WO2001042217A1

WO2001042217A1 - Beta-3 adrenoceptor agonists

Info

Publication number: WO2001042217A1
Application number: PCT/US2000/033222
Authority: WO
Inventors: Kelly Horne Donaldson; Barry George Shearer; David Edward Uehling
Original assignee: Glaxo Group Limited
Priority date: 1999-12-10
Filing date: 2000-12-08
Publication date: 2001-06-14
Also published as: AU1953101A; GB9929118D0

Abstract

The following compounds: 3'-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]-[1,1'-biphenyl]-3-carboxylic acid; 6-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-2-pyridinecarboxylic acid; (R)-3-[3-[[2-(3-chlorophenyl)-2-hydroxyethyl]amino]ethyl]amino]phenyl]-4-pyridinecarboxylic acid; 3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-4-pyridinecarboxylic acid; (R)-3-[3-[[2-(3-chlorophenyl)-2-hydroxyethyl]amino]ethyl]amino]phenyl]-2-pyridinecarboxylic acid; 3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]proyl]amino]phenyl]-2-pyridinecarboxylic acid; and pharmaceutically acceptable derivatives thereof are disclosed. These compounds are useful as agonist for human beta-3 adrenoceptor.

Description

BETA-3 ADRENOCEPTOR AGONISTS

Field of the Invention This invention relates to a new class of chemical compounds and to their use in medicine. In particular, the invention relates to biaryl derivatives, methods for their preparation, pharmaceutical compositions containing them, and their use as agonists at atypical beta-ad renoceptors (also known as beta-3- adrenoceptors).

Background of the Invention

Atypical beta-adrenoceptors belong to the family of adrenoceptors which mediate the physiological actions of the hormones adrenaline and noradrenaline. Such receptors have been described for example by J R S Arch et. al., Nature, 309, 163-165 (1984); C Wilson et. al., Eur. J. Pharmacol., 100, 309-319 (1984); L J Emorine et. al., Science, 245, 1118-1121 (1989); and A. Bianchetti et. al. Br. J. Pharmacol., 100, 831-839 (1990).

Phenethanolamine derivatives having activity at atypical beta-adrenoceptors are disclosed in, for example, European Patent Applications EP-A-0455006 and EP-A-0543662. Sub-types of the adrenoceptors, α,-, α₂-, β_r, β₂- and β₃-(atypical) can be identified on the basis of their pharmacological properties and physiological effects. Chemical agents which stimulate or block these receptors (but not β₃) are widely used in clinical medicine. More recently, emphasis has been placed upon specific receptor selectivity in order to reduce side effects caused, in part, by interactions with other receptors.

Atypical beta-adrenoceptors are known to occur in adipose tissue and the gastrointestinal tract. Atypical beta-adrenoceptor agonists have been found to be particularly useful as thermogenic anti-obesity agents and as anti-diabetic agents. Compounds having atypical beta-adrenoceptor agonist activity have also been described as being useful in the treatment of hyperglycaemia, as animal growth promoters, as blood platelet aggregation inhibitors, as positive inotropic agents and as antiatherosclerotic agents, and as being useful in the treatment of glaucoma.

A UK patent application filed on 13 June 1998 as GB 9812709.5, discloses compounds of Formula (I) and pharmaceutically acceptable derivatives thereof:

(l) wherein R¹ is an aryl, pyridyl, thiazolyl, phenoxymethyl, or pyrimidyl group, optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxy, C-μβalkoxy, C-j.galkyl, nitro, cyano, hydroxymethyl, trifluoromethyl, -NR⁶R⁶, and -NHSO2R⁶, where each R⁶ is independently hydrogen or C-ι_4alkyl; R2 is hydrogen or O^alkyl; X is oxygen, NH, or NC^alkyl; R3 is cyano, tetrazol-5-yl, or CO2 ^ where R⁷ is hydrogen or C,.₆alkyl; R4 and R5 are independently hydrogen, C-j. alkyl, -CO₂H, -C0₂C₁.₆alkyl, cyano, tetrazol-5-yl, halogen, trifluoromethyl, or C^alkoxy, or, when R^ and R^ are bonded to adjacent carbon atoms, R4 and R⁵ may, together with the carbon atoms to which they are bonded, form a fused 5 or 6 membered ring optionally containing one or two nitrogen, oxygen, or sulfur atoms; and Y is N or CH.

