TRICYCLO SUBSTITUTED AMIDES AS GLUCOKINASE MODULATORS

BACKGROUND OF THE INVENTION

The present invention is directed to tri(cyclo) substituted amide compounds. In particular, the present invention is directed to amide compounds substituted i) at the carbonyl carbon with an ethyl attached to a phenyl ring and a carbocyclic ring, and ii) at the amino with a nitrogen bearing heteroaryl ring, which are modulators of glucokinase and are useful in the prophylactic or therapeutic treatment of hyperglycemia and diabetes, particularly type II diabetes. Glucokinase ("GK") is believed to be important in the body's regulation of its plasma glucose level. GK, found principally in the liver and pancreas, is one of four hexokinases that catalyze the initial metabolism of glucose. The GK pathway is saturated at higher glucose levels than the other hexokinase pathways (see R.L. Printz et al., Annu. Rev. Nutr., 13:463- 496 (1993)). GK is critical to maintaining the glucose balance in mammals. Animals that do not express GK die soon after birth with diabetes, while animals that overexpress GK have improved glucose tolerance. Activation of GK can lead to hyperinsulinemic hypoglycemia (see, for example, H.B.T. Christesen et al., Diabetes, 51:1240-1246 (2002)). Additionally, type II maturity-onset diabetes of the young is caused by the loss of function mutations in the GK gene, suggesting that GK operates as a glucose sensor in humans (Y. Liang et al., Biochem. J., 309:167-173 (1995)). Thus, compounds that activate GK increase the sensitivity of the GK sensory system and would be useful in the treatment of hyperglycemia, particularly the hyperglycemia associated with type II diabetes. It is therefore desirable to provide novel compounds that activate GK to treat diabetes, in particular compounds which demonstrate improved properties desirable for pharmaceutical products compared to known GK activators. International Patent Publication No. WO2001/044216 and U.S. Patent No. 6,353,111 describe (£)-2,3-disubstituted-N-heteroarylacrylamides as GK activators. International Patent Publication No. WO2002/014312 and U.S. Patent Nos. 6,369,232, 6,388,088, and 6,441,180 describe tetrazolylphenylacetamide GK activators. International Patent Publication No. WO2000/058293, European Patent Application No. EP 1169312 and U.S. Patent No. 6,320,050 describe arylcycloalkylpropionamide GK activators. International Patent

Publication No. WO2002/008209 and U.S. Patent No. 6,486,184 describe alpha-acyl and alpha-heteroatom-substituted benzene acetamide GK activators as anti-diabetic agents. International Patent Publication No. WO2001/083478 describes hydantoin-containing GK activators. International Patent Publication No. WO2001/083465 and U.S. Patent No. 6,388,071 describe alkynylphenyl heteroaromatic GK activators. International Patent

Publication No. WO2001/085707 and U.S. Patent No. 6,489,485 describe para-amine substituted phenylamide GK activators. International Patent Publication No. WO2002/046173 and U.S. Patent Nos. 6,433,188, 6,441,184, and 6,448,399 describe fused heteroaromatic GK activators. International Patent Publication No. WO2002/048106 and U.S. Patent No. 6,482,951 describe isoindolin-1 -one GK activators. International Patent

Publication No. WO2001/085706 describes substituted phenylacetamide GK activators for treating type II diabetes. U.S. Patent No. 6,384,220 describes para-aryl or heteroaryl substituted phenyl GK activators. French Patent No. 2,834,295 describes methods for the purification and crystal structure of human GK. International Patent Publication No.

WO2003/095438 describes N-heteroaryl phenylacetamides and related compounds as GK activators for the treatment of type II diabetes. U.S. Patent No. 6,610,846 describes the preparation of cycloalkylheteroaryl propionamides as GK activators. International Patent Publication No. WO2003/000262 describes vinyl phenyl GK activators. International Patent Publication No. WO2003/000267 describes aminonicotinate derivatives as GK modulators.

