FIELD OF THE INVENTION

The present invention includes compounds that are glucocorticoid receptor modulators. The present invention also includes compositions and methods of using compounds and compositions.

BACKGROUND OF THE INVENTION

Glucocorticoid receptor modulators are glucocorticoid receptor ligands that are used to treat a variety of conditions because of their powerful anti-inflammatory, antiproliferative and immunomodulatory

The invention also relates to compositions comprising a therapeutically effective amount of a compound of Formula I and a pharmaceutically acceptable carrier. Also provided is a method of contacting a glucocorticoid receptor with a compound of Formula I. Further provided are methods of treating a condition in a subject mediated by glucocorticoid receptor activity by administering to the subject a compound of Formula I. DETAILED DESCRIPTION

This detailed description herein is intended only to acquaint others skilled in the art with the inventions, the principles, and the practical applications so that others skilled in the art may adapt and apply the inventions in their numerous forms, as they may be best suited to the requirements of a particular use. These inventions, therefore, are not limited to the embodiments described in this specification, and may be modified.

A. Definitions

The term "subject" refers to any animal, including mammals, such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, primates, or humans. The term "treating" (and corresponding terms "treat" and "treatment") includes palliative, restorative, and preventative ("prophylactic") treating of a subject. The term "palliative treating" refers to treatment that eases or reduces the effect or intensity of a condition in a subject without curing the condition. The term "preventative treating" (and the corresponding term "prophylactic treating") refers to treatment that prevents the occurrence of a condition in a subject. The term "restorative treating" ("curative") refers to treatment that halts the progression of, reduces the pathologic manifestations of, or entirely eliminates a condition in a subject. Treating can be done with a therapeutically effective amount of compound, salt or composition that elicits the biological or medicinal response of a tissue, system or subject that is being sought by an individual such as a patient, researcher, doctor, veterinarian, or clinician

The terms "pharmaceutically effective" or "therapeutically effective" refer to an amount of a compound herein, or salt thereof, that is sufficient to provide an effective treatment, as discussed above It is understood that what comprises a pharmaceutically or therapeutically effective amount may be a lesser amount of the compound or salt when it is administered in combination with another agent than when utilized alone

B. Compounds

sodium salt of the compound of Formula I

Pharmaceutically acceptable acid addition salts are formed from acids which form non-toxic salts Examples include the acetate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate, stearate, succinate, tartrate, tosylate and trifluoroacetate salts Pharmaceutically acceptable base salts are formed from bases which form non-toxic salts Examples include the aluminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts.

For a review on suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002). A salt may be readily prepared by mixing together solutions of compounds of the present invention and the desired acid or base, as appropriate. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the salt may vary from completely ionized to almost non-ionized.

The compounds of the present invention may be administered as prodrugs. Thus, certain

Also included are acid addition or base salts wherein the counterion is optically active, for example, D-lactate or L-lysine, or racemic, for example, DL-tartrate or DL-arginine.

Isomers may be separated by conventional techniques well known to those skilled in the art. The present invention includes isotopically-labeled compounds of the invention wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed. For the treatment of the conditions referred to below, the compounds of the present invention can be administered. Salts of the compounds of the present invention could also be used.

C. Compositions Compounds or salts of the present invention may be part of a composition. Compositions can also include one or more compounds or salts of the present invention. The composition can also include an enantiomeric excess of one or more compounds of the present invention. Other pharmacologically active substances and carriers can be included in the composition.

One embodiment is a composition comprising a compound of Formula I or a salt thereof. Another

