MXPA06005570A - Isoflavonoid prodrugs, compositions thereof and therapeutic methods involving same - Google Patents

Abstract

Phosphate esters of isoflavonoid compounds are prepared for use as prodrugs, medicaments and in formulations, drinks and foodstuffs.

Description

ISOFLAVONOID PROFORMATORS, COMPOSITIONS THEREOF AND THERAPEUTIC METHODS INVOLVING THEMSELVES Field of the Invention This invention relates to compounds, formulations, beverages, foodstuffs, methods and therapeutic uses that involve, contain, comprise, include and / or to prepare certain prodrugs of isoflvans and analogues thereof. In particular, the invention relates to phosphate esters of isoflavonoids and derivatives, drugs that involve them and therapeutic uses thereof.

BACKGROUND OF THE INVENTION Isoflavones and many derivatives thereof possess a very wide range of important biological properties that include estrogenic effects. Isoflavones such as genistein and daidzein, have been shown to be involved in the modulation or attenuation of levels of estrogenic spheroids in the body. More recently, isoflávenos and in particular dehidroequol have been shown to possess strong chemotherapeutic properties. In some areas of biological activity, there are still some contradictions, for example, some isoflavonoids act as estrogen receptor agonists, while others act as estrogen receptor antagonists. It is believed that there is a correlation between the decrease in levels of biologically active estrogenic spheroids in the body with lower incidences of cancer such as breast cancer and many other diseases and conditions. However, the biological activity of isoflavonoids in animals is not conserved across the spectrum of the isoflavonoid family and therefore, it can not be predicted, especially where bioavailability is involved. In this way, each specific structural variation of the basic isoflavonoid molecule can give a highly individual biological profile in animals ranging from no effect to a potent effect. In addition, it is thought that some conjugates of biologically active molecules, such as phosphate esters of some biologically active estrogenic spheroids, can be largely inactive. There is a strong need to identify new, improved, better and / or alternative pharmaceutical compositions and agents for the treatment, alleviation and prevention of diseases, conditions and disorders. There is also an additional need to provide new isoflavonoid compounds and derivatives for the improved formulation, bioavailability and delivery of these compounds. There is also a need for new and different therapies to be available for both specialists and the general public, to combat the numerous and various types of diseases and disorders which affect members of the population. There is therefore a requirement for the provision of new isoflavonoid compounds and derivatives thereof, which are therapeutically beneficial and which exhibit improved bioavailable and bioavailable properties, alternative or at least comparable, with those of known isoflavonoid compounds.

