RELATED APPLICATION This application claims the benefit of U.S. Provisional Application No.

61/191,639, filed on September 10, 2008, the entire teachings of which are incorporated herein by reference.

GOVERNMENT SUPPORT

This invention was made in part through funding from the United States government through National Institutes of Health grant numbers PHS 5R37 DK 15556, PHS 5R37 CA 25836 and 5R01 CA 18119. The United States Government has certain rights in this invention.

BACKGROUND OF THE INVENTION The estrogen receptor family to date has been shown to include 2 primary subtypes, ERa and ERβ (collectively referred to as "the estrogen receptor"). The estrogen receptor is a ligand modulated transcription factor that is part of the broader family of steroid nuclear hormone receptors. Estrogen acting through the estrogen receptor mediates a large number of physiological events in both males and females. In females, agonist activity mediated through the estrogen receptor is responsible for the development and maintenance of the reproductive system as well as the maintenance of bone mineral density. Estrogens (compounds affecting physiological processes thru one or both of the estrogen receptors) have found utility in a wide variety of pharmaceutical products including, for example, contraception, osteoporosis, oncology, and postmenopausal symptoms (e.g., hot flush, vaginal dryness). Estrogens have also been associated with an anti-inflammatory effect as well as effects on the immune system. For example, it has been reported that estrogens can reduce indices of inflammation in preclinical models of arthritis. Additionally, it has also been reported that estrogens can reduce the mortality in preclinical models of sepsis (Harris, Heather A. Molecular Endocrinology (2007), 21(1), 1-13). Interestingly, it appears that the anti-inflammatory effects can be mediated by estrogen receptor beta since estrogen receptor beta selective agonists have shown efficacy in many animals models of inflammation. The estrogen inflammation nexus has further support in humans since the use of aromatase inhibitors (compounds that block the conversion of the precursor steroids androstenedione and testosterone) is associated with reduced levels of estrogens and increases in joint and muscle pain (Morales, et al. Breast Cancer Research and

Treatment (2007), 104(1), 87-91). Unfortunately, the use of a non-selective estrogen would be generally contraindicated in a patient taking an aromatase inhibitor. However, compounds that demonstrated little or no ERalpha agonism but that retained ERbeta agonism could be useful in such a population due to such a compounds ability to provide an anti-inflammatory effect without mammary tissue proliferation. Accordingly, there is much use for new estrogen receptor ligands, particularly those with subtype selectivity.

SUMMARY OF THE INVENTION Described herein are compounds of structure formula I ₅ compositions containing compounds of formula I and methods of using those compounds.

In some embodiments, this invention includes a compound according to formula I or a pharmaceutically acceptable salt thereof

I wherein:

Ri and R ₁ ' are each independently selected from the group consisting of hydrogen,

Ci _-6 alkyl, Ci _-6 alkanoyl, benzyl and benzoyl; D and G are each independently selected from S or C(R ₆ )(R ₇ ), provided that when D is S then G is C(R ₆ )(R ₇ ) and when G is S then D is C(R ₆ )(R ₇ );

E is (C(R ₆ )(R ₇ )),,; each R ₆ and each R ₇ are independently selected from hydrogen, C ₁ . ₅ alkyl, or C ₁ . ₅ haloalkyl; or R ₆ and R ₇ taken together with the carbon they are attached from a cyclopentyl or cyclohexyl ring; n is 0 or 1 ; and R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from the group consisting of hydrogen, Cj _-3 alkyl, Ci _-3 haloalkyl and halogen.

In certain embodiments, this invention includes a compound according to formula I, or a pharmaceutically acceptable salt thereof, wherein D is S; and Ri and Ri ' are each hydrogen.

In certain embodiments, this invention describes a compound of formula II, or a pharmaceutically acceptable salt thereof

II wherein:

G is C(R ₆ )(R ₇ ) and E is (C(R ₆ )(R ₇ )),,; each Rg and each R ₇ are independently selected from hydrogen, Ci _-5 alkyl, or Ci _-J haloalkyl; or R ₆ and R ₇ taken together with the carbon they are attached from a cyclopentyl or cyclohexyl ring; n is 0 or 1 ; and

R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from the group consisting of hydrogen, C] _-3 alkyl, Cj _-3 haloalkyl and halogen.

In some embodiments, this invention includes a compound of formula I, or a pharmaceutically acceptable salt thereof, wherein D, E and G are each C(R ₆ )(R ₇ ); Ri and Ri ' are each hydrogen; at least one of R ₂ and R ₄ is fluorine or methyl; provided that at least one of D, E and G is CUi; and provided that R ₂ and R ₄ are not both methyl.

In certain embodiments, this invention describes a compound of formula III, or a pharmaceutically acceptable salt thereof

Ill wherein: D, E and G are each C(R ₆ )(R ₇ ); each R ₆ and each R ₇ are independently selected from hydrogen, Ci _-5 alkyl, or Ci _-5 haloalkyl; or R ₆ and R ₇ taken together with the carbon they are attached from a cyclopentyl or cyclohexyl ring;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from the group consisting of hydrogen, C] _-3 alkyl, Cj _-3 haloalkyl and halogen, wherein at least one of R ₂ and R ₄ is fluorine or methyl; provided that at least one of D, E and G is CH ₂ ; and provided that R ₂ and R ₄ are not both methyl.

In certain embodiments, this invention includes a compound of formula I, or a pharmaceutically acceptable salt thereof, wherein D, E and G are each CH ₂ , Ri and Ri' are each independently selected from the group consisting of hydrogen, Ci _-6 alkyl, C] _-6 alkanoyl, benzyl and benzoyl; and at least one of R ₂ and R ₄ is fluorine.

In certain embodiments, this invention describes a compound of formula IV, or a pharmaceutically acceptable salt thereof

IV wherein:

Ri and Ri ' are each independently selected from the group consisting of hydrogen, Ci- ₆ alkyl, Ci _-6 alkanoyl, benzyl and benzoyl;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from the group consisting of hydrogen, Ci _-3 alkyl, Ci _-3 haloalkyl and halogen; and at least one of R ₂ and R ₄ is fluorine.

In some embodiments, this invention includes a compound of formula I, or a pharmaceutically acceptable salt thereof, wherein D, E and G are each C(R ₆ )(R ₇ ); Ri and R] ' are each hydrogen; and at least one of R ₂ and R ₄ is fluorine.

In some embodiments, this invention describes a compound of formula V, or a pharmaceutically acceptable salt thereof:

V wherein: D, E and G are each C(R ₆ )(R ₇ ); each R ₆ and each R ₇ are independently selected from hydrogen, C _1-S alkyl, or Ci _-5 haloalkyl; or R ₆ and R ₇ taken together with the carbon they are attached from a cyclopentyl or cyclohexy] ring;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently selected from the group consisting of hydrogen, Ci _-3 alkyl, Ci _-3 haloalkyl and halogen; and and at least one of R ₂ and R ₄ is fluorine.

In certain embodiments, this invention includes a compound of formula I, or a pharmaceutically acceptable salt thereof, wherein n is 0; one of R ₂ or R ₄ is methyl and the other is hydrogen; Rj and Ri ' are each hydrogen;R ₃ and R ₅ are each hydrogen; D is CH ₂ and G is C(R ₆ )(R ₇ ) or D is C(R ₆ )(R ₇ ) and G is CH ₂ ..

In some embodiments, this invention describes a compound of formula VI, or a pharmaceutically acceptable salts thereof,

VI wherein:

D is CH ₂ and G is C(R ₆ )(R ₇ ) or D is C(R ₆ )(R ₇ ) and G is CH ₂ ; each R ₆ and each R ₇ are independently selected from hydrogen, Ci _-5 alkyl, or Ci _-5 haloalkyl; or R ₆ and R ₇ taken together with the carbon they are attached from a cyclopenlyl or cyclohexy 1 ring; and one of R ₂ or R ₄ is methyl and the other is hydrogen

In some embodiments, this invention includes a compound of formula I or a pharmaceutically acceptable salt thereof, wherein n is 0; one of R ₂ or R ₄ is fluorine and the other is hydrogen; R] and Ri ' are each hydrogen; R ₃ and R ₅ are each hydrogen; and D is CH ₂ and G is C(R ₆ )(R ₇ ) or D is C(R ₆ )(R ₇ ) and G is CH ₂ .