erythema multiforme (Stevens-Johnson syndrome), exfoliative dermatitis, mycosis fungoides, psoriasis, and seborrheic dermatitis; e) allergic states, such as seasonal or perennial allergies, allergic rhinitis, bronchial asthma, contact dermatitis, atopic dermatitis, serum sickness, and drug hypersensitivity reactions; f) ophthalmic diseases and conditions, such as allergic corneal marginal ulcers, herpes zoster ophthalmicus, anterior segment inflammation, diffuse posterior uveitis and choroiditis, chronic uveitis, sympathetic ophthalmia, allergic conjunctivitis, keratitis, chorioretinitis, optic neuritis, iritis and iridocyclitis; g) respiratory diseases, such as symptomatic sarcoidosis, Loeffler's syndrome, berylliosis, fulminating or disseminated pulmonary tuberculosis, and aspiration pneumonitis; h) hematologic disorders, such as idiopathic thrombocytopenic purpura, secondary thrombocytopenia, acquired (autoimmune) hemolytic anemia, erythroblastopenia (Red Blood Cell anemia), and congenital (erythroid) hypoplastic anemia; i) neoplastic diseases, such as leukemia and lymphoma; j) edematous states, such as inducing diuresis or emission of proteinuria in the nephrotic syndrome, without uremia, of the idiopathic type or that due to lupus erythematosus; k) gastrointestinal diseases, such as ulcerative colitis, regional enteritis, inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel syndrome;

I) miscellaneous conditions, such as tuberculous meningitis and trichinosis; and

a) asthma of whatever type, etiology, or pathogenesis, in particular asthma that is a member selected from the group consisting of atopic asthma, non-atopic asthma, allergic asthma, atopic bronchial IgE-mediated asthma, bronchial asthma, essential asthma, true asthma, intrinsic asthma caused by pathophysiologic disturbances, extrinsic asthma caused by environmental factors, essential asthma of unknown or inapparent cause, non-atopic asthma, bronchitic asthma, emphysematous asthma, exercise- induced asthma, allergen induced asthma, cold air induced asthma, occupational asthma, infective asthma caused by bacterial, fungal, protozoal, or viral infection, non-allergic asthma, incipient asthma, wheezy infant syndrome and bronchiolytis; b) chronic or acute bronchoconstriction, chronic bronchitis, small airways obstruction, and emphysema; c) obstructive or inflammatory airways diseases of whatever type, etiology, or pathogenesis, in particular an obstructive or inflammatory airways disease that is a member selected from the group consisting of chronic eosinophilic pneumonia, chronic obstructive pulmonary disease (COPD), COPD that includes chronic bronchitis, pulmonary emphysema or dyspnea associated or not associated with COPD, COPD that is characterized by irreversible, progressive airways obstruction, adult respiratory distress syndrome (ARDS), exacerbation of airways hyper-reactivity consequent to other drug therapy and airways disease that is associated with pulmonary hypertension; d) bronchitis of whatever type, etiology, or pathogenesis, in particular bronchitis that is a member selected from the group consisting of acute bronchitis, acute laryngotracheal bronchitis, arachidic

comprising administering to the subject a compound or salt of the present invention.

The present invention includes a method of treating conditions such as asthma, dermatitis, inflammatory bowel disease, Alzheimer's disease, psychotic major depression, neuropathy, transplant rejection, multiple sclerosis, chronic uveitis, or chronic obstructive pulmonary disease in a subject comprising administering to the subject a compound or salt of the present invention.

The present invention includes a method of treating rheumatoid arthritis in a subject comprising administering to the subject a compound or salt of the present invention. Rheumatoid arthritis is considered a chronic autoimmune and inflammatory disease producing inflamed joints, which eventually swell, become painful, and experience degradation of cartilage, bone, and ligaments of the joint. A result of rheumatoid arthritis is deformity, instability, and stiffness of the joint and scarring within the joint. The joints deteriorate at a highly variable rate. Many factors, including genetic predisposition, may influence the pattern of the disease. People with rheumatoid arthritis may have a mild course, occasional flare-ups with long periods of remission without disease, or a steadily progressive disease, which may be slow or rapid. Rheumatoid arthritis may start suddenly, with many joints becoming inflamed at the same time. More often, it starts subtly, gradually affecting different joints. Usually, the inflammation is symmetric, with joints on both sides of the body affected. Typically, the small joints in the fingers, toes, hands, feet, wrists, elbows, and ankles become inflamed first, followed by the knees and hips.