International Patent Publication No. WO2003/015774 describes compounds as GK modulators. International Patent Publication No. WO2003047626 describes the use of a GK activator in combination with a glucagon antagonist for treating type II diabetes. International Patent Publication No. WO2003/055482 describes amide derivatives as GK activators. International Patent Publication No. WO2003/080585 describes aminobenzamide derivatives with GK activity for the treatment of diabetes and obesity. International Patent Publication No. WO2003/097824 describes human liver GK crystals and their used for structure-based drug design. International Patent Publication No. WO2004/002481 discloses arylcarbonyl derivatives as GK activators. International Patent Publication Nos. WO2004/072031 and WO2004/072066 disclose tri(cyclo) substituted amide compounds as GK activators.

International Patent Application PCT/GB2005/050129 (published after the priority date of the present application) discloses amide compounds substituted i) at the carbonyl carbon with an ethyl/ethenyl attached to a phenyl ring and a carbocyclic ring, and ii) at the amino with a nitrogen bearing heteroaryl or unsaturated heterocyclyl ring, which are modulators of glucokinase and are useful in the prophylactic or therapeutic treatment of hyperglycemia and diabetes, particularly type II diabetes.

The present invention provides novel GK activators which may demonstrate improved properties desirable for pharmaceutical products compared to known GK activators, such as increased potency, increased in vivo efficacy and/or longer half-life.

SUMMARY OF THE INVENTION

Compounds represented by Formula (I):

(I) or pharmaceutically acceptable salts thereof, are useful in the prophylactic or therapeutic treatment of hyperglycemia and diabetes, particularly type II diabetes.

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to compounds of Formula (I):

(I) wherein A is a nitrogen containing heteroaryl ring selected from 5-methylpyrazin-2- yl, 5-methylpyrid-2-yl, 5-chloropyrid-2-yl, pyrid-2-yl, 5-methylisoxazol-3-yl, isoxazol-3-yl, 5-methylthiazol-2-yl and pyrimidin-4-yl; and pharmaceutically acceptable salts thereof.

A is preferably 5-methylpyrazin-2-yl, 5-methylpyrid-2-yl or 5-methylthiazol-2-yl, more preferably 5-methylpyrazin-2-yl.

In one embodiment of the present invention A represents 5-methylpyrazin-2-yl:

In a second embodiment of the present invention A represents 5-methylpyrid-2-yl:

In a third embodiment of the present invention A represents 5-chloropyrid-2-yl:

In a fourth embodiment of the present invention A represents pyrid-2-yl:

In a fifth embodiment of the present invention A represents 5-methylisoxazol-3-yl:

In a sixth embodiment of the present invention A represents isoxazol-3-yl:

N-

In a seventh embodiment of the present invention A represents 5-methylthiazol-2-yl:

In an eighth embodiment of the present invention, A represents 4-pyrimidinyl:

N^N The carbon atom linking the phenyl ring and the cyclopentanone containing sidechain to the amide carbonyl carbon is a chiral centre. Accordingly, at this centre, the compound may

be present either as a racemate or as a single enantiomer in the (R)- or (^-configuration. The (R)-enantiomers are preferred. The carbon atom which is the point of attachment of the cyclopentanone ring to the side chain is also chiral. Accordingly, at this centre, the compound may be present either as a racemate or as a single enantiomer in the (R)- or (^-configuration. The (R)-enantiomers are preferred.

The term "pharmaceutically acceptable salts" includes salts prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like. Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, methanesulfonic, and tartaric acids.

When the compound of the above formulae and pharmaceutically acceptable salts thereof exist in the form of solvates or polymorphic forms, the present invention includes any possible solvates and polymorphic forms. The type of solvent that forms the solvate is not particularly limited so long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone or the like can be used.

Since the compounds of Formula (I) are intended for pharmaceutical use they are preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure, at least 95% pure and especially at least 98% pure (% are on a weight for weight basis).

The invention also encompasses a pharmaceutical composition that is comprised of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable carrier.

Preferably the composition is comprised of a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

Moreover, within this embodiment, the invention encompasses a pharmaceutical composition for the prophylaxis or treatment of hyperglycemia and diabetes, particularly type

II diabetes, by the activation of GK, comprising a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of compound of Formula (I), or a pharmaceutically acceptable salt thereof.