Summary of the Invention The present invention has surprisingly found that the phosphate esters of isoflavonoid compounds show good solubility and aqueous bioavailability and exhibit beneficial biological properties. In particular, phosphate esters, when administered, will exhibit a wide range of therapeutic activities including the ability to direct estrogen levels in the body. While it is not desired to be limited by theory, it is believed that prodrugs of isoplavenes and derivatives thereof, and in particular isoflavonoid phosphate esters of the invention, will result in the reduction in the supply of estrogenic spheroids, reducing the risk of severity of diseases or conditions related to estrogen. It is also thought that the isoflavonoid phosphate esters of the invention will provide regulation of a range of molecular targets in mammalian cells. Typically, these molecular objectives are intimately involved in the processes of signal transduction that are fundamental to critical cellular processes, such as growth, differentiation, migration and cell death. It can be observed therefore, that these surprising biochemical effects have broad and important implications for the health of animals including humans. These and other preferred objects of the invention are described herein. Thus, in accordance with one aspect of the invention, an isoflavonoid phosphate ester compound of the general formula I is provided: in which RIA R2 and Z are independently M2P0-, hydrogen, hydroxy, OR9, OC (O) R? 0 / OS (O) R10? CHO, C (O) R 0, COOH, CO2R 10, CONR 3 4 alkyl, haloalkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkoxyaryl, thio, alkylthio, amino, alkylamino, dialkylamino, nitro or halo, or R 2 is as previously defined, and Ri and Z taken together with the carbon atoms to which they are attached, form a ring of five elements selected from Ri is as previously defined, and R2 and Z taken together with the carbon atoms to which they are attached form a ring of five selected elements W is Ri, and A and B taken together with the carbon atoms to which they are attached, form a ring of six elements selected from wherein R3 is hydrogen, alkyl, aryl, arylalkyl, an amino acid, C (0) Ru wherein R1? is hydrogen, alkyl, aryl, arylalkyl or an amino acid, or C02R? 2 wherein Ri2 is hydrogen, alkyl, haloalkyl, aryl, heteroaryl or arylalkyl, R4 is hydrogen, alkyl or aryl, or R3 and R4 together with the nitrogen to which they are attached, comprise pyrrolidinyl or piperidinyl, R5 is M2P0 ~, hydrogen, C (0) Ru, wherein Rn is as previously defined, or CO2R12 wherein Ri2 is as previously defined, R6 is M2PO4-, hydrogen, alkyl, aryl, amino, thio, NR3R4, COR11 wherein Rn is as previously defined, C02Ri2 wherein R12 is as previously defined or CONR3R, R7 is hydrogen, C (0) Ru wherein Ru is as previously defined, alkyl, haloalkyl, aryl, arylalkyl or Si (R? 3) 3 wherein each Ri3 is independently hydrogen, alkyl or aryl, Rs is M2PO4-, hydrogen, hydroxy, alkoxy or alkyl, R9 is alkyl, haloalkyl, aryl, arylalkyl, C (0) Rn wherein Rn is as previously defined, or Si (R? 3) 3 wherein R13 is as previously defined, Rio is hydrogen, alkyl, haloalkyl, amino, aryl, arylalkyl, an amino acid, alkylamino or dialkylamino, the "" represents either a single bond or a double bond, M is independently hydrogen, an alkyl, alkenyl, alkynyl, alkoxyalkyl, alkylthioalkyl or aminoalkyl, straight or branched, a substituted or unsubstituted cycloalkyl, an aryl, aralkyl or alkylaryl, and a substituted cycloalkyl wherein at least one ring contains one or more than one nitrogen, sulfur, oxygen, phosphorus or silicon heteroatom in at least one ring; T is independently hydrogen, alkyl or aryl, X is 0, NR4 or S, preferably 0, and Y is wherein Ri4 / Ris and Ris are independently M2P04-, hydrogen, hydroxy, 0R9 OC (O) R? 0, OS (0) io, CHO, C (O) R? 0, C00H, C02R? o, CO R3R4, alkyl, haloalkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkoxyaryl, thio, alkylthio, amino, alkylamino, dialkylamino, nitro or halo, and wherein at least one of Ri, R2 / R5, R6, Re / Ri4 , R1S, R16, Z, W or A where they are independently presented are M2P04-, or a pharmaceutically acceptable salt thereof. In a preferred embodiment, the phosphate ester portion may be present as the corresponding salt -0-P0 (0M) / wherein M is hydrogen or a pharmaceutically acceptable counter ion, more preferably Na +, K +, Li +, Mg ++ or NH3 +, more preferably Na +. It has been surprisingly found by the inventors, that the compounds of the general formula I: wherein Ri, R2, W, A, B and Z are as defined above, have particular utility and effectiveness in the treatment, prophylaxis, defense relief against and / or prevention of the following diseases and disorders (for convenience here subsequently called as the "therapeutic indications"). (a) all forms of cancers (pre-malignant, benign and malignant) in all tissues of the body. In this regard, the compounds can be used as the only form of anticancer therapy or in combination with other forms of anti-cancer therapy including but not limited to radiotherapy and chemotherapy. (b) diseases and disorders associated with inflammatory reactions of an abnormal or prolonged nature in any of the tissues of the body, including but not limited to rheumatoid arthritis, tendonitis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, sclerosing cholangitis; (c) papulonodular skin lesions including, but not limited to, sarcoidosis, angiosarcoma, Kaposi sarcoma, Fabrice disease (d) papuloscarbaeous skin lesions including but not limited to psoriasis, Bowen's disease, and Reiter; (e) actinic damage characterized by degenerative changes in the skin including but not limited to solar keratosis, photosensitive diseases and wrinkles; (f) diseases and disorders associated with abnormal angiogenesis that affect any tissue within the body that includes but is not limited to angiomas and telangiectasia; (g) bone marrow proliferative disorders including but not limited to megaloblastic diseases, myelodysplastic syndromes, polytemia vera, thrombosis and myelofibrosis; (h) autoimmune diseases characterized by abnormal immunological responses including, but not limited to, multiple sclerosis, Type 1 diabetes, systemic lupus erythematosus, and biliary cirrhosis; (i) neurodegenerative diseases and disorders characterized by degenerative changes in the structure of the neurological system including, but not limited to, Parkinson's disease, Alzheimer's disease, muscular dystrophy, Lou-Gehrig disease, motor neuron disease; (j) diseases and disorders associated with degenerative changes within blood vessel walls including, but not limited to, atherosclerosis, atheroma, coronary artery disease, stroke, myocardial infarction, hypertensive vascular disease, malignant hypertension, tronboangiitis obliterans , fibromuscular dysplasia; (k) diseases and disorders associated with abnormal immunological responses that include but are not limited to dermatomyositis and scleroderma; (1) diseases and disorders associated with degenerative changes within the eye that include but are not limited to cataracts, macular degeneration, retinal atrophy. In particular, it has surprisingly been found that the isoflavene compounds also have a potent effect on the production and function of reproductive hormones such as estrogens and androgens.

As a result of this, these compounds can be used in the treatment and prevention of the following disorders and diseases: (a) conditions in women associated with the abnormal balance of estrogen / androgen, which include but are not limited to cyclic mastalgia, acne , dysmenorrhea, uterine fibroids, endometriosis, ovarian cysts, premenstrual syndrome, water menopause symptoms, osteoporosis, senile dementia, infertility; and (b) conditions in man associated with abnormal estrogen / androgen balance, which includes but is not limited to benign prostatic hypertrophy, infertility, gynecomastia, hereditary alopecia and various other forms of alopecia. Thus, in accordance with another aspect of the invention, there is provided a method for the treatment, prophylaxis, alleviation, defense against and / or prevention of one or more therapeutic indications, which comprise administering to a subject, a therapeutically effective amount of one or more compounds of formula I as defined above. In accordance with another aspect of the present invention, there is provided the use of compounds of formula I for the manufacture of a medicament for the treatment, alleviation, defense against, prophylaxis and / or prevention of one or more of the therapeutic indications. In accordance with another aspect of the present invention, there is provided the use of one or more compounds of formula I in the treatment, alleviation, defense against, prophylaxis and / or prevention of one or more of the therapeutic indications. In accordance with another aspect of the present invention, there is provided an agent for the treatment, prophylaxis, alleviation, defense against and / or treatment of therapeutic indications, which comprises one or more compounds of formula I either alone or in association with one or more carriers or excipients.

In accordance with another aspect of the present invention, there is provided a therapeutic composition which comprises one or more compounds of formula I in association with one or more pharmaceutical carriers and / or excipients. In accordance with another aspect of the present invention, a beverage or food product is provided, which contains one or more compounds of formula I. In accordance with another aspect of the present invention, there is provided a microbial culture or a food product containing one or more microbial strains, in which the microorganisms produce one or more compounds of formula I. In accordance with another aspect of the present invention , one or more microorganisms are provided which produce one or more compounds of formula I. Preferably, the microorganism is a purified culture, which can be mixed and / or administered with one or more other cultures which produce compounds of formula I. Through this specification and the following claims, unless otherwise required by the text, the word "comprises" and variations such as "comprise" or "comprising" shall be understood as implying the inclusion of a member or stage or group of members or stages but without the exclusion of any other member or stage or group of members or stages.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 represents pharmacokinetic data comparing concentrations of free and total dehydroequol in serum from mice injected i.p. with bolus dosages of DHE bisphosphate prepared in PBS and dosed at 25 mg / kg. Figures 2a and 2b depict pharmacokinetic data comparing concentrations of free and total dehydroequol in serum from mice injected i.p. with bolus dosages of dehydroequol prepared in different formulations. The DHE bisphosphate formulations were prepared in PBS and dosed at 25 mg / kg. The DEG PEG: PBS formulations were prepared in PEG: PBS 1: 1 formulations and dosed at 50 mg / kg. The DHE-HPBCD formulations were prepared in 20% HPBCD (HPBCD prepared in PBS) and dosed at 50 mg / kg (total DHE levels not shown).

