Title Selective β3 Adrenergic Agonists

Field of Invention The present invention is m the field of medicine, particularly in the treatment of Type II diabetes and obesity. More specifically, the present invention relates to selective β3 adrenergic receptor agonists useful in the treatment of Type II diabetes and obesity.

Background of the Invention The current preferred treatment for Type II, non-insulm dependent diabetes as well as obesity is diet and exercise, with a view toward weight reduction and improved insulin sensitivity. Patient compliance, however, is usually poor. The problem is compounded by the fact that there are currently no approved medications that adequately treat either Type II diabetes or obesity. The invention described herein is directed toward an effective and timely treatment for these serious diseases.

One therapeutic opportunity that has recently been recognized involves the relationship between adrenergic receptor stimulation and anti-hyperglycemic effects. Compounds that act as β3 receptor agonists have been shown to exhibit a marked effect on lipolysis, thermogenesis and serum glucose levels m animal models of Type II (non-insulin dependent) diabetes.

The β3 receptor, which is found in several types of human tissue including human fat tissue, has roughly 50% homology to the βi and β2 receptor subtypes yet is considerably less abundant. The importance of the β3 receptor ι=; a relatively recent discovery since the ammo- acid sequence of the human receptor was only elucidated m the late 1980 's. A large number of publications have appeared m recent years reporting success m discovery of agents that stimulate the β3 receptor. Despite these recent developments, there remains a need to develop a

selective β3 receptor agonist which has minimal agonist activity against the βi and β2 receptors.

The present invention provides compounds which are selective β3 receptor agonists. As such, the compounds effectively lead to an increase in insulin sensitivity and are useful in treating Type II diabetes and other ailments implicated by the β3 receptor, without cardiac or tremor- related side effects.

Summary of Invention

The present invention provides compounds of the Formula I :

(I) wherein:

Rl is OH, halo, SO2NHR2, C02R2- CONHR2, NHCOR2 , -NH(optionally substituted aryl), CF3 , or CF2H;

Rl ' is H, halo, C1-C4 alkyl, OH, SO2NHR2 , C02R2- CONHR2, NHCOR2, CF3 or CF2H;

R2 is H, C1-C4 alkyl, or aryl;

R3 is H or C1-C4 alkyl;

R4 is a moiety selected from the group consisting of:

R5 and Rβ are independently C1-C4 alkyl; R7 is an optionally substituted heterocycle or a group selected from the group consisting of:

R8 is independently H, halo or C1-C4 alkyl; R9 is halo, CN, OR10, C1-C4 alkyl, C1-C4 haloalkyl, CO2R2 , CONR11R12 , CONH(Cι-C4 alkyl or C1-C4 alkoxy) , SR2, CSNR2, CSNR11R12, SO2R2 - SO2 R11R12, SOR2, NR11R12, aryl, heterocycle, optionally substituted aryl, optionally substituted heterocycle, or C1-C4 alkyl or C2-C4 alkenyl optionally substituted with CN;

RlO is independently C1-C4 alkyl, C1-C4 haloalkyl, (CH2) _n C3-C8 cycloalkyl, (CH2. _n aryl,

(CH2)nheterocycle, (CH2)n optionally substituted C3-C8 cycloalkyl, (CH2)n optionally substituted aryl, or (CH2)n optionally substituted heterocycle;

RU and R12 are independently H, C1-C4 alkyl, or combine with the nitrogen to which each are bound to form morpholinyl, piperidinyl, pyrrolidinyl, or piperazinyl; Xl is 0 or S;

X2 is absent or a 1 to 5 carbon straight or branched alkylene; m is 0 or 1; n is 0, 1, 2, or 3; o is l, 2, 3, 4, 5, or 6; or a pharmaceutically acceptable salt or solvate thereof.

The present invention also provides a novel process for making compounds of Formula I.

The compounds of the present invention are selective β3 receptor agonists and as such are useful for

treating Type II diabetes and obesity, as well as useful for stimulating the β3 receptor. Therefore, the present invention also provides for methods of treating Type II diabetes and obesity, as well as a method of stimulating the β3 receptor.

In addition, the present invention provides the use of compounds of Formula I for treating Type II diabetes and obesity as well the use of compounds of Formula I for stimulating the β3 receptor.

Detailed Description

For the purposes of the present invention, as disclosed and claimed herein, the following terms are defined below. As they relate to the present invention, the terms below may not be interpreted, individually or collectively, to describe chemical structures that are unstable or impossible to construct.

The term "halo" represents fluorine, chlorine, bromine, or iodine. The term "C1-C4 alkyl" represents a cyclo, straight cr branched chain alkyl group having from one to four carbon atoms such as methyl, ethyl, -propyl, isopropyi. cyclopropyi , n-butyl, isooutyi , sec-butyl, t- butyl and the like. A "haloalkyl" is one sucn aiκyl substituted with one or more halo atoms, preferably one to three halo atoms. An example of a haloalkyl is trifluoromethyi . An "alkoxy" is a alkyl group covalently bonded by an -O- linkage.

The term "1 to 5 carbon straight or branched alkylene" represents a one to five carbon, straight or branched, alkylene moiety. A branched alkylene may have one or more points of branching. A 1 co 5 carbon straignt or branched alkylene may optionally be unsaturated at one or more carbons. Thus, a 1 to 5 carbon straight or branched alkylene includes 1 to 5 carbon alkylene, alkenylene and alkylidene moieties. Examples include methylene, ethylene, propylene, butylene, -CH(CH3 )CH2-

CH(C2H5)CH2-, -CH(CH3 )CH(CH3 ) - , -CH2C (CH3 ) 2~ ■ -CH2CH(CH3)CH2", -C(CH3)2CH=, -CH=CHCH2~, -CH=CH- , and the like.

