 H); 1.18-1.30 (d, 6H) ppm; LRMS (electrospray): m / z [M-H] * 198.

The following compounds, Preparations G2-G3, were prepared from the appropriate pyrazole according to the general method described above for the preparation of G1.

Preparation G2 5-Cyclopentyl-4-nitro-β-pyrazole - 3-carboxylic acid Preparation G3 5-Cyclobutyl-4-nitro-1H-pyrazole-3-carboxylic acid 20

Preparation H1 5-1sopropyl-1-pyrazole-3-carboxylic acid 5-Isopropyl-1H-pyrazole-3-carboxylic acid ethyl ester. (18.9 g, 104 mmol) and a 1 M solution of NaOH (260 mL, 259 mmol) were dissolved in 1,4-dioxane (300 mL) and the reaction mixture was heated to 50 under nitrogen for 3 hours ° C. The reaction mixture was cooled, the pH was adjusted to pH 2 with concentrated hydrochloric acid and the solvent was removed under reduced pressure. The resulting solid was azeotroped with toluene (2 x 30 mL), dissolved in ethyl acetate (500 mL) and washed with water. The aqueous layer was removed and extracted with ethyl acetate (2 x 200 ml), after which the combined organic layers were dried over MgSO 4. The solvent was removed under reduced pressure and the residue was azeotroped with dichloromethane (2 x 50 ml), 600260 ΐ.

55% with which the title compound (14.7 g) was obtained in the form of a white solid.

^ -NMR (400 MHz, DMSO-d 6) δ 12.50-13.30 (b &, 2H); 6.42 (s,

5 (1H); 2.84-2.94 (quintet, 1H); 1.15-1.19 (d, 6H) ppm; LRMS

(electrospray): m / z [M-H] + 153.

The following compounds, Preparations H2-H3, were prepared from the suitable ester according to the general method described above for the preparation of. Hl.

Preparation H 2 5-Cyclopenty1-1H-pyrazole-3-carboxylic acid 15

Preparation H3

5-Cyclobutyl-1 H -pyrazole-3-carboxylic acid Preparation II

5-Isopropyl-1 H -pyrazole-3-carboxylic acid ethyl ester

Hydrazine hydrate (6.6 ml, 134 mmol) was added to a solution of 5-methyl-2,4-dioxo-hexanoic acid ethyl ester (23.8 g, 188 mmol) in ethanol (100 ml) at room temperature under nitrogen. . The reaction was allowed to proceed for 18 hours at room temperature, after which the solvent was removed under reduced pressure. The residue was partitioned between dichloromethane (300 ml) and water (300 ml) and the aqueous layer was removed. The organic layer was washed with water (2 x 200 ml), dried over MgSO 4 and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel eluting with a solvent gradient of pentane: ethyl acetate (which changed from 4: 1 to 2: 1 volume ratio) and to give the title compound (18.9 g) in the A white solid form was obtained.

1026091 - '56' X H NMR (40 O MHz, CDCl 3) δ 10.80-10.95 (bs, 1 H); 6.61 (s, 1H), - 4.3 ^ -4.40 (quartet, 2H); 2.98-3.08 (quintet, 1H); 1.35-1.41 (t, 3H), 1.24-1.32 (d, 6H) ppm; LRMS (electros-pray): m / z [M-H] + 181.

5

The following compounds, Preparations 12-13, were prepared from the appropriate diketone according to the general method described above for the preparation of II.

10

Preparation 12 5 -Cyclopentyl-1H-pyrazole'-3-carboxylic acid ethyl ester Preparation 13 15 5-Cyclobutyl-1H-pyrazole-3-carboxylic acid ethyl ester

Preparation J1 5-Methyl-2,4-dioxo-hexanoic acid ethyl ester

At room temperature, sodium pellets 20 (3.39 g, 148 mmol) were dissolved in ethanol (100 ml) and a solution of diethyl oxalate (20 ml, 147 mmol) in 3-methyl-2 was added for 30 minutes at room temperature. -butanone (18.9 ml, 177 mmol) was added dropwise. The reaction mixture was diluted with ethanol (100 ml), heated to 60 ° C and stirred at this temperature for 2 hours. After cooling to room temperature, the reaction mixture was poured into ice-cold 2 N hydrochloric acid (200 ml) and extracted with diethyl ether (300 ml), and ethyl acetate (300 ml). The combined organic extracts were dried over MgSO 4 and concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel, eluting with a pentane: ethyl acetate solvent gradient (from 99: 1 to 95: 5 by volume) with which the title product (23.8 g) was obtained in the form of a yellow oil; 102 6091 -

> ·. I

♦ 57.0-NMR (400 MHz, CDCl 3) δ 14.40-14.80 (bs, 1H); 6.40 (s, 1 H); 4.30-4.39 (quartet, 2H); 2.60-2.71 (quintet, 1H); 1.35-1.40 (t, 3H), 1.15-1.20 (d, 6H) ppm; LRMS (electros-pray): m / z [M-H] + 185.

5

The following compounds, Preparations J2-J3, were prepared from the appropriate ketone according to the general method described above for the preparation of J1.

10

Preparation J2 4- Cyclopentyl-2,4-dioxo-butanoic acid ethyl ester Preparation J3

4-Cyclobutyl-2,4-dioxo-butanoic acid ethyl ester Preparation KI

5- Amino-1- (1,2-dimethyl-propyl) -1 H -imidazole-4-carboxylic acid amide A mixture of 2-cyano-2-aminoacetamide (200 mg, 2 mmol) and triethyl orthoformate (0.37 ml, 2 , 42 mmol) in dry acetonitrile (5 ml) was heated at 80 ° C for 3 hours.

1,2-dimethylpropylamine (211 mg, 2.4 mmol) was added and the reaction mixture was allowed to cool to room temperature and stirred overnight. The mixture was concentrated and purified with flash chromatography. chromatography (1-3% MeOH / CH 2 Cl 2) to give 221 mg of the title compound as a light brown solid.