The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as a pharmaceutical.

The compounds and compositions of the present invention are effective for treating hyperglycemia and diabetes, particularly type II diabetes, in mammals such as, for example, humans.

The invention also provides a method of prophylactic or therapeutic treatment of a condition where activation of GK is desirable comprising a step of administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

The invention also provides a method of prophylactic or therapeutic treatment of hyperglycemia or diabetes, particularly type II diabetes, comprising a step of administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

The invention also provides a method for the prevention of diabetes, particularly type II diabetes, in a human demonstrating pre-diabetic hyperglycemia or impaired glucose tolerance comprising a step of administering an effective prophylactic amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as a GK activator.

The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for the prophylactic or therapeutic treatment of hyperglycemia or diabetes, particularly type II diabetes. The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for the prevention of diabetes, particularly type II diabetes, in a human demonstrating pre-diabetic hyperglycemia or impaired glucose tolerance.

The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the activation of GK.

The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prophylactic or therapeutic treatment of hyperglycemia or diabetes, particularly type II diabetes. The invention also provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prevention of diabetes, particularly type II diabetes, in a human demonstrating pre-diabetic hyperglycemia or impaired glucose tolerance.

The compounds and compositions of the present invention may be optionally employed in combination with one or more other anti-diabetic agents or anti-hyperglycemic agents, which include, for example, sulfonylureas (e.g. glyburide, glimepiride, glipyride, glipizide, chlorpropamide, gliclazide, glisoxepid, acetohexamide, glibornuride, tolbutamide, tolazamide, carbutamide, gliquidone, glyhexamide, phenbutamide, tolcyclamide, etc.), biguanides (e.g. metformin, phenformin, buformin, etc.), glucagon antagonists (e.g. a peptide or non-peptide glucagon antagonist), glucosidase inhibitors (e.g. acarbose, miglitol, etc.), insulin secetagogues, insulin sensitizers (e.g. troglitazone, rosiglitazone, pioglitazone, etc.) and the like; or anti-obesity agents (e.g. sibutramine, orlistat, etc.) and the like. The compounds and compositions of the present invention and the other anti-diabetic agents or anti-hyperglycemic agents may be administered simultaneously, sequentially or separately. The pharmaceutical compositions of the present invention comprise a compound of

Formula (I), or a pharmaceutically acceptable salt thereof, as an active ingredient, a pharmaceutically acceptable carrier and optionally other therapeutic ingredients or adjuvants. The compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, as well as administration through inhaling, 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 pharmaceutical compositions may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

The pharmaceutical compositions according to the invention are preferably adapted for oral administration.

In practice, the compounds of Formula (I), or pharmaceutically acceptable salts thereof, 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 compounds of Formula (I), or a pharmaceutically acceptable salt thereof, 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. Thus, the pharmaceutical compositions of this invention may include a pharmaceutically acceptable carrier and a compound of Formula (I), or a pharmaceutically acceptable salt thereof. The compounds of Formula (I), or pharmaceutically acceptable salts thereof, can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds. The pharmaceutical compositions of this invention include pharmaceutically acceptable liposomal formulations containing a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

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 or other such excipient. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example, starch, gelatin, or acacia; and lubricating agents, for example, magnesium stearate, stearic acid, or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer time. For example, a time delay material such as glyceryl monostearate, or glyceryl distearate may be used.

In hard gelatin capsules, the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate, or kaolin. In soft gelatin capsules, the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil. 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 cachet or capsule preferably contains from about 0.05mg to about 5g of the active ingredient.

For example, a formulation intended for the 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 between 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 and 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 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 5wt% to about 10wt% of the compound of Formula (I), 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.