In certain embodiments, this invention describes a compound of formula VII, or a pharmaceutically acceptable salt thereof,

VII D is CH ₂ and G is C(R ₆ )(R ₇ ) or D is C(R ₅ )(R ₇ ) and G is CH ₂ ; each R ₆ and each R ₇ are independently selected from hydrogen, C _{1 -5} alkyl, or C ₁ - ₅ haloalkyl; or R ₆ and R ₇ taken together with the carbon they are attached from a cyclopentyl or cyclohexyl ring; and one of R ₂ or R ₄ is fluorine and the other is hydrogen

In certain embodiments, this invention includes a compound according to formula I, wherein said compound is a) 4, 4'-(2-methylpropane-l,2-diyl)diphenol; b) 4-(2-(4-hydroxyphenyl)-2-methylpropyl)-3-methylphenol; c) 4-(2-(4-hydroxyphenyl)-2-methylpropyl)-2-methylphenol; d) 3-chloro-4-(4-hydroxyphenylethyl)phenol; d) 4-(4-hydroxybenzylthio)phenol; e) 2-fluoro-4-(4-hydroxyphenethyl)phenol; f) 4-(l -(4-hydroxyphenyl)propan-2-yl)-3-methylphenol; g) 2-fluoro-4-(2-(4-hydroxybenzyl)butyl)phenol; h) 4,4 '-(propane- 1 ,3-diyl)bis(2-fiuorophenol); i) 4-(3-(4-hydroxyphenyl)-3-methylbutan-2-yl)-3-methylphenol; j) 2-fluoro-4-(l-(4-hydroxyphenyl)hexan-3-yl)phenol; k) 2-fluoro-4-(2-(4-hydroxyphenyl)-2-methylpropyl)phenol;

1) 4-(l-(4-hydroxyphenyl)ethylthio)phenol; m) 2-fluoro-4-( 1 -(4-hydroxyphenyl)-2-methylpropan-2-yl)phenol; n) 4-(l-(4-hydroxyphenyl)pentan-2-yl)-2-methylphenol; o) 2-fluoro-4-(l -(4-hydroxyphenyl)propan-2-yl)phenol; p) 2-fluoro-4-(2-(4-hydroxyphenyl)pentyl)phenol; q) 2-fluoro-4-(l -(4-hydroxy-3-methylphenyl)propan-2-yl)phenol; r) 2-fluoro-4-(3-(4-hydroxyphenyl)propyl)phenol; s) 4-(3-(4-hydroxyphenyl)hexyl)-2-methylphenol; t) 4-(4,4,4-trifluoro- 1 -(4-hydroxyphenyl)butylthio)phenol; u) 2-fluoro-4-(2-(4-hydroxyphenyl)propyl)phenol; v) 2-fluoro-4-(2-(4-hydroxy-3-methylphenyl)propyl)phenol; w)4-(l-(4-hydroxyphenyl)propy!thio)phenol; x) 4-(l -(4-hydroxyphenyl)-2-methylpropylthio)phenol; y) 3 -fluoro-4-(2-(4-hydroxyphenyl)propyl)phenol; z) 4,4'-(propane-l,2-diyl)bis(2-fluorophenol); aa) 4-(2-(4-hydroxyphenyl)propyl-2-methylphenol; bb) 4,4'-(2-methylpropane-l,2-diyl)bis(2-fluorophenol); cc) 2-fluoro-4-(l -(4-hydroxybenzyl)cyclopentyl)phenol; dd) 4,4'-(propane-l,2-diyl)bis(2-fluorophenol); ee) 2-fluoro-4-(2-(4-hydroxy-2-methylphenyl)propyl)phenol; ff) 2-fluoro-4-(l-(3-fluoro-4-hydroxybenzyl)cyclohcxyl)phenol; gg) 2-fluoro-4-(l -(3-fluoro-4-hydroxybenzyl)cyclopentyl)phenol; hh) 4-(2-(4-hydroxyphenyl)propan-2-ylthio)phenol; ii) 4-(33,3-trifluoro-l-(4-hydroxyphenyl)propylthio)phenol; jj) 2-flυoro-4-(3,3,3-trifluoro-l-(4-hydroxyphenyl)propylthio)p henol; kk) 2-fluoro-4-(2-(4-hydroxyphenyl)propan-2-ylthio)phenol;

11) 2-fluoro-4-(l-(4-hydroxy-2-methylphenyl)-2-methylpropan-2-yl )phenol; mm) 2-flυoro-4-(l -(4-hydroxyphenyl)ethylthio)phenol; nn) 2-fluoro-4-(l-(4-hydroxyphenylthio)ethyl)phenol; oo) 2-fluoro-4-(4,4,4-trifluoro- 1 -(4-hydroxyphenyl)butylthio)phenol; and pp) 4-(l -(4-hydroxyphenyl)butylthio)phenol; or a pharmaceutically acceptable salt of any of the foregoing.

In other embodiments, this invention includes a compound according to formula I, wherein said compound is qq) (S)-2-fluoro-4-(2-(4-hydroxyphenyl)propylphenol; or rr) (R)-2-fluoro-4-(2-(4-hydroxyphenyl)propylphenol or a pharmaceutically acceptable salt of any of the foregoing.

In some embodiments, this invention includes a pharmaceutically acceptable dosage form comprising a compound according to any one of the structural embodiments of this invention, or 4-(4-hydroxyphenethyl)-2-methylphenol, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

In certain embodiments, this invention includes a pharmaceutical composition comprising a compound according to any one of the structural embodiments of the invention, or 4-(4-hydroxyphenethyl)-2-methylphenol, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, this invention includes a method of treating a disorder selected from the group consisting of: hot flushes, vaginal or vulvar atrophy, atrophic vaginitis, pruritus, dyspareunia, dysuria, frequent urination, urinary incontinence, endometriosis, alzheimer's disease, cognitive decline, decreased libido, senile dementia, neurodegenerative diseases, depression, anxiety, insomnia, schizophrenia, infertility, prostatic hypertrophy, breast cancer, prostate cancer, prostate neoplasia, uterine leiomyomas, endometrial cancer, endometrial polyps, ovarian cancer, benign breast disorders, colon cancer, hypercholesterolemia, hyperlipidemia, cardiovascular disease, atherosclerosis, restenosis, peripheral vascular disease, inflammatory disorders, autoimmune disorders, inflammatory bowel disease, Crohn's disease, ulcerative colitis, arthritis, rheumatoid arthritis, osteoarthritis, pleurisy, ischemia/reperfusion injury, asthma, psoriasis, multiple sclerosis, systemic lupus erythematosus, sepsis, cataracts, leukocytosis, systemic inflammatory response syndrome, septic arthritis, ischemic colitis, meningitis, multiple organ dysfunction syndrome, and type II diabetes comprising the administration of an effective amount of a compound according to any one of the structural embodiments of this invention, or a pharmaceutically acceptable salt thereof, or an effective pharmaceutical dosage form of the invention, or an effective amount of a pharmaceutical composition of the invention, to a patient in need thereof.

In a specific embodiment, the disorder is selected from: hot flushes; prostatic hypertrophy, prostate cancer, inflammatory disorders, inflammatory bowel disease, Crohn's disease, rheumatoid arthritis, osteoarthritis and sepsis. In an even more specific embodiment, this disorder is sepsis,

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a plot comparing survival of a nicotine treated (positive control) group and a vehicle treated (control) group in a mouse model of LPS-induced sepsis. FIGS. 2A-2D are plots comparing survival of vehicle treated (control) groups and groups treated with test article at 30 mg/kg. DETAILED DESCRIPTION OF THE INVENTION

The compounds of this invention may be prepared by various methods known to those of skill in the art. The methods of preparation provided herein are not intended to be in any way to be limiting but rather representative. The term "alkyl" as used herein refers to both straight and branch chain hydrocarbon radicals, having the number of carbon atoms falling within the specified range. For example, Ci _-6 alkyl means that a hydrocarbon radical is attached that may contain anywhere from 1 to 6 carbon atoms with the remaining valence filled in by hydrogen atoms. The definition also includes separately each permutation as though it were separately listed. Thus, Ci _-2 alkyl includes methyl and ethyl. The term Ci _-3 alkyl includes methyl, ethyl, propyl and 2-propyl. The term Ci- ₄ alkyl includes methyl, ethyl, n-propyl, 2-propyl, n-butyl, 2-butyl, iso-butyl and tert- butyl. The term Ci _-5 alkyl includes methyl, ethyl, 2-propyl, n-butyl, 2-methylbutyl, tert-butyl, n-pentyl, pentan-2-yl, pentan-3-yl, and tert-pentyl, iso-pentyl. The term "halogen" as used herein refers to a fluorine, chlorine, bromine or iodine radical.

The term "haloalkyl" refers to an alkyl radical wherein said alkyl radical is the same as defined for the term "alkyl" except that the alkyl radical additionally has from 1 to 5 halogen atoms attached to the alkyl chain. For example, Ci haloalkyl includes -CH ₂ F, -CHF ₂ , -CF ₃ and the like, C ₂ haloalkyl includes -CH ₂ F, CHF ₂ , CF ₃ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ CF ₃ , -CF ₂ CHF ₂ , -CF ₂ CF ₃ and the like. C,. ₃ haloalkyl is defined to include -CH ₂ F, -CHF ₂ . -CF ₃ , -CH ₂ CF ₃ , -CHFCF ₃ , -CF ₂ CF ₃ , -CHClCH ₃ , - CH ₂ CH ₂ Cl, -CH ₂ CH ₂ CF ₃ , and the like. C _]-4 haloalkyl is defined to include -CH ₂ F, - CHF ₂ , -CF ₃ , -CH ₂ CF ₃ , -CHFCF ₃ , -CF ₂ CF ₃ , -CHClCH ₃ , -CH ₂ CIT ₂ Cl, -CH ₂ CH ₂ CF ₃ , - CH ₂ CH ₂ CH ₂ CF ₃ , CHClCF ₂ CH ₂ CH ₃ , CF ₂ CH ₂ CH ₂ CHF ₂ , CH ₂ CH ₂ CH ₂ CH ₂ F, CH ₂ CH ₂ CH ₂ CH ₂ Cl, and the like.

The compounds of this invention may be present as solids and when so present, may be in an amorphous form or they may be crystalline. When the compounds of this invention are in the crystalline form, they might be present as a single polymorph or a mixture of polymorphs or even as a mixture of amorphous material together with one or more distinct polymorphs - the invention is not limited according to any particular solid or liquid state form.