Detailed Description of the Invention The term "isoflavonoid" is taken generally, to mean benzopyran molecules fused to the ring, which have a pendant phenyl group from the pyran ring based on a 1,2-diphenylpropane system. Thus, the classes of compounds generally referred to as isoflavones, isoflávenos, isoflávanos, isoflavanonas, isoflavanoles and the like, are generically referred to herein as isoflavonoids, isoflavonoid compounds, or isoflavone metabolites or derivatives thereof. The preferred isoflavonoid compounds of the invention are isoflavan-4-ones, isoflavones, isoflavones and isoflavones, which are generally hydrogenated products from the base isoflavones, in which the compounds can also be optionally substituted. The term "alkyl" is taken to include straight chain, branched chain and saturated cyclic alkyl groups (in the case of 5 carbons or greater) of 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secbutyl, tertiary butyl, pentyl, cyclopentyl and the like. The alkyl group is more preferably methyl, ethyl, propyl or isopropyl. The alkyl group may be optionally substituted by one or more of fluorine, chlorine, bromine, iodine, carboxyl, C 1 -C 4 alkoxycarbonyl, C 1 -C 4 alkylaminocarbonyl, di (C 1 -C 4 alkyl) -no-carbonyl, hydroxyl, C 1 -C 4 alkoxy -C4, for iloxy, C1-C4 alkyl carbonyloxy, C1-C4 alkylthio, C3-Ce cycloalkyl or phenyl. The term "alkenyl" is taken to include straight chain, branched chain and cyclic saturated hydrocarbons (in the case of 5 carbons or greater) of 2 to 10 carbon atoms, preferably from 2 to 6 carbon atoms, with at least one double bond such as ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 2-methyl-1-propenyl, 2-methyl-2- propenyl and the like. The alkenyl group is more preferably ethenyl, 1-propenyl or 2-propenyl. The alkenyl groups may be optionally substituted by one or more of fluorine, chlorine, bromine, iodine, carboxyl, C 1 -C 4 alkoxycarbonyl, C 1 -C 4 alkylaminocarbonyl, di- (C 1 -C 4 alkyl) -aminocarbonyl, hydroxyl, C1-C4 alkoxy, formyloxy, alkylcarbonyloxy C? -C, alkylthio Cx-C, C3-C6 cycloalkyl or phenyl. The term "alkynyl" is taken to include both straight chain and branched chain hydrocarbons of 2 to 10 carbon atoms, preferably 2 to 6 carbon atoms, with at least one triple bond such as ethynyl, 1-propynyl, -propynyl, 1-butynyl, 2-butynyl and the like. The alkynyl group is more preferably ethynyl, 1-propynyl or 2-propynyl. The alkynyl group can be optionally substituted by one or more of fluorine, chlorine, bromine, iodine, carboxyl, C 1 -C 4 alkoxycarbonyl, C 1 -C 4 alkylaminocarbon, di- (C 1 -C 4 alkyl) -aminocarbonyl, hydroxyl, C 1 -C 4 alkoxy, formyloxy, C 1 -C 4 alkylcarbonyloxy, C 1 -C 4 alkylthio, C 3 -C 6 cycloalkyl or phenyl. The term "aryl" is taken to include phenyl, biphenyl and naphthyl and may be optionally substituted by one or more C1-C4 alkyl, hydroxy, C1-C4, alkoxycarbonyl, C1-C4, alkylcarbonyloxy C1-C4 alkyl or halo. The term "heteroaryl" is taken to include rings of five elements and six elements, which include at least one oxygen, sulfur or nitrogen in the ring, in which the rings may be optionally fused to other aryl or heteroaryl rings including but They not limited to furyl, pyridyl, pyrimidyl, thienyl, imidazolyl, tetrazolyl, pyrazinyl, benzofuranyl, benzothiophenyl, quinolyl, isoquinolyl, purinyl, morpholinyl, oxazolyl, thiazolyl, pyrrolyl, xantinilo, purine, thymine, cytosine, uracil, and isoxazolyl. The heteroaromatic group can be optionally substituted by one or more fluorine, chlorine, bromine, iodine, carboxyl, C1-C4 alkoxycarbonyl, alkylamino carbonyl C1-C4alkyl, di- (C1-C4alkyl) -amino-carbonyl, hydroxyl, alkoxy C 1 -C 4, formyloxy, C 1 -C 4 alkylcarbonyloxy, C 1 -C 4 alkylthio, C 3 -C 6 cycloalkyl or phenyl. The heteroaromatics may be partially or totally hydrogenated as desired. The term "halo" is taken to include fluorine, chlorine, bromine and iodine, preferably fluorine and chlorine, more preferably fluorine. The reference to, for example, "haloalkyl", will include monohalogenated, dihalogenated and even perhalogenated alkyl groups. Preferred haloalkyl groups are trifluoromethyl and pentafluoroethyl. The term "pharmaceutically acceptable salt" refers to an organic and inorganic portion that bears a charge and can be administered in asstion with a pharmaceutical agent, for example, as a counter cation or a counter anion in a salt. The pharmaceutically acceptable cations, which include the M portion, are known to those skilled in the art and include, but are not limited to, sodium, potassium, calcium, zinc and quaternary amine. Pharmaceutically acceptable anions are known to those of skill in the art and include but are not limited to chloride, acetate, citrate, bicarbonate and carbonate. The term "pharmaceutically acceptable derivative" or "prodrug" refers to a derivative of the active compound which, after administration to the container, is capable of directly or indirectly providing the original compound or metabolite, or having the activity itself. As used herein, the terms "treatment", "prophylaxis" or "prevention", "relief" and the like, are considered in their broader context. In particular, the term "treatment" does not necessarily imply that an animal is treated until full recovery. Accordingly, "treatment" includes alleviation of the symptoms or severity of a particular condition or preventing or otherwise reducing the risk of developing a particular condition. The invention in particular, refers to the compounds of the general formula II and uses thereof: in which R-R-2 Rd / Rβ / Ri 4 / Ris / W and Z are as defined above, the drawing "" rsents either a single bond or a double bond, and more preferably, wherein the pattern "" rsents a double bond.