Pain associated with rheumatoid arthritis is typically somatic nociceptive joint pain. Swollen wrists

proposed indication, the compounds or salts of the invention can be assessed for their biopharmaceutical properties, such as solubility and solution stability (across pH), and permeability. Doses for compounds or salts of the invention range from 0.1 mg to 100 mg for oral administration and doses range from 2 mg or less for inhaled administration. The dose may be administered in single dose or two or more divided doses and may fall outside of the typical range given herein.

The dosages are based on an average human subject having a weight of about 60 kg to 70 kg. Dosing and dosing regimen depend upon subject and a variety of conditions that may affect dosing (age, sex, body weight, etc.). A physician or other medical professional will readily be able to determine doses for subjects whose weight falls outside this range, such as infants and the elderly. Oral Administration

The compounds of the invention and salts thereof may be administered orally. Oral administration may involve swallowing, so that the compound or salt enters the gastrointestinal tract, and/or buccal, lingual, or sublingual administration by which the compound or salt enters the blood stream directly from the mouth.

Formulations suitable for oral administration include solid, semi-solid and liquid systems such as tablets; soft or hard capsules containing multi- or nano-particulates, liquids, or powders; lozenges (including liquid-filled); chews; gels; fast dispersing dosage forms; films; ovules; sprays; and

calcium phosphate dihydrate. Tablets may also optionally comprise surface active agents, such as sodium lauryl sulfate and polysorbate 80, and glidants such as silicon dioxide and talc. When present, surface active agents may comprise from 0.2 weight % to 5 weight % of the tablet, and glidants may comprise from 0.2 weight % to 1 weight % of the tablet.

Tablets also generally contain lubricants such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate with sodium lauryl sulphate. Lubricants generally comprise from 0.25 weight % to 10 weight %, preferably from 0.5 weight % to 3 weight % of the tablet.

Other possible ingredients include anti-oxidants, colorants, flavoring agents, preservatives and taste-masking agents. Exemplary tablets contain up to about 80% drug, from about 10 weight % to about 90 weight % binder, from about 0 weight % to about 85 weight % diluent, from about 2 weight % to about 10 weight % disintegrant, and from about 0.25 weight % to about 10 weight % lubricant.

Solid formulations for oral administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.

Suitable modified release formulations for the purposes of the invention are described in US Patent No. 6,106,864. Details of other suitable release technologies such as high energy dispersions and osmotic and coated particles are to be found in Pharmaceutical Technology On-line, 25(2), 1-14, by

semi-solid, or thixotropic liquid for administration as an implanted depot providing modified release of the active compound. Examples of such formulations include drug-coated stents and semi-solids and suspensions comprising drug-loaded poly(dl-lactic-coglycolic)acid (PGLA) microspheres.

Topical Administration

The compounds or salts of the invention may also be administered topically, (intra )dermally, or transdermal^ to the skin or mucosa. Example 1 could be administered to the skin. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibres, bandages and microemulsions. Liposomes may also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol and propylene glycol. Penetration enhancers may be incorporated - see, for example, J Pharm Sci, 88 (10), 955-958, by Finnin and Morgan (October 1999).

Other means of topical administration include delivery by electroporation, iontophoresis, phonophoresis, sonophoresis and microneedle or needle-free (e.g. Powderject™, Bioject™, etc.) injection.

Formulations for topical administration may be formulated to be immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.

A suitable solution formulation for use in an atomiser using electrohydrodynamics may comprise a compound of the present invention, propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents which may be used instead of propylene glycol include glycerol and polyethylene glycol.

Formulations for inhaled/intranasal administration may be formulated to be immediate and/or modified release using, for example, PGLA. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release. In the case of dry powder inhalers and aerosols, the dosage unit is determined by means of a valve which delivers a metered amount. Units in accordance with the invention are typically arranged to administer a metered dose or "puff which may be administered in a single dose or, more usually, as divided doses throughout the day.

Dose ranges for inhaled administration range from 2 mg to less or 1 mg to less.

Combination

The compounds or salts of the invention may be administered in combination with one or more other therapeutic agents, such as a drug. The compound of the present invention or salt thereof may be administered at the same time or different time as one or more other therapeutic agents.

