WO2006059165A1 - Pyrrolopyridine-2-carboxylic acid amide derivative useful as inhibitor of glycogen phosphorylase - Google Patents

Abstract

The present invention is directed to a novel form of a pyrrolopyridine-2-carboxylic acid amide of formula (I), which is an inhibitor of glycogen phosphorylase, compositions containing it and their use in therapy. The invention is also directed to processes for production of the novel form, novel intermediates used in said processes and processes for the production of said intermediates.

Description

PYRROLOPYRIDINE-2-CARBOXYLIC ACID AMIDE DERIVATIVE USEFUL AS INHIBITOR OF GLYCOG EN PHOSPHORYLASE

BACKGROUND OF THE INVENTION

The present invention is directed to a novel form of a pyrrolopyridine-2-carboxylic acid amide, which is an inhibitor of glycogen phosphorylase, compositions containing it and their use in therapy. The invention is also directed to processes for production of the novel form, novel intermediates used in said processes and processes for the production of said intermediates. Inhibitors of glycogen phosphorylase are useful in the treatment of diabetes, hyperglycemia, hypercholesterolemia, hyperinsulinemia, hyperlipidemia, atherosclerosis or myocardial ischemia.

International Patent Application No. PCT/US2004/016243 (published after the priority date of the present invention) discloses pyrrolopyridine-2-carboxylic acid amide inhibitors of glycogen phosphorylase, including the compound 5-chloro-lH-pyrrolo[2,3-c]pyridine-2-carboxylic acid [1-(S)- (4-fluorobenzyl)-2-(4-hydroxypiperidin- 1 -yl)-2-oxoethyl] amide.

PCT/US2004/016243 describes the synthesis of 5-chloro-lH-pyrrolo[2,3-c]pyridine-2- carboxylic acid [l-(iS)-(4-fluorobenzyl)-2-(4-hydroxypiperidin-l-yl)-2-oxoethyl]amide as the free base by amide coupling of either 2-(S)-[(5-chloro-lH-pyrrolo[2,3-c]pyridine-2-carbonyl)amino]-3-(4- fluorophenyl)propionic acid and 4-hydroxypiperidine or 5-chloro-lH-pyrrolo[2,3-c]pyridine-2- carboxylic acid and 2-(S)-amino-3-(4-fluorophenyl)- 1 -(4-hydroxypiperidin- 1 -yl)propan- 1 -one hydrochloride in dimethylformamide in the presence of diisopropylethylamine.

It is desirable to obtain novel forms of this compound having advantageous pharmacological properties and to provide improved processes for the production of such compounds.

SUMMARY OF THE INVENTION

5-Chloro- lH-pyrrolo[2,3-c]pyridine-2-carboxylic acid [ 1 -(S)-(4-fluorobenzyl)-2-(4- hydroxypiperidin-l-yl)-2-oxoethyl]amide hydrochloride, which is an inhibitor of glycogen phosphorylase, is useful in the prophylactic or therapeutic treatment of diabetes, hyperglycemia, hypercholesterolemia, hyperinsulinemia, hyperlipidemia, hypertension, atherosclerosis or tissue ischemia e.g. myocardial ischemia, and as a cardioprotectant. There are also provided processes for its production.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound which is a hydrochloride salt of 5-chloro-lH- pyrrolo[2,3-c]pyridine-2-carboxylic acid [ 1 -(S)-(4-fluoroberizyl)-2-(4-hydroxypiperidin- 1 -yl)-2- oxoethyl] amide.

In the following, 5-chloro-lH-pyrrolo[2,3-c]pyridine-2-carboxylic acid [l-(S)-(4- fluorobenzyl)-2-(4-hydroxypiperidin-l-yl)-2-oxoethyl] amide may be referred to as the compound of Formula (I):

(I) and accordingly the compound of the invention is a hydrochloride of the compound of Formula (I).

The hydrochloride of the compound of Formula (I) is preferably in crystalline form. The hydrochloride of the compound of Formula (I) may also be amorphous.

The invention also embraces solvates of the hydrochloride of the compound of Formula (I). Solvates include complexes of the hydrochloride of the compound of Formula (I) with physiologically acceptable solvents, and refers in particular to hydrates.

The hydrochloride of the compound of Formula (I) may contain up to about 15% w/w of water. When the hydrochloride of the compound of Formula (I) is a hydrate it preferably comprises from 5 to 15% w/w, e.g. 10 to 15% w/w, of water.

The hydrochloride of the compound of Formula (I) may be an anhydrate. The polymorphic form of the hydrochloride of the compound of Formula (I) may be identified by its characteristic X-ray powder diffraction pattern. A form of the hydrochloride is shown in Figures 2-5 and exhibits characteristic peaks in the

X-ray diffraction pattern. A further form of the hydrochloride is shown in Figure 6 and also exhibits characteristic peaks in the X-ray diffraction pattern. A characteristic peak is one which has a relative intensity above 10% of the largest peak in the powdered XRD pattern. Any one of these peaks alone, or in combination, may be used to identify the form of the hydrochloride. In addition to these characteristic peaks, Figures 2-5 and Figure 6 show other minor peaks not reaching the 10% threshold in all samples. The intensity of these peaks vary with the particular orientation of the polymorph. These additional peaks may be used to confirm the presence of these forms of the hydrochloride, but their absence should not be used to determine that the particular material is not the hydrochloride. As is readily apparent to one skilled in the art, the results of any X-ray powder diffraction may vary. This variance can be due to test sample preparation, the particular model of X-ray diffractometer used, the operator's technique, etc. The term "approximately" if used in defining a position of a characteristic peak in an X-ray powder diffraction pattern is defined as the stated 2Θ value ± 0.2 °2Θ. The invention also provides a pharmaceutical composition comprising a hydrochloride of the compound of Formula (I), in admixture with a pharmaceutically acceptable carrier.

The invention also provides a pharmaceutical composition for the treatment of disease by inhibiting glycogen phosphorylase, resulting in the prophylactic or therapeutic treatment of diabetes, hyperglycemia, hypercholesterolemia, hyperinsulinemia, hyperlipidemia, hypertension, atherosclerosis or tissue ischemia e.g. myocardial ischemia comprising a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of a hydrochloride of the compound of Formula (I).

Since the hydrochloride of the compound of Formula (I) is intended for pharmaceutical use it is preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure especially at least 98% pure (% are on a weight for weight basis). The pharmaceutical compositions of the invention comprise a hydrochloride of the compound of Formula (I) as an active ingredient, a pharmaceutically acceptable carrier and optionally other therapeutic ingredients or adjuvants. The compositions include those suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The compositions are preferably suitable for oral administration. The pharmaceutical compositions may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

In practice, the hydrochloride of the compound of Formula (I) can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g. oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present invention can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion, or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the hydrochloride of the compound of Formula (I) may also be administered by controlled release means and/or delivery devices. The compositions may be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation. The hydrochloride of the compound of Formula (I) can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.

