OA12337A - Crystalline therapeutic agent. - Google Patents

Description

012337

CRYSTALLINE THERAPEUTIC AGENT

The présent invention relates to a polymorph of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7“OXO-2H-pyrazolo(4,3- d]pyrimidin-5-yl3-3- pyridylsulfonyl}-4-ethy!piperazine having the formula (I):

having a PXRD pattern substantially as defined hereinafter:

As detailed in WO 01/27113, 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- dJpyrimidin-5-yl]-3-pyridylsulfonyl}~4-ethylpiperazine is a potent and sélective cGMP PDES inhibitor and is ’especiallyuseful in the treatment of inter alia male erectile dysfunction.

For successful utility within the pharmaceutical industry it is critical that thephysicochemical properties of an active material are either known or can bereasonably predicted throughout the necessary processes during both itsmanufacture and pharmaceutical processing as well as during its shipping,storage and eventual therapeutic use. In some cases compounds can exhibitdésirable médicinal properties which cannot be translated directly into a suitabiepharmaceutical composition because the active compound itself has 012337 unsatisfactory physical properties such as for example poor Chemical orProcessing properties. As such, stable, crystalline materials are highly désirablefor use in the pharmaceutical industry as crystalline materials are, in general,more stable than their amorphous counterparts, they hâve a finite structure which 5 can be reproducibly characterîsed by X-ray diffraction which can be used toidentify the presence of a spécifie polymorphie form.

The présent invention provides the crystal structure of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)~7-oxo-2H-pyrazolo[4,3“ dJpyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpiperazine. ^0 The crystalline material of i-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2- rnethoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpiperazine as defined herein may also be referred to as a non-solvatedproduct of 1 -{6-6thoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyî)-7-oxo-2H- pyrazolo[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpiperazine wherein a non-solvated product as defined herein means a product which has essentially noretained solvent bound within the crystal lattice or otherwise entrapped orengaged within the crystals. Solvent retained on the surface of the crystal latticeis not included within the définition of retained solvent, although it is preferredthat the crystalline material of the présent invention does not retain solvent on the 20 surface. Essentially no retained solvent as defined herein means that there isless than 1%, preferably iess than about 0.85%, more preferably less than about0.3% and especialiy from 0 to 0.25% total retained solvent within the crystallattice structure or entrapped within the crystals.

The crystals of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2~(2-methoxyethyl)-7- 25 oxo-2H-pyrazolo[4,3- d3pyrimidin-5-yl]-3-pyridylsulfonyl}~4-ethylpiperazine havingthe XRD pattern as defined hereinafter hâve a high melting point and earlyinvestigations show that they are non-hygroscopic, are stable and hâve goodformulation properties. 012337

The préparation of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)- 7-oxo-2H-pyrazoio[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpiperazinehaving the formula (I) according to the présent invention may be carried out asillustrated in the Example and Préparations sections hereinafter. $ The XRD pattern of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)- 7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpîperazinehaving the formula (I) can be characterised according to the method detailedhereinafter. X-Rav Diffraction HO The single crystal X-ray diffraction (XRD) pattern for 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-ylî-3- pyridylsuifonyl}-4-ethyipiperazine, having Mpt 162-165°C (as prepared accordingto the process of Example 1) was determined by single crystal X-Ray diffraction.A représentative crystal was surveyed and a 1 Â data set (maximum sin Θ/λ=0.5) Ί5 was collected on a Siemens R4RA/v diffractometer. Atomic scattering factorswere taken from the International Tables for X-Ray Crystallography (InternationalTables for X-Ray Crystallography, Vol. IV, pp. 55, 99, 149 Birmingham: KynochPress, 1974). All crystallographic calculations were facilitated by the SHELXTLSystem (G.M. Sheldrick, SHELXTL, User Manual, Nicolet Instruments Co., 1981). 20 All diffractometer data were collected at room température.

The simulated powder X-ray diffraction (PXRD) pattern, (as reported in Table 1for intensifies above 1%) was generated from single crystal data using Cerius2Diffraction-Crystal Module using Radiation = 1.54178Â with a 0.5 Polarisationfactor and no monochromator. The crystal size was taken to be 500 x 500 x 500 25 Â3 with Lorentzian peak shape. Diffraction peaks were simulated over the two- theta angle range of 2-55°. PCT/ 012337 4

The crystallographic calculations (intensity calculations) were facilitated by use ofthe SHELXTL System (G.M. Sheldrick, SHELXTL, User Manual, NicoletInstrument Co., 1981). The final R-index was 4.60%.

The main peaks (in degrees Θ) of the simulated PXRD pattern are illustrated in5 Table I. Main peaks as defined herein are all those having an intensity above 1%. TABLE 1 d-spacing (À) Intensity _ d-spacing lA) Intensity i%) d-spacing (A) Intensity (%) d-spacing (Â) Intensity (%) 12.2486 1.24 4.2323 19.04 3.061 9.18 2.4041 2.24 9.7301 100 4.1595 1.19 3.0483 2.22 2.3424 1.22 8.5596 36.31 4.0421 1.35 3.0428 1.94 2.299 1.13 7.3923 15.02 3.9186 7.81 2.9929 1.59 2.264 1.23 7.3856 8.35 3.8951 10.26 2.9804 2.1 2.2605 1.29 7.3284 1.69 3.8491 5.34 2.9403 3.46 2.2079 2.65 5.8222 14.49 3.8485 2.46 2.9073 1.74 2.1855 1.9 5.7617 1.96 3.8427 43.14 2.8719 1.68 2.1679 1.92 5.7461 43.25 3.7357 3.1 2.8532 4.7 2.1468 1.35 5.1166 7.17 3.6961 3.99 2.8122 11.19 2.1119 4.52 5.1078 32.71 3.6805 4.51 2.8097 2.82 2.0769 1.15 4.8807 17.47 3.6042 5.28 2.7776 2.17 2.0471 1.12 4.8674 10.85 3.5988 6.09 2.7774 1.46 2.028 1.63 4.865 10.98 3.5819 6.8 2.7162 1.76 1.9935 1.03 4.8425 13.19 3.5721 8.55 2.7041 3.63 1.9883 2.96 4.8018 3.32 3.5355 2.47 2.6875 1.42 1.9181 1.27 4.7632 10.57 3.407 1.2 2.6443 2.19 1.9058 1.03 4.7491 4.16 3.3461 4.91 2.6084 3.75 1.884 1.23 4.641 1.96 3.3318 7.54 2.6025 1.6 1.8797 2.49 4.629 19.73 3.3154 4.36 2.5809 1.31 1.8749 2.81 4.3314 9.53 3.2827 1.7 2.5448 1.21 1.7939 1.57 4.3301 3.65 3.2774 Ï.22 2.5438 3.26 1.7617 1.24 4.3066 26.66 3.2434 19.95 2.4325 2.47 T.7379 1.82 4.2909 51.91 3.224 2.11 2.4212 1.24 1.7194 1.03 4.2798 24.75 3.0663 ï.38 2.4163 4.39 012337

Thus the présent invention provides 1-{6-ethoxy-5-(3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo(4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpiperazine materia, having a PXRD pattern substantially as defined in Table1 when measured according to the method described hereinbefore. 5 As will be appreciated by the skilled crystallographer, whilst the relative intensifies of the various peaks within Table 1 may vary due to a number offactors such as for example orientation effects of crystals in the X-ray beam orthe purity of the material being analysed or the degree of crystallinity of thesampie, the peak positions will remain substantially as defined in Table. 10 The skilled crystallographer will also appreciate that measurements using a different wavelength will resuit in different shifts according to the Braggéquation - ηλ - 2d sin θ.