In another aspect, the invention in particular relates to the compounds of the general formula III and uses thereof: in which Ri, R2, 5 e, Ri4 / 15 / W and Z are as defined above. In another aspect, the invention in particular refers to the compounds of the general formula IV and uses thereof: wherein i / R2 R5 / Re, i4 / Ris / W and Z are as defined above. Particularly preferred compounds of the present invention are the isoflavonoid compounds as follows: lsoflavonoid-0-PO (OM) 2 wherein M is independently hydrogen or a counter cation, and wherein the isoflavene compound or derivative is mono, di or per-phosphorylated and can be derived from the following compounds isoflavanone, isoflavene, isoflavanol and isoflavane containing hydroxyl and derivatives 1-22 as follows: 6 10 11 12 13 14 16 17 18 19 20 21 22 wherein R2 / Ri6 / W and Z are independently H, OH, Cl, Br, Me or OMe, and Ri4 is H, OMe, Me, Cl or Br. In a more preferred embodiment the isoflavonoid compound or derivative is a new mono, di or dihydrodaidzein dihydrogenestein ester, dihydrodaidzein, tetrahydrodaidzein, dehydroequol or equol ester, more preferably it is a dehydroequol phosphate ester. The compounds of the present invention have particular application in the treatment of diseases associated with or resulting from estrogenic effects, androgenic effects, vasodilatory and spasmodic effects, inflammatory effects and oxidative effects. The amount of one or more compounds of formula I which are required in a therapeutic treatment according to the invention, will depend on a number of factors, which include the specific application, the nature of the particular compound used, the condition to be treated , the mode of administration and the condition of the patient. The compounds of formula I can be administered in a manner and amount as conventionally practiced. See, for example, Goodman and Gilman, The Pharmacol ogi cal Basis of Therapeutics, 1299 (7th Edition, 1985). The specific dosage used will depend on the condition to be treated, the condition of the subject, the route of administration and other well-known factors as indicated above. In general, a daily dose per patient may be in the range of 0.1 mg to 2 g; typically from 0.5 mg to 1 g; preferably, from 50 mg to 200 mg. The length of the dosage can vary from a single dose given once every day or two, to doses twice or three times daily given over the course of a week to many months to many years, as required, depending on the severity of the dose. the condition to be treated or relieved. It will further be understood that for any particular subject, the specific dosage regimens must be adjusted during the time in accordance with the individual needs and professional judgment of the person administering or supervising the administration of the compositions. The production of pharmaceutical compositions for the treatment of therapeutic indications described herein, is typically prepared by mixing the compounds of the invention (for convenience subsequently referred to as "the active compounds), with one or more pharmaceutically carriers and / or excipients. or veterinarily acceptable, as well as well known in the art.The carrier must, of course, be acceptable in the sense of being compatible with any of the other ingredients in the formulation and must not be detrimental to the subject.The carrier or excipient may be a solid or a liquid, or both, and is preferably formulated with the compound as a unit dose, for example, a tablet, which may contain from 0.5% to 59% by weight of the active compound, or up to 100% by weight of the active compound. One or more active compounds can be incorporated into the formulations of the invention, which can be prepared by any of the well-known pharmacy techniques, consisting essentially of mixing the components, optionally including one or more accessory ingredients. Formulations of the invention include those for oral, rectal, optical, buccal (e.g., sublingual), parenteral (e.g., subcutaneous, intramuscular, intradermal or intravenous) administration and transdermal administration, although the most appropriate route in any given case it will depend on the nature and severity of the condition to be treated and the nature of the particular active compound which is being used. The formulation suitable for oral administration can be presented in discrete units, such as capsules, pouches, lozenges or tablets, each containing a predetermined amount of the active compound-as a powder or granules.; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil in water or water in oil emulsion. Such formulations can be prepared by any suitable pharmacy method, which includes the step of bringing into association the active compound and a suitable carrier (which may contain one or more accessory ingredients as indicated above). In general, the formulations of the invention are prepared uniformly and intimately by mixing the active compound with a liquid or finely divided solid carrier or both, and then, if necessary, forming the resulting mixture such as to form a unit dosage. For example, a tablet can be prepared by compressing or modulating a powder or granules containing the active compound, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, the free flowing compound such as a powder or granules optionally mixed with a binder, lubricant, inert diluent and / or dispersing / surface active agent (s). The molded tablets can be molded in a suitable machine, the powder compound moistened with an inert liquid binder. Formulations suitable for buccal (sublingual) administration include pellets comprising the active compound in a flavored base, usually sucrose and acacia or tragacanth; and tablets comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia. The compositions of the present invention suitable for pharmaceutical administration, suitably comprise sterile aqueous preparations of the active compounds, in which the preparations are preferably isotonic with the blood of the proposed container. These preparations are preferably administered intravenously, although administration can also be effected by means of subcutaneous, intramuscular or intradermal injection. Such preparations can be conveniently prepared by mixing the compound with water or a glycine buffer and providing the resulting sterile and isotonic solution with the blood. The injectable formulations according to the invention generally contain from 0.1% to 60% w / v of the active compound and are administered at a rate of 0.1 ml / minute / kg. Formulations suitable for rectal or vaginal administration are preferably presented as unit dose suppositories. These can be prepared by mixing the active compound with one or more conventional solid carriers, for example, cocoa butter and then forming the resulting mixture. Formulations or compositions suitable for topical administration to the skin, preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol or oil. Carriers which may be used include, Vaseline, lanolin, polyethylene glycols, alcohols and combination of two or more thereof. The active compound is present in a general manner, at a concentration from 0.1% to 0.5% w / w, for example, from 0.5% to 2% w / w. Examples of such compositions include cosmetic creams for the skin. Formulations suitable for transdermal administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the container for a prolonged period of time. Such patches suitably contain the active compound as an optionally buffered aqueous solution of, for example, a concentration of 0.1 M to 0.2 M with respect to said active compound. Formulations suitable for transdermal administration can also be delivered by iontophoresis (see for example, Pharmaceutical Research 3 (6), 318 (1986)) and typically take the form of an optionally buffered aqueous solution of the active compound. Suitable formulations comprise citrate or bis / tris buffer (pH 6) or ethanol / water and contain from 0.1M to 0.2M of active ingredient. Formulations suitable for inhalation may be supplied as a composition for atomization in the form of a suspension or emulsion solution. The composition for inhalation atomization may further comprise a pharmaceutically acceptable propellant such as carbon dioxide or nitrous oxide. The active compounds can be provided in the form of food products, such as being added to, mixed in, coated, combined or otherwise added to a food product. The term "food product" is used in its broadest possible sense and includes liquid formulations such as beverages, which include dairy products and other foods, such as health bars, desserts, etc. Pharmaceutical formulations containing compounds of the invention can be easily prepared in accordance with standard practices. The compounds of the present invention have potent antioxidant activity and thus, find wide application in veterinary and pharmaceutical uses, in cosmetics such as skin creams to prevent the aging of the skin, in sunscreens, foods, healthy drinks, shampoos, and the like. It has been surprisingly found that the compounds of formula I interact synergistically with vitamin E to protect lipids, proteins and other biological molecules from oxidation. Accordingly, a further aspect of this invention provides a composition comprising one or more compounds of formula I, vitamin E, and pharmaceutically, veterinarily or cosmetically acceptable carriers and / or excipients.