The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

In preparing the compositions for oral dosage form, any convenient pharmaceutical media may be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like may be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like may be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets may be coated by standard aqueous or nonaqueous techniques.

A tablet containing the composition of this invention may be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets may be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. Each tablet preferably contains from about 0.05mg to about 5g of the active ingredient and each sachet or capsule preferably contains from about 0.05mg to about 5g of the active ingredient.

For example, a formulation intended for oral administration to humans may contain from about 0.5mg to about 5g of active agent, compounded with an appropriate and convenient amount of carrier material, which may vary from about 5 to about 95% of the total composition. Unit dosage forms will generally contain from about lmg to about 2g of the active ingredient, typically 25mg, 50mg, lOOmg, 200mg, 300mg, 400mg, 500mg, 600mg, 800mg, or lOOOmg.

Pharmaceutical compositions of the present invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.

Pharmaceutical compositions of the present invention suitable for injectable use include sterile aqueous solutions or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In all cases, the final injectable form must be sterile and must be effectively fluid for easy syringability. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

Pharmaceutical compositions of the present invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, or the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations may be prepared, utilizing a hydrochloride of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, via conventional processing methods. As an example, a cream or ointment is prepared by admixing hydrophilic material and water, together with about 5%w/w to about 10%w/w of the compound, to produce a cream or ointment having a desired consistency.

Pharmaceutical compositions of this invention can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

In addition to the aforementioned carrier ingredients, the pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like. Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient. Compositions containing a hydrochloride of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may also be prepared in powder or liquid concentrate form.

Generally, dosage levels on the order of O.Olmg/kg to about 150mg/kg of body weight per day are useful in the treatment of the above-indicated conditions, or alternatively about 0.5mg to about 7g per patient per day. For example, diabetes and hyperglycemia may be effectively treated by the administration of from about 0.01 to 50mg of a hydrochloride of the compound of Formula (I) per kilogram of body weight per day, or alternatively about 0.5mg to about 3.5g per patient per day. Similarly, hypercholesterolemia, hyperinsulinemia, hyperlipidemia, hypertension, atherosclerosis or tissue ischemia e.g. myocardial ischemia may be effectively treated by the administration of from about 0.01 to 50mg of a hydrochloride of the compound of Formula (I) per kilogram of body weight per day, or alternatively about 0.5mg to about 3.5g per patient per day, e.g. 50mg to lOOOmg.

It is understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy. The hydrochloride of the compound of Formula (I) may be used in the treatment of diseases or conditions in which glycogen phosphorylase plays a role.

Thus the invention also provides a method for the treatment of a disease or condition in which glycogen phosphorylase plays a role comprising a step of administering to a subject in need thereof an effective amount of a hydrochloride of the compound of Formula (I). Diseases or conditions in which glycogen phosphorylase plays a role include diabetes

(including Type I and Type II, impaired glucose tolerance, insulin resistance and diabetic complications such as neuropathy, nephropathy, retinopathy and cataracts), hyperglycemia, hypercholesterolemia, hyperinsulinemia, hyperlipidemia, hypertension, atherosclerosis, tissue ischemia e.g. myocardial ischemia. The invention also provides a method for the treatment of hyperglycemia or diabetes comprising a step of administering to a subject in need thereof an effective amount of a hydrochloride of the compound of Formula (I).

The invention also provides a method for the prevention of diabetes in a human demonstrating pre-diabetic hyperglycemia or impaired glucose tolerance comprising a step of administering to a subject in need thereof an effective prophylactic amount of a hydrochloride of the compound of Formula (I).

The invention also provides a method for the treatment of hypercholesterolemia, hyperinsulinemia, hyperlipidemia, hypertension, atherosclerosis or tissue ischemia comprising a step of administering to a patient in need thereof an effective amount of a hydrochloride of the compound of Formula (I).

The invention also provides a method of cardioprotection e.g. following reperfusion injury, comprising a step of administering to a subject in need thereof an effective amount of a hydrochloride of the compound of Formula (I).

The invention also provides the use of a hydrochloride of the compound of Formula (I) in the treatment of a condition as defined above.

The invention also provides the use of a hydrochloride of the compound of Formula (I) in the manufacture of a medicament for the treatment of a condition as defined above.

The invention also provides a hydrochloride of the compound of Formula (I) for use in the treatment of a condition as defined above. In the methods and uses of the invention the term "treatment" includes both therapeutic and prophylactic treatment.

The hydrochloride of the compound of formula (I) may, particularly for the treatment of type II diabetes, be administered to a patient at night time, e.g. at bed time and preferably after the patient has consumed their last meal of the day such that inhibition of glycogen phosphorylase occurs during the fasting period.

The hydrochloride of the compound of Formula (I) may be administered alone or in combination with one or more other therapeutically active compounds. The other therapeutically active compounds may be for the treatment of the same disease or condition as the hydrochloride of the compound of Formula (I) or a different disease or condition. The therapeutically active compounds may be administered simultaneously, sequentially or separately.

The hydrochloride of the compound of Formula (I) may be administered as polypharmacy with other active compounds for the treatment of diabetes, for example PPAR agonists, biguanides, sulfonylureas and other insulin secretagogues, insulin sensitisers, alpha-glucosidase inhibitors, dipeptidyl peptidase IV inhibitors, glucokinase activators, GLP-I and GLP-I analogues, insulin, insulin analogues, α2 agonists, fatty acid oxidation inhibitors, α-glucosidase inhibitors, β-agonists, phosphodiesterase inhibitors, lipid lowering agents, antiobesity agents, amylin antagonists, lipoxygenase inhibitors, somostatin analogs, glucagon antagonists, insulin signalling agonists, PTPlB inhibitors, gluconeogenesis inhibitors, antilypolitic agents, GSK inhibitors, galanin receptor agonists, anorectic agents, CCK receptor agonists, leptin, CRF antagonists and CRF binding proteins. The hydrochloride of the compound of Formula (I) may also be administered in combination with thyromimetic compounds, aldose reductase inhibitors, glucocorticoid receptor antagonists, NHE-I inhibitors or sorbitol dehydrogenase inhibitors. These additional agents may be formulated and administered by methods known to those skilled in the art.

The hydrochloride of the compound of Formula (I) may exhibit advantageous properties compared to known glycogen phosphorylase inhibitors, for example, the solid compound exhibits improved handling properties which make it easier to isolate and formulate than known forms of the compound of Formula (I). The isolation of the hydrochloride salt of the compound of Formula (I) is improved both chemically and enantiomerically relative to the free base. In crystalline form the hydrochloride of the compound of Formula (I) also exhibits advantageous properties desirable for pharmaceutical active ingredients.