MS 197 (M + H) +; ^ -NMR (400 MHz, DMS0-d6) δ 7.18 (s, 1H); 6.68 (bs, 1 H); 6.58 (bs, 1 H); 5.71 (bs, 2H); 3.8 (m, 1 H), 1.93 (m, 1 H); 1.3 (d, 3H, J = 7.1 Hz); 0.87 (d, 3H, J = 7.1 Hz); 0.7 (d, 3H, J. = 6.6 Hz).

35 '. ·· ... ·

The following compounds, Preparations K2-K11, were prepared from the appropriate amine and orthoformate according to the general method described above. for the preparation of KI.

»

Preparation K2 5-Amino-1- (1 (S), 2-dimethyl-propyl) -1H-imidazole-4-carboxylic acid amide Preparation K3 5-Amino-1- (1 (R), 2-dimethyl-propyl 1 H -imidazole-4-carboxylic acid amide

Preparation K4 5-Amino-1- (1-methyl-butyl) -1H-imidazole-4-carboxylic acid amide Preparation K5 15-Amino-1- (cyclopentyl) -1H-imidazole-4-carboxylic acid amide Preparation K6 5-Amino-1 - (isopropyl) -1H-imidazole-4-carboxylic acid amide Preparation K7 5-Amino-1- (1-isopropyl-2-methyl-propyl) -1H-imidazole-4-carboxylic acid amide

Preparation K8 5-Amino-1- (2-ethyl-propyl) -1H-imidazole-4-carboxylic acid amide Preparation K9 5-Amino-1- (tetrahydrofuran-3-yl) -1H-imidazole-4-carboxylic acid amide 3 °

Preparation K10 5-Amino-1- (tetrahydrofuran-3-yl-methyl) -1H-imidazole-4-carboxylic acid amide Preparation Kil 5-Amino-1-cyclopentyl-2-methyl-1H-imidazole-4-carboxylic acid amide 102 60 9 * * '". · 59

Preparation LI

5-Amino-1-cyclopentyl-1H-pyrazole -4-carboxylic acid amide Concentrated H 2 SO 4 (14.7 ml) was added to 5-amino-1-cyclopentyl-1H-pyrazole-4-carbonitrile (5) , 18 g, 29.4 mmol). The reaction mixture was stirred at room temperature for 44 hours. The reaction mixture was poured out on ice and the pH was adjusted to 11 by the addition of NH 4 OH. The solid was filtered off to give 3.74 g of the title compound in the form of a brown solid.

MS 195 (M + H) +; ^ -NMR (400 MHz, CD 3 OD) δ 7.69 (s, 1H); 4.54 (m, 1 H); 2.08-1.69 (m, 8 H).

15

Preparation M1 -.

5-Amino-1-cyclopentyl-1H-pyrazole-4-carbonitrile To a solution of sodium ethoxide (0.34 g of Na in 30 ml of absolute ethanol) was added cyclopentylhydrazine-20-hydrochloride (2.0 g, 14.6) at room temperature mmol). The reaction mixture was heated at 75 ° C for 2 hours. The reaction mixture was allowed to cool to room temperature and stirred for 12 hours. The solvent was removed in vacuo to give an oily residue that was dissolved in 50 ml of water and then extracted with EtOAc. The combined organic layers were washed with brine, dried (MgSO 4) and concentrated to give 1.69 g of the title compound in the form of an orange solid.

MS 177 (M + H) +; ^ -NMR (400 MHz, CD3OD) δ 7.05 (s, 1H) .; 4.51 (m, 1 H); 2.05-1.57 (m, 8H).

Preparation NI

Cyclopentyl hydrazine hydrochloride

At room temperature, borane-tetrahydrofuran complex (105.4 ml of a 1 M solution in. THF, 105.4 mmol) was added 1026091-60 to N'-cyclopentylidene hydrazine carboxylic acid tert-butyl ester (20.89 g, 105.4 mmol) ) and stirred for 15 minutes while gas evolution occurred. 6 N hydrochloric acid (52.7 ml) was added slowly and the reaction mixture was stirred at room temperature for 10 minutes. The reaction mixture was heated at 80 ° C for 15 minutes, then cooled to room temperature and evaporated. The resulting solid was washed with THF (250 ml) and a white solid was removed by filtration. The filtrate was concentrated to give 12.7 g of the title compound as a white solid.

MS 101 (M + H) +; X H NMR (400 MHz, CD 3 OD) 5 3.57 (m, 1 H); 2.02 (m, 2H); 1.77-1.62 (m, 6H).

Preparation 01 N '- Cyclopentylidene hydrazine carboxylic acid tert-butyl ester t-butyl carbazate (14.9 g, 113 mmol) in hexanes (100 ml) was heated to 75 ° C. Cyclopentanone (10.0 ml, 113 mmol) was added and the reaction mixture was kept at 75 ° C for 1 hour. The reaction mixture was allowed to cool to room temperature. The crystallized product was isolated by filtration to give 20.89 g of the title compound in the form of a white solid.

MS 199 (M + H) +; X H NMR (400 MHz, CDCl 3) δ 2.57 (m, 2H); 2.27 (m, 2H); 1.88 (m, 2 H); 1.79 (m, 2 H); 1.52 (s, 9H).

30

Preparation of PI

5-Amino-1-cyclopentyl-1H- [1,2,3] triazole-4-carboxylic acid amide Sodium ethoxide (prepared from 920 g of sodium in 40 ml of ethanol, 40 mmol) was heated to 70 ° C and a solution 35 of azidocyclopentane (2.2 g, 20 mmol) and cyanoacetamide (1.68 g, 2.0 mmol) in ethanol (5 ml). The reaction mixture was cooled after 2 hours and concentrated 102 609% 61 to remove most of the ethanol. The residue was diluted with a pH 7 buffer and extracted with EtOAc (3x). The combined organic layers were washed with brine, dried (Na 2 SO 4), filtered and concentrated to give 500 mg of the title compound in the form of a light brown solid.

MS 196 (M + H) +; -NMR (400 MHz, CD3 OD) δ 4.6 (m, 1H); 2.1 (m, 4H); 1.95 (m, 2H); 1.7 (m, 2H).