For example, "in combination" includes: simultaneous administration of a combination of

F. Use in the Preparation of a Composition or Medicament In one embodiment, the present invention comprises methods for the preparation of a composition or medicament comprising the compounds or salts of the present invention for use in treating condition mediated by glucocorticoid receptor activity.

In another embodiment, the invention comprises the use of one or more compounds or salts of the present invention in the preparation of a composition or a medicament for inflammation, inflammation related condition, rheumatoid arthritis, dermatitis, Alzheimer's disease. The present invention also includes the use of one or more compounds or salts of the present invention for preparation of a composition or a medicament for treating one or more conditions detailed in the Methods section.

G. Schemes

The compounds of the present invention may be prepared using the methods illustrated in the general synthetic schemes and experimental procedures detailed below. The reactions of the synthetic methods herein are carried out in suitable solvents which may be readily selected by one skilled in the art of organic synthesis, said suitable solvents generally being any solvent which is substantially nonreactive

A-6 A-7

The compound B-1 , 7-bromo-2-ethoxy-3,4,4a,9-tetrahydro-4a-(phenylmethyl)-(4aS) -phenanthrene, may be prepared as described in European Patent Applications EP 1201649 and EP 1201660 (both to Liu et al.) and EP 1201665 (Murry et al.).

Preparation 1 : (S)-4α-benzyl-7-bromo-2-ethoxy-3,4,4α,9-tetrahydrophenanth rene Br ^ ^^ B-1

Starting Material A-8 (450 g; 1.17 moles) was dissolved in ethanol (4.5 L) at ambient temperature. 21 % sodium ethoxide in ethanol (44 ml_; 0.12 moles) was added and the mixture was heated to reflux for three h O h S i M i l A 8 d h i i hill d 2 °C A l

distillation process was repeated twice more. More p-toluenesulfonic acid (6.1 g) was added each time fresh toluene was added. During the reaction, two intermediates (detected by LC) were formed as the substrate was converted into product. The end point of the reaction was an equilibrium point between the two intermediates and the product. Once the endpoint was reached, the mixture was cooled to ambient temperature. The mixture was washed with 0.5 M NaOH (2 L). The phases separated quickly and both were dark with a small rag layer. The mixture was washed with water (2 L). The phases separated very slowly. The mixture was dried by azeotropic distillation. Methanol (4 L) was added to the mixture and solvent (4 L) was distilled from the mixture. The methanol addition and solvent distillation were repeated twice more. Methanol was added to the mixture and precipitation occurred a few minutes later. More methanol (4 L) was added to the mixture and then brought to reflux. After 30 minutes, the mixture was cooled to 0ºC. The mixture was filtered and the solid was washed with chilled methanol (2 X 2L). The solid was dried in a vacuum oven at 65°C. The title compound (882 g; 98 area %) was obtained as a beige solid. ¹ H NMR (DMSO) δ ppm: 1.71 (m, 2H), 2.06 (m, 2H), 2.31 (dd, 1H), 2.39 (m, 1H), 2.68 (d, 1H), 2.77 (m, 1H), 2.86 (dd, 1H), 3.36 (d, 1H), 3.86 (m, 4H), 5.45 (m, 1H), 6.50 (m, 2H), 7.00 (m, 4H), 7.37 (dd, 1H), 7.44 (d, 1H).

3.87 (m, 7H), 5.49 (m, 1H), 6.47 (m, 2H), 6.93 (m, 2H), 7.01 (m, 1H), 7.42 (d, 1H), 7.64 (d, 1H), 7.79 (dd, 1H).