A hydrochloride of the compound of Formula (I) may be prepared, for example, by treating a compound of Formula (I) with HCl. Suitable conditions include dissolution of the compound of Formula (I) in an alcoholic solvent, e.g. methanol, ethanol or propanol, followed by the addition of hydrochloric acid e.g. 2M hydrochloric acid, and the subsequent removal of the solvent under reduced pressure.

A compound of Formula (I), 5-chloro-lH-pyrrolo[2,3-c]pyridine-2-carboxylic acid [l-(S)-(4- fluorobenzyl)-2-(4-hydroxypiperidin-l-yl)-2-oxoethyl] amide, may be prepared by the processes described in PCT/US2004/016243.

Accordingly, in a first process, the compound of Formula (I) may be prepared by coupling a compound of Formula (II), namely 5-chloro-lH-pyrrolo[2,3-c]pyridine-2-carboxylic acid, or a protected or activated derivative thereof, with an amine of Formula (III), namely 2-(5)-amino-3-(4- fluorophenyl)-l-(4-hydroxypiperidin-l-yl)propan-l-one hydrochloride, as shown in Scheme 1.

Scheme 1

Typically, the compound of Formula (II), or a protected or activated derivative thereof, is combined with the compound of Formula (III) in the presence of a suitable coupling agent. Examples of suitable coupling reagents are l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/hydroxybenzotriazole (EDCI / HOBt), 1,1-carbonyldiimidazole (CDI), dicyclohexylcarbodiimide/ hydroxybenzotriazole (DCC / HOBt), O-(lH-benzotriazol-l-yl)-N,N,N',N- tetramethyluronium tetrafluoroborate (R. Knorr et ah, Tetrahedron Lett., 1989, 30, 1927-1930) and polymer supported carbodiimide-1 -hydroxybenzotriazole (for representative procedures, see for example, Argonaut Technical Note 501 available from Argonaut Technologies, Inc., Foster City, California). The couplings may be performed in an inert solvent, preferably an aprotic solvent at a temperature of about O⁰C to about 45⁰C for about 1 to 72h in the presence of a tertiary amine base such as diisopropylethylamine (DIPEA) or triethylamine. Exemplary solvents include acetonitrile, chloroform, dichloromethane, N,N-dimethylformamide (DMF) or mixtures thereof. Use of these coupling agents and appropriate selection of solvents and temperatures are known to those skilled in the art or can be readily determined from the literature. These and other exemplary conditions useful for coupling carboxylic acids are described in Ηouben-Weyl, VoI XV, part II, E. Wunsch, Ed., G. Thieme Verlag, 1974, Stuttgart, and M. Bodansky, Principles of Peptide Synthesis, Springer- Verlag, Berlin, 1984 and The Peptides, Analysis, Synthesis and Biology (Ed., E. Gross and J. Meienhofer), VoIs 1-5, Academic Press NY 1979-1983.

The compound of Formula (II) can be obtained by the synthesis illustrated below in Scheme 2:

(V)

Scheme 2

The compound of Formula (V) may be prepared by condensation of an ortho methyl nitro compound of Formula (IV) with an oxalate ester in a solvent such as diethyl ether in the presence of a base such as potassium ethoxide or DBU. The compound of Formula (VI) is prepared from a compound of Formula (V) under reducing conditions, such as iron powder and ammonium chloride, or by hydrogenation in ethanol using palladium catalysis. The compound of Formula (VI) undergoes ester hydrolysis using aqueous alkali to give the pyrrolopyridine-2-carboxylic acid of Formula (II). Further information on the conversion of the compound of Formula (IV) to the compound of Formula (II) is available in the literature (Kermack, et al, J. Chem, Soc, 1921, 119, 1602; Cannon et ah, J. Med. Chem., 1981, 24, 238; Julian et al, in Heterocyclic Compounds, VoI 3 (Wiley, New York, NY, 1962, R.C. Elderfield, Ed.) p 18. The compound of Formula (IV) is available commercially.

Alternatively, the compound of Formula (II) can also be obtained by the synthesis in Scheme 3:

(VIl) (VIM) (II)

Scheme 3

According to Scheme 3, the compound of Formula (II) is prepared by a Heck coupling of an ortho-iodo aminopyridine (VIII) followed by cyclisation at a temperature of between 100 to 150°C in the presence of catalyst such as palladium acetate and a base such as DABCO in a solvent such as DMF (See Chen et al, J. Org. Chem. 1997, 62, 2676). The ortho-iodo aminopyridines (VIII) can be made by direct iodination of the aminopyridine (VII) using iodine in the presence of silver sulfate in a solvent such as ethanol at ambient temperature (see Sy, W., Synth. Commun., 1992, 22, 3215). The compound of Formula (VII) is available commercially.

Alternatively the compound of Formula (VIII) for use in Scheme 3 may be prepared according to Scheme 4 by deprotection of an N-pivaloyl compound of Formula (X) by heating under reflux using hydrochloric acid. The N-pivaloyl compound of Formula (X) is in turn made by deprotonation of the compound of Formula (EX) with an organolithium such as tert-butyllithium in a suitable solvent such as THF, followed by quenching with iodine at a low temperature. The compound of Formula (EX) may be made by protection of the aminopyridine of Formula (VII) with trimethylacetyl chloride and a base such as triethylamine in a solvent such as dichloromethane.

(VIi)

Scheme 4

A further alternative for the preparation of a compound of Formula (VIII), shown in Scheme 5, is by the deprotection of an N-Boc protected compound of Formula (XII) using an acid such as trifluoroacetic acid in a solvent such as dichloromethane at ambient temperature. The N-BOC compound of Formula (XII) is in turn made by deprotonation of the compound of Formula (XI) with an organolithium such as n-butyllithium in the presence of N,N,N',N'-tetramethylethylenediamine (TMEDA) in a suitable solvent such as ether at temperatures around -70°C followed by the addition of iodine at temperatures around -10°C. The N-Boc aminopyridine of Formula (XI) is routinely made from the aminopyridine of Formula (VII) using di-tert-butyldicarbonate by heating in a solvent such as 1,4-dioxane.

(VII) (Xl) (XII) <^VI")

Scheme 5 The compound of Formula (III) can be obtained by the synthesis described in Scheme 6 below:

Scheme 6

Compounds of Formula (XIII) are readily prepared by known techniques. PG represents a protecting group such as, for example, tert-butyloxycarbonyl (Boc). Compounds of Formula (XIV) are made from carboxylic acids of Formula (XIII) using standard coupling conditions, such as described above for Scheme 1.