Pharmaceutical compositions of this invention can be in a form suitable for inhaled administration. Such administration can be in forms and utilizing carriers described in, for example, 1) Particulate Interactions in Dry Powder Formulations for Inhalation, Xian Zeng et al, 2000, Taylor and Francis, 2) Pharmaceutical Inhalation Aerosol Technology, Anthony Hickey, 1992, Marcel Dekker, 3) Respiratory Drug Delivery, 1990, Editor: P.R. Byron, CRC Press. In addition to the aforementioned carrier ingredients, the pharmaceutical compositions 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 compound of Formula (I), or a pharmaceutically acceptable salt thereof, may also be prepared in powder or liquid concentrate form.

Generally, dosage levels of the order of from about 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 1Og per patient per day. For example, type II diabetes may be effectively treated by the administration of from about 0.01 to lOOmg of the compound per kilogram of body weight per day, or alternatively about 0.5mg to about 7g per patient per day.

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 disease in the particular diabetic patient undergoing therapy. Further, it is understood that the compounds and salts thereof of this invention can be administered at subtherapeutic levels prophylactically in anticipation of a hyperglycemic condition.

The compounds of Formula (I) may exhibit advantageous properties compared to known glucokinase activators, such properties may be illustrated in the assays described herein or in other assays known to those skilled in the art. In particular, compounds of the invention may exhibit improved values for K _1n , V _max , EC ₅ O, maximum activation (glucose concentration = 5mM), maximum blood glucose reduction on basal blood glucose levels and/or reduction of postprandial glucose peak in an oral glucose tolerance test (OGTT), or other advantageous pharmacological properties such as enhanced aqueous solubility and/or enhanced metabolic stability, compared to known GK activators. The compounds of the invention may also demonstrate one or more of the following properties compared to known compounds: reduced neurotoxicity, longer duration of action (e.g. improved half-life/higher plasma protein binding), improved bioavailability, and /or increased potency (e.g. in vitro or in vivo).

EXPERIMENTAL

In accordance with this invention, the compounds of Formula (I) can be prepared following the protocol illustrated in Scheme 1 below:

SCHEME 1

The carboxylic acid II, or an activated derivative thereof, may be condensed with the amine III, or a salt thereof, e.g. the hydrochloride salt, using a variety of coupling conditions known to those skilled in the art. For example, it is possible to condense the enantiopure carboxylic acid II with amine III, or a salt thereof, using a reagent that causes negligible racemisation, e.g. benzotriazol-l-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (J. Coste et al., Tetrahedron Lett., 1990, 31, 205-208), to furnish enantiopure amides of Formula (I). Alternatively the carboxylic acid carboxylic acid II may be treated with (COC1) ₂ and DMF in dichloromethane e.g. at -45°C, followed by the addition of the amine III and pyridine.

Alternatively, a racemic mixture of amides can be prepared from racemic carboxylic acid II and then separated by means of chiral high performance liquid chromatography employing a chiral stationary phase (which can be purchased from, for example, Daicel Chemical Industries, Ltd, Tokyo, Japan) to provide the desired compound of Formula (I).

The amines III are commercially available or are readily prepared using known techniques.

The carboxylic acid II can be prepared following the protocol illustrated in Scheme 2 below (illustrated using the (R)-isomer): SCHEME 2

IV

The compound of formula IV may be converted to the sulfanyl carboxylic acid V by treatment, for example, with aqueous sulfuric acid in dioxane under heating. Conversion of the sulfanyl group to a sulfonyl group may be performed according to methods known to those skilled in the art, for example by oxidation using mCPBA (3-Chloroperoxybenzoic acid) in a solvent such as dichloromethane to provide the sulfonyl carboxylic acid II.

The compound of formula IV can be prepared following the protocol illustrated in Scheme 3 below (illustrated using the (R)-isomer):

SCHEME 3

The reaction of the amide of formula VI with the compound of formula VII may conveniently be performed in a solvent such as dry THF, in the presence of an agent such as Lithium bis(trimethylsilyl)amide.

The compound of formula VII, 7(5>iodomethyl-2(5),3(5>diphenyl-l,4- dioxaspiro[4,4]nonane, may be prepared according to the methods described in

WO2003/095438.