Summary of the Invention Briefly, in one aspect, the present invention provides the following compounds:

3'-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]-[1 , 1 '-biphenyl]-

3-carboxylic acid

6-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-2- pyridinecarboxylic acid (R)-3-[3-[[2-(3-chlorophenyl)- 2-hydroxyethyl]amino]ethyl]amino]phenyl]-4- pyridinecarboxylic acid

3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-4- pyridinecarboxylic acid

(R)-3-[3-[[2-(3-chlorophenyl)-2-hydroxyethyl]amino]ethyl]amino]phenyl]-2- pyridinecarboxylic acid

3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-2- pyridinecarboxylic acid; and pharmaceutically acceptable derivatives thereof. Preferably, the compounds of this invention are agonists for human beta-3 adrenoceptor ("β₃"). More preferably, the compounds of this invention are selective agonists for β₃.

In another aspect, the present invention provides a pharmaceutical formulation comprising a compound of the invention and a pharmaceutically acceptable carrier.

In another aspect, the present invention provides a method for the prevention or treatment of clinical conditions or diseases susceptible to amelioration by administration of an atypical beta-adrenoceptor agonist, comprising administration of an effective amount of a compound or composition of this invention.

Detailed Description of the Invention

As used herein, "a pharmaceutically acceptable derivative" means a pharmaceutically acceptable salt, ester, or salt of such ester, which, upon administration to the recipient, is capable of providing (directly or indirectly) a compound of the present invention or an active metabolite or residue thereof. It will be appreciated by those skilled in the art that the compounds of the present invention may be modified to provide pharmaceutically acceptable derivatives thereof at any of the functional groups in the compounds of the present invention. Of particular interest as such derivatives are compounds modified at the carboxyl function, hydroxyl functions or at amino groups. It will be appreciated by those skilled in the art that the pharmaceutically acceptable derivatives of the compounds of the present invention may be derivatised at more than one position.

Preferred pharmaceutically acceptable derivatives of the compounds of the present invention are pharmaceutically acceptable salts thereof. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulphuric, nitric, perchloric, fumaric, maleic, phosphoric, glycollic, iactic, salicylic, succinic, toluene- p-sulphonic, tartaric, acetic, citric, methanesulphonic, formic, benzoic, malonic, naphthalene-2-sulphonic and benzenesulphonic acids. Other acids such as oxalic, while not in themselves pharmaceutically acceptable, may be useful in the preparation of salts useful as intermediates in obtaining compounds of the invention and their pharmaceutically acceptable acid addition salts. Salts derived from appropriate bases include alkali metal (e.g. sodium), alkaline earth metal (e.g. magnesium), ammonium and NR₄ ⁺ (where R is C^alkyl) salts.

The compounds of the present invention act as agonists at atypical beta - adrenoceptors and as such are useful in the treatment of clinical conditions susceptible to amelioration by administration of an atypical beta-adrenoceptor agonist. Such conditions include hyperglycaemia, obesity, hyperlipemia, irritable bowel syndrome and its associated pain, motility dysfunction, excessive gastrointestinal secretion, non-specific diarrhea, neurogenic inflammation, regulation of intraocular pressure, triglyceridemia, diabetes, e.g. non-insulin- dependent diabetes mellitus (NIDDM or type 2), such as obese NIDDM and non- obese NIDDM, diabetic complications such as retinopathy, nephropathy, neuropathy, cataracts, coronary heart diseases and arteriosclerosis, osteoporosis; and gastrointestinal disorders, particularly inflammatory gastrointestinal disorders. They are also of use in increasing the high-density- lipoprotein (HDL) cholesterol concentration and decreasing the triglyceride concentration in blood serum, especially human blood serum, and are therefore of potential use in the treatment and/or prophylaxis of atherosclerosis. They also may be useful for the treatment of hyperinsulinaemia, depression, muscle wasting, and urinary incontinence. References in this specification to treatment include prophylactic treatment as well as the alleviation of symptoms.

In a further aspect, the invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for the treatment of a condition susceptible of amelioration by an atypical beta-adrenoceptor agonist.

While it is possible that, for use in therapy, a compound of the invention may be administered as the raw chemical it is preferable to present the active ingredient as a pharmaceutical formulation. The invention thus further provides a pharmaceutical formulation comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof together with one or more pharmaceutically acceptable carriers thereof and, optionally, other therapeutic and/or prophylactic ingredients. The carher(s) or excipient(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. The compounds for use according to the present invention may be formulated for oral, buccal, parenteral, rectal or transdermal administration or in a form suitable for administration by inhalation or insufflation (either through the mouth or the nose).