The compound of Formula (III) may be prepared from compounds of Formula (XIV) by removal of the protecting group, where PG = Boc, under acidic conditions using for example trifluoroacetic acid in dichloromethane at temperatures of around 25⁰C. A second process for the production of the compound of Formula (I) is illustrated in Scheme

7:

Scheme 7

The compound of Formula (I) may be prepared according to Scheme 7 by coupling the carboxylic acid of Formula (XV), or a protected or activated derivative thereof, with the amine of Formula (XVI). Examples of suitable coupling agents and conditions are as described above. The amine of Formula (XVI) is commercially available.

The compound of Formula (XV) may be prepared according to Scheme 8, which uses methods analogous to those shown above in Scheme 1.

^{(ll) (}^¹" P⁰" Scheme 8

The carboxylic acid of Formula (II) is reacted with commercially available L-4- fluorophenylalanine under the conditions provided for in Scheme 1. For example, methyl or tert-butyl protected L-4-fluorophenylalanine (XVII) is reacted with the compound of Formula (II) in DMF using a carbodiimide coupling agent. Alternatively, the activated succinimidyl ester of the compound of Formula (II) may first be prepared before subsequent reaction with L-4-fluorophenylalanine (XVII) in the presence of a base.

As a further aspect of the present invention, there is provided a novel process for the production of the compound of Formula (XV). As is discussed below, this process is efficient by virtue of avoiding racemisation of certain key intermediates and by permitting ready purification through use of reagents and products which are readily crystallisable. This process may be employed in the synthesis of 5-chloro-lH-pyrrolo[2,3-c]pyridine-2-carboxylic acid [l-(5)-(4-fluorobenzyl)-2-(4- hydroxypiperidin-l-yl)-2-oxoethyl] amide free base (compound of Formula (I)) and salt forms thereof, including the hydrochloride salt.

Thus, there is provided according to this aspect of the invention a process for preparing a compound of Formula (XV):

^(XV) or an acid addition salt thereof, which comprises reacting a compound of Formula (XVIII):

(XVlIl) or an acid addition salt thereof, with a compound of Formula (XVII):

(XVII) or a protected derivative thereof. The reaction is preferably conducted in a basic aqueous solution.

There is also provided according to the invention a process for preparing a compound of Formula (I):

(I) or an acid addition salt thereof, which comprises reacting a compound of Formula (XVIII):

^(XV"¹⁾ or an acid addition salt thereof, with a compound of Formula (III):

(III) or a protected derivative thereof. The process is preferably conducted in a basic aqueous solution.

According to the above mentioned processes, the compounds of Formula (XVIII) and (XVII) or (XVIII) and (III) are preferably reacted in aqueous solution under Schotten-Baumann conditions (see The Chemistry of Amides, Wiley, New York, 1970 pp 73-185) i.e. in the presence of base.

The basic aqueous solution may suitably contain an organic solvent immiscible with water e.g. THF.

The aqueous solution may suitably be made basic with hydroxide or carbonate e.g. sodium hydroxide and/or sodium carbonate. For example, a suspension of N-5-chloropyrrolo[2,3-c]pyridin-2-carbonyl chloride or a salt thereof, preferably the hydrochloride salt in anhydrous THF can be added dropwise to an aqueous solution of (L)-4-fluorophenylalanine (XVII) containing NaOHZNa₂CO₃. Preferably a slight excess of (L)-4-fluorophenylalanine is used e.g. about 1.2 molar equivalents. Low temperature may be needed to prevent hydrolysis of the acyl chloride and racemization of the amino acid (even in acylated form). Preferably the reaction is carried at 0-5⁰C since lower temperatures may result in stirring difficulties. The reaction mixture may be neutralized and the THF removed under vacuum. The product may be recovered as a precipitate after acidification of the solution (pH=l-2). The addition of EtOAc to the aqueous solution before acidification and filtration may provide a precipitate of better quality.

The compound of Formula (XV) may be purified by crystallization. A preferred solvent system for crystallization is 2M aqueous HCl solution and an alcoholic solvent e.g. methanol, ethanol or propanol, for example about a 2:1 mixture of 2M aqueous HCl solution and 2-propanol, from which the extended acid intermediate (compound of Formula (XV)) can be obtained as a crystalline hydrochloride salt.

A particular advantage of this process for preparing a compound of Formula (XV) is that it can be performed without the need to protect the amino acid (compound of Formula (XVII)). Also the process minimises the opportunity for racemisation of the chiral carbon.

In order to produce a compound of Formula (XVIII), 5-chloropyrrolo[2,3-c]pyridine-2- carboxylic acid may be treated with SOCl₂ in an inert solvent. Reaction may be carried out in suspension, for example in toluene or acetonitrile, with 3-4 equivalent of SOCl₂ at reflux temperature. If the reaction is carried out in toluene, the acid chloride product (XVIII) may be isolated by filtration. If the reaction is carried out in acetonitrile, the solvent may be removed under reduced pressure to recover the acid chloride.

In the preparation of a compound of Formula (I), the compound of Formula (XV) so produced may be reacted with a compound of Formula (XVI):

(XVI) A range of methods for performing this coupling will be apparent to those skilled in the art. To enable the use of the isolated hydrochloride salt of the extended acid without liberation of base, suitable solvents include. Reduced levels of racemisation or side reactions are observed when the solvent used is EtOAc, propanol or THF. Suitable coupling agents include 2-chloro-4,6-dimethyl- 1,3,5-triazine (CDMT) or (DMTMM). The reaction is performed in the presence of base, such as N- methylmorpholine or an excess of 4-hydroxypiperidine, preferably an excess of 4-hydroxypiperidine. Crude compound of Formula (I) may be isolated following the reaction.

The hydrochloride of the compound of Formula (I) may be purified by recrystallization, the recrystallisation conditions may determine the polymorphic form of the material which is obtained. Suitable solvent systems for crystallization are mixtures of 2M aqueous HCl solution and an alcoholic solvent e.g. methanol, ethanol or propanol, for example about a 2: 1 mixture of 2M aqueous HCl solution and 2-propanol, and mixtures of an alcoholic solvent e.g. methanol, ethanol or propanol, and acetonitrile, for example about a 1:5 mixture of methanol and acetonitrile.

Thus according to a further aspect of the invention there is provided a process for the preparation of a hydrochloride of the compound of Formula (I) which comprises recrystallising a hydrochloride of the compound of Formula (I) from a mixture of aqueous HCl and an alcoholic solvent or a mixture of an alcoholic solvent and acetonitrile.

The invention also provides a hydrochloride of the compound of Formula (I) obtainable by such a recrystallisation method. Reaction of the acid chloride of Formula (XVIII) with (XVII) according to the process of the invention to provide the compound of Formula (XV) is more efficient and leads to lower levels of racemisation of the chiral centre than the previous processes of Scheme 8 (for example, the succinimidyl ester of (II) in particular is difficult to handle and decomposes in water). The process of the invention may thereby enable higher yields of the compound of Formula (I) (and hence its hydrochloride salt to be obtained.