Preparation Q1 (2-Allyloxy-5-chloro-phenyl) acetic acid

At room temperature a solution of KQH (2.3 g, 0.04 mol) in ethanol (20 ml) and water (2 ml) was added to (2-allyloxy-5-chloro-phenyl) acetonitrile (1.6 g , 8 mmol). The mixture was heated at 93 ° C for 3.5 hours and then concentrated in vacuo. The residue was dissolved in water (5 ml) and treated with concentrate. 20 hydrochloric acid (4 ml) which was added dropwise at 0 ° C. The mixture was stirred for 15 minutes and then diluted with water and extracted with EtOAc (2x). The organic layers were combined and washed with brine, dried (MgSO 4), filtered and concentrated to give 1.7 g of product in the form of a colorless solid was obtained.

MS 225 (Μ-ΗΓ; X H NMR (400 MHz, acetone-d 6) δ 7.28 (d, 1 H, J - 3 Hz); 7.22 (dd, 1 H, J = 8.6, 3.0 Hz), 6.98 (d, 1H, J * 8.6 Hz), 6.01 (m, 1H), 5.42 (d, 1H, J = 17.5 Hz), 5.2 (d (1H, J = 11.4 Hz), 4.6 (m, 2H), 3.6 (s, 2H).

The following compound, Preparation Q2, was prepared from the appropriate nitrile according to the general method described above for the preparation of Q1.

". "1 02 609"% 62

Preparation Q2 (2-Allyloxy-5-fluoro-phenyl) acetic acid Preparation R1 (2-Allyloxy-5-chloro-phenyl) acetonitrile

At 0 ° C, thionyl chloride (3.6 ml, 50 mmol) was added over a period of 8 minutes to a solution of 2-allyloxy-5-chlorobenzyl alcohol (5.9 g, 30 mmol) in dry THF ( 50 ml) containing 0.2 ml of DMF. The reaction was stopped after 2.5 hours by the careful addition of water. The mixture was extracted with EtOAc (3x) and the combined extracts were dried (MgSO 4), filtered and concentrated to give 7.1 g of the corresponding benzyl chloride. This product was dissolved in dry DMF (80 ml) and sodium cyanide (2 g, 40 mmol) was added. The mixture was heated at 80 ° C for 2 hours, cooled, diluted with water and extracted with EtOAc (3x). The combined extracts were washed with water (3x) and brine, dried (MgSO 4), filtered and concentrated in vacuo. The product was purified by flash chromatography (5- * 30% EtOAc / hexanes) to give 2.0 g of the title compound as a colorless oil.

25 H NMR (400 MHz, CDCl3) δ 7.38 (s, 1H) 7 7.22 (m, 1H); 6.8 (d, 1H, J = 9.1 Hz); 6.01 (m, 1 H); 5.4 (d, 1H, J = 17.5 Hz); 5.3 (d, 1H, J = 10.8 Hz); 4.59 (m, 2 H); 3.65 (s, 2H).

The following compound, Preparation R2, was prepared from the appropriate benzyl alcohol according to the general method described above for the preparation of R1.

Preparation R2 (2-Allyloxy-5-fluoro-phenyl) acetonitrile 1026091- *. *> 63

Preparation SI

2-Allyloxy-5-chloro-berizyl alcohol

Lithium aluminum hydride (1.17 g, 0.031 mol) was added portionwise at 0 ° C to a solution of 2-allyloxy-5-5 chloro-benzoic acid allyl ester (7.4 g, 0.029 mol) in dryness. THF (100 ml). After 1.5 hours the reaction was stopped by the successive addition of water (1.2 ml), 15% NaOH (1.2 ml) and water (3.6 ml). · The mixture was dried (Mg-SO 4) , filtered and concentrated to give 5.9 g of the title compound as a colorless oil.

MS 197 (M-H) *; ^ -NMR (400 MHz, acetone-d 6) δ 7.42 (d, 1 H, J '- 3 Hz); 7.19 (dd, J = 8.5, 3.0 Hz); 6.95 (d, 1H, J = 8.5 Hz); 6.03 (m, 1 H); 5.4 (d, 1H, J = 17.5 Hz); 5.22 (d, 1H, J = 11.4 Hz); 4.63 (m, 2H); 4.6 (m, 2H).

The following compound, Preparation S2, was prepared from the suitable ester according to the general method described above for the preparation of S1.

Preparation S2 2-Allyloxy-5-fluoro-benzyl alcohol Preparation T1 2-Allyloxy-5-chloro-benzoic acid allyl ester

At room temperature, cesium carbonate (20, 8 g, 63.7 mmol) was added to a mixture of 5-chloro-2-hydroxy-benzoic acid (5 g, 29 mmol) and allyl bromide (5.5 mL, 64 30 mmol) in dry DMF (60 ml). After 20 hours, the reaction mixture was diluted with water and extracted with ether (3x). The combined organic extracts were washed with water and brine, dried (MgSO *), filtered and concentrated in vacuo to give 7.4 g of the title compound in the form of an oil.

1 02 60 91 ^ '·' ... · '· 64 MS 253 (M + H) *; X H NMR (400 MHz, acetone-d 6) δ 7.7 (d, 1 H, J = 2.8 Hz) ƒ 7.52 (dd, 1 H, J = 8.5, 2.8 Hz); 7.18 (d, 1H, J = 8.5 Hz); 6.02 (m, 1 H); 5.5 (dd, 1H, J - 17.5, 1.6 Hz); 5.41 (d, 1H, J = 17.5 Hz); 5.22 (d, 2H, J = 10.3 Hz); 4.85 (d, 2H, J = 5.4 Hz); 4.63 (m, 2H).

The following compound, Preparation T2, was prepared from the appropriate acid according to the general method described above for the preparation of T1.

10

Preparation T2

2-Allyloxy-5-fluoro-benzoic acid allylester Preparation UI

[2- (3-Morpholin-4-yl-propoxy) -phenyl] -acetic acid methyl ester

Morpholine (90 μΐ, 1.05 mmol) was added to a mixture of [2- (3-bromo-propoxy) -phenyl] -acetic acid methyl ester (300 mg, 1.05 mmol) and sodium bicarbonate (132 mg, 1, 58 mmol) in dry acetonitrile (3 ml). The mixture was heated to 85 ° C overnight and then concentrated and purified by flash chromatography (1-3% methanol / dichloromethane) to give the title compound as a colorless oil.