For oral administration, the pharmaceutical compositions may take the form of, for example, tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g. pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g. lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc or silica); disintegrants (e.g. potato starch or sodium starch glycollate); or wetting agents (e.g. sodium lauryl sulphate). The tablets may be coated by methods well known in the art. Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g. sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g. lecithin or acacia); non-aqueous vehicles (e.g. almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g. methyl or propyl- p-hydroxybenzoates or sorbic acid). The preparations may also contain buffer salts, flavoring, coloring and sweetening agents as appropriate. Preparations for oral administration may be suitably formulated to give controlled release of the active compound.

For buccal administration the compositions may take the form of tablets or lozenges formulated in conventional manner. The compounds according to the present invention may be formulated for parenteral administration by injection e.g. by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form e.g. in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile pyrogen-free water, before use.

The compounds according to the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, e.g. containing conventional suppository bases such as cocoa butter or other glycerides. In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously, transcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds according to the present invention may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

Suitable therapeutic ingredients which may be formulated with compounds of the invention, together with one or more pharmaceutical carriers or excipients, include ingredients which may be used in the same clinical conditions as those listed herein for atypical beta-adrenoceptor agonists. Such ingredients may include, for example, PPAR-gamma agonists.

A proposed dose of the compounds according to the present invention for administration to a human (of approximately 70kg body weight) is 0.1 mg to 1g, preferably to 1 mg to 100mg of the active ingredient per unit dose, expressed as the weight of free base. The unit dose may be administered, for example, 1 to 4 times per day. The dose will depend on the route of administration. It will be appreciated that it may be necessary to make routine variations to the dosage depending on the age and weight of the patient as well as the severity of the condition to be treated. The precise dose and route of administration will ultimately be at the discretion of the attendant physician.

Atypical beta-adrenoceptor agonists are compounds which demonstrate a pharmacological response mediated at atypical beta-adrenoceptors. This activity has been measured as the ability to stimulate lipolysis by rat adipocytes at sub-micromolar concentrations, in a response that is resistant to blockade by standard beta-adrenoceptor blocking drugs such as propranolol.

Another useful means of identifying an atypical beta-adrenoceptor agonist involves the measurement of agonist activity at atypical beta-adrenoceptors in the rat isolated lower oesophagus. The rat oesophagus assay is based upon that described by Ford et. al., Br. J. Pharmacol., 105(suppl.), 235P, 1992. The relative potency of each test compound (EPMR) is compared to isoprenaline as follows:

EC₅₀ agonist EPMR =

EC₅₀ isoprenaline wherein EC₅₀ is the molar concentration of agonist which produces 50% of the maximum possible response for that agonist.

A particularly useful method for determining agonist activity at human atypical beta-adrenoceptors involves the use of Chinese hamster ovarian (CHO) cells transfected with the human beta-3-adrenoceptor according to Method 1. The cell lines may also be transfected with human beta-1 - and beta-2 adrenoceptor in a similar manner to provide a method of determining the selectivity of the compounds of the invention at the three receptors.

Method 1 - Cell culture

General cell culture guidelines are observed (Fershney, R.A. (1987) Culture of animal cells: A manual of basic technique. Wiley-Liss, Inc., N.Y.). A standard cell culture incubator is used (37°C, 5% CO₂ in air, 95% relative humidity). H- β₃CHO cells are grown in DMEM/F12 (with pyroxidine-HCI, 15 mM HEPES, L- glutamine), supplemented with 10% heat-inactivated FBS, 500 μg/ml G418, 2 mM L-glutamine, 100 units penicillin G and 100 μg streptomycin sulfate. One confluent flask of cells is trypsinised and resuspended in the above medium at a concentration of 30-40,000 cells/100 μl and plated into 96-well flat bottom plates. The cells are then used for assay within 18-24 hours. The medium is aspirated from each well, and replaced with 180 μl DMEM/F12 with 500 mM IBMX. Antagonists, if required, are added at this stage. The plate is then placed back in the incubator for 30 min. Drugs are then added to the wells (20 μl, 10 x required final concentration) for 60 min. Responses were determined by measuring cAMP levels of a 20 ul sample of extracellular media using a scintillation proximity based radio-immunoassay (NEN Flashplates). As used herein, a compound is considered to be an agonist for hβ₃ if the compound stimulates the accumulation of extracellular cAMP with CHO-6CRE- luciferase cells expressing hβ₃. The relative potency of a hβ₃ agonist may be compared to its potency for stimulating the accumulation of extracellular cAMP with CHO-6CRE-luciferase cells expressing hβ₂ and hβ The compounds of this invention have an EC₅₀ of at most 100 nM at hβ₃ and are at least 50 times more potent at hβ₃ than at hβ₂ or hβ