Certain compounds of this invention may contain a stereocenter. When the compounds of this invention are drawn or named in a way that specifies a particular stereochemistry at one or more positions, then that stereochemistry is the required stereochemistry for that position. However, when the stereochemistry is not specified but one or more additional chiral centers are present, then those chiral centers may be present in either the R- or S- configuration, or a combination of the two. Thus, the molecule may be one single enantiomer or a mixture of diastereomers. In certain embodiments, it is actually preferred that the compound of the invention is present as a preponderance of one enantiomer or the other. For example, it may be preferred that one of the enantiomers be present in greater than 50% relative to the other enantiomer. In some embodiments, it is preferred that one of the enantiomers be present in at least 70%, 80%, 90% or 95% (ee) relative to the other enantiomer.

Where compounds of this invention include one or more basic sites such as amines, acid addition salts can be made and this invention includes such acid addition salts. Some representative (non-limiting) acid addition salts include hydrochloride, hydrobromide, hydroiodide, acetate, benzenesulfonate, mesylate, besylate, benzoate, tosylate, citrate, tartrate, sulfate, bisulfate, lactate, maleate, mandelate, valerate, laurate, caprylate, propionate, succinate, phosphate, salicylate, napsylate, nitrate, tannate, resorcinate and the like, including multiprotic salts as well as mixtures of the acid addition salts. In cases where an amine is present, this invention also embraces quaternized ammonium salts of those amines. It should be appreciated that N-oxides of amines are also embraced within the definition of the compounds of this invention. Likewise, where compounds of this invention include one or more acid sites such as carboxylic acids, phenols and the like, basic addition salts can be made and this invention includes such basic addition salts. For example, some representative (non-limiting) acidic compounds of this invention may be present as their lithium, sodium, potassium, ammonium, trialkyammonium, calcium, magnesium, barium and the like.

The compounds of this invention can also be present as solvates and such solvates are embraced within the scope of this invention even where not explicitly described. Such solvates are preferably hydrates but can be solvates comprised of other solvents, preferably where those solvents are considered to be non-toxic or at least acceptable for administration to mammals, preferably humans. The solvates can be stoichiometric or non-stoichiometric, singular or in combination. Some exemplary solvates include water, ethanol, acetic acid and the like. The therapeutic utility of these compounds includes "treating" a mammal, preferably a human where treating is understood to include treating, preventing, or ameliorating the symptoms associated with, or reducing the incidence of, reducing the pathogenesis of, facilitating the recovery from or delaying the onset of the syndrome, illness, malady or condition being considered. The compounds of this invention can also be useful in states or conditions where no clear deficit, illness or malady per se is perceived but rather, where a preferred condition, sensation, performance, capability or state is obtainable through therapeutic intervention with a compound of this invention. The compounds of this invention, when used as therapeutics can be administered by any method known to one of skill in the art such as orally, bucally, intravenously, subcutaneously, intramuscularly, transdermal Iy, intradermal Iy, intravascularly, intranasally, sublingually, intracranially, rectally, intratumorally, intravaginally, intraperitonealy, pulmonary, ocularly and intratumorally. In certain embodiments of this invention, the mode of administration may be via intravenous delivery of the compound. In this mode, the compound may be delivered in a treatment regimen or a prophylactic regimen. For example, a patient considered at high risk for sepsis might be put on an IV drip wherein the compound is included in the IV delivery vehicle. Such an IV administration might begin prior to, during or after a high risk event, for example, a patient entering surgery who is thought to be at elevated risk of sepsis could advantageously be put on an IV drip containing the compound of this invention by itself or together with other agents such as antibiotics, nutritionals, etc.

When administered, the compounds and compositions of this invention may be given once daily or with multiple daily doses such as twice per day, three times per day, four times per day or continuously.

In one embodiment of this invention, the compound is administered orally where it can be formulated for solid dosage administration or liquid dosage administration. Solid dosage administration can be in the form of a tablet, granule, capsule, pill, pellet, powder and the like. Liquid dosage formulations include syrups, solutions, gels, suspensions, elixirs, emulsions, colloids, oils, and the like.

As mentioned previously, the compounds of this invention may be solids and when present as solids, they maybe of defined particle size. Where the compound of this invention is not particularly water soluble, it is sometimes preferable to administer the compound with a certain particle size - a particle size with a preferred range where the average mean particle size diameter is under 100 microns, or 75 microns, or 50 microns, or 35 microns, or 10 microns or 5 microns.

Solid dosage formulations will comprise at least one compound of this invention together with one or more pharmaceutical excipients. Those excipients are known to one of skill in the art and include, by way of non-limiting example diluents (monosaccharides, disaccharides and polyhydric alcohols including starch, mannitol, dextrose, sucrose, microcrystalline cellulose, mallodextrin, sorbitol, xylitol, fructose and the like), binders (starch, gelatin, natural sugars, gums, waxes and the like), disintegrants (alginic acid, carboxymethylcellulose (calcium or sodium), cellulose, crocarmellose, crospovidone, microcrystalline cellulose, sodium starch glycolate, agar and the like), acidic or basic buffering agents (citrates, phoshphates, gluconates, acetates, carbonates, bicarbonates and the like), chelating agents (edetic acid, edetate calcium, edetate disodium and the like), preservatives (benzoic acid, chlorhexidine gluconate, potassium benzoate, potassium sorbate, sorbic acid, sodium benzoate and the like), glidants and lubricants (calcium stearate, oils, magnesium stearate, magnesium trisilicate, sodium fumarate, colloidal silica, zinc stearate, sodium oleate, stearic acid, and the like), antioxidants and/or preservatives (tocopherols, ascorabtes, phenols, and the like) and acidifying agents (citric acid, fumaric acid, malic acid, tartaric acid and the like) as well as coloring agents, coating agents, flavoring agents, suspending agents, dessicants, humectants and other excipients known to those of skill in the art.

In certain embodiments, the pharmaceutically acceptable excipient is not water. The compounds of this invention may be present in a pharmaceutical dosage form. A pharmaceutical dosage form refers to a pill, capsule, tablet, injectable solution in ampule, suppository, loaded syringe, syringe cartridge, etc. Generally, the pharmaceutical dosage forms of this invention will include a compound of this invention together with one or more excipients where the drug plus the one or more excipients comprise the pharmaceutical dosage form.

The solid dosage formulations of this invention can be prepared in different forms including most commonly, tablets and capsules. The tablets can be formulated by a wide variety of methods known to one of skill in the art including, for example, preparing a dry powder mixture of the drug substance in combination with one or more of the excipients granulating the mixture and pressing to together into a tablet and optionally coating the tablet with an enteric or non-enteric coating. The final coat typically includes a light protective pigment such as titanium oxide and a shellac or wax to keep the tablet dry and stable. While not intending to be limited by theory or example, in some instances it might be preferred to prepare the tablets by wet granulating the drug with one or more of the excipients and then extruding the granulated material.

The solid dosage forms of this invention also include capsules wherein the drug is enclosed inside the capsule either as a powder together with optional excipients or as granules containing usually including one or more excipients together with the drug and wherein the granule in turn can be optionally coated, for example, enterically or non-enterically.

In certain embodiments of this invention, the solid dosage formulations of this invention are formulated in a sustained release formulation. Such formulations are known to those of skill in the art and generally rely on the co-formulation of the drug with one or more matrix forming substances that slow the release of an estrogen of this invention thus extending the compound's lifetime in the digestive track and thereby extend the compounds half-life. Some non-limiting matrix forming substances include hydroxypropyl methylcellulose, carbopol, sodium carboxymethylcellulose and the like.

In some embodiments of this invention, the compounds are formulated for delivery other than via a solid oral dosage form. For example, in certain instances it might be preferable to deliver a compound of this invention by a pulmonary route. A pulmonary route of administration typically means that the compound of this invention is inhaled into the lung where it is absorbed into the circulation. Such a route of administration has the advantage of avoiding a first pass liver effect thereby possibly increasing bioavailability as well as decreasing or eliminating undesirable effects on the liver. Formulating a compound of the invention for pulmonary delivery can be accomplished by micronizing the compound of the invention to a very fine size particle, typically with a mean average diameter of less than 20 microns, or less than 10 microns or between 2 and 5 microns. The powder may then be inhaled by itself or more likely mixed with one or more excipients such as lactose or maltose. The powder can then be inhaled in a dry powder inhaling device either once or multiple times per day depending on the particular compound and the patients need. Other types of pulmonary dosage forms are also embraced by this invention. In an alternative to the dry powder delivery, the compound of this invention may be suspended in an aerosolizing medium and inhaled as a suspension through a meter dosed inhaler or a nebulizer. The compounds of this invention can be formulated for transdermal delivery.

Effective advantage of these compounds can be taken through a wide variety of transdermal options. For example, the compounds of this invention maybe formulated for passive diffusion patches where they are preferably embedded in a matrix that allows for slow diffusion of the compound into the treated subject's circulation. For this purpose, the compound is preferably dissolved or suspended in solvents including by way of non-limiting examples one or more of ethanol, water, propylene glycol, and Klucel HF. In some instances, a polymer matrix (e.g. acrylate adhesive) will comprise the bulk of the transdermal formulation. In some instances, the transdermal formulations maybe designed to be compatible with alternate transdermal delivery technologies. For example, some transdermal technologies achieve greater and/or more consistent delivery by creating micropores in the skin using radio frequency, heat, ultrasound or electricity. In some cases, the compounds of this invention can be used with microneedle technology wherein the compound is loaded into very small needles which due not need to penetrate the dermis to be effective.