Preparation H^C B-4

The (S)-methyl 4β-benzyl-7,7-(1 ,2-ethylenedioxy)-4β,5,6,7,8,10hexahydrophenanthrene-2-carb oxylate (201 g; 0 515 moles which may be prepared as described in Preparation 3) and 50 ml of ethylene glycol

Preparation 5: (4bS,8aR)-4b-benzyl-7-oxo-4b,5,6,7,8,8a,9,10-octahydrophenan threne-2-carboxylic acid

(4bS,8aR)-4b-benzyl-7-oxo-4b,5,6,7,8,8a,9,10-octahydrophe nanthrene-2-carboxylic acid (6.5 g, 19.44 mmol) was suspended in water (65 ml_). 2.5 M NaOH (11.7 ml_, 29.16 mmol) was added followed by benzaldehyde (2.16 ml_, 21.38 mmol). Over time (at 50ºC for 4 hours or at 25°C over night) the mixture became homogeneous. The reaction was considered complete when there was less than 2% of (4bS,8af?)-4b-benzyl-7-oxo-4b,5,6,7,8,8a,9,10-octahydrophena nthrene-2-carboxylic acid (Preparation 5) remaining. The mixture was cooled to 25°C if it was not already at 25°C. EtOAc (65 ml_) was added to the mixture followed by aqueous 2 M HCI (29 ml_). Crystallization normally occurred during the acid addition or shortly thereafter. The mixture was stirred for 60 minutes. Heptane (65 ml_) was added and the mixture was stirred for an additional 60 minutes. Do not worry about separating out the aqueous phase; filter the entire mixture and wash the solid with water followed by heptane. A pale yellow solid (6.55 g, 80% yield) was obtained. ¹ H NMR (DMSO): δ 1.70 (m, 1 H), 1.85 (m, 1 H), 2.45 (m, 3H), 2.65 (d, 3H), 2.95 (m, 2H), 3.50 (d, 1 H), 6.15 (d, 2H), 6.25 (d, 1 H), 6.70 (t, 2H), 6.90 (t, 1 H), 7.30 (d, 1 H), 7.50 (m, 5H), 7.70 (d, 2H), 12.75 (s, 1 H).

Preparation 7: (4bR,6E,7S,8aR)-4b-benzyl-6-benzylidene-7-hydroxy-7-phenyl-4 b,5,6,7,8,8a,9,10- octahydrophenanthrene-2-carboxylic acid

O C q y q SO ₄ p maintaining the temperature below 1OºC. A total of 50 ml of the saturated solution was added. Solids formed in the solution and were removed by vacuum filtratration. The filter cake was washed with tetrahydrofuran (40 mL) and water (40 mL). Ethyl acetate (100 mL) was added and the layers separated. The organic layer was washed with saturated ammonium chloride (100 mL), dried over sodium sulfate, and solvent removed at reduced pressure. The aqueous layer was checked by HPLC/MS and did not contain any product. The residue was taken up in methanol (15 ml) and water added. The solution became milky and eventually a precipitate formed. Small amounts of methanol and water were added to improve the quality and quantity of precipitate. The solids were collected by vacuum filtration and air dried for about 2 hours. 3.4 grams of the title compound as a light yellow solid was obtained in 81 % yield. 1 H NMR (400 MHz, DMSO-O ₆ ) d ppm 12.65 (1 H, s), 7.44 - 7.56 (7 H, m), 7.34 - 7.39 (2 H, m), 7.30 (2 H, t, J=7.7 Hz), 7.12 - 7.22 (2 H, m), 6.78 (1 H, t, J=7.4 Hz), 6.50 (2 H, t, J=7.8 Hz), 6.12 (1 H, d, J=8.3 Hz), 5.86 (2 H, d, J=7.3 Hz), 5.44 (1 H, s), 3.61 (1 H, d, J=14.2 Hz), 2.96 (1 H, dd, J=17.6, 7.9 Hz), 2.80 - 2.91 (1 H, m), 2.66 - 2.74 (1 H, m), 2.50 - 2.65 (2 H, m), 2.08 (1 H, t, J=13.3 Hz), 1.82 - 1.93 (1 H, m, J= 13.2 Hz), 1.75 - 1.82 (1 H, m), 1.59 - 1.74 (2 H, m); LC/MS, t _r = 3.77 minutes (5 to 95% acetonitrile/water over 5 minutes at 1 ml/min, at 254 nm, at 50 ºC).