Examples The invention is further illustrated by the following intermediates and examples. All temperatures are in degrees centigrade. HPLC characterization was carried out where specified using a Dynamax-60A C18 83-201 -C, 25cm x 4.6mm column, eluting with 5-40% CH₃CN in H₂0 with 0.1 % TFA buffer, with a program time of 30.0 min and flow rate of 1.5mL/min). Retention times are expressed as t_r in minutes. Optical rotation values are expressed as [a]Q values. Mass spectra (MS) were obtained using electrospray (positive or negative ion) analysis. 1 H nmr was carried out in deuterated choloroform, unless otherwise indicated.

Intermediate 1

6-bromopicolinic acid methyl ester To a suspension of bromopicolinic acid (6.0 g) in methanol (54 mL) was added 48% hydrobromic acid in water (6 mL). The reaction heated at reflux overnight. As the reaction cooled to room temperature a white solid fell out. The mixture was partitioned in dichloromethane (100 mL) and a saturated aqueous solution of sodium bicarbonate (100 mL). The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure to yield a white solid. The solid was absorbed on silica gel and eluted with 4:1 hexane:ethyl acetate. The title compound (6.0 g) was obtained as a white solid. TLC: R_f (3:1 hexane:ethyl acetate)=0.44

Intermediate 2

3'-Nitro-[1 ,1'-biphenyl]-3-carboxylic acid methyl ester

A flask flushed with nitrogen gas and equipped with a reflux condenser was charged with 3-bromobenzoate (21 g, 98 mmol) and anhydrous toluene (250 mL). Under positive nitrogen pressure the contents of the flask were cooled to -78 °C in a dry ice/acetone bath. Tetrakis(triphenyiphosphine)palladium (3.76 g), 2 M aqueous sodium carbonate (75 mL) and a slurry of 3-nitrophenylboronic acid (20 g, 120 mmol) in methanol (60 mL) were added in that order. The mixture was allowed to warm to ambient temperature and then brought to 85 °C. The mixture was heated at 85 °C for 5 h, then allowed to cool to ambient temperature. The mixture was partitioned between a mixture of 2 M aqueous sodium carbonate (150 mL) and ammonium hydroxide (15 mL) and methylene chloride (350 mL). The organic layer was separated, dried over sodium sulfate, filtered and concentrated to afford the crude product. Purification by flash silica gel chromatography (eluting with 10:1 hexane: ethyl acetate followed by 8:1 hexane: ethyl acetate) and concentrating the relevant fractions afforded the title compound (13.27 g). Fractions showing impure product were concentrated, resubjected to chromatography and relevant fractions were concentrated to provide material which was then recrystallized to supply additional product (1.91 g. m.p., 88-90 °C).

Similarly prepared were:

Intermediate 3

6-(3-Nitrophenyl)-2-pyridinecarboxylic acid methyl ester as a pale orange solid (330 mg); TLC: R_f (7:3 hexane: ethyl acetate) = 0.22; from 6-bromopicolinic acid methyl ester (1.0 g), 3-nitrophenylboronic acid (940 mg).

Intermediate 4

3-(3-Nitrophenyl)-N-phenyl-2-pyridinecarboxamide as a pale yellow solid (420 mg); mp=152-155 °C; from 3-iodopicolinanilide (1.40 g, prepared according to the method of Epsztajn, Syn. Comm., 1997, 27 (6), 1075) and 3- nitrophenylboronic acid (865 mg).

Intermediate 5

3-(3-Nitrophenyl)-4-pyridinecarboxylic acid methyl ester

To a stirred mixture of 3-iodomethylisonicotinate (455 mg, prepared according to the method of Epsztajn, Syn. Comm., 1997, 27 (6), 1075) in toluene (8 mL) was added a solution of 3-nitrophenylboronic acid (304 mg) in ethanol (2 mL) at -78 °C. After the solution was degassed [1 ,1 '- bis(diphenylphosphino)ferrocene]dichloropalladium(ll) complex with dichloromethane (1 :1 , 44.6 mg) then 1 N aqueous sodium carbonate (2 mL) were added to the mixture. The mixture was warmed to room temperature, then heated at 90 °C for five hours. The reaction was allowed to cool to room temperature overnight and poured into water (50 mL). The mixture was extracted three times with ethyl acetate (50 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The residue was absorbed onto silica and purified by silica gel chromatography eluting with 7:3 hexane:ethyl acetate to provide the title compound (282 mg) as a white solid. mp=98-100 °C.