MS 294 (M + H) +; X H NMR (400 MHz, CD 3 OD) 6 7.22 (dd, 1H, J = 8.4, 7.6 Hz); 7.18 (d, 1H, J = 7.6 Hz); 6.9 (t, 1H, J = 7.6 Hz); 6.83 (d, 1H, J = 8.4 Hz); 4.0 (t, 2H, J = 5.8

Hz); 3.75 (bs, 4H); 3.65 (s, 3H); 3.6 (s, 2H); 2.5 (bs, 6H); 1.98 (bs, 2 H).

30

Preparation VI

[2- (2-Morpholin-4-yl-ethoxy) -phenyl] -acetic acid 10% Pd / C (2g) was added to a solution of [2- (2-morpholin-4-yl-ethoxy) -phenyl] -acetic acid-benzyl ester (35 g, 28.2 mmol) in absolute ethanol (100 ml) and shaking under 40 psi of hydrogen for 4 hours. The reaction mixture was purged with nitrogen and filtered. The 1 02 60 91 ^ '-'> 65 ''. ·. . The filtrate was concentrated and the product was crystallized from ethanol / ether to give 7.0 g of the title product as a colorless solid.

• '5' MS 266 {M + H) 1 H-NMR (400 MHz, CD 3 OD) δ 7.19 (m, 2H); 6.93 (m, 2H); 4.3 (m, 2H); 3.9 (m, 4H); 3.5 (s, 2H); 3.22 (m, 2H); 3.01 (m, 4H).

Preparation W1 [2- (2-Diethylaminoethoxy) -phenyl] -acetic acid-benzyl ester cesium carbonate (1 g, 3.1 mmol) was added to a solution of (2-hydroxy-phenyl) -acetic acid-benzyl ester (3.00 mg, 1 (2 mmol) and N, N-diethyl-2-chloroethylamine hydrochloride (200 mg, 1.2 mmol) in dry DMF (10 ml) and the mixture was stirred at room temperature for 48 hours. The mixture was concentrated in vacuo and purified by flash chromatography (1- 2 * 4% MeOH / .CH 2 Cl 2) to give 350 mg of the title compound as a light brown oil.

MS 343 (M + H) +; 1 H NMR (400 MHz, ace ton-d 6). δ 7.38 (m, 5H) / 7.22 (πν, 2H); 6.98 (d, 1H, J = 7.9 Hz)? 6.88 (t, 1H, J = 7.1 Hz); 5.12 (s, 2H); 4.0 (m, 2H); 3.7 (s, 2H); 2.78 (m, 2H); 2.58 (m, 4H); 1.0 (m, 6H).

Preparation Π [2- (2-Morpholin-4-yl-ethoxy) -phenyl] -acetic acid-benzyl ester Diethyl azodicarboxylate (12.3 ml, 77.5 mmol) was added dropwise to a solution of (2-hydroxy-phenyl) acetic acid benzyl ester (15 g, 62 mmol), triphenyl phosphine (20.3 g, 77.5 mmol) and; 2-morpholin-4-yl-ethanol (11.25 ml, 92.7 mmol) in dry THF (100 ml) which was in a cold water bath. The mixture was stirred at room temperature for 20 hours and then concentrated to approximately 50 ml. The residue was acidified with 1 N hydrochloric acid and washed with ether (6 x 200 ml). The organic layers were discarded 102 60 91 - * 66 and the aqueous layer was made basic at pH 10 with solid Na 2 CO 3 and extracted with ether (2 x 200 ml). The ether layers were combined, dried (MgSOe), filtered and concentrated. Flash chromatography (5% THF / CH 2 Cl 2 containing 0.5% saturated NH 3 / MeOH) gave 17 g of the title compound in the form of an oil.

MS 356 (M + H) +; 1 H NMR (400 MHz, CDCl 3) δ 7.4-7.2 (m, 7H) / 6.92 (t, 1H, J = 7.5 Hz); 6.83 (d, 1H, J = 7.5 Hz); 5.1 (s, 10 2H); 4.05 (m, 2 H); 3.65 (m, 6H); 2.7 (m, 2 H); 2.51 (m, 4 H).

The following compound, Preparation X2, was prepared from the appropriate alcohol according to the general method described above for the preparation of X1.

Preparation X2 [2- (3-Bromo-propoxy) -phenyl] -acetic acid methyl ester Preparation Y1 (2-Hydroxy-phenyl) -acetic acid-benzyl ester

A solution of (2-hydroxy-phenyl) -acetic acid (15 g, 0.1 mol) in dry DMF (120 ml) was treated at room temperature with benzyl bromide (13 ml, 0.11 mol) and sodium bicarbonate (12 , 5 g, 0.15 mol). After stirring for 18 hours, a further 3 ml of benzyl bromide and 5 g of sodium bicarbonate were added and stirring was continued for 20 hours. The mixture was diluted with water (300 ml) and extracted with ethyl acetate. (3 x 100 ml). The combined organic layers were washed with a solution of NaH-CO 3 (1 x) and brine, dried (Na 2 SO 4), filtered and concentrated. After the addition of hexanes, the product crystallized, which was filtered off and dried, yielding 21 g of the title compound as colorless crystals.

1026091 · =, * 67 MS 243 (M + H) +; ^ -NMR (400 MHz, CDCl 3) δ 7.38 (m, 5H); 7.2 (t, 1H, J = 7.1 Hz); 7.08 (d, 1 H, J = 7.1 Hz); 6.95 (d, 1H, J = 7.6 Hz); 6.7 (t, 1H, J = 7.6 Hz); 5.18 (s, 2H); 3.71 (S, 2H).

5.

Preparation Z1 trans- (2-allyloxy-cyclohexyl) acetic acid

At room temperature, sodium hydroxide (1 ml of a 15% solution) was added to a solution of (2-allyloxy-cyclohexyl) acetic acid allyl ester (322 mg, 1/37. Mmol) in methanol (6 ml). The mixture was heated at 65 ° C for 1.5 hours, after which it was cooled and concentrated. The residue was treated with 1 N hydrochloric acid and extracted with EtOAc (2x). The combined extracts were dried (MgSO 4), filtered and concentrated to give 262 mg of the title compound as a colorless oil.