The "acyl" moiety, alone or in combination, is derived from an alkanoic acid containing from one to seven carbon atoms. The term "acyl" also includes moieties derived from an aryl carboxylic acid.

The term "aryl" represents an optionally substituted or unsubstituted phenyl or naphthyl . The term (CH2)n ^ar "yl ^1S preferably benzyl or phenyl.

The term "optionally substituted" as used herein means an optional substitution of one to three, preferably one or two groups independently selected from halo, C1-C4 haloalky _^ , hyαroxy, carboxy, tetrazoly__, acyl, COOR2 , CONR11R12, CONH(C -C4 alkoxy) , cyano, C1-C4 alkoxy, C1-C4 alkyl, pnenyl, benzyl, nitro, NR 1R12, NHCO.C1-C4 alkyl) , NHCO (benzyl) , NHCO (phenyl) , SR2 , S.C1-C4 alkyl) , OCO(Cι-C4 alkyl) , SO2 ( R11R12 ) > SO2 (C1-C4 alkyl ¹ , or SO2 (phenyl) ; provided that sucn substitution does not entirely destroy biological activity, as defined m this speci ication.

R and R12 are independently H, C1-C4 alkyl, or combine with the nitrogen to whicn each is bound to form morpnolinyl, piperidmyl, pyrronαinyi, or piperazmyl The term "heterocycle" represents a stable, optionally substituted or unsuostituteα, saturated or unsaturated 5 or 6 membered ring, said ring having from one to four heteroatoms that are the same or different and that are selected from the group consisting of sulfur, oxygen, and nitrogen; and when heterocycle contains two adjacent carbon atoms, the ad acent carbon atoms may be structured to form a group of the formula -CH=CH-; provided that (1^ when the neterocyclic ring contains 5 members, the heteroatoms comprise not more tnan two sulfur or two oxygen atoms but not both; and (2) when tne heterocyclic ring contains 6 members and is aromatic, sulfur ana oxygen are not present. The heterocycle may be attached at any carbon or nitrogen wnicn affords a stable structure. The

heterocycle may oe optionally substituted. Examples of an heterocycle include pyrazole, pyrazolme, lmiαazole, isoxazole, tπazole, tetrazole, oxazole, 1, 3-dioxolone, thiazole, oxadiazole, thiadiazole, pyridme, pyrimidine, piperazme, morpholme, pyrazme, pyrrolidine, piperidine, oxazolidone, oxazolidinedione, lmidazolidmone.

The term "leaving group" as used in the specification is understood by those skilled in the art. Generally, a leaving group is any group or atom that enhances the electrophilicity of the atom to which it is attached for displacement. Preferred leaving groups are p- nitrobenzene sulfonate, triflate, mesylate, tosylate, imidate, chloride, bromide, and iodide.

The term "pharmaceutically effective amount", as used herein, represents an amount of a compound of the invention that is capable of stimulating the β3 receptor mammals. The particular dose of the compound administered according to this invention will, of course, be determined by the particular circumstances surrounding the patient, including the compound administered, tne route of administration, the particular condition being treated, and similar considerations.

The term "unit dosage rorm" refers to pnyεicall" discrete units suitable as unitary αosages for numan subjects and other mammals, eacn unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, association with a suitable pharmaceutical carrier.

The term "treating, " as used herein, describes the management and care of a patient for the purpose of combating the disease, condition, or disorder and includes the administration of a compound of present invention to prevent the onset of the symptoms or complications, to alleviate symptoms or complications, or to eliminate the disease, condition, or disorder.

The term "selective" means preferential agonism of the β3 receptor over agonism of tne βi or β2 receptor.

In general, the compounds of the present invention demonstrate at a minimum a twenty fold differential (preferably over a 50x differential) the dosage required to behave as an agonist to the β3 receptor and the dosage required for equal agonism of the β and β2 as measured m the Functional Agonist Assay. The compounds demonstrate this differential across the range of doses. Thus, β3 selective compounds behave as agonists for the β3 receptor at much lower concentrations with lower toxicity by virtue of their minimal agonism of the other receptors.

As previously noteα, the present invention provides compounαs of the Formula I.

Preferred compounds are those of Formula la:

H (la) wherei :

Rl is F, OH, -S02NR2 -ITHCOR; :F2H;

R»

R8 is independently H, halo or C1-C4 alkyl;

R9 is OR10, tetrazolyi, CONR11R 2 , or SO2 R11R12;

RlO is ⁽ CH2>n ^ar yl _' ⁽ CH2 ; _n heterocycle, said aryl or heterocycle being optionally substituted with tetrazolyi, CN, O2R2. CONR11R1 , or ΞO2 P 1R12;

Rll and R12 are independently H, C1-C4 alkyl, or combine with the nitrogen to whicn each is oound to form morphol yl, piperidmyl, pyrrolidmyl or piperazmyl;

X2 is 1 to 2 carbon alkylene; n is 0 , 1 , 2 , or 3;

or a pharmaceutically acceptable salt

Other preferred compounds are those of Formula lb:

(lb) wherein:

Rl is F, -SO2NR2, -NHCOR2, or CF2H;

P.7 is an optionally substituted heterocycle or

R8 is independently H, halo or C1-C4 alkyl;

R9 is OR10, CO R11R12 , or SO2 R11R12;

RlO is independently ⁽ CH2)n ^ar Yl- (CH2 ) nheterocycle, said aryl or heterocycle being optionally substituted with tetrazolyi, CN, O2R2 , CONR11R12, or ΞO2NR11R12;

Rll and R12 are independently H, C -C4 alkyl, c: combine with the nitrogen 10 which each is bound ;o form morpholinyl, piperdinyi, pyrrolid yi or piperaz yl ; n is 0, 1, or j o is , 3, 4, or t> ; or a pharmaceutically acceptable salt or solvate thereof.