The amide of formula VI may be prepared following the protocol illustrated in Scheme 4 below:

SCHEME 4

The phenyl acetic acid of formula VIII may be reacted with trimethylacteylchloride in a solvent such as acetone and in the presence of potassium carbonate, before the addition of 1 (R),2(R))-(-)-pseudoephedrine to produce the compound of formula VI.

The phenyl acetic acid of formula VIII may be readily prepared by those skilled in the art, for example from ethyl (4-cyclopropylsulfanylphenyl)oxoacetate according to the methods described in WO2004/0181067.

Further details for the preparation of the compounds of Formula (I) are found in the examples.

During the synthesis of the compounds of Formula (I), labile functional groups in the intermediate compounds, e.g. hydroxy, oxo, carboxy and amino groups, may be protected.

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.

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.

EXAMPLES

Abbreviations and acronyms: Ac: Acetyl; tBME: tert-Butylmethylether; ATP: Adenosine 5 '-triphosphate; DMF: Dimethylformamide; Et: Ethyl; GK: Glucokinase; GIc:

Glucose; G6P: Glucose-6-phosphate; G6PDH: Glucose-6-phosphate dehydrogenase; GST- GK: Glutathione S-transferase-Glucokinase fusion protein; NADP(H): β-Nicotinamide adenine dinucleotide phosphate (reduced); rt: Room temperature.

Preparation 1: (4-Cyclopropylsulfanylphenyl)oxoacetic acid

2M aqueous NaOH (163mL) was added to a solution of ethyl (4- cyclopropylsulfanylphenyl)oxoacetate (40.62g, 162.5mmol) in EtOH (20OmL) and the stirred mixture heated at 60 ⁰ C for 2h. After cooling, the mixture was concentrated to 15OmL and washed with ether (2xl00mL). Sufficient concentrated HCl was then added to adjust the pH to 1 and the resulting precipitate was extracted into EtOAc (2x300mL). The combined organic phases were washed with water (3xl00mL), brine (20OmL) and dried (MgSOzi). Removal of the solvent afforded the title compound: mlz (ES ^" ) = 221.0 [M- H ⁺ ] ^" .

Preparation 2: (4-Cyclopropylsulfanylphenyl)acetic acid

Hydrazine hydrate (14.19g, 283.5mmol) was cooled to -50 ⁰ C and (4- cyclopropylsulfanylphenyl)oxoacetic acid (Preparation 1, 12.6g, 56.7mmol) added in one portion. The vigorously-stirred slurry was warmed firstly to rt and then at 80 ⁰ C for 5min. Solid KOH (8.76g, 156.5mmol) was added in four equal portions and the resulting solution heated at 100 ⁰ C for 2Oh. On cooling to room temperature, water (25mL) was added and the aqueous phase washed with Et ₂ O (2OmL). The ethereal phase was itself washed with water (2x15mL) and sufficient concentrated HCl added to the combined aqueous phases to adjust the pH to 1. The resulting precipitate was then extracted into EtOAc (2x30OmL) and the combined organic phases washed with water (3x10OmL), brine (20OmL) then dried (MgSOzi).

Evaporation of the solvent afforded the title compound: mlz (ES ^" ) = 207.1 [M- H ⁺ ] ^" .

Preparation 3 : 2-(4-Cyclopropylsulfanylphenyl)-λ ^/ -(2(λ)-hydroxy-l (λ)-methyl-2- phenylethylJ-λ ^f -methylacetamide

Anhydrous acetone (148mL) was added to (4-cyclopropylsulfanylphenyl)acetic acid

(Preparation 2, 16.41g, 78.8mmol) and K ₂ CO ₃ (32.67g, 236.4mmol) to form a slurry which was cooled to -10 ⁰ C with stirring. Neat trimethylacetyl chloride (10.2mL, 82.74mmol) was

introduced dropwise, ensuring the temperature did not exceed -10 ⁰ C during the addition. The reaction mixture was stirred at -10 ⁰ C for 20min, warmed to 0 ⁰ C for 20min then cooled to -15°C and solid (l(R),2(R))-(-)-pseudoephedrine (19.53g, 118.2mmol) was added in one portion. After 1 Omin, the reaction mixture was brought to rt, where stirring was continued for 1.5h. Water (10OmL) was added and the mixture extracted with EtOAc (50OmL). The organic phase was washed with water (2xl00mL) and the combined aqueous layers back-extracted with EtOAc (2x250mL). The combined organic layers were then washed with brine (10OmL) and dried (MgSOzi). The solvent was removed and the solid yellow residue recrystallized from EtOAc-IH to afford the title compound: mlz (ES ⁺ ) = 356.1 [M + H] ⁺ .