MS 199 (M + H) +; ^ -NMR (400 MHz, acetone-d 6) δ 5.9 (m, 1H); 5.22 (dd, 1H, J - 17.5, 2.1 Hz); 5.06 (dd, 1H, J - 10.5, 2.1 Hz); 4.1 (dd, 1H, J = 12.9, 5.4 Hz); 3.9 (dd, 1H, J = 13.2, 5.4 Hz); 3.0 (m, 1 H); 2.81 (d, 1 H, J = 13.2 Hz); 2.77 (dd, 1H, J = 15.3, 4.2 Hz); 2.1-1.55 (m, 5H); 1.3-1.0 (m, 4H).

25

The following compound, Preparation Z2, was prepared from the suitable ester according to the general method for the preparation of Z1.

Preparation Z2

(trans) -2- (Azido-cyclohexyl) -acetic acid Preparation AAI

(2-Allyloxy-cyclohexyl) acetic acid allyl ester Sodium hydride (95%, 273 mg, 10.8 mmol) was added at 0 ° C

added to a solution of trans- (2-hydroxy-cyclohexyl) acetic acid (814 mg, 5.14 mmol) in dry DMF (25 ml).

1026091-

* J

68

Allyl bromide (1 ml, 11.5 mmol) was added after 15 minutes and the reaction was allowed to warm to room temperature. After 4 hours, an additional 0.5 ml of allyl bromide was added to the gelatinous mixture and the reaction mixture was stirred overnight. The reaction was stopped by the addition of a saturated solution of NaHCC> 3 and extracted with EtOAc (3x). The combined organic layers were washed with water and brine, dried (MgSO 4), filtered and concentrated. The product was purified by flash chromatography (5- * 1 0- * 2 0%

EtOAc / hexanes) to give 322 mg of the title compound in the form of a colorless oil.

MS 239 (M + H) *; 1 H NMR (400 MHz, acetone-d 6) δ 5.9 (m, 2H); 5.3 (d, 1H, J = 17.5 Hz); 5.2 (m, 2H); 5.03 (d, 1H, J = 10.5 Hz); 4.52 (d, 2H, J = 5.3 Hz); 4.08 (dd, 1H, J = 13.2, 5.4 Hz); 3.84 (dd, 1H, J = 13.2, 5.8 Hz); 2.98 (m, 1 H); 2.8 (d, 1H, J = 13.7 Hz); 2.7 (dd, 1H, J = 15.0, 4.5 Hz); 2.2-1.58 (m, 5H); 1.3-1.0 (m, 4H).

20

Preparation BBl (trans) -2- (Azido-cyciohexyl) -acetic acid ethyl ester At room temperature, sodium azide (993 mg, 7.58 mmol) was added to a solution of cis- (2-methanesulfonyl-oxy-cyclohexyl) acetic acid ethyl ester (668 mg, 2.53 mmol) in dry DMF (15 ml). The mixture was heated at 97 ° C for 16 hours, cooled, diluted with water and extracted with EtOAc (2x). The combined organic layers were washed with water (2x) and brine, dried (MgSO 4), filtered and concentrated to give 488 mg of the title compound in the form of a brown oil.

^ -NMR (400 MHz, acetone-d 6) δ 4.08 (kw, 2H, J = 7.0 Hz); 3.15 (m, 1 H); 2.6 (m, 1 H); 2.18 (m, 1 H); 1.8-1.0 (m, 4H); 1.2 (t, 3H, J = 7.0 Hz).

10260 91 - • / 69 • »*. ". .

Preparation CC1

Cis- (2-methanesulfonyloxy-cyclohexyl) -acetic acid ethyl ester Methanesulfonyl chloride (0.25 ml, 3.2 mmol) was added at 0 ° C to a solution of cis- (2-hydroxy-5-cyclohexyl) -acetic acid ethyl ester ( 497 mg, 2.7 mmol), pyridine (650 μΐ, 8 mmol) and DMAP (489 mg, 4 mmol) in dry CH 2 Cl 2 (8 ml). The mixture was allowed to warm to room temperature and stirred for 20 hours. The mixture was diluted with CH 2 Cl 2 and washed with 1 N hydrochloric acid (3x) and brine, dried (MgSO 4), filtered and concentrated to give 668 mg of the title compound in the form of a brown oil.

MS 265 (M + H) +; ^ -NMR (400 MHz, acetone-d 6) δ 4.9 (m, 1H); 4.04 (kw, 2H, J = 7.0 Hz); 3.1 (s, 3H); 2.42 (dd, 1H, J = 16.2, 7.0 Hz); 2.23 (dd, 1H, J = 16.2, 7.4 Hz); 2.1 (m, 2H); 1.7-1.3 (m, 7H); 1.2 (t, 3 £, J = 7.0 Hz).

ORGANIC PROTOCOLS

The usefulness of the compounds of Formula (I), the stereoisomers and prodrugs thereof, and the pharmaceutically acceptable salts of the compounds, stereoisomers and prodrugs in the treatment or prevention of diseases (as described in this specification) in Animals, in particular mammals (e.g. humans) can be demonstrated by their activity in use; actual assays known to one skilled in the art, including the in vitro and in vivo assays described below. Such assays also provide the ability to compare the activity of the compounds of Formula (I) with the activity of other known compounds.

1.. Activity of Inhibition of Phosphodiesterase (PDE) Preferred PDE compounds suitable for use in the art. use. in accordance with the present invention, potent inhibitors of cGMP are PDE9. The activity of the 102609H.

70.

"I

inhibition of PDE against cyclic guanosine 3 ', 5' monophosphate (cGMP) and cyclic adososine 3 ', 5' monophosphate (cAMP) phosphodiesterases in vitro is determined by measuring their ic50 values (the concentration of the compound required for 50% inhibition of erizyme activity).

Phosphodiesterase-9 can be obtained from full-length human recombinant clones transfected into SF9 cells, as described in Fisher et al., Jour-10: nal of Biological Chemistry, 1998, 273, 15559-15564.