Such further PXRD patterns of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4- 15 ethylpiperazine generated by use of alternative wavelengths are considered to bealternative représentations of the PXRD pattern of the crystalline material of theprésent invention and as such are within the scope of the présent invention.

The same compound, as defined by the XRD pattern described in Table 1,when made via alternative routes (as detailed in the Examples section 2o hereinafter) can hâve a melting point in the range of from 155-165°C (measuredusing a Perkin Eimer DSC7/TGA7 at a heating rate of 20°C/minute).

Expérimente indicate that the crystalline compound as defined hereintends to exist in one polymorphie form as detailed hereinbefore and asexemplified hereinafter. Monomorphic compounds are particularly désirable for 25 pharmaceutical purposes. »c·· 012337 6

The crystalline compound of the présent invention, hereinafter referred toas “the compound” can be administered alone but, in human therapy willgeneraliy be administered in admixture with a suitable pharmaceutical excipientdiluent or carrier selected with regard to the intended route of administration andstandard pharmaceutical practice.

For example, the compound can be administered orally, buccally orsublingually in the form of tablets, capsules (including soft gel capsules), ovules,élixirs, solutions or suspensions, which may contain fiavouring or colouringagents, for immédiate-, delayed-, modified-, or controlled-release such assustained-, dual-, or pulsatile delivery applications. The compound may also beadministered via intracavernosal injection. The compound may also beadministered via fast dispersing or fast dissolving dosages forms or in the form ofa high energy dispersion or as coated particles. Suitable pharmaceuticalformulations of the compound may be in coated or un-coated form as desired.

Such tablets may contain excipients such as microcrystalline cellulose,lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, glycineand starch (preferably corn, potato or tapioca starch), dîsintegrants such assodium starch glycollate, croscarmellose sodium and certain complex silicates,and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia.Additionally, lubricating agents such as magnésium stéarate, stearic acid,glyceryl behenate and talc may be included.

Solid compositions of a similar type may also be employed as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, a cellulose, milk sugar or high molecular weight polyethylene glycols. For aqueous suspensions and/or élixirs, the compound may be combined with various sweetening or fiavouring agents, colouring matter or dyes, with emulsifying 012337 7 and/or suspending agents and with diiuents such as water, éthanol, propyleneglycol and glycerin, and combinations thereof.

Modified release and pulsatile release dosage forme may containexcipients such as those detailed for immédiate release dosage forms together 5 with additional excipients that act as release rate modifiers, these being coatedon and/or inciuded in the body of the device. Release rate modifiers include, butare not exclusiveiy limited to, hydroxypropylmethyl cellulose, methyl cellulose,sodium carboxymethyicellulose, ethyl cellulose, cellulose acetate, polyethyleneoxide, Xanthan gum, Carbomer, ammonio méthacrylate copolymer, 30 hydrogenated castor oil, carnauba wax, paraffin wax, cellulose acetate phthaiate,hydroxypropylmethyl cellulose phthaiate, methacrylic acid copolymer andmixtures thereof. Modified release and pulsatile release dosage forms maycontain one or a combination of release rate modifying excipients. Release ratemodîfying excipients maybe présent both within the dosage form i.e. within the 35 matrix, and/or on the dosage form i.e. upon the surface or coating.

Fast dispersing or dissolving dosage formulations (FDDFs) may containthe following ingrédients: aspartame, acesulfame potassium, citric acid,croscarmellose sodium, crospovidone, diascorbic acid, ethy, acrylate, ethylcellulose, gelatin, hydroxypropylmethyl cellulose, magnésium stéarate, mannitol, 2o methyl méthacrylate, mint flavouring, polyethylene glycol, fumed silica, Silicondioxide, sodium starch glycolate, sodium stearyl fumarate, sorbitol, xylitol. Theterms dispersing or dissolving as used herein to describe FDDFs are dépendentupon the solubility of the drug substance used i.e. where the drug substance isinsoluble a fast dispersing dosage form can be prepared and where the drug 25 substance is soluble a fast dissolving dosage form can be prepared.

The compound can also be administered parenterally, for example, intracavernosally, intravenously, intra-arterially, intraperitoneally, intrathecaily, intraventricularly, intraurethrally intrasternally, intracranially, intramuscularly or PCT*. 012337 8 subcutaneously, or they may be administered by infusion or needleless injectiontechniques. For such parentéral administration they are best used in the form ofa stérile aqueous solution which may contain other substances, for example,enough salts or glucose to make the solution isotonie with blood. The aqueoussolutions should be suitably buffered (preferably to a pH of from 3 to 9), ifnecessary. The préparation of suitable parentéral formulations under stérileconditions is readily accomplished by standard pharmaceutical techniques well-known to those skilled in the art.

For oral and parentéral administration to human patients, the daily dosagelevel of the compound will usually be from 10 to 500 mg (in single or divideddoses).

Thus, for exampie, tablets or capsules of the compound may contain from5 mg to 250 mg of active compound for administration singly or two or moreat a time, as appropriate. The physician in any event will détermine the actualdosage which will be most suitable for any individual patient and it will var’ withthe âge, weight and response of the particular patient. The above dosageu areexemplary of the average case. There can, of course, be individual instanceswhere higher or lower dosage ranges are merited and such are within the scopeof this invention. The skilled person will also appreciate that, in the treatment ofcertain conditions (including MED and FSD), the compound may be taken as asingle dose on an “as required” basis (i.e. as needed or desired).

Example Tablet Formulation

In general a tablet formulation could typieally contain between about 0.01 mg and500mg of the compound whilst tablet fill weights may range from 50mg to1000mg, An example formulation for a 10mg tablet is illustrated: vo 9 01233 Inaredient %w/w Free acid, Free base or Sait of Compound 10.000* Lactose 64.125 Starch 21.375 Croscarmellose Sodium 3.000 Magnésium Stéarate 1.500 * This quantity is typically adjusted in accordance with drug activity.

The tablets are manufactured by a standard process, for example, directcompression or a wet or dry granulation process. The tablet cores may becoated with appropriate overcoats.

The compound can also be administered intranasally or by inhalation andare conveniently delivered in the form of a dry powder inhaler or an aérosol sprayprésentation from a pressurised container, pump, spray or nebuliser with the useof a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, a hydrofluoroalkanejjgch as 1,1,1,2-tetrafluoroethane(HFA 134A [trade mark] or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EA [trademark]), carbon dioxide or other suitable gas. In the case of a pressurisedaérosol, the dosage unit may be determined by providing a valve to deliver ametered amount. The pressurised container, pump, spray or nebuliser maycontain a solution or suspension of the active compound, e.g. using a mixture oféthanol and the propellant as the solvent, which may additionally contain alubricant, e.g. sorbitan trioleate. Capsules and cartridges (made, for example,from gelatin) for use in an inhaler or insufflator may be formulated to contain apowder mix of a compound of the invention and a suitable powder base such aslactose or starch. 012337 10 Aérosol or dry powder formulations are preferably arranged so that eachmetered dose or “puff” contains from 1 to 50 mg of a compound of the inventionfor delivery to the patient. The overall daiiy dose with an aérosol will be in therange of from 1 to 50 mg which may be administered in a single dose or, moreusually, in divided doses throughout the day.

The compound may also be formulated for delivery via an atomiser.Formulations for atomiser devices may contain the following ingrédients assolubilisers, emulsifiers or suspending agents: water, éthanol, glycerol,propylene glycol, low molecular weight polyethylene glycols, sodium chloride,fluorocarbons, polyethylene glycol ethers, sorbitan trioleate, oleic acid.