Preparation 4: 2(λ)-(4-Cyclopropylsulfanylphenyl)-3-(3(λ)-oxocyclopentyl) propionic acid

LHMDS (162mL of a IM solution in THF, 162mmol) was diluted with anhydrous

THF (16ImL) and cooled to -20 ⁰ C with stirring. A solution of 2-(4- cyclopropylsulfanylphenyl)-N-(2(R)-hydroxy-l(R)-methyl-2-phe nylethyl)-N-methylacetamide (Preparation 3, 3Og, 84.4mmol) in anhydrous THF (245mL) was added via cannula over lOmin, ensuring the reaction temperature remained below -15°C throughout the addition. The reaction was allowed to warm to -7°C over 30min then cooled to -12°C and a solution of 7(5)- iodomethyl-2(5),3(5)-diphenyl-l,4-dioxaspiro[4,4]nonane (27g, 64.2mmol) in a mixture of anhydrous THF (11 ImL) and DMPU (18.9ml) added via cannula over lOmin, ensuring the reaction temperature remained below -7°C throughout. The reaction was warmed to 2°C and stirred for 4.5h before being poured into a mixture of toluene (77OmL) and 20% aqueous NH ₄ Cl (55OmL). After stirring vigorously, the organic layer was separated and washed with

20% aqueous NH ₄ Cl (55OmL) and brine (10OmL). The aqueous phases were combined and extracted with EtOAc (50OmL) which, after separation, was washed with brine (10OmL). The combined organic phases were dried (MgSO ₄ ), filtered, evaporated and the resulting oil purified by flash chromatography (IH-EtOAc, 9:1 changing incrementally to 1:1) to afford 2(R)-(4-cyclopropylsulfanylphenyl)-3-(2(5),3(5)-diphenyl-l,4 -dioxaspiro[4.4]non-7(R)-yl)-N-

(2(R)-hydroxy-l(R)-methyl-2-phenylethyl)-N-methylpropionamid e: mlz (ES ⁺ ) = 648.3 [M + H] ⁺ . A stirred solution of this amide (30.7g, 47.38mmol) in 1,4-dioxane (62mL) was diluted with 4.5M aqueous H ₂ SO ₄ (61.5mL) and the resulting mixture heated under gentle reflux for 18h. After cooling on ice, water (162mL) was added and the mixture extracted with EtOAc (25OmL). The aqueous layer was separated and extracted further with EtOAc (2x15OmL) and the combined organic phases washed with water (3x200mL), ensuring the final wash was pH neutral, and brine (10OmL). After drying (MgSO ₄ ) and filtering, the solvent was removed and the residue purified by flash chromatography (CH ₂ Cl ₂ then CH ₂ Cl ₂ -THF, 5:1 changing to 3:1) to afford the title compound: mlz (ES ⁺ ) = 305.1 [M+ H] ⁺ .

Preparation 5: 2(λ)-(4-Cyclopropanesulfonylphenyl)-3-(3(λ)-oxocyclopentyl )propionic acid

A stirred solution of 2(R)-(4-cyclopropylsulfanylphenyl)-3-(3(R)- oxocyclopentyl)propionic acid (Preparation 4, 5.Og, 16.43mmol) in CH ₂ CI ₂ (25OmL) was cooled to 1°C on ice and 70% mCPBA (8.099g, 32.85mmol) added portionwise, maintaining the temperature below 3°C. After 6h the solvent was removed and the residue purified by flash chromatography (1 %ACOH in CH ₂ CI ₂ then THF) to afford the title compound: mlz (ES ⁺ ) = 337.1 [M + H] ⁺ .