The methods, uses and therapeutic compositions can be for administration to humans or animals, such as domestic and companion animals (such as dogs and cats), birds (such as chickens, turkeys, ducks), livestock animals (such as cattle, sheep, pigs and goats), for use in aquaculture and similar applications. Prodrugs and isoflavonoid derivatives can also be co-administered with other active materials that do not impair the desired action, or with materials that supply the desired action, such as antibiotic, antifungal, anti-inflammatory or antiviral compounds. The active agent can comprise two or more isoflavones or derivatives thereof in combination or synergistic mixture. The active compounds can also be administered with lipid lowering agents such as probucol and nicotinic acid; inhibitors of platelet aggregation such as aspirin; antithrombotic agents such as cu dinate; calcium channel blockers such as verapamil, diltiazem, and nifedipine; inhibitors of the enzyme that converts angiotensin (ACE) such as captopril and enalapril, and β-blockers such as propanolol, terbutalol and labetalol. The compounds can also be administered in combination with non-steroidal anti-inflammatories such as ibuprofen, indomethacin, aspirin, fenoprofen, mefenamic acid, flufenamic acid and sulindac. The compounds can also be administered with corticosteroids. The co-administration can be simultaneous or sequential. The simultaneous administration can be effected by the compounds being in the same unit dose, or in individual and discrete unit doses administered at the same or similar time. Sequential administration may be in any order as required, and typically will require a progressive physiological effect of the first active or initial agent to be current when the second or last active agent is administered, especially where a synergistic or cumulative effect is desired. The isoflavone compounds are suitable starting materials for the synthesis of the isoflavonoid compounds of formula I and these isoflavone starting materials can be prepared by standard methods known to those skilled in the art. Suitable methods can be found in, for example, International Patent Applications WO 98/08503 and WO 00/49009, which are incorporated herein by reference in their entirety. The protection of the chemical functional group, deprotection, synthons and other techniques known to those skilled in the art can be used where appropriate in the synthesis of the compounds of the present invention. The derivatization of hydroxy substituted isoflavones to form the conjugates of the present invention can be carried out by any suitable method, as is known to one skilled in the art. The isoflavone starting materials can also be obtained in the form of concentrates or extracts from plant sources. Again, those skilled in the art will readily be able to identify suitable plant species, however, plants of particular utility include legume plants. More preferably, the isoflavone extract can be obtained from chickpeas, lentils, beans, red cloves or species of underground cloves and the like. The aqueous solubility of isoflavonoids is important for its formulation in pharmaceuticals, foodstuffs and cosmetics, many of which are water-based systems. Low solubility is also frequently an impediment to efficient bioavailability in orally administered products. The low solubility is a particularly serious impediment to the formulation of intravenous medications, which are more frequently supplied in aqueous medium. The isoflavonoid phosphate esters of the invention, are presented in forms which have increased bioavailability, especially improved aqueous solubility relative to the unmodified compounds, while substantially retaining the active properties of such unmodified compounds. The phosphate ester is useful as a pro-drug having a polar leaving group (solubilizer), which can be easily hydrolyzed under physiological conditions to produce the corresponding isoflavonoid compound. In preferred embodiments, an alcohol functionality of an isoflavonoid is esterified using a phosphoric acid group to provide a phosphate ester. In general, absorptive and digestive gastrointestinal tract fluids, other acids, and various enzymes, are capable of hydrolyzing the esterified isoflavonoid to the starting isoflavonoid. The phosphate ester is preferably a (OH) 2P02 group due to the presence of two polar groups, and which are a good solubilizer and have high biological compatibility. Where the M group for M2P04- is not hydrogen, it may generally be expected that the solubility may be less for the compound and may therefore be less favorable. Where M is an alkyl group, for example, the non-polar group is preferably selected to be small. It is also contemplated to employ metal salt complexes of the esterified isoflavones, especially Li +, Na +, iC \ Mg + and ammonium salts, including NH4 + and low molecular weight mono or poly-alkylammonium counter ions. The following examples are not considered to limit the invention as described.