Alternatively, the compound can be administered in the form of asuppository or pessary, or they may be applied topically in the form of a gel,hydrogel, lotion, solution^ cream, ointment or dusting powder. The compoundmay also be dermally administered. The compound may also be transdermallyadministered, for example, by the use of a skin patch. The compound may alsobe administered by the ocular, pulmonary or rectal routes.

For ophthalmic Use, the compound can be formulated as micronisedsuspensions in isotonie, pH adjusted, stérile saline, or, preferably, as solutions inisotonie, pH adjusted, stérile saline, optionally in combination with a preservativesuch as a benzylalkonium chloride. Alternatively, the compound may beformulated in an ointment such as petrolatum.

For application topically to the skin, the compound of the invention can beformulated as a suitable ointment containing the active compound suspended ordissolved in, for example, a mixture with one or more of the following: minerai oil,liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylenepoiyoxypropylene compound, emulsifying wax and water. Alternatively, it can beformulated as a suitable lotion or cream, suspended or dissolved in, for example, 012337 11 a mixture of one or more of the following: minerai oil, sorbitan monostearate, apolyethylene glycoi, iiquid paraffin, polysorbate 60, cetyl esters wax, cetearylalcohol, 2-octyldodecanoI, benzyl alcohol and water.

The compound may also be used in combination with a cyclodextrin. 5 Cyclodextrins are known to form inclusion and non-inclusion complexes with drugmolécules. Formation of a drug-cyclodextrin compiex may modify the solubility,dissolution rate, bioavailability and/or stability property of a drug molécule. Drug-cyclodextrin complexes are generally usefu, for most dosage forms andadministration routes. As an alternative to direct complexation with the drug theq cyclodextrin may be used as an auxiiiary additive, e.g. as a carrier, diluent orsolubiliser. Alpha-, beta- and gamma-cyciodextrins are most commonly usedand suitable examples are described in WO-A-91/11172, WO-A-94/02518 and WO-A-98/55148.

Generally, in humans, oral administration the compound is the preferred5 route, being the most convenient and, for example in MED, avoiding the well-known disadvantages associated with intracavernosal (i.c.) administration. Apreferred oral dosing regimen in MED for a typical man is from 25 to 250 mg ofcompound when required. In circumstances where the récipient suffers from aswailowing disorder or from impairment of drug absorption after oral >0 administration, the drug may be administered parenterally, sublingually orbuccally.

For veterinary use, the compound, or a veterinarily acceptable sait thereof,or a veterinarily acceptable solvaté or pro-drug thereof, is administered as asuitably acceptable formulation in accordance with normal veterinary practice >5 and the veterinary surgeon wilt détermine the dosing regimen and route ofadministration which will be most appropriate for a particular animal. 012337 12

Thus the invention provides a pharmaceutical composition comprising thecrystalline compound of the présent invention together with a pharmaceuticallyacceptable diluent or carrier.

It further provides a veterinary formulation comprising the crystallineg compound of the présent invention together with a veterinarily acceptable diluent or carrier.

The crystalline material as defined herein may be prepared according tothe préparations and Examples hereinafter.

Préparation of Starting Materials for Example 1 10 Préparation 1 A/-i3-Carbamovl-5-ethvl-1-(2-methoxvethvl)-1Z-/-pvrazol-4-vl}-2-ethoxv-5-(4-ethvl- 1-piperazinvl sulfonvl) nicotinamide. o

(a) 2-Hydroxv-5-sulfonicotinic acid 2-Hydroxynicotinic acid (27Kg, 194.2mol) was added portionwise to 30% oleum (58.1 Kg) at 50°C over 1hr. This caused an exotherm to 82°C. The réaction 15 012337 r 13 mixture was heated further to 140°C. After maintaining this température for 12hrsthe reactor contents were cooled to 15C and filtered. The fiiter cake was then re-siurried wîth acetone (33Kg) at room température, filtered and dried to afford thetitle compound (35.3Kg, 83%) as a white solid. Décomposition pt 273°C. δ 5 (DMSOde): 7.93 (1 H, d), 8.42 (1H, d). m/z (Found:220 [M+H]+, 100%. C6H6NO6Srequires 220.17). (b) Ethvl 2-hvdroxv-5-suifonicotinoate 2-Hydroxy-5-sulfonicotinic acid (500g, 2.28mol) was dissolved in ethanoi (2.5L)with stirring and heated to 80°C. After 30mins 0.5L of solvent was distiiled off, ,0 then repiaced with fresh éthanol (0.5L) and taken back to 80°C. After a further60mins 1.0L of solvent was distiiled off, then repiaced with fresh éthanol (1.0L)and taken back to 80°C. After a further 60mins 1 .OL of solvent was distiiled off,the reaction cooled to 22°C and stirred for 16hr. The precipitated product wasfiltered, washed with éthanol (0.5L) and dried at 50°C under vacuum to afford the U title compound (416g, 74%) as a white solid. Décomposition pt 237°C. δ(DMSOd6): 1-25 (3H, t), 4.19 (2H,q), 7.66 (1H, d), 8.13 (1H, d). m/z (Found:248[M+Hf, 100%. C8H10NO6S requires 248.22). (c) Ethvl 2-chloro-5-chlorosulfonicotinoate

Ethyi 2-hydroxy-5-sulfonicotioate (24.7g, 0.1 mol) was slurried in thionÿl chloride ,0 (238g, 2.0mol) and dimethylformamide (1 .OmL) with stirring. The reaction mixture was then heated to reflux for 2.5hr. The bulk of the thionyl chloride was removedunder vacuum with residual thionyl chloride removed with a toluene azeotrope toafford the crude title compound (30.7g, 108%) as a yellow oil. d (CDCI3): 1.46(3H, t), 4.50 (2H, q), 8.72 (1H, d), 9.09 (1H, d). This was taken directly onto the 2c next step. (d) Ethvl 2-chloro-5-(4-ethyl-1 -piperazinylsulfonvDnicotinoate 4 0/233^' 14