Additional preferred compounds are those of the Formula Ic:

(Ic) wnerem:

Rl is F. -S02NR2, -NHCOR2 , or CF2H; R9 is ORl0» C0 R11R12- SO2NR11R12 , S0R2 , NR11R12, optionally substituted aryl, optionally substituted aryloxy, or optionally substituted heterocycle; Rio is (CH2)naryl, (CH2)nheterocycle, said aryl or heterocycle being optionally substituted with tetrazolyi, CONR11R12 , or SO2 R11R12;

Rll and R12 are independently H, C1-C4 alkyl, or combine with the nitrogen to which each is bound to form morpholinyl, piperidinyl, pyrrolidinyl, or piperazinyl; Xl is 0 or S;

or a pharmaceutically acceptable salt or solvate thereof.

Particularly preferred compounds are those of Formula Ia, lb or Ic, wherein RG is CONH2 and OR10, wherein RlO is a optionally substituted aryl, particularly phenyl, or optionally substutituted heterocycle, particularly pyridine.

Additional preferred compounds include the following :

(All lsomers of: ) 5-{3- [2-hydroxy-3- (2-fluorophenyloxy • - propyiammo] -2-methylbutyl} -thiophene-2-suifonamide

(All lsomers of: ) 5- (2-fluoro-4- {3- [2-hydroxy-3- (2- fluorophenyloxy) -propylamino] -2-methylbutyl} -phenyl) -1H- tetrazole

(All isomers of:) 4- {3- [2-hydroxy-3- (3-hydroxyphenyioxy) - propyiammo] -2-methylbutyl}benzamide

(All isomers of: ! 4-{3-[2-hydroxy-3- ^■ :2-fluorophenyloxy ι - propylamino] -2-methylbutyl}-3-methylbenza ide

(All isomers of:) 4-{3- [2-hydroxy-3- (3-acetylammo- phenyloxy) -propylamino] -2-methylbutyl}benzamide

By virtue of their acidic moieties, some of the compounds of Formula I include the pharmaceutically acceptable base addition salts thereof. Such salts include those derived from inorganic bases such as ammonium and alkali and alkaline earth metal hydroxides, carbonates, bicarbonates, and the like, as well as salts derived from basic organic amines such as aliphatic and aromatic amines, aliphatic diam es, hydroxy alka ineε, and the like. Such bases useful preparing the salts of this invention thus include ammonium hydroxide, potassium caroonate, sodium bicaroonate, calcium hydroxide, metnylamme, diethylamme, ethylenediamme, cyclohexylam e, ethano amme and the like. Because of the basic moiety , some of the compounds of Formula I can also exist as pharmaceutically acceptable acid addition salts. Acids commonly employed to form such salts include inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, as well as organic acids such as para- toluenesulfonic, methanesulfonic, oxalic, para- promophenyisulfonic, carbonic, succ ic, citric, benzoic, acetic acid, and related inorganic and organic aciαs Ξucn pharmaceutically acceptable salts tnus include εulfate, pyrosulfate, biεulfate, sulfite, oisulfite, pnosphate, mono-hydrogenphosphate, dihydrogenphospnate, metaphosphate, pyropnosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, 2-butyne- 1,4 dioate, 3-hexyne-2, 5-dιoate, oenzoate, chlorooenzoate, hydroxybenzoate, methoxybenzoate, phthalate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hippurate, β- hydroxybutyrate, glycollate, maleate, cartrate, methanesulfonate, propanesulfonate, naphthalene-1-

εulfonate, naphtnalene-2-εulfonate, mandelate and tne like salts.

It is recognized that various stereoisomic forms of the compounds of Formulas I may exist. The compounds may be prepared as racemates and can be conveniently used as such. Therefore, the racemates, individual enantiomerε, diastereomers, or mixtures thereof form part of the present invention. Unless otherwise specified, whenever a compound is described or referenced this specification all the racemates, individual enantiomers, diastereomers, or mixtures thereof are included in said reference or description.

The compounαs of Formula I are prepared as described m tne following Schemes and examples.

Scheme I

ci :ι:

In Scheme I, Pi, P ' and P nave the same meaning aε previously described. The reaction of Scheme I is carried out under conditions appreciated in the art for the animation of epoxides . For example, the epoxide (Hi may be combined with the amme (III m an alcohol, preferably, ethanol at room temperature to the reflux temperature of the reaction mixture. Preferably, the reaction is carried out under conditions generally described m Atkins et al. , Tetrahedron Lett. 27:2451 (1986, . These conditions include mixing the reagents the presence of tπmethylsilylacetamide m a polar aprotic so_vent such aε acetonitrile, dimethylformamide (DMF> , acetone, dimethylsulfoxide (DMSO) , dioxane, diethylene glycol methyl ether (diglyme) , tetrahydrofuran 'THF) , or other polar

aprotic solvents which the reagents are soluble. Preferably, the solvent is DMSC. The reaction lε carried out at temperatures ranging from about 0 ^* C to reflux.

The compounds of the present invention can be prepared by a novel combinatorial/parallel synthesis. This synthesis s described m Scheme II.

Scheme II

Formula I

IV) (V)

In Scheme II, Ri , Rl ' , R2, and R4 have the same meaning as previously described. The reaction of Scheme II is preferably carried out by adding to a glass vial: a non-reactive solvent such as methanol, DMF, methylene chloride or acetomtrile, amine (IV) , and ketone (V) . The solution is shaken to allow for imme formation and treated with Amberlite IRA400 borohydride resin (Aldπch.. The slurry is then shaken an additional 24 hours to effect reduction to the secondary am e . Methylene chloride and polystyrene-linked benzaidehyde resin 'Frecnet, J.M. et al., ..T. Am Chem. Soc. .22:492 (1971 is added to the vial, m order to scavenge excess primary amme starting material. The slurry is shaken, preferably overnight. The slurry is then filtered through a cotton plug, and the residual solids rinsed with methanol. Evaporation under a flow of air, followed by drying for several hours at room temperature in a vacuum oven yields the deεired product of sufficient purity.