EXAMPLES The isoflavonoid phosphate esters of the invention can be prepared by standard chemical processes known to those skilled in the art from available starting materials and direct synthetic methods. In this form, various modalities of the inventive subject matter can be prepared and characterized. These examples fall within the group of pro-compounds of formula I have at least one group of the formula M2P04-. These new phosphate esters are all water soluble and easily hydrolysable in vivo, yet are generally generally stable in aqueous solutions in vitro at normal pH at room or body temperature, and are more stable as solids.

Example 1 Esters of Dehydroequol Phosphate A solution of dehydroequol (120 mg, 0.5 mmole) and di-tert-butyl phosphoramidite (330 ul, 1.0 mole) in DMF (1 mL) was stirred under argon while added by drip IH -tetrazole (210 mg in 0.5 ml of DMF, 3.0 mmol). The solution was cooled to -20 ° C, then a solution of m-chloroperbenzoic acid (260 mg in 0.5 ml of methylene chloride, 1.5 mmol) was added dropwise. After warming to room temperature, the mixture was diluted three times with ethyl acetate, then washed with 10% sodium metabisulfite and 10% sodium bicarbonate. The ethyl acetate solution, which contains the butyl esters of dehydroequol phosphates, was washed with 1M HCl and dried over sodium sulfate. After removal of the solvent in vacuo, the residue was treated with 30% TFA in acetic acid for 90 minutes at room temperature. The solvents were removed in vacuo, and the residues were recovered in ethanol and neutralized with sodium hydroxide to pH 5.5. Removal of the solvent in vacuo gave a mixture of sodium salts of dehydroequol phosphates, 130 mg. The analysis of the phosphate mixture indicates the presence of 4'-phosphate, 7-phosphate and the derivatives of 4 ', 7-diphosphate. Where the esterification of the compounds of the invention provides mixtures of phosphate esters, they can be separated into individual components by standard separation techniques including fractional crystallization, column chromatography and HPLC. The isoflavonoid phosphate esters prepared by the above methods include: Similarly, phosphate esters of dihydrodaidzeines, tetrahydrodaidzeines and equol were synthesized to provide the following compounds. (cís) 29 (cis) 30 and pharmaceutically acceptable salts thereof.

Example 2 Deshi droequol-1-phosphate or Dehydroequol with its protected hydroxy group on the 4'-pendant phenyl deposition, is subjected to reaction according to Example 1 to provide the corresponding 7-phosphate derivative. Any suitable protecting group can be employed including MOM or MEM ethers and benzyl ethers. These groups can optionally be removed after phosphorylation. The protecting groups in which they are used can be incorporated in the synthesis of the isoflavonoid starting materials by following any of the methods referred to herein, or they can be attached at a later time taking advantage of synthons, chemical reactivity, polarity, electronic consideration or spherical conditions in or near any of the target hydroxy groups. By these methods, mono-, di- and per-phosphorylated derivatives of compounds 1-22 described herein were synthesized. Phosphorous acids and pharmaceutically acceptable salts thereof are thus prepared. Proton or carbon magnetic resonance spectrum, IR and / or mass spectrum were used to characterize the synthesized compounds.

Example 3 The bioavailability of the isoflavonoid phosphoric esters of the invention was tested by in vitro hydrolysis of dehydroequol phosphate by various enzymes and biological media. The results were determined by measuring the amount of free dehydroequol by HPLC. The serum and medium used include human serum, human blood, rat blood, alkaline phosphatase type VII-S (intestinal bovine mucosa) and alkaline phosphatase type XXIV (human placenta). The bioavailability and conversion rate of the ester depends on a number of factors including the nature of the phosphate ester and substitutions thereof, the medium, any of the enzymes present, the temperature and pH. By controlling these various parameters, it was found that some degree of regulation or control can be obtained by altering the half-life of the ester prodrug to better match the desired bioavailability ratio.

Example 4 Esterified isoflavonoids were found to be readily converted to free isoflavonoids in biological medium such as gastrointestinal fluid and blood. Among other things, gastrointestinal fluids often have enzymes and pH high enough to hydrolyze ester bonds, and the blood generally contains enzymes such as phosphatases which can hydrolyse phosphate ester bonds.

Pharmacokinetic experiments Two separate PK experiments were conducted using dehydroequol (DHE) -bisphosphate formulated in PBS by i.p. and oral Three animals were assigned by time point with 5 time points (15 minutes, 30 minutes, 1 hour, 4 hours and 24 hours) (15 mice per study). The objective was to determine if the PK profile is comparable when i.p. is supplied. against oral.

Administration protocol i.p. 1. Female nude mice were maintained on an isoflavone-free diet for at least one week to remove background levels of isoflavone in plasma. 2. One day before the experiment, 3 mice were assigned by time point and marked with unique identifiers. Each mouse was weighed to determine the density of DHE-bisphosphate required by i.p. to achieve a dose of 50 mg / kg for each mouse. It was prepared in light excess of formulated DHE-bisphosphate and the powder mass adjusted accordingly. The remaining solution was stored at -20 ° C by QA analysis. 3. Each mouse was injected into the left or lower right quadrant of the abdomen, ensuring that the needle does not enter the vessel or bow to the intestine. Once DHE-bisphosphate was administered, the mice were placed in a cage until each point of time (15 minutes, 30 minutes, 1 hour, 4 hours, 24 hours). 4. Each mouse was sacrificed by cervical dislocation, then blood was collected via the thoracic cavity as per SOP BD-009 using a 20 gauge needle. 5. Blood was allowed to clot, then centrifuged at top speed for 3 minutes using a bench top mini-fogger at room temperature. 6. The serum was aspirated in an appropriately labeled eppendorf tube and stored at -20 ° C until analyzed. The animal serum was dosed with control vehicle and the DHE-bisphosphate formulated was stored at -20 ° C together with 200 ul aliquots of the vehicle and DHE-bisphosphate formulated for analysis.