Crude ethyl 2-chloro-5-chlorosu,fonicotinoate (30.7g, 0.1 mol assumed) wasdissolved in ethyl acetate (150mL) with stirring then ice cooled. To this wasadded a solution of N-ethylpiperazine (11.4g, 0.1 mol) and triethyiamine (22.5g,0.22mol) in ethyl acetate (50mL), carefully over 30mins, keeping the internai 5 température below 10°C. Once the addition was complété the reaction wasallowed to warm to 22°C and stir for 1hr. The solid was filtered off and theremaining filtrate was concentrated under vacuum to afford the crude titlecompound (37.1g, 103%) as a crude yellow gum. § (CDCI3): 1.10 (3H, t), .1.42(3H, m), 2.50 (2H, m), 2.60 (4H, m), 3.19 (4H, m), 4.43 (2H, q), 8.40 (1H, d), 8.80 jq (1H, d). m/z (Found:362 [M+Hf, 100%. C14H21CIN3O4S requires 362.85). (e) Ethvl 2-ethoxv-5-(4-ethvl-1 -piperazinvlsulfonvhnicotinoate A solution of Ethyl 2-chloro-5-(4-ethyl-1-piperazinylsulfonyl)nicotinoate (36.1g,0.1 mol) in éthanol (180mL) was cooled to 10°C with stirring. Sodium ethoxide(10.2g, 0.15mol) was added portionwise keeping the température below 20°C.The reaction mixture was then stirred at ambient température for 18 hours. Theprecipitate was filtered off and water (180mL) added to the filtrate. The filtratewas then heated to 40°C for 1 hour. Ethanol (180mL) was then distilled off atambient pressure and the remaining aqueous solution allowed to cool to ambienttempérature. The precipitated product was then filtered off, washed with water 20 and dried under vacuo at 50°C to afford the title compound (12.6g, 34%) as alight brown solid. M.p. 66-68°C. δ (CDCI3): 1.04 (3H, t), 1.39 (3H, t), 1.45 (3H, t), 2.41 (2H, q), 2.52 (4H, m), 3.08 (4H, m), 4.38 (2H, q), 2.57 (2H, q), 8.38 (1H, d), 8.61 (1H, d). m/z (Found:372 [M+H]+, 100%. C16H26N3O5S requires 372.46). (f) 2-Ethoxv-5-(4-ethvl-1-piperaz?nvlsulfonyi)nicotinic acid 25 Ethyl 2-ethoxy-5-(4-ethyl-1-piperazinylsulfonyl)nicotinoate (10.2g, 0.0275mol)was dissolved in toiuene (50mL) and a solution of sodium hydroxide (1.1g,0.0275mol) in water (20rnL) added to it. This two phase mixture was then stirred 012337 15 vigorously at ambiant température overnight. The aqueous phase was separatedoff and adjusted to pH=5.6 by addition of c. hydrochloric acid. The precipitatedproduct was slurried with ice cooling for 15minutes, filtered, water washed anddried under vacuo at 50°C to afford the title compound (4.1g, 43%) as an off- 5 white solid. Mpt 206-207°C. δ <CDCl3): 1.25 (3H, t), 1.39 (3H, t), 2.82 (2H, q),3.03 (4H, m), 3.25 (4H, m), 4.50 (2H, q), 8.25 (1H, d), 8.56 (1 H, d). m/z(Found:344 [M+H]+, 100%. C14H22N3O5S requires 344.38). (g) Ethvi 3-ethvl-7H-pyrazole-5-carboxvlate

To a stirred solution of 2,2-dimethoxybutane (10 g, 84.7 mMol) in CH2Cl2 (50 mL) 10 under a nitrogen atmosphère at 0°C was added pyridine (13.7 mL, 169.5 mMol).The reaction mixture was maintained at 0°C and a solution of trichloroacetylchloride (18.9 mL, 169.5 mMol) in CH2CL2 (35 mL) was added over 1 hour withconstant stirring. The yellow-orange solution begins to precipitate a white solidas the reaction progresses. The reaction mixture is allowed to warm to room 15 température over 20 h. The reaction mixture was diluted with éthanol (150 mL)and re-gcoled to 0°C before treatment with hydrazine hydrate (8.2 mL, 169.5mMol) as a solution in éthanol (35 mL) over 30 min. The reaction was heated to50°C and solvent was distilled at atmospheric pressure. The température wasincreased until the head température reached 78°C. Reflux was maintained for a 20 further 2 h, before cooling to room température. The reaction mixture was dilutedwith water (250 mL) and éthanol was removed by évaporation at redûcedpressure. The résultant mixture was extracted with CH2CI2 (3 x 200 mL). The 012337 ., 16 combined organics were dried (MgSO4), filtered and evaporated at reducedpressure to afford the title compound as a brown oil, 12.05 g, 85%. 1H NMR (300 MHz, CDCfe): 5= 1.20 (3H, t), 1.28 (3H, t), 2.67 (2H, q), 4.29 (2H,q), 6.55(1 H, s), 12.56(1 H, s). 5 LRMS m/z= 167.1 [M-H]+, C8Hi2N2O2 requires 168.2. (h) Ethvl 3-ethvl- 7B-pyrazole-5-carboxvlic acid

Aqueous sodium hydroxide solution (10M; 100 ml, 1.0 mol) was added dropwiseto a stirred suspension of the title compound of Préparation 1 (g) (66.0 g, 0.39mol) in methanol and the resulting solution heated under reflux for 4 hours. Theto coo, reaction mixture was concentrated under reduced pressure to ca. 200 ml,diluted with water (200 ml) and this mixture washed with toluene (3 x 100 ml).

The resulting aqueous phase was acidified with concentrated hydrochloric acid topH 4 and the white precipitate collected and dried by suction to provide the titlecompound (34.1 g), δ (DMSOde): 1.13 (3H,t), 2.56 (2H,q), 6.42 (1H,s). 15 fi) 4-Nitro-3-n-propyl-1 H-pvrazole-5-carboxvlic acid

Fuming sulphuric acid (17.8 ml) was added dropwise to stirred, ice-cooled fumingnitric acid (16.0 ml), the resulting solution heated to 50°C, then 3-n-propyl-1 H-pyrazole-5-carboxylic acid (Chem. Pharm. Bull., 1984, 32, 1568; 16.4 g, 0.106mol) added portionwise over 30 minutes whilst maintaining the reaction 20 température below 60°C. The resulting solution was heated for 18 hours at60°C, allowed to cool, then poured onto ice. The white precipitate was collected, 'CT/ 012337 η washed with water and dried by suction to yield the title compound (15.4 g), m.p.170-172°C. Found: C, 42.35; H, 4.56; N, 21.07. C7H9N3O4 requires C, 42.21; H,4.55; N, 21.10%. δ (DMSOd6): 0.90 (3H,t), 1.64 (2H,m), 2.83 (2H,m), 14.00(1H,s). (i) 3-Ethvl-4-nitro-1 H-pyrazoie-5-carboxvlic acid

Obtained from the title compound of Préparation 1(h), by analogy with theprocess of Préparation 1(i), as a brown solid (64%). δ (DMSOde): 1.18 (3H,t), 2.84 (2H,m), 13.72(1 H, s). (k) 4-Nitro-3-n-propyi-1 H-pyrazole-5-carboxamide 0 A solution of the title compound of Préparation 1(i) (15.4 g, 0.077 moi) in thionylchloride (75 ml) was heated under reflux for 3 hours and then the cool reactionmixture evaporated under reduced pressure. The residue was azeotroped withtetrahydrofuran (2 x 50 ml) and subsequently suspended in tetrahydrofuran (50ml), then the stirred suspension ice-cooled and treated with gaseous ammoniafor 1 hour. Water (50 ml) was added and the resulting mixture evaporated underreduced pressure to give a solid which, after trituration with water and drying bysuction, furnished the title compound (14.3 g), m.p. 197-199°C. Found: C, 42.35; H, 5.07; N, 28.38. C7H10N4O3requires C, 42.42; H, 5.09; N, 28.27%. δ (DMSOds): 0.90 (3H,t), 1.68 (2H,m), 20 2.86 (2H,t), 7.68 (1 H,s), 8.00 (1 H,s). (l) 3-Ethvl-4-nitro-1 H-pyrazole-5-carboxamide 012337 18

Obtained from the title compound of Préparation 1 (j), by anaiogy with Préparation1(k), as a white soiid (90%). δ (DMSOd6): 1.17 (3H,t), 2.87 (2H,m), 7.40 (1H,s), 7.60 (1H,s), 7.90 (1H,s).LRMS: m/z 185 (M+1)+. 5-Ethvl-1 -f2-methoxvethvl)-4-nitro-1 H-pyrazole-3-carboxamide.