The tests are performed using a modification of the batch method of WJ, Thompson et al. (Biochem., 1979, 18, 5228) or a scintillation proximity determination for the direct detection of AMP / GMP using a change is made to the protocol described by Amersham plc under product code TRKQ7090 / 7100. In summary, the effect of PDE9 inhibitors is investigated by measuring a fixed amount of enzyme in the presence of variable concentrations of the inhibitor and low substrate concentrations (in a 3: 1 ratio of unnoticed to [3 H] -labeled cGMP at a concentration of about 1/3 K *,) such that IC 50 “Ki. The final volume of the assay is adjusted to 100 μΐ with test buffer [20 mM Tris-HCl pH 7.4, 5 mM MgCla, 1 mg / ml bovine serum albumin]. The reactions are started with enzyme, incubated for 30-60 minutes at 30 ° C with <30% of the substrate being converted, and the reactions are stopped with 50 μΐ yttrium silicate SPA-30 beads (containing 3 mM from the respective contain unlabeled cyclic nucleotides for PDE9 and PDE11). The plates are resealed and shaken for 20 minutes, after which 30 minutes are waited in the dark until the beads have settled, after which a TopCount plate reader 35 (Packard, Meriden, CT) is counted . The units of radioactivity are converted to a percentage of activity of an unrestrained control (100%), plotted against the concentration of the inhibitor and the IC 50 values of the inhibitor are obtained with the "Fit Curve" Microsoft Excel extension.

2. Effect of specific PDE9 inhibitors on metabolic syndrome in animals - Effects on plasma levels of glucose, triglyceride, insulin and cGMP in oJb / oJb mice. Biological data a. Test compounds; The PDE9 inhibitor compounds being tested are dissolved in 10% DMSO / 0.1% Pluronic® P105 Block Copolymer Surfactant (BASF Corporation, Parsippany, NJ) in 0.1% physiological saline without the pH being adjusted and are adjusted via a gavage administered with oral 15 feeding needles for mice (20 gauge, Popper & Sons,

Ine., Ney Hyde Park, NY). A volume of 4 ml / kg body weight is administered for each dose. Alternatively, the compounds are administered in powdered mouse food (Mouse Breeder / Auto-JL K20 mouse food, PMI Feeds, Ine., St. Louis, MO that can be supplied specifically for the y customer by Research Diets, Ine., New

Brunswick, NJ) in the form of a mixture of compound and feed; the compounds are mixed with the feed in such a way that the animal will consume specified dosages. The compounds are tested in a dosage range of 1-500 mg / kg / day.

b. Test animals:

Male and female oJb / ob mice are obtained from Jackson Laboratories (Bar Harbor, ME) and are used in the studies when they are 6 to 10 weeks old. Mice are housed at five per cage and have unlimited access to Dl1 mouse food (Purina, Brentwood, MO) and water.

35 1026091-72; .

. »'C. Test protocol:

The mice are allowed to acclimate for one week prior to the start of the study in Pfizer's animal facilities. When the compounds are administered in the powdered mouse feed, the mice are transferred to the powdered diet three days prior to the start of the dosing period to allow them to get used to the changed diet. At this time, the mice are randomly assigned to groups of ten with five mice per cage. On day 1, blood samples were taken behind the eye socket and plasma glucose was determined as described below. On day 1, for mice in which the compounds were administered in the form of a mixture with the feed, the feed was replaced with the mixture of the compound with the feed and this is refreshed every day during the study. On day 5, the mice were bled from the sinus behind the eye socket at about 8 o'clock in the morning to prepare plasma for the glucose and triglyceride analysis described below. Terminal plasma samples are then taken immediately after bleeding from the sinus behind the eye socket as described below.

On day 1, mice, to which the compounds are administered via a gastric probe, are dosed only in the afternoon with the carrier or with a test PDE9 inhibitor compound. The mice are then dosed twice a day on days 2-4, in the morning and in the afternoon. On day 5, the mice receive a dose in the morning and are bled 3 hours later to prepare plasma for the glucose and triglyceride analysis described below. Terminal plasma samples are taken on day 5 after bleeding through the sinus behind the eye socket as described below. The body weight is determined on days 1 and 5 of the study and the food consumption is determined over the 5 day period.

'' 02 60914 '• ♦ 73 <' t d. Terminal bleeding and tissue collection:

On the morning of the last day of the study, mice to which the compound was administered as a mixture with the feed were bled at about 8 o'clock in the morning through the sinus behind the eye socket and then immediately after terminal plasma withdrawal performed as described below. On the morning of the last day of the study, the mice, to which the compounds were administered via a tube probe, are dosed with test compound or carrier at about 8 o'clock in the morning. 3 hours after the last dose, 25 μΐ of blood is collected via the sinus behind the eye socket and this is added to 100 μΐ 0.025% heparinized physiological saline in Denville Scientific micro tubes 15 (Denville Scientific Ine., Metuchen, NJ). The tubes are centrifuged for 2 minutes at the highest speed in a Beekman Microfuge 12 (Beekman Coulter Ine., Fuller-ton, GA). The plasma is collected for the determination of glucose and triglyceride in plasma. The mice are then sacrificed by decapitation and approximately 1 milliliter of blood is collected in Becton-Dickineon Microtainer tubes for plasma separation with lithium heparin (Bect-on-Dickinson Ine., Franklin Lakes, NJ). The tubes are centrifuged in the Beekman Microfuge 12 for 5 minutes at the highest speed. The plasma is collected in 1.5 ml Eppendorf tubes that are frozen in liquid nitrogen. The plasma samples are stored at -80 ° C until they are analyzed.

30 e. Analysis of metabolites and hormones;

Glucose, triglycerides and cholesterol in plasma are measured with the Roche / Hitachi 912 Clinical Chemistry Analyzer (Roche Diagnostics Corp., Indianapolis, IN) using Roche kits. Free fatty acids are measured on the same device using a kit from Wako Chemical (Richmond, VA). plasma cGMP is measured with a Biotrak enzyme immunoassay system from Ameraham 102 6091 - 74 (Piscataway1, NJ). Measurements of insulin in plasma are determined by a similar method in which the Mercodia ELISE Insulin kit from ALPCO (Uppsala, Sweden) is used. All determinations are carried out according to the supplier's instructions.

f. Results:

The PDE9 inhibitor has more than 40% inhibition against PDE9 at a concentration of 1 μΜ. In some compounds, the PDE9 inhibitor has an IC 50 of less than 500 nM. In other compounds, the PDE9 inhibitor has an IC5 <> of less than 50 nM. In some compounds, the PDE9 inhibitor has a selectivity for PDE9 over PDE1 of more than 10. In other compounds, the PDE inhibitor has a selectivity for PDE9 over PDE1 of more than 50, and in others. compounds more than 100.