Preparation 8: (4bR,7R,8aR)-4b-benzyl-7-hydroxy-6-oxo-7-phenyl-4b,5,6,7,8,8 a,9,10-

q grams of a orange brown oil was obtained. HPLC/MS obtained. The mother liquor was purified utilizing preparative reverse phase chromatography to afford an additional 22g of the title compound for a total recovery of 27.5g in 85% yield. MH ⁺ [m/z] 501 M+Na [m/z] 523 ; 1 H NMR (400 MHz, DMSO-Cf ₆ ) δ ppm 2.00 (s, 4 H) 2.10 - 2.21 (m, 3 H) 2.51 (q, 3 H) 2.73 - 2.85 (m, 4 H) 2.96 - 3.10 (m, 3 H) 6.15 (dd, 1 H) 6.60 (dd, 2 H) 7.09 - 7.15 (m, 3 H) 7.25 (d, 1 H) 7.30 (d, 1 H) 7.34 - 7.40 (m, 4 H) 7.69 (s, 1 H).

Preparation 9: (4bR,7R,8aR)-4b-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-6- oxo-7-phenyl- 4b,5,6,7,8,8a,9,10-octahydrophenanthrene-2-carboxamide

(4bR,7R,8aR)-4b-benzyl-7-hydroxy-6-oxo-7-phenyl-4b,5,6,7, 8,8a,9,10-octahydrophenanthrene-2- b li id (21 49 2 l) 3 i 2 i li (5 8 52 2 l) d 1 th li id l (20 l

added Mel_i ^* l_iBr (0.1 mL). After 1 hour the LCMS indicated that starting material remained so an additional 0.15mL was added. The reaction stirred to room temperature over 2 hours. By HPLC, 2.5% of the starting material remained. To the reaction was added NH ₄ CI slowly and off gassing was observed. The reaction was diluted to 125mL with acetonitrile and water. The reaction was purified by reverse phase chromatography then was lyophilized. The resulting dried powder was dissolved in acetonitrile and water again with two drops of concentrated HCI. The solution was lyophilized to dryness. This afforded the title product (231.4mg) as the HCI salt in 73% yield. LRMS ES ⁺ 533.1 1 H NMR (300 MHz, METHANOL-O ₄ ) δ ppm 1.23 - 1.28 (m, 3 H) 1.53 - 1.62 (m, 1 H) 1.90 - 2.20 (m, 3 H) 2.70 - 2.74 (m, 3 H) 2.84 (d, J=14.90 Hz, 1 H) 2.92 - 3.22 (m, 2 H) 3.28 - 3.40 (m, 4 H) 3.91 (d, J=12.08 Hz, 1 H) 6.50 (d, J=8.26 Hz, 2 H) 6.84 - 6.92 (m, 2 H) 6.99 - 7.10 (m, 3 H) 7.13 - 7.29 (m, 3 H) 7.45 (dd, J=8.15, 1.91 Hz, 1

H) 7.56 - 7.65 (m, 2 H) 7.77 (d, J=1.81 Hz, 1 H) 7.88 (dd, J=8.26, 5.84 Hz, 1 H) 8.51 - 8.63 (m, 2 H).

Example 1 : (4bR,6R,7R,8aS)-4b-benzyl-6,7-dihydroxy-6-methyl-N-(2-methyl pyridin-3-yl)-10-oxo-7- phenyl^b^δJ^δa^iO-octahydrophenanthrene^-carboxamide