Intermediate 6 3'-Amino-[1 ,1 '-biphenyl]-3-carboxylic acid methyl ester

To a stirred solution of 3'-nitro-[1 ,1'-biphenyl]-3-carboxylic acid methyl ester (4.47 g) in anhydrous tetrahydrofuran (125 mL) under a blanket of nitrogen was added 10% palladium on activated charcoal (860 mg). The reaction was placed under a hydrogen atmosphere and stirred overnight. The reaction mixture was filtered through Celite and the solvent was removed under reduced pressure to yield a gray oil (4.4 g). The residue was purified by silica gel chromatography eluting with 3:1 hexane:ethyl acetate. Concentration of the relevant fractions provided the title compound as a white solid (3.5 g). n.m.r. δ values include 3.83 (s, 2H), 3.93 (s, 3H), 6.70 (d, 1 H), 6.93 (d, 1 H), 7.00 (d, 1 H), 7.25-7.21 (m, 1 H), 7.47 (t, 1 H), 7.73 (d,1 H), 7.98 (d, 1 H), 8.23 (d, 1 H).

Intermediate 7

6-(3-Aminophenyl)-2-pyridinecarboxylic acid methyl ester

To a stirred solution of 6-(3-nitrophenyl)-2-pyridinecarboxylic acid methyl ester (334 mg) in anhydrous ethyl acetate (20 mL) under a blanket of nitrogen was added 10% palladium on activated charcoal (300mg). The reaction was evacuated and placed under a hydrogen atmosphere and stirred for 1.25 hours. The reaction mixture was filtered through Celite and the solvent was removed under reduced pressure to yield a yellow oil (290 mg). The oil was purified by silica gel chromatography eluting with 1 :1 hexane:ethyl acetate to obtain the title compound (280 mg) as a white film. TLC: R_f (1 :1 hexane: ethyl acetate)=0.34.

Similarly prepared were:

Intermediate 8

3-(3-Aminophenyl)-4-py dinecarboxylic acid methyl ester as a yellow oil (240 mg); TLC: R_f (1 :1 hexane:ethyl acetate)= 0.12; from 3-(3-nitrophenyl)-4- pyridinecarboxylic acid methyl ester (275 mg).

Intermediate 9

3-(3-Aminophenyl)-N-phenyl-2-pyridinecarboxamide as a yellow solid (278 mg); Electrospray MS (positive ion): (M+H) 290; mp= 156-161 °C; from 3-(3- nitrophenyl)-N-phenyl-2-pyridinecarboxamide (400 mg).

Intermediate 10

3'-[[2R-[[2-(3-Chlorophenyl)-2R-[[(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]propyl]amino]-[1 ,1 '-biphenyl]-3-carboxylic acid methyl ester To a stirred solution of 3'-amino-[1 ,1'-biphenyl]-3-carboxylic acid methyl ester (844 mg) and [2R-(tert-butoxycarbonyl)-[2R-(tert-butyldimethylsilanoxy)-2-(3- chlorophenyl)ethyl]amino]-propionaldehyde (1.73 g, prepared according to the method described in WO95/33724) in anhydrous dichloromethane (15 mL) was added acetic acid (2 drops). After stirring for twenty-five minutes, sodium triacetoxyborohydride (1.71 g) was added and the mixture was stirred for 3 days. The reaction was partitioned between saturated aqueous sodium bicarbonate and dichloromethane. The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure to yield a white foam. The residue was purified by silica gel chromatography eluting with 10:1 hexane:ethyl acetate followed by 5:1 hexane:ethyl acetate to provide the title compound as a white foam (1.90 g). TLC: R_f (5:1 hexane:ethyl acetate)=0.53.

Similarly prepared were:

Intermediate 11

6-[3-[[2R-[[2-(3-Chlorophenyl)-2R-[r(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]propyl]amino]phenyl]-2-pyridinecarboxylic acid methyl ester as a colourless oil (725 mg);_Electrospray MS (positive ion): (M+H) 654; from 6-(3-aminophenyl)-2-pyridinecarboxylic acid methyl ester (280 mg) and [2R-(tert-butoxycarbonyl)-[2R-(tert-butyldimethylsilanoxy)-2-(3- chlorophenyl)ethyl]amino]-propionaldehyde (1.3 g).