Oral administration protocol 1. Female BALB / c mice were maintained on an isoflavone-free diet for at least one week to remove background levels of isoflavone in plasma. 2. One day before experimentation, 3 'mice were assigned by time point and marked with unique identifiers. Each mouse was weighed to determine the density of DHE-bisphosphate required to dose animals at 50 mg / kg. 3. Each mouse is fed restricted and forced into an appropriate volume of DHE-bisphosphate to achieve a dose of 50 mg / kg. Once DHE-bisphosphate was administered, the mice were placed in a cage until the point of time (15 minutes, 30 minutes, 1 hour, 4 hours, 24 hours). The control animals were force-fed with 200 μl of 1% control CMC. The control animals were eliminated at time points of 15 minutes, 30 minutes, 1 hour, 4 hours, 24 hours. 4. At the designated time points, each mouse was sacrificed by cervical dislocation, then the blood was collected via the chest cavity as per SOP BD-009 using a 20 gauge needle. 5. The blood was allowed to clot, then centrifuged at high speed top for 3 minutes using a bench top mini-microfuge at room temperature. 6. The serum was aspirated into an appropriately labeled eppendorf tube and stored at -20 ° C until analyzed. The animal serum was dosed with control vehicle and the DHE-bisphosphate formulated was stored at -20 ° C together with 200 ul aliquots of the vehicle and DHE-bisphosphate formulated for analysis. 7. The three remaining animals were force-fed with formulation vehicle at time zero and were removed at the time of 30 minutes. The serum was stored with other samples.

Results When dosed at 25 mg / kg in mice, the DHE-bisphosphate molecule was metabolized to the DHE-free form with serum blood concentrations averaging 98.6 uM 15 minutes after the i.p injection. The drug was rapidly excreted at a rate of 62 μM / hr with serum levels decreased to 12 μM 1 hour after administration, total DHE concentrations (conjugate + free), reached 120 μM 15 minutes after administration and excreted (120 μM / hr) reaching a serum concentration of 30.85 1 hr after administration (Table 1 and Figure 1).

Table 1: Bisphosphate: free against total The comparison of free and total dehydroequol concentrations in serum was taken from mice dosed ip with DHE-bisphosphate and a formulation of DHE-PEG: PBS revealed that approximately equal concentrations of the free form of the drug were achieved in serum 15 minutes later to administration, however, half of the DHE-bisphosphate dosages were required (25 mg / kg) to achieve this result, compared to the formulation DHE-PEG: PBS (50 mg / kg) (74.4 μM against 62 uM respectively) (Table 1, Table 2 and Figure 2a). Interestingly, the free: total proportions observed for the preparation of DHE-bisphosphate and the DHE PEG: PBS formulation were some 5-fold different with more total DHE appearing in the plasma 15 minutes after the administration of rats dosed with the PEG formulation: PBS, when compared to the preparation of DHE-bisphosphate (120.8 μM phenoxodiol against 511.6 μM dehydroequol). Plasma concentrations of free dehydroequol were 1.8 times (DHE-bisphosphate) and 2.2 times (DHE-PEG: PBS) lower than those achieved in mice 15 minutes after administration of an HPBCD formulation of dehydroequol (50 mg / kg) (Fig. 2b, Table 3).

Table 2: PEG: free PBS against total Table 3: DHE Formulation Free HPBCD The uses of esterified isoflavonoids include any of the uses recently discovered or currently known for isoflavonoids or derivatives thereof, which include those listed above or described in the literature. Esterified isoflavonoids are found to be indicated in the treatment of osteoporosis and other symptoms of estrogen deficiency in postmenopausal women. Also, the compounds of the present invention are used to prevent osteoporosis and consequent fractures resulting from osteoporosis, which are major contributors to morbidity and mortality in the elderly. Still further, the esterified isoflavones are used prophylactically to provide UV protection and in other forms, to improve the overall health of the skin, to stimulate the immune system and reduce the undesirable effects of oxidation (ie, provide antioxidant benefits). Importantly, the compounds of the invention are used to treat cancer, which includes cancers of the breast, ovary and prostate. The esters of isoflavonoid phosphates of the invention, show almost unexpectedly, some beneficial and / or marked activity in the subjects to be treated. This comparison shows the particular usefulness and effectiveness of conjugated isoflavonoid compounds of the invention, and in particular, those conjugates of compounds 1 to 34 described above. Genistein phosphates were found to have poorer pharmacokinetic properties and profiles compared to the isoflavonoid counterparts described and exemplified above. Thus, modalities and specific applications of esterified isoflavonoid compounds have been described. It should be apparent, however, to those skilled in the art that many more modifications in addition to those already described are possible without departing from the inventive concisions of this document. The inventive subject matter, therefore, will not be restricted except in the spirit of the appended claims. Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications different from those specifically described. It will be understood that the invention includes such variations and modifications. The invention also includes all steps, features, compositions and compounds referred to, or indicated in this specification individually or collectively, and any and all combinations of any two or more of said steps or features. Reference to any prior art in this specification is not, nor should it be taken as an acknowledgment or any form of suggestion that the prior art forms part of the common general knowledge in the field of intent.