Together, our test results in a hyperglycaemic insulin-resistant ob / ob mouse suggest that selective PDE9-20 inhibition improves metabolic parameters associated with metabolic syndrome.

1026091-

Claims (8)

1. RL (VLV) 30. N-V 2 => 9) 7 R; or, (h) R 3; P, including the carbon atoms to which it is attached, is (C 3 -C 8) cycloalkyl, (C 3 -C 8) heterocycloalkyl, aryl or heteroaryl; optionally and independently substituted with 1 to 3 substituents independently selected from each other from halogen, (C1 -C5) alkyl, (C1 -C5) alkoxy and trifluoromethyl; J is O, S, -N (R 15) -, -N (R 15) CO -, -CN (R 15) -, -SO 2 N (R 15) -, or -N (R 15) SO 2 -; 5 x is 0, 1, 2, 3, 4, 5 or 6; R10 is -C02 H, -CONR30 R31, -NR30 R31 or -N (R15) SO2 R40; R 1 and R 2 are independently H or (C 1 -C 3) alkyl; R 3 is (C 1 -C 8) alkyl, (C 3 -C 8) cycloalkyl, (C 3 -C 8) cycloalkyl -10 methyl, (C 3 -C 8) heterocycloalkyl, (C 3 -C 8) heterocycloalkyl-methyl, aryl or heteroaryl; optionally and independently substituted with 1 to 3 substituents independently selected from halogen, hydroxy, oxo, (C 1 -C 5) alkyl and (C 1 -C 5) alkoxy; is; R 15 is H or (C 1 -C 5) alkyl; R30 and R31 are separately and independently H, (C3 -C5) alkyl, (C3 -C8) cycloalkyl, (C3 -C8) heterocycloalkyl, aryl or heteroaryl, wherein R30 and R31 are optionally and independently substituted with 1 to 3 substituents independently selected from halogen, oxo, (C 1 -C 5) alkyl, -CO 2 R 40; -COR40, -OR40, -CONRS0R51, -NRS0R51 and -SO2R40; or R30 and R31 together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocycloalkyl ring, which ring optionally has 1 additional heteroatom independently selected from N, O and S, the 5- to 8- Each heterocycloalkyl ring is optionally and independently substituted with 1 to 3 substituents independently selected from halogen, (C 1 -C 5) alkyl, -CO 2 R 40, -COR 40, -OR 4, -CONRS-R51. -NR 50 R 51 and -SO 2 R 4 °; R 40 is H, (C 1 -C 5) alkyl, (C 3 -C 8) cycloalkyl, (C 3 -C 8) heterocycloalkyl, aryl, heteroaryl> R 50 and R 51 are separated and independently H, (C 1 -C 8) alkyl, (C 3 -C8) cycloalkyl, (C3 -C8) heterocycloalkyl, aryl or heteroaryl; or R50 and RS1 together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocycloalkyl ring. t, which ring optionally has 1 additional heteroatom independently selected from N, 0 and S. A compound of Formula (I), (η 4, Jn (CH 2) x R 10 (I) a stereoisomer or prodrug thereof, or a pharmaceutically acceptable salt of said compound, stereoisomer or prodrug, wherein: VVS, (a> R3; (b) R3; (C) R3; OO r1 O hn hn- ^ n ^ "- \ W, vW VW · .. (d) r3I (e) R3; (f) R3 ; A compound according to claim 1 wherein: The compound of claim 2 wherein: A is (a) or (b). A compound according to claim 1 which: 1- {[2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl } pyrrolidine-2-carboxylic acid; 20 - {[2- (1-cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetyl} -pyrrolidine-2 (S carboxylic acid; 3-isopropyl-5- [2- (2-oxo-2-piperazin-1-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one-trifluoroacetate ; 1-cyclopentyl-6- [2- (2-oxo-2-piperazin-1-yl-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d] pyrimidine-4-one; 3-isopropyl-5- [2- (2-morpholin-4-yl-2-oxo-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; 3-isopropyl-5- [2- (2-oxo-2-pyrrolidin-1-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; 5- {2- [2- (4-ethyl-piperazin-1-yl) -2-oxo-ethoxy] -benzyl} -3-isopropyl-1,6-dihydro-pyrazolo [4,3-d] pyrimidine- 7-on; 35 N, N-diethyl-2-12- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetamide; 1026091-L. 1- {[2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl} -pyrrolidine- 2-carboxylic acid methyl ester; 4- {[2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-5 d] pyrimidin-5-ylmethyl) -phenoxy] -acetyl-piperazine-1-carboxylic acid - tert-butyl ester; N- (2-dimethylamino-ethyl) -2- [2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -phenoxy] - acetamide; 10 - {[2- (1-cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetyl} -pyrrolidine-2 - carboxylic acid methyl ester; 4- {[2- (1-cyclopentyl-4-oxo-4,5-dihydro-1H-pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetyl} -piperazine-1-carboxylic acid tert-butyl ester; 1-cyclopentyl-6- [2- (2-oxo-2-pyrrolidin-1-yl-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d] pyrimidine-4-one; 1-cyclopentyl-6- [2- (2-morpholin-4-yl-2-oxo-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d] pyrimidine-4-one; 2- [2- (1-Cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -N- (2-dimethyl-tin-ethyl-ethyl) -acetamide; 1-cyclopentyl-6 - {[2- [2- (4-ethyl-piperazin-1-yl) -2-oxo-ethoxy] -benzyl} -1,5-dihydro-pyrazolo [3,4-d] pyrimidine 25 4-one; 2- [2- (1-cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -N, N-diethyl-acetamide; [2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3- d] pyrimidin-5-ylmethyl) -phenoxy] -acetic acid; . [2- (1-cyclopentyl-4-oxo-4,5-dihydro-1 H -pyrazolo [3,4-d] pyrimidin-6-ylmethyl) -phenoxy] -acetic acid; 3-isopropyl-5- [2- (5-chloro-2-morpholin-4-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; 3-isopropyl-5- [2- (2-pyrrolidin-1-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; 1026091-v * 3-isopropyl-5- [2- (2-morpholin-4-yl-ethoxy) -cyclohexylmethyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; ". 