Examples

2(R)-(4-Cyclopropanesulfonylphenyl)-3-(3(R)-oxocyclopenty l)propionic acid (Preparation 5) was coupled with amines selected from 2-amino-5-methylpyrazine, 2-amino- 5-methylpyridine, 3-aminoisoxazole, 2-amino-5-methylthiazole and 4-aminopyrimidine using the following procedure to provide Examples 1-5.

CH ₂ Cl ₂ (6OmL) and DMF (0.08mL, 1.064mmol, 1.2 eq) were cooled to -10 ⁰ C and oxalylchloride slowly added (0.09mL, 0.465mol, 1.2 eq). After stirring for 15 min the reaction mixture was cooled to -30 ⁰ C and (2R)-2-(4-cyclopropanesulfonylphenyl)-3- (tetrahydropyran-4-yl)propionic acid (Preparation 8, 0.300g, 0.886mmol, 1.0 eq) was added. The reaction was stirred at -30 ⁰ C for 45min then pyridine (1.395mol, 0.3ImL in ImL CH ₂ Cl ₂ , 4.5eq) and the amine (between 1.2 and 5.0eq) were slowly added in parallel at -40 ⁰ C. The reaction mixture was stirred for 15min then the ice bath removed. The reaction mixture was stirred for 2h until it reached rt. The solvent was removed under partial vacuum and the crude mixture dissolved in EtOAc (1OmL) and aqueous HCl (1.5mL). The layers were separated and the aqueous phase extracted with EtOAc (5mL). The organic fractions were combined and washed with H ₂ O (1OmL), saturated aqueous NaHCθ3 (2 x 10 mL), water (5mL) and brine (5mL) and dried (Mg ₂ SOzi). Purification was by flash chromatography (EtOAc:heptane, 2:1) and/or recrystallisation.

2(R)-(A- δ _c : 1.4, 6.5, 30.1, 33.3, 35.8, 38.9,

Cyclopropane- 39.3, 45.2, 52.2, 128.7, 129.7, sulfonylphenyl)-N- 129.8, 131.5, 136.6, 140.7, 143.9,

(5 -methylpyridin- 147.2, 150.3, 174.0, 219.9

2-yl)-3-(3(R)- ES ⁺ = 427 [M+H] ⁺ oxocyclopentyl)- propionamide

2(R)-(A- δ _H : 0.90-1.00 (m, 2H), 1.18-1.30 Cyclopropane- (m, 3H), 1.46-1.58 (m, IH), 1.80- sulfonylphenyl)-N- 2.45 (m, 8H), 3.75-3.85 (m, IH), (isoxazol-3-yl)-3- 7.00 (d, IH), 7.53 (d, 2H), 7.80 (d,

Q(R)- 2H), 8.30 (d, IH), 10.33 (s, IH) oxocyclopentyl)- ES ⁺ = 425 [M+Na] ⁺ propionamide

2(R)-(A- δ _H : 0.80-1.30 (m, 6H), 1.45-1.60

Cyclopropane- (m, IH), 1.75-1.85 (m, IH), 1.98- sulfonylphenyl)-N- 2.18 (m, 3H), 2.22-2.40 (m, 6H),

(5-methylthiazol- 3.70-3.80 (m, IH), 7.05 (d, IH),

2-yl)-3-(3(R)- 7.41 (d, 2H), 7.75 (d, 2H) oxocyclopentyl)- ES ⁺ = 433 [M+H] ⁺ propionamide

2(R)-(A- δ _H : 0.95-1.05 (m, 2H), 1.23-1.32

Cyclopropane- (m, 3H), 1.45-1.60 (m, IH), 1.80- sulfonylphenyl)-N- 2.45 (m, 8H), 3.85-3.95 (m, IH),

(pyrimidin-4-yl)- 7.54 (d, 2H), 7.83 (d, 2H), 8.19 (d,

3-(3(R)- IH), 8.59 (d, IH), 8.95 (s, IH), 9.12 oxocyclopentyl)- (br s, IH) propionamide ES ^' = 412 [M-Hf

2(R)-(4-Cyclopropanesulfonylphenyl)-3-(3(R)-oxocyclopenty l)propionic acid (Preparation 5) may also be coupled with amines selected from 2-amino-5-chloropyridine, 2- aminopyridine and 3-amino-5-methylisoxazole using the procedure described above to provide Examples 6-8.