product as its parent compound. The resulting solid was crystallized by the following procedure: The solid was slurried with MeOH (7 mL), the solids collected after 1 hour and washed with an additional 2 mL of MeOH to furnish the title compound 1802 mg, 79%. Analytical data as follows: ¹ H NMR (500 MHz, D ₆ DMSO) δ ppm 1.18 (s, 3 H) 1.51 (dd, J=12.66, 1.96 Hz, 1 H) 1.99 (d, J=14.96 Hz, 1 H) 2.40 (dd, J=18.88, 4.85 Hz, 1 H) 2.43 - 2.49 (m, 1 H) 2.49 (br. s., 3 H) 2.56 (t, J=12.82 Hz, 1 H) 2.61 (d, J=12.53 Hz, 1 H) 2.75 (d, J=15.12 Hz, 1 H) 2.92 (dd, J=18.59, 12.74 Hz, 1 H) 4.05 (d, J=12.45 Hz, 1 H) 6.65 (d, J=8.27 Hz, 1 H) 6.79 (dd, J=7.69, 1.59 Hz, 2 H) 7.05 - 7.13 (m, 3 H) 7.15 - 7.21 (m, 1 H) 7.25 (t, J=7.69 Hz, 2 H) 7.49 (dd, J=7.94,5.10Hz, 1 H) 7.62 (dd, J=7.44, 1.25 Hz, 2 H) 7.87 (dd, J=8.23, 2.13 Hz, 1 H) 8.01 (dd, J=7.78, 1.00Hz, 1 H) 8.46 (dd, J=5.01, 1.50Hz, 1 H) 8.54 (d, J=2.09 Hz, 1 H) 10.18 - 10.58 (m, 1 H, NH). Dependant upon the water content of DMSO, two OH protons can be visible at 4.78 and 5.44 ppm. HRMS m/z 547.2619 (C ₃₅ H ₃₅ N ₂ O ₄ : calcd for M+H, 547.2591 ).

I. BIOLOGICAL DATA Lipopolysaccharide (LPS)-lnduced Human Whole Blood

Venous blood from human donors was collected as 10 ml aliquots in tubes containing sodium heparin (BD Vacutainer from Becton Dickinson and Company, Franklin Lakes, NY). Blood was added to sterile polystyrene round bottom 96-well tissue culture plates (Corning Costar) at 180 μl/well. Blood was placed in a humidified 37C incubator with 5% CO while compounds were prepared (nearly 60 minutes)

( , g p , , Sigma-Aldrich). Calibrators for standard curves were prepared in human plasma/serum assay diluent as a 1/5 serial dilution to achieve final concentrations ranging from 50000 pg/ml to 3.2 pg/ml. Samples were added at 10 to 20 μl/well and calibrators were added at 20 μl/well, then incubated at room temperature with gentle shaking for 2 hours. Plates were again washed 3x with wash buffer. Detection antibody was diluted in human plasma/serum antibody diluent to 1 μg/ml and added to the plate at 20 μl/well. Plates were incubated as before for 2 hours and washed again. Read Buffer T (4x) was diluted 1 :1 with mqH ₂ O to 2x concentration and 150 μl added to each well. Plates were read on the SECTOR Imager 6000 (Meso Scale Discovery) to generate raw signal values. Individual sample signal values were compared to positive and negative controls (vehicle treated blood with LPS and vehicle treated blood without LPS, respectively) to generate % inhibition. Triplicate values were averaged for each donor. The values for three or four donors were averaged and graphed using A- parameter fit curves in the LabStats plug-in for the Microsoft Excel application.

Prednisolone was obtained from Sigma-Aldrich (Saint Louis, MO).

Table 1. Mean Values of Prednisolone Inhibition

Comparator A is (4βS,7S,8αf?)-4β-benzyl-7-hydroxy-Λ/-((2-methylpyridin-3 -yl)methyl)-7-(3,3,3- trifluoropropyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene- 2-carboxamide, the synthesis for which is described as Example No. 771 C-3 on page 241 of WO 00/66522 (Dow et al.), and has the following structure:

The following comparative compounds, Comparators B, C and D, can be prepared by methods described herein, those known in the art and Scheme D, below.

Comparator S 1 ,2,3,4,4a,9,10,10a-octahydrophenanthrene-2,3 diol, the synthesis of which is described as Example 32 on page 107 of international application WO 2004/005229 (Chantigny et al.) and has the following structure:

stimulated human whole blood. Prednisolone, a full agonist of GR, demonstrates full inhibition of IFN ,

TNF and IL-1 . Example 1 also inhibits cytokine release in a concentration-dependent manner. Being a partial agonist/antagonist, Example 1 does not show inhibition to the extent of prednisolone. Furthermore, the level of inhibition observed for Example 1 is different between cytokines measured,that is to say,. IFN =73%, TNF =43%, and IL-1 =38%. In contrast to Example 1 , Comparator F and Comparator G do not significantly inhibit TNF or IL-1 (less than 20% at 300OnM) and show a very much less efficacious inhibition of IFN (only 22% or 41 % inhibition at 3000 nM, respectively). Thus, while Example 1 , Comparator F, Comparator G and prednisolone bind to the same receptor, they demonstrate markedly different activities.