The compounds of this invention may be employed alone or in combination with other therapeutic agents. By way of non-limiting example, the compounds of this invention can be used in combination with anti-lipidemics (statins, fibrates, omega-3 oils, niacinates and the like), bone anti-resorptives (bisphosponates, estrogens, selective estrogen receptor modulators (SERMs), calcitonin, and the like), bone anabolic agents (PTH and fragments e.g. teriparatide, PTHRP and analogues e.g. BaO58), anti-diabetics (e.g. insulin sensitizers, glucose absorption and synthesis inhibitors (e.g. metformin)), anti-anxiety agents, antidepressants, anti-obesity agents, contraceptive agents, anti-cancer agents, PPARγ agonists (e.g. pioglitazone), and the like. When used in combination, the compounds of this invention may be co- formulated or co-administered wherein said co-administration does not require dosing at exactly the same time but rather indicates that the patient is undergoing treatment with one or more of the additional agents during the timeframe of treatment with the estrogens of this invention. Thus, the additional drug(s) for combination treatment can be administered concomitantly, sequentially or separately from the compounds of this invention.

The compounds of this invention may be administered according to different dosage scheduling and the dosage may be adjusted as deemed necessary by the subject or preferably by the subject in consultation with a qualified practitioner of medicine. Dosing of the compounds of this invention can take place by multiple routes and consequently, the dosing schedule and amounts are dependent not only on the particular subject's weight, sex, age, therapy contemplated, etc but also by the route of the drug chosen. By way of non-limiting example, the compounds of this invention may be dosed by the oral route in a once daily, twice daily, three times daily or more than three times per day depending on the particular needs of that subject, the formulation of the drug, etc. The dosage will typically fall between 0.01 mg and 500 mg of drug per daily dosage, or 0.1 mg and 250 mg, or 1 mg and 150 mg, or between 5 mg and 100 mg.

The compounds of this invention can be prepared by a variety of synthetic routes and techniques known to those of skill in the art. The processes disclosed herein should not be construed as limiting the examples or scope of the invention in any way but rather are provided as just some of the representative ways that the compounds of this invention can be or were prepared.

In some cases, protective groups are employed in the synthesis of the compounds of this invention and it should be appreciated that there are a diverse array of protective groups and strategies that can be employed in organic synthesis (T.W.Green and P.G.M.Wuts (2006) Greene's Protective Groups in Organic Synthesis, herein incorporated by reference in its entirety) and that where a protective group is referred to generically, any appropriate protective group should be considered.

In some instances, leaving groups are employed in the synthesis of compounds of this invention. Where a specific leaving group is referred to, it should be appreciated that other leaving groups might also be used. Leaving groups typically include those groups that can stabilize an anion. In the case of nucleophilic aromatic substitutions, the leaving group may be an anion or a neutrally charged group. In some cases, the leaving group for nucleophilic aromatic substitution may be a group that is not typically considered to be a stabilized anion (e.g. fluoride or hydride). While not intending to be bound by theory or the examples, some typical nucleophilic leaving groups include halogens, sulfonates (O-mesylates, O-tosylates, etc), hydrides, quaternized amines, nitro, and the like. Additional discussion and examples can be found in leading textbooks on organic chemistry including, for example, March ^' s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5 ^l Edition, which is herein incorporated in its entirety.

Scheme 1

Scheme 1. (a) R ₆ MgCI or R ₇ MgCI, THF; (b) Et ₃ SiH, BF ₃ OEt ₂ , CH ₂ CI ₂ and (c) BBr ₃ , CH ₂ CI ₂ or base

Scheme 2

Scheme 2. (a) KOt-Bu, R ₆ IZR ₇ I, 18-crown-6, THF; (b) Et ₃ SiH, CF ₃ COOH; (c) BBr ₃ , CH ₂ CI ₂ Scheme 3

n = 1 n = 2

Scheme 3. (a) NaH, Br(CH ₂ ) _n+3 Br, THF; (b) Et ₃ SiH, CF ₃ COOH; (c) BBr ₃ , CH ₂ CI ₂ or base

Biphenyls General procedure for the Grignard addition to desoxyanisoin (cpd II, scheme

1)

Desoxyanisoin II (400 mg, 1.56 mmol) was dissolved in 10 mL THF and the

Grignard reagent (4.68 mmol) was added. The mixture was refluxed for 6 h (unless stated otherwise), then cooled to 0 °C, quenched with water (10 mL) and extracted with EtOAc (3 x 15 mL), The organic extracts were dried over Na ₂ SO ₄ and the solvent was removed under vacuum.

General procedure for the dehydroxylation of 1 ,2-Bis-(4-methoxy-phenyl)- alkan-2-ols (cpd III in scheme 1)

The starting material was dissolved in CH ₂ Cl ₂ and the mixture was brought to 0 ⁰ C. Et ₃ SiH was added and after 2 min BF ₃ OEt was added dropwise. The reaction was stirred for 30 min at 0 ⁰ C, quenched with water and extracted three times with EtOAc. The organic extracts were dried over Na ₂ Sθ4 and the solvent was removed under vacuum.

General procedure for the deprotection of methoxy groups (cpd IV where R ₁ and R ₁ ' are each -CH ₃ ) with BBr ₃

The starting material was dissolved in CH ₂ Cl ₂ and brought to 0 ⁰ C. BBr ₃ was added dropwise and the mixture was brought to rt and stirred for 17 h. The reaction was brought to 0 ⁰ C, quenched with water and extracted three times with EtOAc. The organic extracts were dried over Na ₂ SO ₄ and the solvent was removed under vacuum.

General procedure for the alkylation of desoxyanisoin (II in scheme 2)

KO-tBu and 18-crown-6 were added to THF at rt and the suspension was stirred for 15 min. A solution of the alkyl iodide and starting material were added at rt and stirring was continued for 30 min. The mixture was filtered and the filter cake was washed with EtOAc. The filtrate was absorbed on silica gel and the solvent was removed under vacuum. The silica gel was charged on a column and flash chromatography was performed as usual.

General procedure for the deoxygenation with Et ₃ SiHZTFA

The starting material was dissolved in TFA, and Et ₃ SiH was added dropwise at rt. The reaction was stirred for 24 h, quenched with water and extracted three times with diethyl ether. The organic extracts were dried over Na ₂ SO ₄ and the solvent was removed under vacuum.

Individual Compound Data Example 1

4,4'-(2-methylpropane-l,2-diyl)diphenol

¹ H NMR (500 MHz, acetone-t/ ₆ j δ 8.07 (s, OH), 8.02 (s, OH), 7.12 (d, J = 8.8, 2H), 6.74 (d, J = 8.8, 2H), 6.66 (d, J = 8.6, 2H), 6.59 (d, J = 8.6, 2H), 2.73 (s, 2H). 1.23 (s, 6H): ¹³ C NMR (500 MHz, acetone-^; δ 156.39, 155.94, 140.71,

132.08, 130.66, 127.95, 115.35, 1 15.00, 50.81, 38.65, 28.72; MS (EI) m/z 242 (M ⁺ , 3). HRMS (EI) calcd for Ci ₆ Hi ₈ O ₂ : 242.1307, found 242.1307. Anal. Ci ₆ H ₁₈ O ₂ : Calcd. C, 79.31; H, 7.49. Found C, 78.90; H, 7.51. Example 2 4-(2-(4-hydroxyphenyl)-2-methylpropyl)-3-methylphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.27 (s, 6H), 1.88 (s, 3H), 2.73 (s, 2H), 6.45 (d, IH), 6.53 (s, I H), 6.60 (d, IH), 6.73 (m, 2H), 7.05 (m, 2H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 20.29, 28.89, 39.37, 46.80, 1 12.64, 115.30, 117.35, 127.98, 128.95, 133.02, 139.15, 140.80, 155.98, 156.15

Example 3 4-(2-(4-hydroxyphenyl)-2-methylpropyl)-2-methylphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.23 (s, 6H), 2.08 (s, 3H), 2.70 (s, 2H), 6.46 (d, IH), 6.59 (m, 2H), 6.76 (m, 2H), 7.12 (m, 2H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 16.17, 28.69, 38.55, 50.85, 114.41, 115.27, 123.52, 127.90, 129.29, 130.59, 133.67,140.86, 154.23, 155.83

Example 4

3-chloro-4-(4-hydroxyphenylethyl)phenol

1H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 2.74 (m, 2H) ₅ 2.86 (m, 2H), 6.70 (m, IH),

6.74 (m, 2H), 6.88 (s IH), 7.02 (m, 2H), 7.08 (m, IH) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 35.92, 36.25, 1 15.10, 115.90, 116.75, 130.15, 130.79, 132.15, 133.19, 134.37, 156.42, 157.29 Example 5 4-(4-hydroxyphenethyl)-2-methylphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 2.15 (s, 3H), 2.73 (m, 4H), 6.69 (m, IH), 6.73 (m, 2H), 6.83 (m, IH), 6.92 (m, IH), 7.02 (m, 2H) ¹³ C-NMR (125 MHz (CDs) ₂ CO) 5 16.18, 38.23, 38.25, 1 15.21, 115.80, 124.53, 127.32, 130.13, 131.63, 133.58, 133.68, 154.20, 156.26