5- [5-fluoro-2- (2-morpholin-4-yl-ethoxy) -benzyl] -3-5. isopropyl-1,6-dihydro-pyrazolo [4,3-d] pyrimidin-7-one; 3-cyclopentyl-5- [5-Fluoro-2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; 3-isopropyl-5- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,6-dihydro-pyrazolo [4,3-d] pyrimidine-7-one; 9- (1,2-dimethyl-propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one; 2- (2- (2-Morpholin-4-yl-ethoxy) -benzyl] -9- (tetrahydrofuran-3-yl) -1,9-dihydro-purine-6-one; 5- [2- ( 2-diethylamino-ethoxy) -benzyl] -3-isopropyl-1,6-15 dihydro-pyrazolo [4,3-d] pyrimidin-7-one; 3-cyclopentyl-5- [2- (2-morpholine-4) -yl-ethoxy) -benzyl] - 1,6-dihydro-pyrazolo [4,3-d] pyrimidin-7-one; 3-cyclobutyl-5- [2- (2-morpholin-4-yl-ethoxy) -benzyl] - 1,6-dihydro-pyrazolo [4,3-d] pyrimidin-7-one; 9- (1 (R)> 2-dimethylpropyl) - {2- (2-morpholin-4-yl-ethoxy-benzyl] - 1,9-dihydro-purine-6-one; 9- (2-methyl-butyl) -2- [2- (2-morol-4-yl-ethoxy) -benzyl] -1,9-dihydro- purine-6-one; 9-cyclopentyl-2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -25 1,9-dihydro-purine-e-one; 5- [2- (2 -morpholin-4-yl-ethoxy) -benzyl] -3-pyridin-3-yl-1,6-dihydro-pyrazolo [4,3-d] pyrimidin-7-one; 9- (1,2-dimethyl) propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one; 9-isopropyl-2- [2- (2-morpholine) 4-yl-ethoxy) -benzyl] - 1,9-dihydro-purin-6-one; 2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -9- (tetrahydrofuran-2-ylmethyl) ) -1 9-dihydro-purine-6-one; 9- (1-isopropyl-2-methyl-propyl) -2- [2- (2-morpholin-4-35 yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one; 9- (1-ethyl-propyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one; 026091-1- [9-cyclopentyl-8-methyl) -2- [2- (2-morpholin-4-yl-ethoxy) -benzyl] -1,9-dihydro-purine-6-one; 3-cyclopentyl-5- [2- (2-morpholin-4-yl-ethoxy) -benzyl] - 3,6-dihydro- [1,2,3] triazolo [4,5-d] pyrimidine-7-one ; 5 1-cyclopentyl-6- [2- (2-morol-4-yl-ethoxy) -benzyl] -1,5-dihydro-pyrazolo [3,4-d] pyrimidine-4-one; 9-cyclopentyl-2- [2- (3-morpholin-4-yl-propoxy) -benzyl] -1,9-dihydro-purine-6-one; N - [(1 R, 2 S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-yl] - 2-pyrrolidin-1-yl-acetamide; N - [(1 R, 2 S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-yl] -2 -morpholin-4-yl-acetamide; 2-diethylamino-N - [(1 R, 2 S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1] -yl] -acetamide; 1- {[(1R, 2S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1 H -pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-yl-ylcarbamoyl] -methyl} -pyrrolidine-2 (S) -carboxylic acid methyl ester; 2-cyclobutylamino-N - [(IR, 2S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1-- yl] -acetamide; 2-cyclopropylamino-N - [(IR, 2S) 2- (3-isopropyl-7-oxo-6,7-dihydro-1H-pyrazolo [4,3-d] pyrimidin-5-ylmethyl) -cyclohex-1] -yl] -acetamide; a stereoisomer or prodrug thereof or a pharmaceutically acceptable salt of that compound, stereoisomer or prodrug. 30 5. Pharmaceutical preparation that. a compound according to claim 1, a stereoisomer or prodrug thereof or a pharmaceutically acceptable salt of said compound, stereoisomer or prodrug and a pharmaceutically acceptable carrier, vehicle or diluent. 1026091-: .. 1 ' A is (a), (b), (c) or (h); R1 and R2 are H; R 3 is (C 3 -C 5) alkyl or (C 3 -C 5) cycloalkyl; P is (C 3 -C 6) cycloalkyl or aryl; J is O or S; and is 10 x 1, 2 or 3. 6. Method for the treatment of a condition, disease or symptom selected from the group consisting of type 1 diabetes, type 2 diabetes, hyperglycemia, dyslipemia, impaired glucose tolerance, metabolic syndrome and cardiovascular disease, which method administering to a mammal in need of such treatment or prevention, a therapeutically effective amount of a compound according to claim 1, a stereoisomer or prodrug thereof, or a pharmaceutically acceptable salt of said compound, stereo isomer or prodrug; or a pharmaceutical composition comprising said compound according to claim 1, stereoisomer or prodrug thereof, or a pharmaceutically acceptable salt of said compound, stereoisomer or prodrug. 15 The method of claim 6 wherein said condition, disease or symptom is diabetes or cardiovascular disease. A method for inhibiting the activity of phosphodiesterase 9 in a mammal in need of such inhibition, said method comprising the administration of a phosphodiesterase 9 inhibitory amount of a compound according to claim 1, a stereoisomer or prodrug thereof or a pharmaceutically acceptable salt of that compound, stereoisomer or prodrug; or a pharmaceutical composition comprising a compound of Formula (I), a stereoisomer or prodrug thereof or. a pharmaceutically acceptable salt of said compound, stereoisomer or prodrug and a pharmaceutically acceptable carrier, vehicle or diluent. 1 026081 -.