Intermediate 12 (R)-3-[[2-rr2-(3-Chlorophenyl)-2-[r(tert-butyl)dimethylsilyl]oxy1ethyl][(tert- butoxy)carbonyl]amino]ethyl]amino]-[phenyl]-4-pyridinecarboxylic acid methyl ester as a pale yellow oil (384 mg); Electrospray MS (positive ion): (M+Na) 662; from 3-(3-aminophenyl)-4-pyridine-carboxylic acid methyl ester (1 11.6 mg) and (R)-[(tert-butoxycarbonyl)-[2-(tert-butyldimethylsilanloxy)-2-(3- chlorophenyl)ethyl]amino]-acetaldehyde (249 mg, prepared according to the method described in WO95/33724).

Intermediate 13

3-[3-[[2R-[[2-(3-Chlorophenyl)-2R- [(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]propyl]amino]phenyl]-4-pyridinecarboxylic acid methyl ester as a yellow oil (305 mg); TLC: R_f (1 :1 hexane: ethyl acetate)=0.66; from 3-(3-aminophenyl)-4-pyridinecarboxylic acid methyl ester (120 mg) and [2R-(tert-butoxycarbonyl)-[2R-(tert-butyldimethylsilanoxy)-2-(3- chlorophenyl)ethyl]amino]-propionaldehyde (398 mg).

Intermediate 14

(R)-3-[3-[[2-[[2-(3-Chlorophenyl)-2-[[(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]ethyl]amino]phenyl]-N-phenyl-2-pyridinecarboxamide as a yellow foam (201 mg); Electrospray MS (positive ion): (M+Na) 723; from 3-(3-aminophenyl)-N-phenyl-2-pyridinecarboxamide (135 mg) and (R)- [(tert-butoxycarbonyl)-[2-(tert-butyldimethylsilanoxy)-2-(3- chlorophenyl)ethyl]amino]-acetylaldehyde (428 mg).

Intermediate 15

3-r3-[[2R-[r2-(3-Chlorophenyl)-2R-[[(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]propyl]amino]phenyl]-N-phenyl-2-pyridinecarboxamide as a yellow foam (301 mg); Electrospray MS (positive ion): (M+H) 715; from 3-(3-aminophenyl)-N-phenyl-2-pyridinecarboxamide (135 mg) and [2R-(tert- butoxycarbonyl)-[2R-(tert-butyldimethylsilanoxy)-2-(3-chlorophenyl)ethyl]amino]- propionaldehyde (442 mg).

Intermediate 16

3'-r[2R-[[2-(3-Chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]-[1 , 1 '- biphenyl]-3-carboxylic acid methyl ester hydrochloride

The 3'-[[2R-[[2-(3-chlorophenyl)-2R-[[(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]propyl]amino]-[1 , 1 '-biphenyl]-3-carboxylic acid methyl ester (1.90 g) was dissolved in 4 N hydrogen chloride in dioxane (10 mL) and the mixture was stirred for 5 h. Diethyl ether (100 mL) was added and the resulting white precipitate was collected, washed with additional ether (20 mL) and dried en vacuo to supply the title compound (1.35 g) as a white solid. Electrospray MS (positive ion): (M+H) 438.4.

Intermediate 17

6-[3-[[2R-[[2-(3-Chlorophenyl)-2R-hydroxyethyl]aminojpropyl]amino]phenyl]-2- pyridinecarboxylic acid methyl ester A solution of 6-[3-[[2R-[[2-(3-chlorophenyl)-2R-[[(tert- butyl)dimethylsilyl]oxy]ethyl][(tert-butoxy)carbonyl]amino]propyl]amino]phenyl]-2- pyridinecarboxyiic acid methyl ester (685 mg) in 4N hydrochloric acid in dioxane (10 mL) was stirred overnight. The mixture was concentrated under reduced pressure to a residue. The residue was dissolved in chloroform:methanol:ammonium hydroxide (9:1 :0.1 ) and absorbed onto silica gel. Silica gel chromatography eluting with chloroform:methanol:ammonium hydroxide (9:1 :0.1 ) provided the title compound as a yellow oil (305 mg). Electrospray MS (positive ion): (M+H) 440.