Example 6

2-fluoro-4-(4-hydroxyphenethyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 2.76 (m, 4H), 6.73 (m, 2H), 6.82 (m, IH), 6.86 (m, IH), 6.93 (m, IH), 7.02 (m, 2H)

Example 7

4-(l-(4-hydroxyphenyl)propan-2-yl)-3-methylphenol

^{1 1} H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.13 (d, 3H), 2.12 (s, 3H), 2.64 (m, IH), 2.74 (m, IH), 3.1 1 (m, I H), 6.57 (s. IH), 6.64 (m, IH), 6.70 (m, 2H), 6.93 (m, 2H),

7.1 1 (m, IH) 1 ^U 3C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 19.62, 21.24, 36.84, 44.39, 113.72, 1 15.59, 117.50, 127.16, 130.75, 132.75, 136.83, 137.14, 155.71, 156.25

Example 8 4-(3-(4-hydroxyphenyl)-3-methylbutan-2-yl)-3-methylphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.96 (d, 3H), 1.16 (s, 3H), 1.28 (s, 3H), 2.14 (s, 3H), 3.20 (m, IH), 6.55 (m, 2H), 6.72 (m, 2H), 6.82 (m, IH), 7.09 (m, 2H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 17.67, 20.80, 24.60, 28.18, 42.02, 44.22, 1 12.84, 1 15.08, 117.24, 128.50, 129.67, 134.43, 138.61, 140.59, 155.69, 155.89

Example 9

2-fluoro-4-( 1 -(4-hydroxyphenyl)pentan-2-yl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.76 (t, 3H), 1.13 (m, 2H), 1.50 (m, 2H), 2.74 (m, 3H), 6.65 (m, 2H), 6.76 (m, IH), 6.87 (m, 4H).

Example 10 2-fluoro-4-(2-(4-hydroxyphenyl)-2-methylpropyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.24 (s, 6H), 2.76 (s, 2H), 6.48 (m, 2H), 6.75 (m, 3H), 7.12 (m, 2H).

Example 1 1

2-fluoro-4-(l-(4-hydroxyphenyl)-2-methylpropan-2-yl)pheno l

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.24 (s, 6H), 2.74 (s, 2H), 6.61 (m, 2H), 6.67 (m, 2H), 6.91 (m, 2H), 6.99 (m, IH). Example 12

4-(l-(4-hydroxyphenyl)pentan-2-yl)-2-methylphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.78 (t, 3H), 1.12 (m, 2H), 1.52 (m, 2H), 2.15 (s, 3H), 2.65 (m, IH), 2.73 (m, 2H), 6.67 (m, 3H), 6.77 (m, IH), 6.89 (m, 3H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 14.32, 16.32, 21.27, 38.73, 43.76, 47.82, 92.11, 115.11, 115.53, 124.36, 126.60, 130.77, 132.69, 136.98, 154.16, 156.11.

Example 13

2 -fluoro-4-( 1 -(4-hydroxyphenyl)propan-2-yl)phenol 1H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.15 (d, 3H), 2.65 (m, IH), 2.75 (m, IH), 2.89 (m, IH), 6.69 (m, 2H), 6.84 (m, 2H), 6.93 (m, 3H).

Example 14

2-fluoro-4-(2-(4-hydroxyphenyl)pentyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.79 (t, 3H), 1.13 (m, 2H), 1.56 (m, 2H), 2.74 (m, 3H), 6.73 (m, 5H), 6.95 (m, 2H)

Example 15 2-fluoro-4-( 1 -(4-hydroxy-3 -methylphenyl)propan-2-yl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.15 (d, 3H), 2.13 (s, 3H), 2.61 (m, IH), 2.74 (m, IH), 2.88 (m, IH), 6.65 (m, 1H),6.72 (m, IH), 6.85 (m, 3H), 6.94 (m, IH)

Example 16 2-fluoro-4-(2-(4-hydroxyphenyl)propyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.16 (d, 3H), 2.68 (m, 2H), 2.77 (m, IH) ₅ 2.90 (m,lH), 6.73 (m, 3H), 6.84 (m, 2H), 7.01 (m, 2H).

Examples 16a and 16b

(S)-2-fluoro-4-(2-(4-hydroxyphenyl)propylphenol (R)-2-fluoro-4-(2-(4-hydroxyphenyl)propylphenol

A racemic mixture of example 16 was separated by chiral HPLC according to the following conditions:

Chiral HPLC method:

Chiral column: Chiralpak AD-H, 250 x 4.6 mm, 5 μ Mobile phase A: n-Hexane Mobile phase B: Ethanol Isocratic: A:B (90: 10) Flow rate: l.OOml/min The first compound to elute was Example 16a and the second sample was 16b. The ee for each enantiomer was greater than 90%. The absolute stereochemistry was not assigned for each compound. Example 17 2-fluoro-4-(2-(4-hydroxy-3-methylphenyl)propyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.15 (d, 3H), 2.16 (s, 3H), 2.65 (m, IH), 2.77 (m, IH), 2.85 (m, IH), 6.70 (m, IH), 6.74 (m, IH), 6.84 (m, 3H), 6.94 (s, IH)

Example 18

3-fluoro-4-(2-(4-hydroxyphenyl)propyl)phenol

1H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.16 (d, 3H), 2.72 (m, 2H), 2.90 (m, IH),

6.51 (m, 2H), 6.73 (m. 2H), 6.90 (m, IH), 7.02 (m, 2H).

Example 19

3-fluoro-4-(l-(4-hydroxyphenyl)propan-2-yl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.15 (d, 3H), 2.67 (m, IH), 2.80 (m, IH), 3.18 (m, IH), 6.48 (m, IH), 6.58 (m, IH), 6.69 (m, 2H), 6.93 (m, 2H), 7.08 (m, IH).

Example 20

4,4' -(propane- 1 ,2-diyl)bis(2-fluorophenol)

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.19 (d, 3H), 2.70 (m, IH), 2.78 (m, IH), 2.92 (m, IH), 6.73 (m, IH), 6.84 (m, 4H), 6.94 (m, IH). Example 21 4-(2-(4-hydroxyphenyl)propyl)-2-methylphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.13 (d, 3H), 2.13 (m, IH), 2.59 (m, IH), 2.74 (m. IH) ₅ 2.86 (m, IH), 6.66 (m, IH), 6.73 (m, 3H), 6.85 (s, IH), 7.03 (m, 2H) ¹³ C-NMR (125 MHz (CDs) ₂ CO) δ 16.19, 21.76, 41.90, 45.01, 115.03, 115.75, 124.31 , 128.04, 128.67, 132.28, 132.63, 138.94, 154.17, 156.24.

Example 22 4,4'-(2-methylpropane-l,2-diyl)bis(2-fluorophenyl)

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.26 (s, 6H), 2.76 (s, 2H), 6.52 (m, 2H), 6.76 (m, IH), 6.92 (m, 2H), 7.02 (m, IH).

Example 23

2-fluoro-4-(l-(4-hydroxybenzyl)cyclopentyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.65 (m, 2H), 1.79 (m, 4H), 1.89 (m, 2H), 2.72 (s, 2H), 6.48 (m, 2H), 6.56 (m, 2H), 6.72 (m, IH), 6.77 (m, IH), 6.84 (m, IH). Example 24

4,4'-(propane-l,2-diyl)bis(2-fluorophenol)

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.17 (d, 3H), 2.73 (m, 2H), 2.93 (m, IH), 6.72 (m, IH), 6.84 (m, 4H), 6.94 (m, IH).

Example 25 2-fluoro-4-(2-(4-hydroxy-2-methylphenyl)propyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.13 (d, 3H), 2.11 (s, 3H), 2.67 (m, IH), 2.75 (m, IH), 3.14 (m, III), 6.57 (s, IH), 6.65 (m, IH), 6.73 (m, IH), 6.81 (m, 2H), 7.10 (m, 2H).

Example 26 2-fluoro-4-(l-3-fluoro-4-hydroxybenzyl)cyclohexyl)phenol

¹ H=NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.31 (m, 3H), 1.53 (m, 5H), 2.66 (s, 2H), 6.30 (m, 2H), 6.70 (m, IH), 6.78 (m, IH), 6.86 (m, 2H).

Example 27

2-fluoro-4-(l-(4-hydroxy-2-methylphenyl)-2-methylpropan-2 -yl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.28 (s, 6H), 1.91 (s, 3H), 2.75 (s, 2H), 6.47 (m, I H), 6.55 (m, IH), 6.60 (m, IH), 6.90 (m, 3H). 1,3-Diarylpropanes Synthesis of Chakones

Biarylpropanes can be prepared according to the scheme below

X = H or F

(E)-l,3-bis(3-fluoro-4-methoxyphenyI)prop-2-en-l-one: l-(3-Fluoro-4- methoxyphenyl) ethanone (2 g, 1 1.9 mmol) and 3-fluoro-4-methoxybenzaldehyde (1.8 g, 11.9 mmol) were dissolved in ethanol (33mL) and a 6M solution of NaOH (4mL, 23.8 mmol) was added. The mixture was stirred at room temperature. Ethanol (25mL) and H ₂ O (25mL) were added and the mixture stirred for 20 minutes. The solid precipitate was filtered and washed with water (5OmL) followed by ethanol (75mL) to obtain pure product, 86% yield.