A mixture of 3-ethyl~4-nitro-fH-pyrazole-5-carboxamide (2.5 kg, 13.6 Mol),sodium carbonate (1.8 Kg, 17.0 Mo,) and 2-bromoethyl methyl ether (1.98 kg, 14.2 Mol) in THF (22.5 L) and water (2.5 L) was heated under reflux and stirredfor 20 hours. The mixture was cooied to ambient température and CH2CI2 (67.5L) and water (22.5 L) were added. The résultant organic and aqueous iayerswere separated. The aqueous phase was extracted with CH2CI2 (22.5 L) and thecombined organic solution was distilled under atmospheric pressure andreplaced with ethyl acetate (33 L) to a final volume of 17 L. The cooied mixturewas granulated at ambient température for 2 hours, fiitered and washed withethy, acetate (2.5 L). This afforded 5-ethy,-1-(2-methoxyethyl)-4-nitro-1H-pyrazole-3-carboxarnide as a white crystalline solid, 2.10 kg, 57%. m.p. = 140°C. 012337 · 19

Found: C, 44.46; H, 5.79; N, 23.01. C9H14N4O4 requires C, 44.63; H, 5.79; N,23.14%. 5{CDCl3): 1.18 (3H, t), 2.98 (2H, q), 3.22 (3H, s), 3.77 (2H, t), 4.28 (2H, q), 6.03{1 H, s), 7.36 (1 H, s). 5 LRMS: m/z = 243 (M+1)+ (m)(ii) 5-Ethv1-1 -(2-methoxvethvl)-4-nitro-1 H-pvrazole-3-carboxamide. A mixture of 3-ethyl-4-nitro-1H-pyrazole-5-carboxamide (25 g, 0.136 Mol),sodium carbonate (18 g, 0.17 Moi) and sodium iodide (20.4 g, 0.136 Moi) weresuspended in ethyl methyl ketone (125 mL) at room température. 2-bromoethyl iO methyl ether (12.8 mL, 0.142 Mol) was added and the mixture was heated toreflux and stirred for 70 hours. The mixture was cooied to ambient températureand water (250 mL) was added. The résultant slurry was warmed to reflux andheld at that température for 30 min before cooling to room température. Therésultant precipitate was granulated at room température for 3 h, filtered and •J5 vacuum dried to afford 5-ethyl-1-(2~methoxyethyl)-4~nitro-1H-pyrazoIe-3-carboxamide as a yeliow crystalline solid 24.3 g, 74%. Data as reported forPréparation 1(m)(i). (ni 4-Amino-5-ethvl-1 -(2-methoxvethvl1-1 H-pyrazole-3-carboxamide,

A mixture of 5-ethyl-1-(2-methoxyethyl)-4-nîtro-1H-pyrazole-3-carboxamide (20 g,20 82.6 mMol) and 5%Pd/C (1 g) in methanol (200 mL) was pressurised at 012337 20 50psi/25°C in a sealed vessel and stirred for 15 hours. At the end of the reactionthe mixture was filtered through arbocel and the filter cake was washed withmethanol. The methanolic solution was distilied at atmospheric pressure andreplaced with ethyl acetate to a final volume of 100 mL. The cooled mixture was 5 granulated at ambient température for 2 h filtered and washed with ethyl acetate(20 mL) to afford 4-amino-5-ethyl-1-(2-methoxyethyl)-1 W-pyrazole-3-carboxamideas a white crystalline solid, 15 g, 88%. m.p. = 131°C. Found: C, 50.75; H, 7.62;N, 26.38. C9H16N4O2 requires C, 50.94; H, 7.55; N, 26.42%. ô(CDCl3): 1.20 (3H, t), 2.63 (2H, q), 3.32 (3H, s), 3.74 (2H, t), 3.95 (2H, s), 4.15w (2H, t), 5.27 (1 H, s), 6.59 (1 H, s). LRMS: m/z = 213 (M+1)+ (o) /V-f3-Carbamovl~5-ethvl-1 -(2-methoxvethv0-1 H-pyrazol-4-vh-2-ethoxv-5-(4- ethvl-1-piperazinvl sulfonvl) nicotinamide.

route (a) 2-ethoxy-5-(4-ethyl-1-piperazinylsulfonyl)nicotinic acid (2.31 kg, 6.73 Mol) wassuspended in ethyl acetate (16.2 L) and 1,1-carbonyldimidazole (1.09 kg, 6.73 15 012337 21

Mol) was added at room température. The reaction mixture was heated at 45°Cfor 40 minutes and then the reaction was stirred for a further 40 minutes at reflux.After cooling to ambient température 4-amino-5-ethyl-1-(2-rnethoxyethyl)-1/7-pyrazole-3-carboxamide (1.5 kg, 7.06 Mol) was added to the cooled mixture, andthe reaction stirred for a further 15 hours under reflux. The mixture was cooledfiltered and the filter cake was washed with 90% water Z 10% ethyl acetate, (2 mL/g) to afford /V-[3-carbamoyl-5-ethyl-1-(2-methoxyethyl)-1H-pyrazol-4-yl}-2-ethoxy-5-(4-ethyl-1-piperazinyl sulfonyl) nicotinamide as an off white crystallinesolid, 3.16 kg, 88%. m.p. = 156°C. Found: C, 51.33; H, 6.56; N, 18.36.C23H35N7O6S requires C, 51.40; H, 6.53; N, 18.25%. Ô(CDCI3): 1.04 (3H, t), 1.22 (3H, t), 1.60 (3H, t), 2.44 (2H, q), 2.54 (4H, m), 2.96(2H, q), 3.12 (4H, m), 3.36 (3K, s), 3.81 (2H, t), 4.27 (2H, t), 4.80(2H, q),5.35(1 H, s), 6.68 (1H, s), 8.66 (1H, d), 8.86 (1H, d), 10.51 (1 H, s). LRMS: m/z = 539 (M+1)+ route (b) 2-ethoxy-5-(4-ethyl-1-pipèrazinylsulfonyl)nicotinic acid (27.5g, 0.08 Mol) wassuspended in ethy, acetate (0.193 L) and 1,1-carbonyldimidazole (13.3g, 0.082Mol) was added at room température. The reaction mixture was heated at 45°Cfor 15 minutes and then the reaction was stirred for a further 60 minutes at reflux.After cooling to ambient température 4-amino-5-ethyl-1-(2-methoxyethyl)-1H-pyrazole-3-carboxarnide (16.5 g, 0.078 Mol) was added to the cooled mixture,and the reaction stirred for a further 17 hours under reflux. The solution wascooled and water (0.07 L) and ethyl acetate (0.17 L) added. The slurry waswarmed to 40°C and the organic separated. The organic was concentrated to byevaporating 0.23 L solvent. The slurry was cooled, granulated at 0- -10°C filteredand the filter cake was washed with 90% water /10% ethyl acetate, (2 mL Zg) toafford /V-[3-carbamoyl-5-ethyl-1 -(2-methoxyethyl)-1 H-pyrazol-4-yl}-2-ethoxy-5-{4- 012337 22 ethyl-1 -piperazinyl sulfonyl) nicotinamide as an off white crystalline solid, 38.7g,89%. m.p. = 159-161°C. ô(CDCI3): 1.04 (3H, t), 1.22 (3H, t), 1.61 (3H, t), 2.42 (2H, q), 2.54 (4H, m), 2.96(2H, q), 3.12 (4H, m), 3.36 (3H, s), 3.81 (2H, t), 4.27 (2H, t), 4.78(2H, q), 5 5.35(1 H, s), 6.68 (1 H, s), 8.66 (1 H, d), 8.86 (1 H, d), 10.51 (1 H, s). LRMS: m/z =538 (M+1)+

Préparation of Startinq Materials for Example 2 a) Pvridine-2-amino-5-sulphonic acid

o=s=o

OH 2-Aminopyridine (80g, 0.85mol) was added portionwise over 30 minutes to oleum10 (320g) and the resulting solution heated at 140°C for 4 hours. On cooiing, the reaction was poured onto ice (200g) and the mixture stirred in an ice/salt bath fora further 2 hours. The resulting suspension was fiitered, the solid washed with icewater (200ml) and cold IMS (200ml) and dried under suction to afford the titie compound as a solid, 111.3g; LRMS : m/z 175 (M+1)+. 15 b) Pvridine-2-amino-3-bromo-5-sulphonicacid HH,