A modification of Scheme II is necessary when the amme hydrochloride salt is used. Addition of resin- bound base to the initial reaction mixture prior to reduction or εcavenging allows the desired reaction to proceed. Imme formation using amme hydrochloride salts,

an aldehyde or ketone, ana a resin bound amme base may be carried out using two different resins: poly(4- vmylpyrid e) , commercially available from Aldrich, and resin (VIII) , synthesized by the reaction of Memfield resin with piperidine (Scheme Ila) :

Scheme Ila

(VI) (VII: (VIII)

In Scheme Ila, PS is polysytrene. Both the poly (4- vmylpyridmeJ and resin (VIII) promote imme formation.

Scheme II can also be carried out by utilization of traditional techniques. Reductive animations described in scheme II are well known in the art. T e are typically performed by mixing the amme and ketone starting materials in a solvent and adding a reducing agent. Solvents typically include lower alcoholε, DMF, and the like. A wide variety of reducing agentε can be utilized, moεt commonly utilized are sodium ooronydride and sodium cyanoborohydride . The reaction is typically performed at room temperature to the reflux temperature of the solvent. Products are isolated by techniques well Known the art. Many of the ketone and ammo starting materials utilized Scheme II can be prepared by techniques recognized and appreciated to one skilled m the art. The syntheεiε of additional starting materials is generally described Schemes III and IV.

Scheme III

: ιx )

(XII

In Scheme III, Xi, Ri ' and P.i are the same as previously defined. Equimoiar amounts of the aromatic compound (Compound IX) and (2S) - (÷ . -glycidyl 3- nitrobenzenesulfonate (Compound X) are dissolved in an inert solvent such as acetone and treated with 1.1 equivalents of a non-reactive acid scavenger, such as K2CO3. The suεpenεion lε then heated at reflux for 16-20 hours with stirring. The solvent is removed in vac c . The residue is partitioned between chloroform or other organic solvent and water. The organic layer is dried over Na2S04 and concentrated in vacua to give the epoxi e (XI r. sufficient purity (-95%) and yield (85-100%) . The epoxide (XI, is dissolved in an alcohol, preferably methanol, and treated with one equivalent cf dibenzylamine. The solution is preferably stirred at reflux for three to four hours and then cooled to ambient temperature. Approximately 10 equivalents cf ammonium formate are added to the flask, followed by 10% palladium on carbon, and the suspension stirred vigorously at reflux for 30-45 minutes. The reaction mixture ic then filtered through Celite, concentrated in vacua to a minimum volume and treated with 1.1 equivalents cf a 1.0 M anhydrous solution of HCl in ether. The solution is concentrated to drynesε. The solid residue is triturated with pentane to

yield products of sufficient purity ι^97%) and yieJd i 60- 100%) . If desired, further purification may be carried out by passing over a short plug of silica, eluting with CHCI3, then 95:5 CHCl3/MeOH, then 25:5:1 CHCI3 /MeOH/NH4θH. Alternatively, the epoxide (XI) is treated with a solution of methanol saturated with ammonia gas and stirred at room temperature in a sealed tube for 16 hours. This solution is then evaporated, and the residue subjected to standard purifications such as column chromatography or recryεtallization. The HCl salt is then optionally produced by the addition of HCl gas in ether.

The reaction of Scheme III is further described m Beedie et al . , U.S. patent 5,013,761 and reference cited therein. U.S. patent 5,013,751 s herein incorporated by reference .

The ketone moieties utilized in Scheme II that are either unknown in the art or not commercially available are prepared in accordance with Scheme IV.

Scheme IV

In Scheme IV, X_2 , F-4 > P-5. F. ,, and PT are the same as previously defined. Preferably, R4 s a substituted phenyl. The reaction described in Scheme IV is referred to as a Heck reaction and is described A.J. Chalk et al . , . Orσ. Chem. 41: 1206 (1976 . The reaction is achieved by treating compound (XIII) with an arylpalladium reagent. The aryipalladium reagent is generated in situ by treating Compound (XIV' with a palladium- riarylphosphine complex. The reaction is generally carried out in under conditions appreciated m the ar .

Another embodiment of the current invention is a process of preparing a compound cf Formula I which comprises :

In step 1, reacting an epoxide of the formula:

with an amme cf formula (B)

H-_κ\

and in step 2, reacting the produ: cr step _ :o rorm an acid addition salt.

Starting materials for the compounds described in Schemes I, II, III, and IV are either commercially available, known in the art, or can be prepared by methods known in the art cr described herein.

The following examples and preparations are provided merely to further illustrate the invention. The scope cf the invention iε not conεtrueα as merely consisting of tne following examples. In tne following examples and preparations, melting point, nuclear magnetic resonance spectra, masε spectra, high pressure liquid chromatography over silica gel, gas chromatography, N,N- dimethylformamide, palladium on charcoal, tetrahydrofuran, ethyl acetate, thin layer chromatography and elemental analysis are abbreviated M.Pt., NMR, MS, HPLC, GC, DMF, Pd/C, THF, EtOAc , TLC and EA respectively. The terms "EA" , "NMR", and "MS" indicate that the data was consistent with the desired structure.