ASSAYS

In vitro GK activity

Using a protocol similar to that described in WO2000/58293, GK activity was measured by coupling the production of G6P by GST-GK to the generation of NADH with G6PDH as the coupling enzyme.

The assay was performed at room temperature (23 ⁰ C) in clear flat bottom 96-well plates in a total volume of lOOμl consisting of 25mM Hepes (pH 7.4), 25mM KCl, 5mM D- glucose, ImM ATP, ImM NADP, 2mM MgCl ₂ , ImM dithiothreitol, 0.2μg purified GST-GK derived from human liver GK and a range of activator concentrations in a final concentration of 5 % DMSO. The incubation time was 15min at which time the reaction has been shown to be linear. The generation of NADH, as an indirect determination of GK activity, was measured at OD ₃₄₀ in a SpectraMAX 190 microplate spectrophotometer (Molecular Devices Corp).

Typically compounds were tested over a range of 10 dilutions from lOOμM to 0.004μM in a final DMSO concentration of 5%. The degree of activation was calculated as a ratio over a control reaction with 5% DMSO only. Values quoted represent the concentration of compound required to produce a 2-fold activation of GK derived from a dose response curve constructed using a 4-parameter logistic model. Additionally, maximum fold activation and an EC ₅ O (concentration required to produce half the maximum fold activation) was calculated from the same dose response curve.

Representative examples of the compounds of Formula (I) had an EC ₅ Q of <500nM.

In vivo GK activity (I)

Following a 4.5 h fasting period, C57BL/6 mice were dosed orally via gavage with GK activator at lOmg/kg body weight followed by a glucose load of 2 g/kg. Blood GIc determinations were made 3 times during the 2.5h post-dose study period.

Mice (n = 9) were weighed and fasted for 4.5h before oral treatment. GK activators were dissolved in Gelucire 44/14-water (1 :9 v/v) at a concentration of lmg/mL. Mice were dosed orally with 1OmL formulation per kg of body weight to equal a 10mg/kg dose. Fifteen min prior to dosing, a pre-dose blood GIc reading was acquired by snipping off a small portion of the animals' tails (<lmm) and collecting 20μL blood for analysis. After GK

activator treatment, further blood GIc readings were taken at 0.5, 1.0, and 2.5h post-dose from the same tail wound. Results were interpreted by comparing the mean blood GIc values of the vehicle treated mice with the the GK activator treated mice over the study duration. Representative examples of the compounds of Formula (I) exhibited a statistically significant decrease in blood GIc compared to vehicle for 2 consecutive assay time points following compound administration.

In vivo GK activity (II)

The antihyperglycaemic effects of examples of the GK activators of the invention were evaluated in oral glucose tolerance tests in 7-8 week old male C57B1/6 oblob mice.

Briefly, mice (n = 6) were weighed and their basal blood glucose levels determined from 20μL of blood withdrawn from a tail cut (T - 27h). After 22h (T - 5h), food was removed and the mice were placed in fresh cages with access to water ad libitum. The blood glucose levels were determined at T - 0.75h from 20μL of blood withdrawn from the tail wound. The GK activators were dissolved in a Gelucire 44/14-water (1 :9 v/v) mixture at a concentration of lmg/mL, then, at T - 0.5h, the mice were dosed orally with 1OmL formulation per kg of body weight to equal a lOmg/kg dose. At T = 0 h, the mice were bled (20μL) for analysis of blood glucose levels, then immediately dosed orally with glucose (2g/kg). Further blood samples (20μL) were taken from each animal at T = +0.5, +1.0, +1.5, +2.0, +3.0, and +4.Oh for the analysis of glucose levels. Representative examples of the compounds of Formula (I) typically reduced the area under the glucose curve by at least 20% in the 2h following administration of glucose.