Br Ο=ψ=Ο

OH

Bromine (99g, 0.62mol) was added dropwise over an hour, to a hot solution ofthe product from stage a) (108g, 0.62mol) in water (600ml) so as to maintain asteady reflux. Once the addition was complété the reaction was cooled and theresulting mixture fiitered. The solid was washed with water and dried under 012337 23 suction to afford the title compound, 53.4g; δ (DMSOd6> 300MHz): 8.08 (1H, s), 8.14 (1H, s); LRMS : m/z 253 (M)+. c) Pvridine-3-bromo-2-chloro-5-sulphonvl chlorideci

Br o=s=o

I

CI A solution of sodium nitrite (7.6g, HO.Ommol) in water (30ml) was added

5 dropwise to an ice-cooled solution of the product from stage b) (25.3g,lOO.Ommol) in aqueous hydrochloric acid (115ml, 20%), so as to maintain thetempérature below 6°C. The reaction was stirred for 30 minutes at 0°C and for afurther hour at room température. The reaction mixture was evaporated underreduced pressure and the residue dried under vacuum at 70°C for 72 hours. A <10 mixture of this solid, phosphorus pentachloride (30.0g, 144mmol) andphosphorus oxychloride (1ml, 10.8mmol) was heated at 125°C for 3 hours, andthen cooied. The reaction mixture was poured onto ice (100g) and the resultingsolid fiitered, and washed with water. The product was dissolved indichloromethane, dried (MgSO4), and evaporated under reduced pressure to ^5 afford the title compound as a yellow solid, 26.58g; δ (CDCI3, 300MHz) : 8.46(1H, s), 8.92 (1H, s). d) 3-Bromo-2“Chloro-5-(4-ethvlpiperazin-1-vlsulphonvl)pvridine

Cl N, A solution of 1-ethylpiperazine (11.3ml, 89.0mmol) and triethylamine (12.5ml,89.0mmol) in dichloromethane (150ml) was added dropwise to an ice-cooled 012337 < 24 solution of the product from stage c) (23.0g, 79.0mmol) in dichloromethane(150ml) and the reaction stirred at 0°C for an hour. The reaction mixture wasconcentrated under reduced pressure and the residual brown oil was purified bycolumn chromatography on silica gel, using an elution gradient of 5 dichloromethane:methanol (99:1 to 97:3) to afford the title compound as anorange solid, 14.5g; δ (CDCU, 300MHz) : 1.05 (3H, t), 2.42 (2H, q), 2.55 (4H, m),3.12 (4H, m), 8.24 (1 H, s), 8.67 (1 H, s). e) 3-Bromo-2-ethoxv-5-f4-ethvlpiperazin-1“Visulphonvl)pvridine

A mixture of the product from stage d) (6.60g, 17.9mmol) and sodium ethoxideΊΟ (6.09g, 89.55mmol) in éthanol (100ml) was heated under reflux for 18 hours,then cooled. The reaction mixture was concentrated under reduced pressure, theresidue partitioned between water (100ml) and ethyl acetate (100ml), and thelayers separated. The aqueous phase was extracted with ethyl acetate (2x100ml), the combined organic solutions dried (MgSO4) and evaporated underi5 reduced pressure to afford the title compound as a brown solid, 6.41g; Found : C, 41.27; H, 5.33; N, 11.11. C^HgoBrNsOsS requires C, 41.35; H, 5.28; N,10.99%; δ (CDCIs, 300MHz) : 1.06 (3H, t), 1.48 (3H, t), 2.42 (2H, q), 2.56 (4H, m),3.09 (4H, m), 4.54 (2H, q), 8.10 (1H, s), 8.46 (1H, s); LRMS : m/z 378, 380(M+1)+. 20 f) Pvridine 2“ethoxv-5-(4-ethvlpiperazin-1-vlsulphonvh-3-carboxvlic acid ethylester 012337

A mixture of the product from stage e) (6.40g, 16.92mmol), triethylamine (12ml,86.1 mmol), and palladium (0) tris(triphenylphosphine) in éthanol (60ml) washeated at 100°C and 200 psi, under a carbon monoxide atmosphère, for 18hours, then cooled. The reaction mixture was evaporated under reducedpressure and the residue purified by column chromatography on silica gel, usingan elution gradient of dichloromethane:methanol (100:0 to 97:3) to afford the titlecompound as an orange oil, 6.2g; δ (CDCI3, 300MHz) : 1.02 (3K, t), 1.39 (3H, t), 1.45 (3H, t), 2.40 (2H, q), 2.54 (4H, m), 3.08 (4H, m), 4.38 (2H, q), 4.55 (2H, q),8.37 (1H, s), 8.62 (1 H, s); LRMS : m/z372 (M+1)+· 10 g)

Pvridine 2-ethoxv-5~(4-ethylpiperazin-1 -ylsulphonvn-3-carboxylic acid

A mixture of the product from stage f) (4.96g, 13.35mmol) and aqueous sodiumhydroxide solution (25ml, 2N, 50.0mmol) in éthanol (25ml) was stirred at roomtempérature for 2 hours. The réaction mixture was concentrated under reducedpressure to half it’s volume, washed with ether and acidified to pH 5 using 4N 012337 26 hydrochloric acid. The aqueous solution was extracted with dichloromethane(3x30ml), the combined organic extracts dried (MgSO4) and evaporated underreduced pressure to afford the title compound as a tan coloured solid, 4.02g; δ(DMSOde, 300MHz) ; 1.18 (3H, t), 1.37 (3H, t), 3.08 (2H, q), 3.17-3.35 (8H, m), 5 4.52 (2H, q), 8.30 (1 H, s), 8.70 (1 H, s). h) 4-f2-Ethoxv-5-(4-ethvlpiperazin-1-vlsulphonvl)pyridin-3-vlcarboxamidol· 1H-3-ethvlpyrazole-5-carboxamide

A solution of 4-amino-3-ethyl-1H-pyrazole-5-carboxamide (WO 9849166,préparation 8) (9.2g, 59.8mmol) in Ν,Ν-dimethylformamide (60ml) was added to 10 a solution of the product from stage g) (21.7g, 62.9mmol), 1- hydroxybenzotriazole hydrate (10.1g, 66.0mmol) and triethylamine (13.15ml,94.3mmol) in dichloromethane (240ml). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (13.26g, 69.2mmol) was added and the reactionstirred at room température for 6 hours. The dichloromethane was removed 15 under reduced pressure, the remaining solution poured into ethyl acetate(400ml), and this mixture washed with aqueous sodium bicarbonate solution(400ml). The resulting crystalline precipitate was filtered, washed with ethylacetate and dried under vacuum, to afford the title compound, as a white powder,22g; δ (CDCI3+1 drop DMSOd6) 0.96 (3H, t), 1.18 (3H, t), 1.50 (3H, t), 2.25-2.56 20 (®H, m), 2.84 (2H, q), 3.00 (4H, m), 4.70 (2H, q), 5.60 (1H, br s), 6.78 (1H, br s), 8.56 (1H, d), 8.76 (1H, d), 10.59 (1H, s), 12.10-12.30 (1H, s); LRMS: m/z 480(M+1)+. 27 012337 i) 2-Methoxyethvl-442-Othoxv-5-(4-ethylpiperazin-1-v!sulphonvl)pvridin-3- vlcarboxamido1-3-ethvlpvrazole-5-carboxamide