The acid chloride of the compound of Formula (XVIII) will advantageously be utilized in the form of a hydrochloride. Under such conditions the compoundof Formula (XV) is produced as an acid addition salt with HCl, which may be crystallised and easily purified. The final product is therefore obtained at a high purity. In contrast, since the compound of Formula (III) prepared using in Scheme 3 is obtained an oil, the compound of Formula (I) prepared according to the Scheme 1 is generally contaminated with several by products having similar polarity and therefore this route is less favoured. One of the advantages of the process of the invention defined above is that protection of labile functional groups in the intermediate compounds is unnecessary. However during the other processes described above labile functional groups in the intermediate compounds, e.g. hydroxy, carboxy and amino groups, may be protected. 5-Chloropyrrolo[2,3-c]pyridine-2-carboxylic acid may be protected in the 1 -position e.g. with an arylmethyl, acyl, alkoxycarbonyl, sulfonyl or silyl group. The protecting groups may be removed at any stage in the synthesis of the compounds of Formula (I) or may be present on the final compound of Formula (I). A comprehensive discussion of the ways in which various labile functional groups may be protected and methods for cleaving the resulting protected derivatives is given in for example, Protective Groups in Organic Chemistry, T.W. Greene and P.G.M. Wuts, (1991) Wiley-Interscience, New York, 2^nd edition.

The novel process of the present invention may additionally be utilised in the production of analogues of the compound of Formula (I). In particular, analogues are contemplated wherein the 4- hydroxypiperidin-1-yl moiety is replaced with a moiety R³ which is (i) a 4-8-membered nitrogen containing heterocyclyl group (linked via a ring nitrogen atom to the amino acid carbonyl), said heterocyclyl group being optionally substituted with 1-3 independent halogen, cyano, Ci_-4alkyl, fluoromethyl, difluoromethyl, trifluoromethyl,

-C₀-₄alkylNR⁷R⁸, - C(O)R⁹, C^alkoxyCo^alkyl-, -COOCo^alkyl, -C₀^alkylNHC(O)R⁹, -C₀^alkylC(O)N(R¹⁰)₂, -C₁. 5 ₄alkoxyCi_-4alkoxy, hydroxyC₀-₄alkyl-, -NHSO₂R¹⁰,

-SO₂NR¹¹R¹², 5- to 6-membered heterocyclyl, phenylC_0-2alkoxy, or phenylC₀-₂alkyl substituents, wherein phenyl is optionally substituted with 1-2 independent halogen, cyano, Ci_-4alkyl,

- SO₂Ci_-4alkyl, -S0₂N(C₀-₄alkyl)(Co-₄alkyl), hydroxy, fluoromethyl, difluoromethyl, or trifluoromethyl substituents, or two bonds on a ring carbon of the heterocyclyl group optionally can form an oxo ( =0 0 ) substituent; or (ii) a moiety -NR^-Co^alkylR⁵) in which

R⁴ is Co-₃alkyl, -C_2-3alkyl-NR⁷R⁸, C_3-6cycloalkyl optionally substituted by hydroxyC₀-₄alkyl- further optionally substituted by hydroxy, Ci_-2alkoxyC_2-4alkyl-, or Ci_-2alkyl-S(O)_n-C_2-3alkyl-; n is O, 1, or 2; and

R⁵ is hydrogen, hydroxyC_2-3alkyl-, Ci_-2alkoxyC₀-₄alkyl-, or aryl, hetaryl, or heterocyclyl; 5 wherein a heterocyclic nitrogen-containing R⁵ ring optionally is mono-substituted on the ring nitrogen with Ci_-4alkyl, benzyl, benzoyl, Ci_-4alkyl-C(O)-,

-SO₂Ci_-4alkyl, -S0₂N(C₀-₄alkyl)(Co-₄alkyl), Ci_-4alkoxycarbonyl or aryl(Ci_-4alkoxy)carbonyl; and wherein the R⁵ rings are optionally mono-substituted on a ring carbon with halogen, cyano,

C(O)-,

or 0 Co^alkylcarbamoyl-, provided that no quaternised nitrogen is included; or two bonds on a ring carbon of the heterocyclyl group optionally can form an oxo ( =0 ) substituent;

R⁷ and R⁸ are independently C₀-₄alkyl, C_3-6cycloalkyl, or

R⁹ is or C_3-6cycloalkyl;

R¹⁰ is Co-₄alkyl, or C_3-6cycloalkyl; 5 R¹¹ and R¹² are independently C₀-₄alkyl or together with the nitrogen to which they are attached may form a 4- to 6-membered heterocycle.

Such analogues may be prepared in the form of pharmaceutically acceptable salts thereof, e.g. the HCl salts.

Specific examples of nitrogen containing heterocyclyl groups which may replace the 4- O hydroxypiperidin- 1 -yl moiety of the compound of Formula (I) include azetidin- 1 -yl, pyrrolidin- 1 -yl, piperidin-1-yl, 1,4-diazapan-l-yl, piperazin-1-yl, morpholin-4-yl, thiomorpholin-4-yl, 1,1-dioxo- thiomorpholin-4-yl, or thiazolidin-3-yl; which groups may be optionally substituted as described above. Preferred substituent groups for 4-hydroxypiperidin-l-yl replacement include -Ci_-4alkoxy, hydroxy and oxo. 5 Even more preferably the replacement group is pyrrolidin- 1-yl or piperidin-1-yl optionally substituted with hydroxyl, e.g. 3-(S)-hydroxypyrrolidin-l-yl.

Thus analogues of the compound of Formula (I) defined by Formula (IA):

(IA) 0 in which R³ is as defined above, may be prepared by reacting a compound of Formula (XVIII) with a compound of Formula (XVII) in an aqueous basic condition and then reacting the product of said reaction with an amine compound of Formula R³-H. Suitable conditions include conditions similar to those described above for the reaction of a compound of Formula (XV) with a compound of Formula (XVI).

Any novel intermediates as defined above are also included within the scope of the invention. The invention also provides 5-chloropyrrolo[2,3-c]pyridine-2-carbonyl chloride, or an acid addition salt thereof, in particular 5-chloropyrrolo[2,3-c]pyridine-2-carbonyl chloride hydrochloride.

All publications, including, but not limited to, patents and patent application cited in this specification, are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as fully set forth. The invention will now be illustrated with reference to the following examples.

EXAMPLES Analytical methods

Thermogravimetric analysis was performed as follows: A sample of the material (ca. 4-5 mg) was heated in a platinum vessel using slow inert gas (nitrogen) flow with 5°C/min heating rate using Derivatograph C instrument.

X ray diffraction measurements were performed using Philips PW 3710/PW 1050 diffractometer (Cu Ka 40 kV, 35 mA).