TNFa repression after a single ED ₈₀ dose (determined from the acute LPS endotoxemia mouse model). In order to repress TNFa significantly over a 24 hour period, it was determined that prednisolone required b.i.d. dosing. DAGR compounds vary in their required frequency of doses.

Body weights are measured on the first and last day of each experiment. Blood samples are obtained after ~3 weeks of dosing for steady-state pharmacokinetic analysis. To assess compound effects on LPS-induced TNF-a, all mice receive an intraperitoneal injection of LPS (Salmonella Typhosa , Sigma, St. Louis, L-7895) 2.5 hr after the last dose on day 28. Mice are sacrificed 90 min. after LPS administration Serum samples are assayed for osteocalcin and TNFa using the Linco multiplex assay (St. Charles, MO) and Luminex 100 (Austin, TX). Samples are diluted 1 :20 and the assay is run according to manufacturer's instructions. An osteocalcin standard is purchased separately from Biomedical Technologies Incorporated (Stoughton, MA). Serum insulin is measured to assess compound effects on insulin resistance. Insulin was measured using the Ultrasensitive Mouse EIA kit from Alpco Diagnostics (Salem, NH) following the manufacturer's protocol.

Cortical Bone Histomorphometry

During the in-life portion of each study, mice received two IP injections of the fluorochrome calcein (C- 0875; Sigma-Aldrich; 20 mg/kg; 200 μL/mouse), dissolved in 2% sodium bicarbonate, on days 1 and 26 for bone histomorphometry measurements. Fluorochrome labels incorporate into the bone mineral and allow measurement of bone formation rate During the tissue harvest the left tibia is excised and cleaned

injection and 5μL was injected in the LC/MS/MS system for analysis. Concentrations below the limit of quantitation (LOQ) are reported as zero (0) and are used in the evaluation of mean concentrations and the estimation of AUC. The area under the concentration-time curve from time zero to time of the last quantifiable concentration (t) [AUC(O-t)] is determined using the linear trapezoidal method.

Statistical Evaluation

ED ₅₀ and ED ₈₀ values are obtained for the various parameters using four-parameter logistic fits of data. For each experiment/dose group, outliers are detected by calculating the number of standard deviations each mouse's value was from the mean of their group and then dividing by the standard deviation of the group. The means and standard deviations used in this calculation omitted the value being examined so that, if it were an outlier, it would have no influence. If the value being examined is more than 2.5 standard deviations from the mean, it was not used in the rest of the calculations.

Percent inhibition values are then calculated for each animal using the means of the vehicle and 10 mpk prednisone control groups. The individual mouse percent inhibition values are then fit to a four- parameter logistic model using the area under curve mean for each group. Since all four parameters are estimated and the lower plateau is not fixed at 0% and the upper plateau was not fixed at 100%, the ED ₅₀ and ED ₈₀ values are calculated by using an inverse calibration formula for a response equal to 50% or 80%. The designation "nd" means not determined.

effects. Dissociation indices were calculated using clinically relevant biomarkers that could be utilized in early clinical development. Serum osteocalcin and LPS-induced serum TNFα are accepted clinically as predictive for bone formation and anti-inflammatory efficacy, respectively. The dissociation index was based on the following tenets:

1) Dissociation required a dose-margin between biomarkers of inflammation and side-effects [such as osteocalcin (OC), insulin, or bone formation rate], and was defined by the formula, using osteocalcin suppression (OC) as the side effect example:

Dl = Side-effect endpoint

Anti-inflammatory endpoint

2) The Dl of a compound can be considered relative to that observed with prednisolone, its clinical comparator. The corrected or normalized Dl was defined as compound Dl divided by prednisolone Dl.