Hydrogenation of Chalcones

Example procedure provided

X = H or F

3-(3-fIuoro-4-methoxypfaenyl)-l-(4-methoxyphenyl)propan-l -one: (E)-3- (3-Fluoro-4-methoxyphenyl)-l -(4-methoxyphenyl)prop-2-en-l-one (3.5g, 12.2 mmol) was hydrogenated with 5%-Pd/C (0.218g, 6.25% by wt) in presence of ethanol (4OmL) at 40psi of H ₂ for 6 hrs. Filter the Pd/C and evaporate the solvents to get the crude product. The crude product is passed through a short silica flash column (100% CH ₂ Cl ₂ ), 76% yield.

Alkylation of Diaryl propan-1-ones

Example procedure provided below

X = H or F R = Me, Et, n-Pr Y = Br or I 2-(3-fluoro-4-methoxybenzy!)-l-(4-methoxyphenyI)biitan-l-oι ie: 3-(3-

Fluoro-4-methoxy phenyl)- l-(4-methoxyphenyl)propan-l -one (1.Og, 3.5 mmol) was dissolved in THF (3OmL) and ethyl iodide (0.36mL, 4.5raol) was added to it. The sodium hydride (60% in mineral oil, 0.69g, 17.4 mmol) was added all at once to the premixed solution and stirred overnight at room temperature. After the reaction is complete, monitored by NMR, the solid was filtered and the solvent concentrated. Add water and extract with methylene chloride to get the crude product, 98% yield. The crude product was used without further purification.

Deoxygenation of butanones

Example procedure provided below

X = H or F

2-fluoro-l-methoxy-4-(2-(4-methoxybenzyl)butyl)benzene: 2-(3-Fluoro-4- methoxybenzyl)-l -(4-methoxyphenyl)butan-l-one (l .lg, 3.5 mmol) was added to TFA (2.6 mL, 34.7 mmol) followed by addition Of Et ₃ SiH (1.22 mL, 7.6 mmol) slowly for 10 minutes. The reaction mixture was stirred overnight at room temperature, neutralized with 10% Na ₂ CO ₃ and extract with dichloromethane. The crude product was isolated after rotary evaporation, 75% yield, and used without further purification.

Preparation of l-alkyϊ substituted systems

Example procedure provided below

l-methoxy-4-((lE,3Z)-3-(4-methoxyphenyl)hexa-l,3-dienyl)-2- methylbenzene and l-methoxy-4-((lE,3E)-3-(4-raethoxyphenyl)hexa-l,3- dienyl)-2-methylbenzene: Propyltriphenylphosphonium bromide (5.4 g, 14.1 mmol) was dissolved in THF (2OmL) and to it added BuLi (8.66 mL of 1.6M solution, 13.8 mmol) at -78 ⁰ C. The reaction mixture was allowed to stir at -78 ⁰ C for 20 minutes then at room temperature for 90 minutes. Then the reaction mixture was again cooled down to -78 ⁰ C and added (E)-3-(4-methoxy-3-methylphenyl)-l- (4-methoxyphenyl)prop-2-en-l-one (2.0 g, 7.1 mmol) slowly for 20 minutes. The reaction mixture was allowed to raise the temperature to 22 ⁰ C (room temperature) and stirred for another lhr. After the reaction is complete as monitored by TLC, quenched with MeOH (1OmL) and evaporated all the solvent at 40 ⁰ C. Added 200 mL of diethyl ether and filtered the solid. Evaporate the ether layer and pass through a short flash column to get the desire product as crude mixture, which is good for the next step.

Hydrogenation of 1-substituted systems:

Example procedure provided below

l-methoxy-4-(3-(4-methoxyphenyl)hexyl)-2-methylbenzene: A mixture of l-methoxy-4-((lE,3Z)-3-(4-methoxyphenyl)hexa-l,3-dienyl)-2-m ethylbenzene and 1 -methoxy-4-(( 1 E,3E)-3 -(4-methoxyphenyl)hexa- 1 ,3 -dienyl)-2-methylbenzene (1.0 g, 3.2 mmol) was hydrogenated with 5%-Pd/C (0.10 g, 10% by wt) in presence of ethanol (3OmL) at 40 psi for 6 hrs. Filter the Pd/C and evaporate the solvents to get the crude product. The crude product is passed through a short silica flash column (95:5 Hex:EtOAc) to get the pure product in the 1 ^st fraction, 78% yield. Boron tribromide deprotection of diary! butanes

Example procedure provided below X = H or F

2-fluoro-4-(2-(4-hydroxybenzyI)butyl)phenol 2-Fluoro- 1 -methoxy-4-(2-(4- methoxy benzyl)butyl)benzene (1.4 g, 4.5 mmol) was dissolved in methylene chloride (4OmL) and BBr ₃ , 1.0M Solution (11.7 mL, 11.7 mmol) was added to it slowly at room temperature. The reaction mixture was stirred for 3 hrs at room temperature and the progress of the reaction was monitored by TLC. After the reaction is complete, add saturated solution OfNaHCO ₃ , separate the layer and wash with methylene chloride. Combine the organic layers, evaporate and dry to get the crude mixture. The crude mixture was passed through a short column (30% EtO Ac/70% Hexane) to obtain pure material, 68% yield.

Example 28 4,4'-(2-methylpropane-l,3-diyl)diphenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.76 (d, 3H), 1.90 (m, IH), 2.28 (m, 2H), 2.58 (m, 2H), 6.75 (m, 4H), 6.98 (m, 4H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 19.23, 38.40, 42.94, 115.76, 130.78, 132.80, 156.30,

Example 29 2-fluoro-4-(2-(4-hydroxybenzyl)butyl)phenol

¹ H-NMR (400 MHz CD ₃ Cl) δ 0.87 (m, 5H), 1.77 (m, 5H), 6.75 (m, 3H), 6.87 (m, 2H), 7.02 (m, 2H) Example 30

4,4 ' -(propane- 1 ,3 -diyl)bis(2-fluorophenol)

¹ H-NMR (400 MHz CD ₃ Cl) δ 2.55 (m, 6H), 6.88 (m, 6H)

Example 31

2-fluoro-4-(3-(4-hydroxyphenyl)propyl)phenol

¹ H-NMR (500 MHz CD ₃ Cl) δ 1.86 (m, 2H) ₅ 2.56 (m, 4H), 6.75 (m, 2H), 6.85 (m, IH), 6.92 (m, 2H), 7.05 (m, 2H).

Example 32

4-(3-(4-hydroxyphenyl)hexyl)-2-metby]phenol

1H-NMR (500 MHz CD ₃ Cl) δ 0.81 (t, 3H), 1.14 (m, 2H), 1.54 (m, 2H), 1.78

(m, IH), 1.90 (m, IH), 2.20 (s, 3H), 2.33 (m, 2H), 2.48 (m, IH), 6.67 (m, IH), 6.82 (m, 4H), 7.02 (m, 2H) ¹³ C-NMR (125 MHz CD ₃ Cl) δ 14.08, 15.72, 20.54, 32.85, 38.98, 39.39, 44.38, 114.70, 115.10, 123.44, 126.72, 128.74, 130.93, 134.93, 138.01, 151.52, 153.40.

Benzylthioethers

(4-methoxybenzyl)(4-methoxyphenyl)sulfane: 4-Methoxybenzene thiol (10.3g, 73.2 mmol) was dissolved in 200 mL of acetone. 4-Methoxybenzyl chloride (1 1 mL, 81 mmol) and potassium carbonate (16.9 g, 122.6 mmol) were added sequentially. The solution was refluxed for 24 hours and then cooled to room temperature at which time the solids were filtered. The volume of the solution was reduced to approximately 50 mL and 150 mL of hexane was added. A white precipitate came out of solution. The solid was allowed to stand overnight and the crystals were filtered, 71% yield.

Benzylic alkylation

Example procedure for monosubstitued alkyl systems shown below

R = Me, CH ₂ CH ₃ , CH ₂ CH ₂ CH ₃ , CH(CH ₃ ) ₂ , CH ₂ CH ₂ CF ₃

(4-methoxyphenyl)(l-(4-methoxyphenyl)butyI)suIfane: (4- Methoxybenzyl)(4~methoxyphenyl)sulfane (2.8Og, 10.8 mmol) is dissolved in 18 mL of tetrahydrofuran and cooled to -78 ⁰ C, a solid precipitate forms. n-Butyl lithium, 2.5 M solution in hexanes (5 mL, 12.5 mmol) is added carefully to the mixture. After 5 minutes the cold bath is removed and the reaction allowed to warm up. As the solution warmed the precipitate dissolved and the solution turned orange. After 5 minutes 1 -bromopropane (2.5 mL, 27.5 mmol) was added and the reaction allowed to warm to r.t. and stir overnight (21 hours). Water (10 mL) was added and the organic layer extracted with ethyl acetate (2 x 35 mL). The combined organic layers were washed with brine (2 x 50 mL), dried with sodium sulfate and the solvent was removed by rotary evaporation. The material was recrystallized from ethanol/water, 85% yield.