Claims (19)

NOVELTY OF THE INVENTION Having described the present is considered as a novelty, and therefore, it is claimed as property contained in the following: CLAIMS

1. A flavonoid phosphate ester compound of the general formula I: characterized in that Ri, R2 and Z are independently M2P04-, hydrogen, hydroxy, OR9, OC (O) R10, OS (O) R10, CHO, C (O) R? 0, COOH, C02Ro, CONR3R4, alkyl, haloalkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkoxyaryl, thio, alkylthio, amino, alkylamino, dialkylamino, nitro or halo, or R2 is as previously defined, and Ri and Z taken together with the carbon atoms to which are united, they form a ring of five elements selected from

Ri is as previously defined, and R2 and Z taken together with the carbon atoms to which they are attached form a ring of five selected elements

W is Ri, and A and B taken together with the carbon atoms to which they are attached, form a ring of six elements selected from wherein R3 is hydrogen, alkyl, aryl, arylalkyl, an amino acid, C (0) Rn wherein Ru is hydrogen, alkyl, aryl, arylalkyl or an amino acid, or C02R? 2 wherein Ri2 is hydrogen, alkyl, haloalkyl, aryl , heteroaryl or arylalkyl, R4 is hydrogen, alkyl or aryl, or R3 and R4 taken together with the nitrogen to which they are attached, comprise pyrrolidinyl or piperidinyl, R5 is M2P04-, hydrogen, C (0) Rn, where Ru is like is previously defined, or CO2R12 wherein Ri2 is as previously defined, Re is M2P04-, hydrogen, alkyl, aryl, amino, thio, NR3R4, CORn where Rn is as previously defined, C02R? 2 wherein R12 is as is previously defined or CONR3R4, R7 is hydrogen, C (0) Rn wherein Rn is as previously defined, alkyl, haloalkyl, aryl, arylalkyl or Si (R? 3) 3 wherein each Ri3 is independently hydrogen, alkyl or aryl , Rs is M2PO4-, hydrogen, hydroxy, alkoxy or alkyl, R9 is alkyl, haloalkyl, aryl, arylalkyl,

C (0) Rü where Rn is as previously defined, or Si (R? 3) 3 wherein RX3 is as previously defined, Rio is hydrogen, alkyl, haloalkyl, amino, aryl, arylalkyl, an amino acid, alkylamino or dialkylamino, the "" represents either a single bond or a double bond, M is independently hydrogen, an alkyl, alkenyl, alkynyl, alkoxyalkyl, alkylthioalkyl or aminoalkyl, straight or branched, a substituted or unsubstituted cycloalkyl, an aryl, aralkyl or alkylaryl, and a substituted cycloalkyl wherein at least one ring contains one or more than one nitrogen, sulfur, oxygen, phosphorus or silicon heteroatom in at least one ring; T is independently hydrogen, alkyl or aryl, X is 0, NR4 or S, preferably O, and Y is wherein R14, Ri5 and Rx6 are independently MP04-, hydrogen, hydroxy, 0R9, OC (0) R? o, OS (0) R? o, CHO, C (0) R? o, COOH, C02R? o / CONR3R4, alkyl, haloalkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkoxyaryl, thio, alkylthio, amino, alkylamino, dialkylamino, nitro or halo, and wherein at least one of Ri, R2, R5, Re, Rβ R? Ris / Rid Z, W or A wherein M2P04- are independently present, or a pharmaceutically acceptable salt thereof. 2. An isoflavonoid phosphate ester according to claim 1 of general formula II: characterized in that Ri, R2, R5, e 14 / R-15 / W and Z are as defined in accordance with claim 1, and the "" pattern represents either a single bond or a double bond. 3. An isoflavonoid phosphate ester according to claim 2, characterized in that the drawing "" represents a double bond. 4. An isoflavonoid phosphate ester according to claim 1 of general formula III: characterized in that Ri, R2, R5, Rs, Ri4, R15 W and Z are as defined according to claim 1.

5. An isoflavonoid phosphate ester according to claim 1 of general formula IV: characterized in that Ri, R2, R5, Re, Ri4, 15 W and Z are as defined according to claim 1.

6. An isoflavonoid compound according to any of claims 1 to 5, characterized in that the ester portion of phosphate is presented as a corresponding salt of M2P04-, where M is a pharmaceutically acceptable cation.

7. An isoflavonoid compound according to claim 6, characterized in that the corresponding salt of M2P04- is NaO (HO) P (O) 0-.

8. An isoflavonoid compound, characterized in that it is selected from (cis) 29 (cis) 30 or a pharmaceutically acceptable salt thereof.

9. A method for the treatment, prophylaxis, alleviation, defense against, and / or prevention of one or more of the therapeutic indications as defined herein, characterized in that it comprises administering to a subject a therapeutically effective amount of one or more compounds of formula I

10. Use of one or more compounds of formula I, for the manufacture of a medicament for the treatment, alleviation, defense against, prophylaxis and / or prevention of one or more therapeutic indications as described herein above.

11. An agent for the treatment, prophylaxis, alleviation, defense against and / or treatment of one or more of the therapeutic indications as defined herein, characterized in that it comprises one or more compounds of formula I, either alone or in association with one or more excipients or carriers.

12. A method for the treatment, prophylaxis, alleviation, defense against and / or prevention of conditions in a subject associated with the abnormal balance of estrogen / androgen, characterized in that the method includes the step of administering one or more compounds of formula I and either alone or in association with one or more carriers or excipients.

13. A method according to claim 12, characterized in that the subject is a woman.

14. A method according to claim 12, characterized in that the subject is a man.

15. Use of one or more compounds of formula I, for the manufacture of a medicament for the treatment, alleviation, defense against, prophylaxis and / or prevention of conditions in a subject, associated with abnormal estrogen / androgen balance.

16. A therapeutic composition, characterized in that it comprises one or more compounds of formula I, in association with one or more pharmaceutical carriers and / or excipients.

17. A beverage or food product, characterized in that it contains one or more compounds of formula I.

18. A composition, characterized in that it comprises one or more compounds of formula I, vitamin E and optionally carriers and / or excipients pharmaceutically, veterinarily or cosmetically acceptable.

19. Use of a phosphate ester portion to increase or improve the bioavailability of an isoflavan-4-one, isoflavene, isoflavan-4-ol or isoflavane compound or pharmaceutically acceptable salt thereof.