BRIEF DESCRIPTION OF THE FIGURES:

Figure 1 shows thermogravimetric analysis of a hydrochloride salt of the invention (Example

1).

Figures 2-5 show X-ray diffraction patterns for a hydrochloride salt of the invention.at different states of hydration (Example 1). Figure 6 show X-ray diffraction pattern for a hydrochloride salt of the invention (Example 2) overlayed with that of Example 1.

EXAMPLE 1

Synthesis of 5-chloropyrrolor2,3-c1pyridine-2-carboxylic acid ri-β7-4-fluorobenzyl)-2-(4- hydroxypiperidin- 1 -yl)-2-oxoethyl1amide hydrochloride

(a) Preparation of 5-chloropyrrolor2,3-c1pyridine-2-carbonyl chloride hydrochloride

Method A: 5-Chloropyrrolo[2,3-c]pyridine-2-carboxylic acid (39.3g, 0.20mol) was suspended in acetonitrile and heated to reflux. Thionyl chloride (44mL, 71.4g, 0.60mol) was added dropwise over 20 min at reflux temperature. The resulting suspension was heated at reflux for a further 3 h (TLC monitoring: n-butanol-acetic acid- water 4: 1: 1, UV visualised. Sample was prepared by quenching into methanolic NH₃ solution). The reaction mixture was evaporated to dryness under reduced pressure and the crude product used in the next step without further purification. Yield 49.3g (98.0%).

Method B: A slurry of 5-chloro-lH-pyrrolo[2,3-c]-pyridin-2-carboxylic acid (300g, 1.52mol) in acetonitrile (3.75L) was heated to reflux. Thionyl chloride (363g, 3.052mol, 223mL) was added dropwise to the mixture and the reaction monitored by tic and hlpc. After completion of the reaction excess thionyl chloride and acetonitrile was distilled off under diminished pressure to obtain a thick slurry. Toluene (2L) was added to the residue, and solvents evaporated under diminished pressure. The product was filtered off under nitrogen and washed with toluene (0.2L) and hexane (0.2L). The product was dried in vacuo at 45-50⁰C over potassium hydroxide to obtain the title c coommppoouunndd.. Y Yiieelldd 336688gg ( (9966%%)).. I IRR ( (KKBBrr)) 11775500 c cmm^""11 ( (ialso 2436 br, 1981, 1869, 1631, 1588, 1529, 1447, 1389, 1340, 1289, 1203, 1140 and 1001 cm ¹).

(b) Preparation ofN-(5-chloropyrrolor2,3-c1pyridin-2-carbonyl)-Z-4- fluorophenylalanine hydrochloride

Method A: To a solution of NaOH (9.41g, 0.235mol, 1.2eq) and Na₂CO₃ (62.3g, 0.588mol, 3.0eq) in deionized water (24OmL) was added Z-4-fluorophenylalanine (43.1g, 0.235mol, 1.2eq) followed by THF (24OmL). The resulting solution was cooled to 0-5⁰C and a suspension of 5- chloropyrrolo[2,3-c]pyridine-2-carbonyl chloride hydrochloride (49.3g, 0.196mol, l.Oeq) in dry THF was added (~30 min). The reaction mixture was stirred at 0-5⁰C for 15 min (HPLC monitoring, direct analysis of the sample). The temperature was maintained at 0-5 ⁰C while the pH of the reaction mixture was adjusted to ~7 by the addition of cone, hydrochloric acid and THF was removed under reduced pressure. EtOAc (5OmL) was added to the remaining aqueous solution and the pH adjusted to 1-2 by the addition of cone, hydrochloric acid (~80mL altogether). The resulting suspension was stirred for 30 min at 0-5⁰C. The precipitate was then filtered, washed with EtOAc (2xl00mL) and dried in vacuo at 4O⁰C. Crude yield 67.6g (86.6%). The crude product was crystallised from a mixture of 2M HCl (54OmL) and 2-propanol (27OmL). Yield 60.9g (78.0 %). ¹H-NMR (DMSO): 13.02 (br s, IH), 9.2 (d, IH), 8.80 (s, IH), 7.95 (s, IH), 7.48 (s, IH), 7.34 (dd, 2H), 6.96 (dd, 2H), 4.81 (m,lH), 3.29 (dd, IH), 3.16 (dd, IH).

Method B: To a solution of NaOH (73.Og, 1.82mol) and Na₂CO₃ (486g, 4.58mol) in deionized water (1.90L) was added Z-4-fluorophenylalanine (336g, 1.82mol) followed by THF (2.80L). The resulting solution was cooled to 0-5⁰C and a suspension of 5-chloropyrrolo[2,3- c]pyridine-2-carbonyl chloride hydrochloride (383g, 1.52mol) in dry THF was added (~30 min). The reaction mixture was stirred at 0-5⁰C for 30 min (HPLC monitoring, direct analysis of the sample). The temperature was maintained at 0-5⁰C while the pH of the reaction mixture was adjusted to ~7 by the addition of cone, hydrochloric acid (23OmL) and THF was removed under reduced pressure. EtOAc (3.0L) was added to the residue and the pH adjusted to 1-2 by the addition of cone, hydrochloric acid (0.6L). The resulting slurry was stirred for 30 min at 0-5⁰C. The precipitate was then filtered, washed with EtOAc (2x500mL) and dried in vacuo at 40-50⁰C. (95% purity by HPLC).

(c) Preparation of 5-chloropyrrolo[2,3-c^"|pyridine-2-carboxylic acid [l-^-4-fluorobenzyl)-2-(4- hydroxypiperidin- 1 -yl)-2-oxoethyl^"lamide hydrochloride Method A: N-(5-Chloropyrrolo[2,3-c]pyridine-2-carbonyl)-L-4-fluorophenylalanine hydrochloride (60.9g, 0.153mol) was suspended in dry THF (46OmL) and the mixture was stirred at room temperature. 4-Hydroxypiperidine (35.7g, 0.353mol) was added portionwise (slight exotherm) and the mixture stirred at room temperature for 10 min. 4-(4,6-Dimethoxy-l,3,5-triazin-2-yl)-4- methylmorpholinium chloride (51.2g, 0.185mol, prepared according to the method of Kunishima et al, Tetrahedron Letters, 1999, 40, 5327-5330) was then added in one portion. The reaction mixture was stirred at room temperature for 1 h (HPLC monitoring, direct sample analysis). The solvent was removed under reduced pressure and the residue partitioned between EtOAc (50OmL) and saturated Na₂CO₃ solution (500mL)-water (60OmL) mixture. The organic layer was separated and the aqueous layer extracted with EtOAc (2xl50mL), the combined organic layers was washed with brine, dried over Na₂SO₄ and evaporated. Crude yield (base) 70.9g. The crude product was crystallised from a mixture of 2M HCl (42OmL) and 2-propanol (21OmL) to give 35.1g (47.7%) of a light yellow crystalline material (water content 13.2% and >98% optical purity). A second crystallisation from the same mixture gave 21.4g (29.1%) pure product with >99% optical purity. ¹H-NMR (DMSO): 13.2 (br s, IH), 9.24 (dd, IH), 8.90 (s, IH), 7.95 (s, IH), 7.50 (s, IH), 7.28 (dd, 2H), 6.96 (dd, 2H), 5.25 (qa, IH), 3.12 (m, IH), 1.85-1.115 (m, 9H).