^■ 1 ^"

Preparation 1 .S) -3- (2-fluorophenyl) -1 , 2-epoxypropane

A solution of 2-fluorophenol (865 mg,

7.72mol) and (2S) - ( _* ) -glycιdyl-3- nitrobenzeneεulfonate (2.0 g, 7.72 mmol) m 50 mL of acetone waε treated with 1.1 equivalentε of K2CO3 (1.17 g, 8.5 mmol) and stirred at reflux for 18 hours. The suspenεion waε cooled to ambient temperature, the εoiidε filtered, and the filtrate concentrated in vacuo to dryneεε. The resulting solids were partitioned between chloroform and water, and the aqueous layer extracted once with chloroform. The organic layers were combined and dried over Na2S04 and concentrated in vacuo to a colorlesε oil which slowly crystallized at 0"C to 1.11 g (86%) white needleε. TLC (Rf = 0.5, CHCI3 , and NMR indicated >95% purity, so the material was used without further purification.

Preparation 2

(S) (2-fluorophenyloxy; -2-hydroxypropylam onium chloride

S-3- (2-fluorophenyl ■ -1, 2-epoxypropane '1.08 g, 6.4 mmol) was disεolved in 50 mL of methanol and treated with dibenzylamine (1.22 , 6.4 mmol, d=1.026) . The mixture waε stirred at reflux for 3 hours and then cooled to ambient temperature. A vast excess

of ammonium formate (3.0 g, 47.6 mmol) waε added followed by 10% palladium on carbon (350 mg) , and the suspension was stirred at reflux for 45 minutes. After cooling the suspension, tne reaction mixture waε filtered through Celite and the filtrate concentrated in vacuo to a colorless oil. The oil was redissolved 10 mL of methanol and treated with a 1.0 M anhydrous solution of HCl ether (7.0 mL, 7 mmol) and reduced m vacuo to dryness . The residue waε triturated pentane and the solids filtered to yield 1.4 g (99%) of a dry white powder. NMR. EA.

Preparation ." (S) -3- (2-fluorophenylthioj -2-hydroxypropyla ine

( 2S) - (- i -glycidyl-3-nitrooenzenesulfonate (3.0 g, 11.5 mmol) and potassium carbonate .1.8 g, 13 mmol' acetone (50 mL. was sparged witn nitrogen for 5 minutes. 2-fluorothiophenci (1.5 g, 11.6 mmol) was added ana the mixture stirred at ambient temperature under a blanκet of nitrogen for 4 hours. The acetone was removed m vacuo and the residue partitioned between water'ethyl acetate. The organic layer was dried (MgSθ4 ) and concentrated to give 1.6 g (75%) of the intermediate epoxide aε a colorieεs oil and was used without further purification. The intermediate epoxide 1.2 g, 6.5 mmol was dissolved in metnancl '10 mL . and cooled to 0°C using an ice bath. The solution was εaturated with ammonia gaε and the reaction vessel was sealed and allowed to stir at ambient temperature for 16 hours. The reaction waε opened at 0°C and the ammonia allowed to evaporate before the

mixture waε concentrated in vacuo . The residue was purified by flash chromatography on silica gel using 25:5:1 CHCI3 :MeOH:NH4θH to give 980 mg of a colorless oil (75%) which rapidly crystallized under vacuum to give a white solid, mp 50-53 °C. NMR. MS. EA.

Preparation 4 4- [ (2-oxocyclohexyl)methyl]benzonitrile

A mixture cf methyl cyclohexanone-2-carboxylate (11.0 g, 70 mmol, from Fluka) , α-bromo-p-tolumtrile (12.3 g, 63 mmol) , potassium carbonate (10.5 g, 76 mmol) in THF (200 mL) was refluxed for 24 hours. The progress of the reaction was followed by GC. The reaction waε diluted with water and the THF was removed under reduced pressure. The aqueous portion was extracted with EtOAc, dried (MgSθ4) to give 19.3 g of a white solid that was 74% pure by gaε chromatrophy . The εolid waε recryεtallized from hexane/EtOAc to give 7.75 g white crystals that were 100% pure by glc. A second crop of 3.65 g was obtained by adding more hexane to the filtrate. Overall, 11.4 g (67%) of 1- [ (4-cyanophenyl)methyl] -I-methoxycarbonyl-2- oxocyciohexane carboxylate, waε obtained; mp S2-84°C. NMR. MS.

Under a blanket of nitrogen, a mixture of l-[ (4- cyanophenyl)methyl] -l-methoxycarbonyl-2-oxocyclohexane carboxylate (7.6 g, 28 mmol) , sodium cyanide (2.1 g, 42 mmol) and DMSO (100 mL) was heated at 115°C for 1.5 hours. The progress of the reaction was monitored by glc. The reaction was cooled and partitioned between water, EtOAc and brine. The organic layer was washed with water and dried (MgS04) . After concentration, crude product was obtained as a tan oil. Purification by plug filtration

(200 g silica gel, 15% EtOAc/hexane gave 3.3 g 'RRSl product as colorless oil. NMR. MS.

Preparation 5 4- [ (2-oxocyclohexyl ; ethyl]benzamide

A DMSO (20 mL) solution of the compounα of Preparation 28 (2.5 g, 11.7 mmol) was cooled in an ice oath. Solid K2CO3 (500 mg) was added foiloweα immediately by 30% H2O2 (3 mL) . After 20 minutes, TLC 3 ~ EtOAc πexane εhowed a trace of starting material remameα. The ice bath waε removed and the reaction waε stirred and room temperature for 1 hour. The reaction waε diluted with 500 mL water and the white solid collected and dried to give 2.44 g

(90%) desired amide. The product was recrystallized from 1/9 EtOAc/hexane to give 2.02 g cf tne titled product aε white cryεtals, mp 167-170°C. NMP . MS.