Preparation of bis-benzyl substituted systems

Example procedure shown below 3

(4-methoxypheny!)(2-(4-methoxyphenyl)propan-2-yl)sulfane: Zinc dust

(3.3g, 50.0 mmol) was mixed with EtOAc (36 mL) and warmed to 40 ⁰ C. The solution was stirred for 5 minutes and a mixture of AcOH (1.4 mL) and water (0.44 mL) was carefully added. The temperature was increased to 60 ⁰ C. After reaching 60 ⁰ C the solution stirred for 5 minutes and then 4-methoxybenzene-l-sulfonyl chloride (2.5 g, 12.2 mmol) in EtOAc (7.5 niL) was added over 10 minutes. The solutions was warmed to 85 ⁰ C, generating a vigorous reflux, and allowed to stir for 10 minutes. Dichlorodimethyl silane (4.5 ml, 37.4 mmol) was carefully added over 20 minutes. The solution stirred for 1 hour under reflux. 2-(4- methoxyphenyl)proρan-2-ol (2.1 g, 12.4 mmol) was added over 10 minutes. The solution was stirred for 1.5 hours and then cooled to r.t. at which time 20 niL of water was added. The mixture was washed with water (2 x 50 mL) and the water back extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with a saturated bicarbonate solution (2 x 50 mL) and brine (2 x 50 mL), dried over sodium sulfate and the solvent removed by rotary evaporation. The material was purified by flash column chromatography (30% EtOAc, 70% hexane), 66% yield.

Use of 3-fluoro-4-methoxybenzenethiol: Example procedure shown below

(3-fluoro-4-methoxyphenyI)(3,3,3-trifluoro-l-(4- methoxyphenyl)propyl)sulfane: 3,3,3-trifluoro-l-(4-methoxyphenyl)propan-l-ol (1.5 g, 6.9 mmol) was dissolved in dichloromethane (30 mL) and zinc iodide (1.3 g, 4.1 mmol) was added to the solution. After stirring for 5 minutes 3-fluoro-4- methoxybenzenethiol (1.5 g, 9.50 mmol) was added to the mixture. The reaction mixture was stirred overnight (~ 16 hours) at which time water (50 mL) was added to the solution. The solution was extracted with dichloromethane (4 x 100 mL). The combined organic layers were washed with 5% sodium hydroxide (1 x 50 mL), water (Ix 100 mL) and brine (1 x 100 mL). The product was purified by flash chromatography (25% EtOAc, 75% Hexane), 90% yield. Boron tribromide deprotection

Example procedure shown below

4-(4-hydroxybenzylthio)phenol: (4-Methoxybenzyl)(4- methoxyphenyl)sulfane (1.6g, 6.1 mmol) was dissolved in 3 mL of dichloromethane and cooled to 0 ⁰ C. 16 mL of a 1.0M solution of boron tribromide was added over 10 minutes. The reaction was kept at 0 ⁰ C for 3.5 hours. 10 mL of saturated sodium bicarbonate was added to the solution. The organic layer was extracted with ethyl acetate ( 2 x 100 mL) and the combined organic layers were washed with brine (2 x 100 mL), dried with sodium sulfate and rotovaped. The product was purified by recrystallization from ethyl acetate/hexane or by flash chromatography (silica gel 40% EtO Ac/60% Hexane), yield 49%.

Example 33

4-(4-hydroxybenzylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 3.93 (s, 2H), 6.75 (m, 4H). 7.06 (m, 2H), 7.19 (m, 2H), ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 40.86 115.82, 116.68, 125.85, 129.89, 130.89, 134.72, 157.21, 157.82

Example 34 4-(l -(4-hydroxyphenyl)butylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.82 (t, 3H), 1.26 (m, 2H), 1.80 (m, 2H), 3.98 (m, IH), 6.71 (m, 4H), 7.03 (m, 2H), 7.12 (m, 2H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 13.92, 21.31, 38.75, 54.31, 1 15.68, 116.44, 124.83, 129.74, 133.96, 136.58, 157.04, 158.15 Example 35 4-(4,4,4-trifluoro- 1 -(4-hydroxyphenyl)butylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 2.08 (m, 2H), 2.27 (m, 2H), 4.08 (m, 2H), 6.76 (m, 4H) ₅ 7.07 (m, 2H), 7.17 (m, 2H)

Example 36

4-( 1 -(4-hydroxyphenyl)propylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.83 (t, 3H), 1.80 (m, IH), 1.90 (m, IH), 3.90 (m, IH), 6.73 (m, 4H), 7.04 (m, 2H), 7.14 (m, 2H) ¹³ C-NMR (125 MHz (CDs) ₂ CO) δ 12.45, 56.32, 115.71, 116.47, 124.85, 129.79, 133.68, 136.59, 157.08, 158.16

Example 37

4-( 1 -(4-hydroxyphenyl)-2-methylpropylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 0.85 (d, 3H), 1.09 (d, 3H), 2.07 (m, IH), 3.82 (m, IH), 6.69 (m, 4H), 7.03 (m, 2H), 7.09 (m, 2H) ¹³ C-NMR (125 MHz (CD ₃ ) ₂ CO) δ 20.62, 21.59, 34.14, 62.79, 115.36, 116.42, 125.26, 130.44, 133.26, 136.12, 156387, 157.

Example 38 4-(2-(4-hydroxyphenyl)propan-2-ylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.56 (s, 6H), 7.01 (m, 4H), 7.03 (m, 2H), 7.24 (m, 2H) ¹³ C-NMR (125 MHz (CDs) ₂ CO) 5 30.07, 50.74, 115.27, 116.13, 123.39, 128.53, 138.13, 139.35, 156.69, 159.14

Example 39 4-(3,3,3-trifluoro-l-(4-hydroxyphenyl)propylthio)phcnol

¹ H-NMR (500 MHz (CD ₃ ) ₂ SO) δ 2.60 (m, IH), 3.01 (m, IH), 4.31 (m, IH), 6.67 (m, 4H), 7.15 (m, 4H)

Example 40 2-fluoro-4-(3,3,3-trifluoro-l-(4-hydroxyphenyl)propylthio)ph enol

¹ H-NMR (500 MHz (CD ₃ ) ₂ SO) δ 2.59 (m, IH), 3.02 (m, I H), 4.42 (m, IH), 6.75 (m, 2H), 2.87 (m, IH), 6.99 (m, IH), 7.15 (m, 3H)

Example 41

2-fluoro-4-(2-(4-hydroxyphenyl)propan-2-ylthio)phenol

I ¹ ₁ H-NMR (500 MHz (CD ₃ ) ₂ SO) δ 1.52 (s, 6H), 6.67 (m, 2H), 6.83 (m, 3H), 7.19 (m, 2H)

Example 42 2-fluoro-4-( 1 -(4-hydroxyphenyl)ethylthio)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ SO) δ 1.48 (d, 3H), 4.31 (q, III), 6.68 (m, 2H), 6.84 (m, I H) ₅ 6.92 (m, IH), 7.06 (m, 3H)

Example 43 2-fluoro-4-( 1 -(4-hydroxyphenylthio)ethyl)phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ CO) δ 1.48 (d, 3H), 4.18 (q, IH), 6.75 (m, 2H), 6.87 (m, 2H), 7.00 (m, IH), 7.17 (m, 2H)

Example 44

2-fluoro-4-(4,4,4-trifluoro-l-(4-hydroxyphenyl)butylthio) phenol

¹ H-NMR (500 MHz (CD ₃ ) ₂ SO) δ 2.02 (m, 3H), 2.28 (m, IH), 4.22 (m. IH) 6.69 (m, 2H), 6.79 (m, IH), 6.92 (m, IH), 7.06 (m, 3H)

BIOLOGICAL METHODS ER binding

The compounds of this invention were tested for affinity to the estrogen receptor α and the estrogen receptor β. Two different methods were performed for determining whether the compounds of this invention were competitive ligands for ER. One assay was the fluorescent polarization assay which was performed as specified by the manufacturer Green (Invitrogen, Madison, WI). Briefly, 1 μl of 1OmM compound was added to 500μl of ES2 screening buffer in a 1.5ml Eppendorf tube to make a 2x10 ^"5 M stock. 10-fold serial dilutions of the test compounds were prepared ranging in concentration from 10 ^" M to 10 ^{" 2} M. 25μl of each dilution in triplicate was added to a black 384-microtiter plate. The final reaction volume was 50μl per well. The test compounds were diluted 2-fold in the final reaction. 2x ER- Fluormone™ complex was prepared with 2nM Fluormone ES2™ and 2OnM ERβ. 25μl of 2x complex was added to each reaction well. The plate was sealed with foil cover and incubated in the dark at room temperature for 4 hours. Polarization values for each well were measured. The polarization values were plotted against the concentration of the test compound. The concentration of the test compound that results in half-maximum shift equals the IC ₅₀ of the test compound. As a control, a competition curve for estradiol was performed for each assay. Curve Fitting was performed using GraphPad Prism® software from GraphPad™ Software Inc.