Similarly prepared was:

Intermediate 18

3-[3-[[2R-[[2-(3-Chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-4- pyridinecarboxylic acid methyl ester as a purple oil (193 mg);

TLC: R_f (9:1 :0.1 chloroform/methanol/ammonium hydroxide) = 0.63; from 3-[3-[[2R-[[2-(3-chlorophenyl)-2R-[[(tert-butyl)dimethylsilyl]oxy]ethyl][(tert- butoxy)carbonyl]amino]propyl]amino]phenyl]-4-pyridinecarboxylic acid methyl ester (305 mg).

Example 1 3'-[[2R-[[2-(3-Chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino1-ri , 1 '- biphenyl]-3-carboxylic acid

A flask was charged with 3'-[[2R-[[2-(3-chlorophenyl)-2R- hydroxyethyl]amino]propyl]amino]-[1 ,1 '-biphenyl]-3-carboxylic acid (1.35 g), methanol (16 mL) and water (5.5 mL). Lithium hydroxide monohydrate (562 mg) was added and the mixture was stirred at ambient temperature for 7 h. Concentration with a rotary evaporator and purification of the residue by silica gel chromatography (eluting with 76:15:1 chloroform/methanol/concentrated ammonium hydroxide followed by 60:15:1 chloroform/methanol/concentrated ammonium hydroxide) gave a white residue. This material was triturated with water and the resulting solid was dried en vacuo to supply 1.12 g of a white solid. Further purification by flash silica gel chromatography using the above solvent system supplied a residue which was triturated with water and dried en vacuo to afford the title compound (866.6 mg) as a white solid. Electrospray MS (positive ion): (M+H) 426.4; Assay found C 65.15; H 5.94; N 6.29. C^H^N- ^O requires C 65.08; H 6.14; N 6.32.

Similarly prepared were: Example 2

6-[3-[[2R-[[2-(3-Chlorophenyl)-2R-hydroxyethyl]amino]propyl]aminolphenyll-2- pyridinecarboxylic acid as a light brown solid (161 mg).

Electrospray MS (positive ion): (M+H) 426;

Assay Found: C 58.12; H 5.05; N 8.62%;

LiCI requires C

58.09; H 5.09; N 8.84%; _ from 6-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-

2-pyridinecarboxyiic acid methyl ester (385mg).

Example 3

3-f3-[[2R-[[2-(3-Chlorophenyl)-2R-hydroxyethyllamino]propyllaminolphenyl]-4- pyridinecarboxylic acid as a white solid (87.3 mg); Electrospray MS (positive ion ): (M+H) 425.7; Assay Found: C 63.33; H 5.94; N 9.50%; C^H^C^N O.SδH requires C 63.33; H 5.81 ; N 9.63%; from 3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]- 4-pyridinecarboxylic acid methyl ester (193 mg).

Example 4

(R)-3-[3-[[2-(3-Chlorophenyl)-2-hydroxyethyllamino]ethyl]amino1phenyπ-4- pyridinecarboxylic acid

A solution of (R)-3-[[2-[[2-(3-chlorophenyl)-2-[[(tert- butyl)dimethylsilyl]oxy]ethyl][(tert-butoxy)carbonyl]amino]ethyl]amino]-[phenyl]-4- pyridinecarboxylic acid methyl ester (316.6 mg) in methylene chloride was treated with trifluoroacetic acid (1.5 mL). The mixture was stirred for 3 h and concentrated with a rotary evaporator to leave a residue that was taken up in 4 N hydrogen chloride in dioxane (5 mL) and stirred at ambient temperature for 16 h. The solvent was decanted and the resulting residue was triturated with ether and placed under high vacuum to supply 330 mg of a pale yellow solid. This material was dissolved in 3:1 methanol :water (4 mL) and lithium hydroxide monohydrate (166 mg) was added. The mixture was stirred at ambient temperature for 3 h and concentrated with a rotary evaporator to leave a residue which was taken up in a few mL 30:15:1 chloroform:methanol:concentrated ammonium hydroxide. The insoluble material was removed by filtration and the filtrate was subjected to flash silica gel chromatography eluting with 71 : 15:1 chloroform:methanol:ammonium hydroxide. Concentration of the relevant fractions supplied 180.7 mg of a pale yellow material which was triturated with water. The residue was dried en vacuo to give the title compound (111 mg) as a pale yellow amorphous solid. Electrospray MS (positive ion): (M+H) 412; Assay found C, 58.74, H, 5.42, N, 10.27; C^H^CI^A .25H₂00.25NH₄CI requires C, 59.01 , H, 5.74, N, 10.17.