Preparation -

2-Tetraione-6-carDoxylιc aciα, et _^ ene Ketai 6-bromo-2-tetraione 2.0 g, _, . C ^J mmol was dissolved in toluene (50 mL and treated witn excess ethylene glycol (4.88 mL, 88.9 mmol) and catalytic -toluenesulfonic acid (15 mg) . The εolution was stirred at reflux 16 nourε , and water was removed from the reaction mixture using a Dean- Stark condenser. After cooling to ambient temperature, the toluene solution was washed 2 x IN NaOH, 1 :■: water, 1 x brine, dried over a2Sθ4 and concentrated m vacua to give 2.23 g (93%; of 6-bromo-2-tetralone ethylene κetal as a brown oil whicn was used without furtner purification.

6-bromo-2-tetralone ethylene ketal (2.2 g, £.15 mmol was dissolved in anhydrous THF (30 mL) , cooled to -78°C and treated with tert-butyllithium (12.05 mL, 20.4 mmol, 2.7M

m pentane ) under an atmospnere of nitrogen. After stirring for 30 minutes, anhydrous carbon dioxide gas waε passed through the reaction mixture for 20 minutes at -78°C. The suεpension waε then allowed to warm to ambient temperature The εolution waε quenched with water and acidified with IN HCl, then extracted 2 x EtOAc. The organic extracts were washed with brine, dried over Na2Sθ4 and concentrated in vacuo to a pale brown oil . The oilv reεidue waε applied to a silica flasn chromatography column and eluted with 30%-50% EtOAC m hexaneε to yield 2- tetralone-6-carboxylιc acid, ethylene ketal 1.06 g (55%) a slowly crystallizing solid. NMP MS.

Preparation ^~~ 2-Tetralone-6-carboxamιde

2-tetralone-6-carboxylιc acid, ethylene ketal '395 mg, 2.07 mmol was co-disεolveα m CH CI2 (50 mL. with N-hydroxyεuccinimide (260 mg, 2.76 mmol at 0°C and treated with a slight excesε of 1, 3-dicycionexyicaroodiimide 502 mg, 2.50 mmol The mixture was allowed to warm to amoient temperature over 30 minutes, during whicn time a fine white precipitate formed. Ammonium chloride 333 mg, 6.23 mmol) and triethvlamme .1.58 mL, 12.5 mmol, were added and the solution stirred at ambient temperature for 16 hours. The suspended urea and salts were filtered awav and the solution concentrated m vacuo to a colorless oil. The oil was applied to a silica flash cnromatograpny column and eluted with 50-10C°- EtOAc m hexanes to yield 250 mg (64% of 2-tetralone-5-carboxamιde, ethyiene ketal aε a white solid, clean by NMF , TLC.

2-tetralone-6-carboxamιde, ethyiene ketal (250 mg, 1.07 mmol) and catalytic p-tolueneεulfonic acid were

stirred m acetone (50 mL) at amoient temperature for 48 hours. The volatiles were removed in vacuo and the reεidue triturated m ethyl acetate. The εolids were filtered, washed and dried to yield 77.5 mg .38%) of 2-Tetralone-6- carboxamide as a white powder, pure by NMR, TLC. MS.

Preparation 8 2-Tetralone-6-morpnolmamιde

2-tetralone-5-carDoxyiιc acid, ethyiene ketal

(395 mg, 2.37 mmol waε cooissoived in CH2Ciι 50 ) witn N-hydroxysuccmimide (260 mg, 2. 6 mmol at 0°C and treated with a slight excess of 1, 3-dicyclohexylcarbodnmide (502 mg, 2.50 mmol) . The mixture was allowed to warm to ambient temperature over 30 minutes, during which time a fine white precipitate formed. Morpholme ⁽ 0.91 mL, 10.4 mmol, d=0.998) was added and the solution stirred at ambient temperature for 16 hours. The suspended urea was filtered away and the solution concentrated m vacuo to a colorless oil. The oil was applied to a silica flash cnromatograpny column and eiuteo with 53-100% EtOAc hexaneε to yieia 323 mg (51%) of 2-Tetraione-ό-morpnclmamιde, ethyiene ketal as a slowly crystallizing solid, clean by NMF , TLC. 2-Tetralone-6-morpholmamιd , ethylene κetal (323 mg, 1.06 mmol) and catalytic p-tclueneεulfonic acid were stirred acetone (50 mL) at ambient temperature for 48 hours. TLC indicated a mixture cf 2-tetralone-6- morpholinamide, ethylene ketal and desired product, so the εolution was heated to reflux for 16 hours. Tne volatiles were removed in vacuo and the residue applied to a silica flash chromatography column and eluted with 50-100% EtOAc m hexanes to yield 27 mg (10%) of 2-tetralone-6- morphol amide a slowly crystallizing solid, pure by NMR, TLC. MS.

Preparation 9

5- [2-fluoro-4- (2- phenyl] -tetrazole

4-bromo-2-fluorobenzonitrile (5.3 g, 26.5 mmol) and 3-methyl-3-buten-2-ol (3.5 g, 40 mmol) were dissolved m N-methylpyrrolidinone (30 mL) and treated with catalytic palladium diacetate '115 mg, 0.5 mmol) , tπs- vo-tolyl) - phosphme .300 mg, 1.3 mmol I , and NaHC03 ,2." g, 32 mmol, . The mixture was stirred at 120 'C for one hour. The solution was cooled to ambient temperature and partitioned between H2θ/ethyi acetate, dried (MgS04 ) and concentrated in vacuo . The residue was applied to a silica chromatography column and eluted with 4:1 hexane/ethyl acetate to yield 2.3 g of a pale yellow oil (43%)

The oil (1.3 g, 6.3 mmol) was dissolved in DMF (30 mL) and treated with sodium azide (455 m , " mmol) and ammonium chloride , ^' 375 mg, ~ ^~ mmol and εtirred at 90 'C for 16 hourε . The reaction mixture was concentrated m vacua and partitioned between 3N NaOH 'diethyl ether. The aqueous layer was acidified with cone. HCl and cooled, and then extracted with diethyl ether, dried (MgS0 ₄ , and concentrated m vacuo to a pale brown oil. The reεidue waε applied to a silica chromatography column and eluted with 25:5:1 CHCI3/MeOH/NH4θH to yield 140 mg of a white solid (9%) .