The second binding method employed was an estrogen receptor β radiolabeled binding assay. Briefly, 7μl of 1OmM compound was added to 500μl of DMSO and then mixed 1 : 1 with Protein Resuspension Buffer (PRB, 0.05M Tris, 10% glycerol, 0.3mg/ml ovalbumin, 0.0 IM 2 mercaptoethanol at pH 8.0) in a 1.5ml Eppendorf tube to make a 7xlO ^"5 M stock. 10-fold serial dilutions of the test compounds were prepared ranging in concentration from 7x10 ^"5 M to 7x10 ^{~10 5} M. The tracer, ³ H- estradiol (Amersham, Piscataway, NJ, 80uCi/mmol), is diluted in PRB to be 7x10 ^{" 8} M. Estrogen receptor-β (Invitrogen, Madison, WI) was diluted to 5nM in PRB. lOμl of each dilution, l Oμl of the tracer and then 50μl of protein was added in duplicate to filter microtiter plates (Fisher Scientific, Pittsburgh, PA). The final reaction volume is 70μl per well. The final competitor concentrations range 1x10 ^{" 5} M to lxlO ^{"!O 5} M. Plates were covered with acetate plate sealer and incubated at room temperature for overnight. The next day, 25μl HAP slurry was added to each well and incubated at room temperature with occasional mixing. The plate was then placed on the vacuum manifold to remove reaction mixture. Each well was washed twice with 200μl of cold HAP wash buffer (0.05Tris, pH 7.3). Then 200μl of cold HAP wash buffer was added to each well and 150μl was removed and put into scintillation vials. The samples were then counted on Beckman Coulter LS6500. The CPM values were plotted against the concentration of the test compound. The concentration of the test compound that results in half-maximum CPM equals the IC ₅₀ of the test compound. As a control, a competition curve for estradiol was performed for each assay. Curve Fitting was performed using GraphPad Prism® software from GraphPad™ Software Inc. With the exception of example 42, all of the examples of this invention demonstrated binding ERβ ICso' _s of less than 1 μM when analyzed by at least one of the two binding methods described. Compound example 42 demonstrated IC _{50 s} of less than 10 μM when tested according to either method. EXPERIMENTAL MODEL OF SEPSIS

Compound example numbers 5, 16 and 30 were tested in a lipopolysaecharide (LPS) induced sepsis murine model. The activity of the compounds as was measured by their ability to reduce fatality as shown in FIGS. 1 and 2A-2D. Included for comparison are nicotine which is known to have activity and the known anti-inflammatory ERbeta selective compound 2-(3-fluoro-4- hydroxyphenyl)-7-vinylbenzo[d]oxazol-5-ol (compound 117 in J. Med. Chem. 2004, 47(21), 5021-5040; "ERB-041"). The objective of this study was to evaluate the effect of example numbers 5, 16, 30 and ERB-041 administered by subcutaneous (sc) injection at a concentration of 30 mg/kg each, on survival in a murine model of lipopolysaecharide (LPS) induced sepsis. Dosing of the test articles was performed once a day from day -3 to day 6. Sepsis was induced on day 0. Nicotine (0.4 mg/kg) was administered as a positive control according to Wang et al., 2004. In a pilot study, sepsis was induced using an intraperitoneal (ip) injection of 75 mg/kg lipopolysaecharide (Sigma L8274 LPS from E.coli O26:B6). This dose was chosen to result in less than 20% survival in BALB/c mice 3 days after injection. In this study, the untreated animals showed 0% survival by day 2. This was more severe than the sepsis induced by 75 mg/kg LPS in a pilot study but within the range of variability observed with this model. Nicotine showed efficacy in the prevention of sepsis-induced mortality resulting in a survival rate of 20% by day 7. All of the test articles were effective in the treatment of sepsis and all were more effective than the nicotine control treatment. Example 5 was the most effective in the treatment of sepsis induced by LPS (70% survival by day 7). Example 30 treatment resulted in 55% survival by day 7 and both example 16 and ERB-041 treatments resulted in 50% survival by day 7. STUDY OBJECTIVE

The objective of this study was to examine the activities of several ERβ ligands in a standard model of LPS-induced sepsis in BALB/c mice. MATERIAL AND METHODS Location of Study Performance

The study was performed at Biomodels AAALAC accredited facility in Watertown MA. Approval for this study (approval number 06-0510-1) was obtained from Biomodels' IACUC. Animals

One hundred and twenty (120) Balb/c male mice aged 5-6 weeks with average body weight of 17.7g were obtained from Charles River Laboratories (Wilmington, MA). Mice were acclimatized for 14 days prior to study commencement. During this period, the animals were observed daily in order to reject animals that present in poor condition.

Housing The study was performed in animal rooms provided with HEPA filtered air at a temperature of 70°F± 5 ⁰ F and 50% ± 20% relative humidity. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour. The room was on an automatic timer for a light/dark cycle of 12 hours on and 12 hours off with no twilight. Sterile Bed-O-Cobs bedding was used. Bedding was changed a minimum of once per week. Cages, tops, bottles, etc. were washed with a commercial detergent and allowed to air dry. A commercial disinfectant was used to disinfect surfaces and materials introduced into the hood. Floors were swept daily and mopped a minimum of twice weekly with a commercial detergent. Walls and cage racks were sponged a minimum of once per month with a dilute bleach solution. A cage card or label with the appropriate information necessary to identify the study, dose, animal number and treatment group marked all cages. The temperature and relative humidity were recorded during the study, and the records retained. Diet

Animals were fed with a sterile Purina Labdiet® 5061 rodent diet and sterilized water was provided ad libitum. Animal Randomization and Allocations.

Animals were randomly and prospcctively divided into six (6) treatment groups of twenty (20) animals per group prior to induction. Each animal was identified by an ear punch corresponding to an individual number. A cage card was used to identify each cage or label marked with the study number, treatment group number and animal numbers. Vehicle and Drug Formulation

The vehicle was 5% DMSO/95% Corn oiliSigma, St. Louis, MO). Test articles were given in 5% DMSO/95% Corn oil (Sigma, St. Louis, MO). Both the vehicle and drug were administered by sc injection once daily from day -3 to day 6.

The test articles were dissolved in DMSO at 108 mg/ml and these stocks were diluted 1 :20 in corn oil for a final concentration of 5.4 mg/ml. Mice were dosed based on body weight at 0.1ml per 18g. Nicotine Positive Control

Nicotine was dissolved in saline at 72 μg/mL and dosed at 0.1 ml per 18g of body weight by IP injection.

Sepsis Induction

Sepsis was induced using an ip injection of 75 mg/kg lipopolysaccharide (Sigma L8274 LPS from E.coli O26;B6) in saline. LPS was dissolved in saline to create a stock solution of 40 mg/ml. This stock solution was diluted 1 :14.8 in saline to give a dosing solution of 2.7 mg/ml, which was given by IP injection at 0.5 ml per

18g of body weight.

Experimental Design One hundred and twenty (120) male BALB/c mice were randomly divided into 6 groups of 20 mice per group. Beginning on day -3 and continuing once daily to day 6, animals in the vehicle control and the test estrogens received a single subcutaneous injection at 9:00 am (groups 1, 3-6).

Animals in group 2 received an ip injection of nicotine (0.4 mg/kg) 30 minutes prior to LPS injection on day 0. Dosing with nicotine resumed 20 hours after the LPS injection and the animals were dosed with nicotine twice a day from day 1 to day 3.

All animals received an ip injection of 75 mg/kg lipopolysaccharide (LPS) on day 0. Every day of the study, animals were weighed and monitored for pain (head bobbing, writhing and lameness). Animal survival was noted on a daily basis

(see Appendix). Those animals exhibiting pain were treated with 0.05 mg/kg buprenorphine. Animal survival was monitored daily for all groups. On day 7 all remaining animals will be euthanized by CO ₂ inhalation.

Animals were monitored daily for inability to ambulate to soft food placed in bottom of cage. Animals received soft food with buprenorphine (0.5 mg/kg PO Q

24hr) beginning 24 hours after LPS injection. To achieve this 0.3 mg of buprenorphine was added to 20 ml of soft food. Each for 2 ml the animals eats the dose will be 0.5 mg/kg. Our experience indicates the animals eat 1-2 ml immediately and then over the course of 24hr eat 2 ml more. The animals can not consume enough to overdose. This food was supplied once a day until the animal was euthanized. Animals were euthanized when they could not ambulate to the soft food. It is our experience with a variety of models that hunched posture; ruffled coat are not reliable indicators of death within 24 hrs. The inability to obtain food however is the indicator of death in these severely debilitated animals within 24 hrs. This gives a reproducible endpoint for statistical analysis. On day 7 all remaining animals were euthanized by CO ₂ inhalation. Study Design

* For Nicotine treatment, Hie first dose was given 30 minutes prior to LPS injection.

Sepsis Evaluation

Efficacy in treating sepsis was based on improvement in survival observed through day 6. Survival was evaluated on a daily basis. Animals were monitored several times during the course of each study day. An animal found dead at 7:00 am or an animal that died during the day from 7:00 am to 7:00 pm was recorded as having died on that day.

Survival statistics were evaluated using Kaplan-Meier Log Rank analysis. Individual group survival was compared with the vehicle control group. Survival

The survival data for all groups is shown in FIGS. 1 and 2A-2D of the invention. All mice received LPS at 75 mg/mg on day 0. This resulted in the death of all vehicle treated control mice by day 2 (0% survival). The rapidity and extent of animal death in the vehicle control group was slightly higher than observed in recent studies, but within the range of variability that is associated with this animal model. The animals treated with nicotine (FIG. 1) demonstrated a significant increase in survival relative to vehicle treated animals (P=0.018). In FIGS. 2A-2D, the effect of the different test articles on survival relative to vehicle is demonstrated.

The animals treated with the test articles at 30 mg/kg sc all demonstrated significant improvement in survival. In FIG. 2A, treatment with ERB-041 resulted in a 50% survival by day 6 (P=0.001). In FIG. 2B, treatment with Example 16 resulted in a 50% survival by day 6 (P<0.001). In chart FIG. 2C _; treatment with example 30 resulted in a 55% survival by day 6 (P<0.001). In FIG. 2D, treatment with Example 5 resulted in a 70% survival by day 6 (PO.001 ). While this invention has been particularly shown and described with references to example embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.