Method B: N-(5-Chloropyrrolo[2,3-c]pyridine-2-carbonyl)-L-4-fluorophenylalanine hydrochloride (45Og, 1.13mol) was suspended in dry THF (3.40L) and the mixture cooled to 20-25⁰C. 4-Hydroxypiperidine (264g, 2.60mol) was added portionwise (slight exotherm) and the mixture stirred at 20-25⁰C for 5-10 min. 4-(4,6-Dimethoxy-l,3,5-triazin-2-yl)-4-methylmorpholinium chloride (380g, 1.37mol, prepared according to the method of Kunishima et al, Tetrahedron Letters, 1999, 40, 5327- 5330) was then added. The reaction mixture was stirred at 20-25⁰C (HPLC monitoring, direct sample analysis). The reaction mixture was poured into a stirred solution of sodium carbonate (70Og) in deionised water (7L), EtOAc (50OmL) was added and the mixture stirred for 10 min. The organic layer was separated and the aqueous layer extracted with EtOAc (IxIL and Ix 50OmL), the combined organic layers was washed with brine (2.0L) and dried over Na₂SO₄ (7Og) and activated carbon (15g) overnight before the solvent was evaporated. The crude product was dissolved in methanol (2.0L) and 2M HCl (2.50L), Celite (1Og) and activated carbon (1Og) added. The resulting slurry was stirred for 30 min. The mixture was filtered and the methanol removed under reduced pressure. The crystal slurry was cooled overnight to 4-5⁰C, filtered, washed with 2M HCl (0.20L) and dried in vacuo at 5O⁰C. The product was recrystallised from a mixture of 2M HCl (2.10L) and 2-propanol (0.9L) and the product dried over KOH in vacuo at 5O⁰C.

Analytical methods

Thermal stability data on the product of Example 1 were obtained by the thermogravimetic method described above. The resultant trace is shown in Figure 1. The substance appeared to be stable below 19O⁰C. The transition at around 4O⁰C corresponds to vaporization of possible solvent or water.

The properties of the product of Example 1 were studied using X-ray scattering following the methods described above. Analysis was performed at 4 stages of the purification process: Figure 2 shows the trace corresponding to crude product obtained in wet form after washing.

Figure 3 shows the trace corresponding to crude product obtained after washing with EtOAc.

Figure 4 shows the trace corresponding to final product after drying. The wet product (50- 60% volatile content) was dried at 3O⁰C (8 h) then the temperature was increased gradually to 5O⁰C over 1O h and the drying continued at this temperature for 24 h. Figure 5 shows the trace corresponding to final product after drying as described above then storing under air above wet silica gel for 12 h resulting in the production of material with equilibrium moisture content and substantially crystalline structure.

EXAMPLE 2 Recrystallisation of 5-chloropyrrolor2,3-c1pyridine-2-carboxylic acid ri-β7-4-fluorobenzyl)-2-(4- hvdroxypiperidin-l-yl)-2-oxoethyl1amide hydrochloride from methanol:acetonitrile

5-Chloropyrrolo[2,3-c]pyridine-2-carboxylic acid [ 1 -($-4-fluorobenzyl)-2-(4- hydroxypiperidin-l-yl)-2-oxoethyl]amide hydrochloride (1Og, material obtained according to Example 1 but without recrystallisation) was dissolved in methanol (2OmL) at 5O⁰C and under continuous stirring acetonitrile (10OmL) was added. The product started to precipitate at the end of the addition of acetonitrile. The mixture was warmed to 4O⁰C to get homogenous solution. After addition of the acetonitrile the suspension was cooled to O⁰C under continuous stirring. The product was crystallized for 12 h at O⁰C. The precipitate was filtered on a sintered glass filter. The filter cake was washed with of acetonitrile (1OmL) and the product was dried at 45⁰C in vacuum yielding product with >99% optical purity. Mp 77-78⁰C. Water content 4.5-5.5% w/w. Analytical methods

The properties of the product of Example 2 were studied using X-ray scattering following the methods described above. Figure 6 shows the trace corresponding to this product overlayed with that obtained for the material of Example 1.

In vitro GP activity Materials α-D-Glucose-1 -phosphate (disodium salt), Glycogen, D-Glucose, Malachite Green Hydrochloride, Ammonium Molybdate tetrahydrate, BSA, HEPES and rabbit muscle phosphorylase a (P 1261) were purchased from Sigma. All other reagents were analytical grade. Method Glycogen phosphorylase assay in vitro:

An assay for glycogen phosphorylase activity in the reverse direction was developed based on the method described by Engers et ah, Can. J. Biochem.,1910,. 48, 746-754]. Rabbit muscle glycogen phosphorylase a (Sigma) was reconstituted at a stock concentration of lOOμg/mL in 25mM Tris/HCl. The pH was measured in a 96-well plate in a final volume of lOOμL containing 5OmM Hepes pH 7.2, 7.5mM glucose, 0.5mM glucose- 1 -phosphate and lmg/mL glycogen. After incubation at 3O⁰C for 30 min, the inorganic phosphate released from glucose- 1 -phosphate was measured by the addition of 150μL of malachite green/molybdate solution prepared as follows: 5mL of 42% ammonium molybdate in 4N HCl, 15mL of 0.045% malachite green, 50μL of Tween 20. Following a 30 min incubation at room temperature, the absorbance was measured at 620nm. For IC₅₀ determination, lOμL of a serial dilution of compound (lOOμM to 0.004μM) in DMSO was added to each reaction in duplicate with the equivalent concentration of DMSO added to the control uninhibited reaction. Dose response curves were then obtained by plotting % inhibition versus logio compound concentration.

IC₅₀ is defined as the concentration of compound achieving 50% inhibition under the assay conditions described. The compound of Formula (I) has an IC₅₀ of < ImM.

Claims

1. A compound which is a hydrochloride salt of the compound of Formula (I):

or a solvate thereof.

2. A compound according to claim 1 in crystalline form.

3. A compound according to claim 1 or 2 which is a hydrate.

4. A compound according to any one of the preceding claims comprising up to 15% w/w of water.

5. A compound according to claim 1 in amorphous form.

6. A compound according to claim 1 characterised by an X-ray diffraction pattern corresponding substantially to that shown in any one of Figures 2-5.