Preparation 10 5- (2-methyl-3-oxobutyl) - hιophene-2-εulfonamide

5-bromothιophene-2-εulfonamιde (5.2 g, 21.5 mmol) and 3-methyl-3-buten-2-ol (2.3 g, 32.2 mmol) were dissolved in N-methylpyrrolidinone (40 mL) and treated with catalytic palladium diacetate (96 mg, 0.43 mmol) , tris-(o- tolyl) -phosphme (262 mg, 0.86 mmol , and NaHC03 (2.2 g, 25.3 mmol) . The mixture was stirred at 160'C for 48 hours. TLC indicated the reaction was ca. 50% completed at that time. The solution was cooled to ambient temperature and partitioned between H2θ/ethyl acetate, dried (MgS04 ) and concentrated in vacuo to a dark brown oil. The residue waε applied to a silica chromatography column and eluted with 2:3 hexane/ethyl acetate to yield 220 mg of a pale brown oil (4.1%) .

Preparation 11 4- (2-methyl-3-oxobutyl . benzamide

4-bromobenzonitrile (9.1 g, 50 mmol and 3- methyl-3-buten-2-ol (6.5 g, 75 mmol) were dissolved in N- methylpyrrolidmone (40 mL) and treated with catalytic palladium diacetate (225 mg, 1.0 mmol) , tπs- _' o-toiyl ) - phosphme '610 mg, 2.0 mmol) , and NaHC03 '5.0 g, 60 m ol) . The mixture was stirred at 120'C for three hours. The solution was cooled to ambient temperature and partitioned between H2θ/ethyl acetate, dried (MgS0 ) and concentrated in vacuo . The residue was applied to a silica

chromatography column and eluted with 3:1 hexane/ethyl acetate to yield 6.2 g of a pale yellow oil (66%)

The oil (4.6 g, 24.6 mmol) was dissolved in DMSO (20 mL) and treated with K2CO3 (1.0 g) and H2O2 (6 mL, 30% w/w) and stirred at 0'C for 10 minutes. The reaction mixture was diluted with 500 mL water and saturated with NaCl, and then extracted with ethyl acetate. The organic extractε were dried (MgS04) , and concentrated in vacuo to a white solid. The solid was recrystallized from 1:1 ethyl acetate/hexane to yield 3.22 g of a white solid (64%) . M.P 112-115.

The following compounds were prepared in a manner analogous to the schemeε and/or preparations described herein or by techniques appreciated m the art

Example l

A 5x8 grid of 4 mL screw cap vials was arranged. To each of the eight rows of vials m the grid was added 33 μmol of ketone (from preparations 4-34, or commercially available) , one ketone per row, as a stock solution m methanol (0.5M, 65 μl .. If solubility was a problem, acetonitrile/methanol or DMF was used. To each column of vials the grid was added 50 μmol of amme hydrochloride, one amme hydrochloride (or amme) (from preparations 2, 3, 36-41 or commercially available) per column, as a stock solution in methanol (0.5M, 100 μl) . To each vial was then added resm VIII (18-20 mg) , 1.01 meq/g, 70-90μeq base) . Teflon lined caps were then placed on each vial. The slurries were then shaken for 24 hours, at which time each vial was treated with approximately 30 mg (2.5 mmol BH4-/g resm, 75 μmol) of Amberlite IRA400 borohydride resm

(Aldrich Chemical) . The caps were replaced, and the vials were shaken for an additional 24 hours, then 150 μl methylene chloride and 40 mg (1 mmol/g resm, 0.4 mmol) polystyrene-linked benzaldehyde resm (Frechet, J.M.; Schuerch, C.J. Am. Chem. Soc. 1971, £3_, 492.) in order to scavenge excess primary amme starting material were added to the vial, and the slurry was shaken for 1 day. Each vial was then filtered through a cotton plug. The residual res was rinsed with three small portions of methanol (approximately 200 μl) . The resulting solutions were then treated with 20μl of cone. HCl (120 μmol) to ensure formation of the HCl salt of the product amine, then each vial was diluted to a volume of approximately 4 mL, and 1 mL of each solution was transferred to a tared 4 mL screw

cap vial. This solution was allowed to evaporate in a fume hood under an air stream until dry, then placed m a vacuum oven for 24 hours at room temperature. The resulting residues were then weighed and submitted directly for testing with no further purification. The bulk of the material (^5%) was similarly evaporated.

The following matrices list additional examples 2-81. These compounds were prepared using combinatorial/parallel techniques in accordance with the present invention. All reaction conditions were the same from plate to plate and m substantial accordance with Scheme 2 and Example 1. The scaffold for each plate was the same and is depicted at tne top corner of the 5x8 matrix. Tne variable functional groups are illustrated in the rows and columns. The ketones and the amines depicted on each plate were prepared in accordance with the schemes and preparations described herein or by techniques known m the art .

As previously noted, the compounds of the present invention are potent, selective β3 adrenergic receptor agonists. This pharmacological activity was determined in the functional agonist β3 assay.