2-ethoxy-5-(4-ethyl-1-piperazinylsulfonyl)nicotinic acid (2.31 kg, 6.73 Mol) wassuspended in ethyl acetate (16.2 L) and 1,1-carbonyldimidazole (1.09 kg, 6.73 5 Mol) was added at room température. The reaction mixture was heated at 45°Cfor 40 minutes and then the reaction was stirred for a further 40 minutes at reflux.After cooling to ambient température 4-amino-5-ethyI-1-(2-methoxyethyl)-1H-pyrazole-3-carboxamide (1.5 kg, 7.06 Mol) was added to the cooled mixture, andthe reaction stirred for a further 15 hours under reflux. The mixture was cooled ΊΟ filtered and the filter cake was washed with 90% water /10% ethyl acetate, (2 mL/g) to afford /V-[3-carbamoyl-5-ethyl-1-(2-methoxyethyl)-1 /7-pyrazol-4-yl}-2-ethoxy-5-(4-ethyl-1-piperazinyl sulfonyl) nicotinamide as an off white crystallinesolid, 3.16 kg, 88%. m.p. = 156°C. Found: C, 51.33; H, 6.56; N, 18.36.CgaHasNyOeS requires C, 51.40; H, 6.53; N, 18.25%. 15 Ô(CDCI3): 1.04 (3H, t), 1.22 (3H, t), 1.60 (3H, t), 2.44 (2H, q), 2.54 (4H, m), 2.96(2H, q), 3.12 (4H, m), 3.36 (3H, s), 3.81 (2H, t), 4.27 (2H, t), 4.80(2H, q),5.35(1 H, s), 6.68 (1H, s), 8.66 (1H, d), 8.86 (1H, d), 10.51 (1H, s). LRMS: m/z = 539 (M+1)+ 012337 28 f

Préparation 3 1-(6-Ethoxv-5-r3-ethvn-6.7-dihvdro-2-(2-methoxvethvl)-7-oxo-2H-pyrazolef4,3- cflpvrimidin-5“Vn-3-Pvridvlsulfonvl)-4-ethvlpiperazine«ethvl acetate solvaté.

route (a) 5 To préparé the compound of Préparation 3 a mixture of the title compound ofPréparation 1, W'[3-carbamoyl-5-ethyl-1-(2-methoxyethyl)-1H-pyrazol-4-yl)-2-ethoxy-5-(4-ethyl-1-piperazinyl sulfonyi) nicotinamide, (1.18 kg, 2.2 Mol),potassium ferf-butoxide (500 g, 4.4 moles) and ethyi acetate (193 g) in éthanol(11.8 L) was heated at 120°C for 20 hours. The reaction mixture was theni0 concentrated under reduced pressure, in total approx. 10 L of solvent weredistilled. To the residue water (2.9 L) was added and the mixture stirred at roomtempérature while aqueous HCl was added until pH 7.5 was obtained. Ethyiacetate (7.5 L) was added and the two phase mixture was warmed to 55°C. Theorganic phase was separated and the aqueous phase was extracted with further 15 ethyi acetate (3.0 L). The combined organic phases were distilled at atmosphericpressure to a final volume of 4L. The precipitated solids were granulated at 5°Cfor 1 h, filtered and washed with ethyi acetate (1.2 L) and dried under vacuum.This afforded 1 -(6-Ethoxy-5-[3-ethyl]-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazole[4,3-d}pyrimidin-5-yl]-3-pyridylsulfonyl)-4-ethylpiperazine as a light yellow 012337 U.··* « 29 crystalline solid, 877 g, 78%. m.p. = 157°C. Found: C, 52.65; H, 6.46; N, 17.76.C23H33N7O5S. 0.2 C2H5CO2CH3 requires C, 53.21; H, 6.49; N, 18.25%. Ô(CDCI3): 1.07 (3H, t), 1.42 (3H, t), 1.61 (3H, t), 2.44 (2H, q), 2.57 (4H, m), 3.08(2H, q), 3.15 (4H, m), 3.32 (3H, s), 3.92 (2H, q), 4.48 (2H, q), 4.77 (2H, q), 8.65(1 H, d), 9.06 (1 H, d). The spectrum also has signais that correspond to a solvatéwith ethyl acetate. LRMS: m/z = 520 (M+1)+ route (b)

To préparé the compound of Préparation 3 a mixture of the title compound of 10 Préparation 1, A/-[3-carbamoyl-5~ethyl-1-(2-methoxyethyl)-1H-pyrazol-4-yl}-2-ethoxy-5-(4-ethyl-1-piperazinyl sulfonyl) nicotinamide, 34.6g, 0.064 Mol),potassium ferf-butoxide (14.8 g, 0.132 moles) and ethyl acetate (8.5g 0.096moles) in éthanol (0.26 L) was heated at 105-107°C for 17 hours. The reactionmixture was then concentrated under reduced pressure, in total approx. 0.16 L of 15 solvent were distilled. To the remaining oil, water (0.28 L) was added and themixture stirred at room température whiie aqueous KCI was added until pH 7.5was obtained. The precipitated soiids were granulated at -10°C to 0°C for 1 h,filtered and washed with éthanol (0.4 L) and dried under vacuum. This afforded1-(6-Ethoxy-5-[3-ethyl3-6,7-dihydro-2"(2-methoxyethyl)-7-oxo-2H-pyrazole[4,3- 20 dJpyrimidin-5-yl]-3"pyridyisulfonyl)-4-ethylpiperazine as a light yellow crystallinesolid, 23.3 g, 69.7%. m.p. = 168-170°C. Ô(CDCI3): 1.07 (3H, t), 1.42 (3H, t), 1.61 (3H, t), 2.44 (2H, q), 2.57 (4H, m), 3.08(2H, q), 3.15 (4H, m), 3.32 (3H, s), 3.92 (2H, q), 4.48 (2H, q), 4.77 (2H, q), 8.64(1H, d), 9.05(1 H, d). 072337 t 30 LRMS: m/z = 520 (M+1)+

Example 1 1-{6-ethoxv-5-F3-ethvl-6.7-dÎhvdro-2-(2-methoxvethvl)-7-oxo-2H-pyrazolof4,3- dlpvrimidin-5-vn-3-pvridylsulfonvl)-4-ethvlpiperazine

10g (0.019 moi) of the title compound of Préparation 3(a), 1-{6-ethoxy-5-[3-ethyl- 6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d3pyrimidin-5-yl]-3- pyridylsulfonyl}-4-ethylpiperazine ethyl acetat© solvaté, was charged followed by12ml/g (120mls) of 16% water in ethyl alcohoi. The slurry was heated to reflux toyield a solution and 6ml/g (60mls) distilled off at atmospheric pressure. Thesolution was then cooled to room température with crystallisation occurring at40°C. The slurry was then cooled to 5-10°C and granulated for 30 minutesfollowing which it was filtered and washed with 2ml/g ethyl alcohoi (20 mis). Thedamp solid was dried in vacuo overnight at 55-60 °C to yield a white crystallinesolid. (Yield 7.6g, 76%).Melting Point 162-165°C. δ (CDG,g): 1.05 (3H,t), 1.42 (3H,t), 1.58 (3H,t), 2.43 (2H,q), 2.57 (4H,t), 3.09 (2H, t), 3.15 (4H,t), 3.30 (3H,s), 3.93 (2H,t), 4.48 (2H,t), 4.90 (2H,q), 8.65 (1K,d), 9.05 (1H,d), 10.65 (1H,s). 15 0 72337. 31