7. A pharmaceutical composition comprising a compound according to any one of the preceding claims in admixture with a pharmaceutically acceptable carrier.

8. A pharmaceutical composition according to claim 7 presented as a discrete unit suitable for oral administration.

9. A pharmaceutical composition according to claim 7 or 8 presented as a solid dosage form.

10. A pharmaceutical composition according to any one of claims 7 to 9 in the form of a tablet, cachet or capsule.

11. A method of prophylaxis or treatment of hyperglycemia or diabetes, which comprises a step of administering a compound or composition according to any one of claims 1 to 10.

12. A method of prevention of diabetes in a human demonstrating pre-diabetic hyperglycemia or impaired glucose tolerance, which comprises a step of administering a compound or composition according to any one of claims 1 to 10.

13. A method or prophylactic or therapeutic treatment of hypercholesterolemia, hyperinsulinemia, hyperlipidemia, atherosclerosis or myocardial ischemia, which comprises a step of administering a compound or composition according to any one of claims 1 to 10.

14. A method of cardioprotection, which comprises a step of administering a compound or composition according to any one of claims 1 to 10.

15. A process for preparation of a hydrochloride of the compound of Formula (I) as defined in any one of claims 1 to 6 which comprises treating a compound of Formula (I) with HCl.

16. A process for preparation of a hydrochloride of the compound of Formula (I) as defined in any one of claims 1 to 6 which comprises recrystallising a hydrochloride of the compound of Formula (I) from a mixture of aqueous HCl and an alcoholic solvent or a mixture of an alcoholic solvent and acetonitrile.

17. A process for preparation of a hydrochloride of the compound of Formula (I) as defined in any one of claims 1 to 6 which comprises reacting a compound of Formula (II):

YYVf

(") or an activated derivative thereof, with a compound of Formula (III):

(III) and treating the product of the reaction with HCl to form the hydrochloride of the compound of Formula (I).

18. A process for preparation of a hydrochloride of the compound of Formula (I) as defined in any one of claims 1 to 6 which comprises reacting a compound of Formula (XV):

(XV) or an activated derivative thereof, with a compound of Formula (XVI): HO

(XVI) and treating the product of the reaction with HCl to form the hydrochloride of the compound of Formula (I).

19. A process for preparing a compound of Formula (XV):

(XV) or an acid addition salt thereof, which comprises reacting a compound of Formula (XVIII):

(XVIII) or an acid addition salt thereof, with a compound of Formula (XVII):

(XVII) or a protected derivative thereof.

20. A process for preparing a compound of Formula (I), or a pharmaceutically acceptable salt thereof, which comprises preparing a compound of Formula (XV) according to the process of claim 19 and then reacting it with a compound of Formula (XVI):

(XVI)

21. A process for preparing a compound of Formula (I):

(I) or an acid addition salt thereof, which comprises reacting a compound of Formula (XVIII):

(XVIII) or an acid addition salt thereof, with a compound of Formula (III):

(III) or a protected derivative thereof.

22. The process according to any one of claims 19 to 21 which is conducted in a basic aqueous solution.

23. A process according to claim 20 or 22 which process involves the further step of treating the product of the reaction of compounds of Formula (XV) and (XVI) or of the compounds of Formula (XVIII) and (III) with HCl to form the hydrochloride of the compound of Formula (I).

24. A process for preparing a compound of Formula (IA):

(IA) wherein R³ represents (i) a 4-8-membered nitrogen containing heterocyclyl group (linked via a ring nitrogen atom to the amino acid carbonyl), said heterocyclyl group being optionally substituted with 1-3 independent halogen, cyano, Ci_-4alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, -C₀- ₄alkylNHC(O)O(Ci_-4alkyl), -C₀-₄alkylNR⁷R⁸, -C(O)R⁹,

-C_{0- 4}alkylNHC(O)R⁹, -C₀-₄alkylC(O)N(R¹⁰)₂,

hydroxyCo_-4alkyl-, -NHSO₂R¹⁰, - SO₂(Ci_-4alkyl), -SO₂NR¹¹R¹², 5- to 6-membered heterocyclyl, phenylC_0-2alkoxy, or phenylC₀-₂alkyl substituents, wherein phenyl is optionally substituted with 1-2 independent halogen, cyano, Ci_-4alkyl,

hydroxy, fluoromethyl, difluoromethyl, or trifluoromethyl substituents, or two bonds on a ring carbon of the heterocyclyl group optionally can form an oxo ( =0 ) substituent; or (ii) a moiety -NR⁴(-C₀-₄alkylR⁵) in which

R⁴ is Co-₃alkyl, -C_2-3alkyl-NR⁷R⁸, C_3-6cycloalkyl optionally substituted by hydroxyCo-₄alkyl- further optionally substituted by hydroxy, Ci_-2alkoxyC₂-₄alkyl-, or Ci_-2alkyl-S(O)_n-C_2-3alkyl-; n is 0, 1, or 2; and

R⁵ is hydrogen, hydroxyC₂-₃alkyl-, Ci_-2alkoxyC₀-₄alkyl-, or aryl, hetaryl, or heterocyclyl; wherein a heterocyclic nitrogen-containing R⁵ ring optionally is mono-substituted on the ring nitrogen with Ci_-4alkyl, benzyl, benzoyl, Ci_-4alkyl-C(O)-, -SO₂Ci_-4alkyl, -S0₂N(Co-₄alkyl)(C₀-₄alkyl), Ci_-4alkoxycarbonyl or aryl(Ci_-4alkoxy)carbonyl; and wherein the R⁵ rings are optionally mono-substituted on a ring carbon with halogen, cyano,

C(O)-,

or Co^alkylcarbamoyl-, provided that no quaternised nitrogen is included; or two bonds on a ring carbon of the heterocyclyl group optionally can form an oxo ( =0 ) substituent; R⁷ and R⁸ are independently C₀-₄alkyl, C_3-6cycloalkyl, or

R⁹ is or C_3-6cycloalkyl;

R¹⁰ is Co-₄alkyl, or C_3-6cycloalkyl;

R¹¹ and R¹² are independently C₀-₄alkyl or together with the nitrogen to which they are attached may form a 4- to 6-membered heterocycle, or a pharmaceutically acceptable salt thereof, which comprises preparing a compound of Formula

(XV) according to the process of claim 19 and then reacting it with a amine compound of Formula R³- H.

25. A process according to any of claims 19 to 24 wherein the compound of Formula (XVIII) is employed as its HCl acid addition salt.

26. A compound of Formula (XVIII):

(XVIII) or an acid addition salt thereof.

27. A compound of Formula (XVIII) in the form of its HCl acid addition salt.