Functional Agonists

&3_ Assev

Cell Lines

The hβ2 DNA was expressed from a plasmid 57537 obtained from American Type Culture Collection, hβi and hβ3 adrenergic receptors were cloned from human genomic libraries using the polymerase chain reaction method with degenerate probes. Full length receptors were cloned, expressed and sequenced to verify identity according to published sequences (hβι _: T. Frielle et. al . (1993)

Molecular Pharmacology 44 : 264-270) . These receptors were then expressed in the DXB-11 variant of CHO cells using a vector restoring tetrahydrofolate reductase and hygromycin resistance. Rat β3 receptor expressing CHO cell line is known in the art. Mol. Pharm. , Vol 40, pp. 895-99 (1991) . CHO cells were grown in 10% dialyzed FBS./high glucose DMEM/0.1% proline.

cAMP Assay Cell membranes were harvested from the above cell line using hypotonic 25 mM Hepes (pH 7.4) , 1 mM EDTA, 20 μg/mL leupeptin, 1 mM PMSF buffer with scraping followed by differential centrifugation. Membranes were incubated in 25 mM Tris (pH 7.6), 0.2% BSA, 2.6 mM Mg, 0.8 mM ATP, 0.1 mM GTP, 5 mM creatine phosphate, creatine kinase 50 U/mL, 0.2 mM IBMX at 32 'C. Agonists were added and incubation continued for 15 m. cAMP produced was assayed using a fluorescent tracer-immuno assay method.

Intact cell assays were performed using suspended cells removed from culture flasks by trypsin treatment. Cells were preincubated with 0.5 mM IBMX at 37 'C. Agonists were added and incubation continued for 15

m. Incubation was stopped by heating suspension in boiling water. cAMP or cGMP in these and the soleus incubations were assayed by RIA (Amersham) .

The compounds of the invention are agonists of the β ₃ receptor. isoproterenol is accepted in the art as a non- selective β ₃ agonist and is widely used as a comparator in evaluating the activity of compounds. See Trends in Pharm. Sci . 15: 3 (1994) . In the Functional Agonist β ₃ assay, the compounds demonstrated at least 30%, preferably 50% and most preferably over 85% of isoproterenol ' ε response at a single dose of 50μmol. Dose response titrations on the agonists described reveal EC ₅₀ values of < 10 μM, preferably < ImM. In the functional assay, dose titration furnishes an EC ₅₀ for isoproterenol of 1. l±O .5 mM.

When screened against the βi and β ₂ receptors in the functional assay, dose titration experiments indicate that greatly reduced or no receptor stimulation is observed with the compounds of the invention. This is defined by measuring the intrinsic activity (maximal response achieved) as compared to isoproterenol. The claimed compounds of Formula I are selective β ₃ receptor agonists and have an intrinsic activity of < 3% of isoproterenol ' s response. Thus, the compounds of the invention are selective β ₃ adrenergic receptor agonists.

As agonists of β3 , the compounds are useful in treating conditions in a mammal in which the β3 receptor has been demonstrated to play a role. The prefered mammal of treatment is a human. The relationship between modulating the β3 receptor and treatment of diseases, such

Type II diabetes and obesity, is well established in the art. Other conditions recognized in the art include: asthma, depression, and gastrointestinal disorders such as gastrointestinal motility. Thus, the present compounds are useful in the treatment of inflammatory bowel disease (Crohn's disease or ulcerative colitis) , irritable bowel

syndrome, non-specific diarrhoea dumping syndrome, asthma, and depression.

In treating non-human mammals, the compounds of the present invention are useful for increasing weight gain and/or improving the feed utilization efficiency and/or increasing lean body mass and/or decreasing birth mortality rate and increasing post/natal survival rate.

The compounds of Formula I are preferably formulated prior to administration. Therefore, yet another embodiment of the present invention is a pharmaceutical formulation comprising a compound of Formula I and one or more pharmaceutically acceptable carriers, diluents or excipients.

The present pharmaceutical formulations are prepared by known procedures using well-known and readily available ingredients. In making the compositions of the present invention, the active ingredient will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of a capsule, sachet, paper or other container. When the carrier serves as a diluent, it may be a solid, semisolid or liquid material which acts as a vehicle, excipient or medium for the active ingredient. Thus, the compositions can be m the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosol (as a solid or in a liquid medium), soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

Some examples of suitable carriers, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, algmates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvmylpyrrolidone, cellulose, water syrup, methyl cellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate and mineral oil. The formulations can additionally include

lubπcating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents. The compositions of the invention may be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient.

The compositions are preferably formulated in a unit dosage form, each dosage containing from about 0.1 to about 500 mg, preferably about 5 to about 200 mg, of the active ingredient. However, it will be understood that the therapeutic dosage administered will be determined by the physician in the light of the relevant circumstances including the condition to be treated, the choice of compound to be administered and the chosen route of administration, and therefore, the above dosage ranges are not intended to limit the scope of the invention m any way. The compounds can be administered by a variety of routes including the oral, rectal, transdermal, subcutaneous, topical, intravenous, intramuscular or mtranasal routes. For all indications, a typical daily dose will contain from about 0.05 mg/kg to about 20 mg/kg of the active compound of this invention. Preferred daily doses will be about 0.1 to about 10 mg/kg, ideally about 0.1 to about 5 mg/kg. However, for topical administration a typical dosage is about 1 to about 500 μg compound per cm ² of an affected tissue. Preferably, the applied amount of compound will range from about 30 to about 300 μg^c ² , more preferably, from about 50 to about 200 μg/cm ² , and, most preferably, from about 60 to about 100 μg/cm ² . The following formulation example is illustrative only and is not intended to limit the scope of the invention m any way.

Formuiation 1

Hard gelatin capsules are prepared using the following ingredients:

Quantity (mg/capsule)

4-{3-[2-hydroxy-3- (3- hydroxyphenyloxy) -propylamino] -2- 25 methylbutyl}benzamide starch, dried 425 magnesium stearate 10

Total 460 mg

The above ingredients are mixed and filled into hard gelatin capsules in 460 mg quantities.

The principles, preferred embodiments and modes of operation of the present invention have been described in the foregoing specification. The invention which is intended to be protected herein, however, is not to be construed as limited to the particular forms disclosed, since they are to be regarded as illustrative rather than restrictive. Variations and changes may be made by those skilled in the art without departing from the spirit of the invention.