Example 5 (R)-3-[3-[[2-(3-Chlorophenyl)-2-hydroxyethyl3amino]ethyl]amino]phenyl]-2- pyridinecarboxylic acid

To a solution of the (R)-3-[3-[[2-[[2-(3-chlorophenyl)-2-[[(tert- butyl)dimethylsilyl]oxy]ethyl][(tert-butoxy)carbonyl]amino]ethyl]amino]phenyl]-N- phenyl-2-pyridinecarboxamide (201 mg) in dichloromethane (10 mL) was added trifluoroacetic acid (1 mL). The mixture was stirred for 1.25 h and the solvent was removed under reduced pressure. Dilute aqueous hydrochloric acid (6%) was added and the reaction stirred for 16 h at 109 °C. Concentrated hydrochloric acid (1.5 mL) was added and the mixture was heated for 16 h. The mixture was concentrated to a residue and purified by flash silica chromatography eluting with 6:3:0.1 chloroform:methanol:ammonium hydroxide to afford, after concentrating the relevant fractions, a solid. The residue was triturated with methanol and water. The title compound was obtained as a beige solid (44.4 mg).

Electrospray MS (positive ion): (M+H) 412;

Assay Found: C 60.57; H 5.34; N 9.68%; C^H^C^N O.Sδ LiCI 0.55H₂O requires C 60.58; H 5.34; N 9.63%.

Example 6

3-[3-[[2R-rr2-(3-Chlorophenyl)-2R-hydroxyethyl]amino3propyl]amino]phenyl]-2- pyridinecarboxylic acid To a solution of the 3-[3-[[2R-[[2-(3-chlorophenyl)-2R-[[(tert- butyl)dimethylsilyl]oxy]ethyl][(tert-butoxy)carbonyl]amino]propyl]amino]phenyl]-N- phenyl-2-pyridinecarboxamide (301 mg) in dichloromethane (10 mL) was added trifluoroacetic acid (1 mL). The mixture was stirred for 16 h and the solvent was removed under reduced pressure. Aqueous hydrochloric acid (6 %) was added and the reaction was heated to 109 °C. The mixture was allowed to cool to ambient temperature, then concentrated hydrochloric acid (2.5 mL) was added and the reaction was stirred overnight at 109°C. The mixture was concentrated to a residue. Purification by silica gel chromatography eluting with 6:3:0.1 chloroform/methanol/ammonium hydroxide afforded a solid which was triturated with diethyl ether and two small portions of water. The title compound was obtained as a beige solid (91 mg). Electrospray MS (positive ion): (M+H) 426;

Assay Found: C 63.08; H 5.68; N 9.58%;

δ LiCIO.35H₂0 requires C 63.07; H 5.68; N 9.59%

The application of which this description and claims forms a part may be used as a basis for priority in respect of any subsequent application. The claims of such subsequent application may be directed to any aspect or feature of the invention or combination of such aspects or features of the invention as described herein. They may take the form of, but are not limited to, composition of matter, product, composition, formulation, process, use, or method of treatment claims and may include, by way of example, and without limitation, one or more of the following claims:

Claims

What is claimed is:

1. A compound selected from the group consisting of: 3'-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]-[1 , 1 '-biphenylj-

3-carboxylic acid;

6-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-2- pyridinecarboxylic acid;

(R)-3-[3-[[2-(3-chlorophenyl)- 2-hydroxyethyl]amino]ethyl]amino]phenyl]-4- pyridinecarboxylic acid;

3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-4- pyridinecarboxylic acid;

(R)-3-[3-[[2-(3-chlorophenyl)- 2-hydroxyethyl]amino]ethyl]amino]phenyl]-2- pyridinecarboxyiic acid; 3-[3-[[2R-[[2-(3-chlorophenyl)-2R-hydroxyethyl]amino]propyl]amino]phenyl]-2- pyridinecarboxylic acid; and pharmaceutically acceptable derivatives thereof.

2. A compound of Claim 1 wherein said compound is an agonist for human beta-3 adrenoceptor.

3. A compound of Claim 1 wherein said compound is a selective agonist for human beta-3 adrenoceptor.

4. A pharmaceutical composition comprising a compound of Claim 1 and pharmaceutically acceptable carrier.

5. A method for the prevention or treatment of clinical conditions or diseases susceptible to amelioration by administration of an atypical beta-adrenoceptor agonist, comprising administration of an effective amount of a compound of Claim 1.