Example 2 146-ethoxv-5-T3-ethvl-6,7-dihvdro-2-(2-methoxvethvl)-7-oxo-2H-pvrazolo[4.3- dlpvrimidin-5-vll-3-pyridvlsulfonvlM-ethvlpiperazine

10g (0.019 mol) of the title compound of Préparation 3(b), 1-{6-ethoxy-5-[3-ethyl- 5 6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyI}-4-ethylpiperazine ethyl acetate solvaté, was charged followed by12m!/g (120mls) of 16% water in ethyl alcohol. The slurry was heated to reflux toyield a solution and 9.5 ml/g (60mls) distilled off at atmospheric pressure. Thesolution was then cooled to room température with crystallisation occurring at 10 40°C. The slurry was then cooled to 0- -10°C and granulated for 30 minutes following which it was filtered and washed with 2ml/g ethy, alcohol (20 mis). Thedamp solid was dried in vacuo overnight at 55-60 °C to yield a white crystallinesolid. (Yield 9.4g, 94 %). Melting Point 158-160°C. δ (CDCI3): 1.05 (3H,t), 1.42 (3H,t), 1.58 (3H,t), 2.43 (2H,q), 2.57 (4H,t), 3.09 (2H, 15 t), 3.15 (4H,t), 3.30 (3H,s), 3.93 (2H,t), 4.48 (2H,t), 4.90 (2H,q), 8.65 (1H,d), 9.05 (1 H,d), 10.65 (1H,s). 012337 ' 32

In the processes of the Examples water and pharmaceutically acceptablealcohols such as methanol, éthanol, propanol preferably éthanol or rso-propanoland mixtures thereof can be used. These processes can also be used for therecrystallisation of the title compound.

As will be appreciated by the skilled chemist solvatés may be prepared by avariety of standard techniques as are known in the art.

Further solvatés of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yl3-3-pyridylsulfonyl}-4-ethylpiperazine can beprepared from either the ethyl acetate solvaté (the compound of Préparation 3) orfrom the compounds of Examples 1 or 2. Such further solvatés may be preparedfor example by: (a) slurrying the starting compound over an extended period (such as forexample about 2 weeks) with the solvent to be tested (e.g. acetonitrile),followed by filtration of solid material and subséquent PXRD analysis (asdefined hereinbefore) of said “wet” i.e. non-dried solid material (solvated)material; or (b) dissolution of the starting compound in the solvent to be tested (e.g.acetonitrile) followed by recrystallisation of the desired solvaté from saidsolution by any appropriate means such as for example, cooling orévaporation techniques and subséquent analysis (using standard PXRDtechniques or infra red analysis as are known in the art) of said “wet” i.e. non-dried solid (solvated) material.

The crystalline compound as defined hereinbefore can be generated fromsolvatés thereof via de-solvation using techniques as are standard in the art suchas for example via heat treatment (at preferably from about 100°C up to about150°C) or using vacuum techniques (from ambient température and above) orvia slurrying in a non-solvating solvent (e.g. as iilustrated in Examples 1 and 2). 012337 33

The skilled chemist may utilise alternative drying techniques to generate thecrystalline compound as defined hereinbefore.

Further solvatés of 1-{6-©thoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yi]-3-pyridylsulfonyl)>4-ethylpiperazine which hâvebeen prepared include : n-butanol, methylethylketone (MEK), acetonitrile,tetrahydrofuran, toluene. The presence of soivated material can be qualified viaPXRD.

For example an acetonitrile solvaté of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- dlpyrimidin-5-yll-3-pyridylsulfonyl}-4-ethylpiperazine was prepared by firstly dissolving the material in hot acetonitrilefollowed by slow cooling and subséquent granulation at ambient températureovernight. The solution was then filtered under vacuum and the résultant solidmaterial (the solvaté) allowed to dry at ambient température and pressure. Thepresence of the solvaté was detected using PXRD. An alternative method fordétection of the solvaté .is by monitoring the weight loss by TGA.. The solvatéfrom acetonitrile is présent as the hemisolvate although other solvatés may bepossible.

These soivated materials hâve been demonstrated to generate the amorphousfree base of 1~{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazolo[4,3- d]pyrimidin-5-yl]-3-pyridylsulfonyl}-4-ethylpiperazine when dried atabout 150°C as illustrated by Differential Scanning Calorimetry DSC).

The DSC experiments herein were performed using a (measured using a PerkinElmer DSC7/TGA7 at a heating rate of 20°C/minute). 012337 34

Figure 1 hereinafter illustrâtes the DSC trace of: the crystalline material havingPXRD pattern as illustrated in Table 1 (the compound of Example 1); the DSCtrace of the ethyl acetate solvaté (the compound of Préparation 3); the DSC traceof the acetonitrile solvaté (of the compound of Example 1, wherein said solvatéwas prepared as detailed hereinbefore).

Claims (2)

1. 012337 35 CLAIMS A polymorph of 1-{6-ethoxy-5-[3-ethyl-6,7-dihydro-2-(2-methoxyethyl)-7-oxo-2H-pyrazoio[4,3- d]pyrimidin-5-yl]-3-pyridylsulfony!}-4-ethylpiperazinehaving a simulated powder X-ray diffraction pattern with main peakssubstantially as defined below: d-spacing &amp; intensity i%î J d-spacing LA). intensity d-spacing (Â) Intensity (%) d-spacing (Â) intensity (%) 12.2486 1.24 4.2323 _ 19.04 3.061 9.18 2.4041 IA- 1 ___J 2.24 9.7301 100 4.1595 1.19 3.0483 2.22 2.3424 1.22 8.5596 36.31 4.0421 1.35 3.0428 1.94 2.299 1.13 7.3923 15.02 3.9186 7.81 2.9929 1.59 2.264 1.23 7.3856 8.35 3.8951 10.26 2.9804 2.1 2.2605 1.29 7.3284 1.69 3.8491 5.34 2.9403 3.46 2.2079 2.65 5.8222 14.49 3.8485 2.46 2.9073 1.74 2.1855 1.9 5.7617 1.96 3.8427 43.14 2.8719 1.68 2.1679 1.92 5.7461 43.25 '3.7357 3.1 2.8532 4.7
2. 2.Ï468 1.35 5.1166 7.17 3.6961 3.99 2.8122 11.19 2.1119 4.52 5.1078 32.71 3.6805 4.51 2.8097 2.82 2.0769 1.15 4.8807 17.47 3.6042 5.28 2.7776 2.17 2.0471 1.12 4.8674 10.85 3.5988 ïï.09 2.7774 1.46 2.028 ï.63 4.865 10.98 3.5819 6.8 2.7162 1.76 1.9935 1.03 4.8425 13.19 3.5721 8.55 2.7041 3.63 1.9883 2.96 4.8018 3.32 3.5355 2.47 2.6875 1.42 ï.9181 1.27 4.7632 10.57 3.407 1.2 2.6443 2.19 ï .9058 1.03 4.7491 4.16 3.3461 4.91 2.6084 3.75 1.884 1.23 4.641 1.96 3.3318 7.54 2.6025 1.6 1.8797 2.49 4.629 19.73 3.3154 4.36 2.5809 1.31 ï.8749 2.81 4.3314 9.53 3.2827 1.7 2.5448 1.21 1.7939 Î.57 4.3301 3.65 3.2774 1.22 2.5438 3.26 1.7617 1.24 4.3066 26.66 3.2434 19.95 2.4325 2.47 1.7379 1.82 4.2909 51.91 3.224 2.11 2.4212 1.24 1.7194 1.03 4.2798 24.75 3.0663 1.38 2.4163 4.39 wherein said PXRD pattern is generated using copper K-aipha1 X-rays having awaveiength of 1.54178 Angstroms.