WO2006091187A1

WO2006091187A1 - Isoflavonoids for preventing radiation- and chemotherapy- induced weight loss

Info

Publication number: WO2006091187A1
Application number: PCT/US2005/005351
Authority: WO
Inventors: Michael R. Landauer
Original assignee: Landauer Michael R
Priority date: 2005-02-22
Filing date: 2005-02-22
Publication date: 2006-08-31

Abstract

Methods to ameliorate weight loss and facilitate weight gain during recovery from chemotherapy, radiation therapy (or both chemotherapy and radiation therapy) treatment regimens are provided.

Description

ISOFLAVONOIDS FOR PREVENTING RADIATION- AND CHEMOTHERAPY- INDUCED WEIGHT LOSS

FIELD

[0001] The present invention relates to methods for protecting mammals, in particular humans, from preventing radiation- and chemotherapy-induced weight loss induced by sublethal radiation, chemotherapy or both. The invention further relates to the use of isoflavones, in particular genistein, to prevent or treat weight loss due to radiation exposure or chemotherapy exposure.

GOVERNMENT RIGHTS

[0002] This invention was made with United States Government support. The Government may have certain rights in this invention.

BACKGROUND

[0003] Radiation therapy plays a critical role in the treatment of cancer, but its clinical use is limited by adverse effects on normal tissues. A frequent side effect of radiotherapy is the reduction of body weight, a condition that has been demonstrated to be a prognostic indicator of reduced survival rates (Beaver, M.E. et al, 2001, Otolaryngol Head Neck Surg 125: 645-8). The degree of radiation-induced side effects is dependent on the area irradiated, as well as the dose and duration of radiotherapy (Thiel H. J., Fietkau R, and Sauer, R., 1988, Recent Results Cancer Res 108: 205-26). For example, radiation to the head and neck can cause anorexia, taste changes, dry mouth, inflammation of the mouth and gums, swallowing problems, jaw spasms, or infection. These conditions can result in a significant reduction in body weight due to reduced tooα mtaKe ana aenyaration (αe Ia Maza M.P. et al, 2001, JAm Coll Nutr 20: 637-642), leading to a reduction in the quality of life of the patient.

[0004] One of the most common symptoms of cancer and a frequent side effect of cancer treatment is unintentional weight loss. Many patients experience a loss of appetite and significant reduction in their weight. Cancer can cause a cachexia wasting syndrome in some patients. This clinical syndrome includes weight loss, decreased appetite, fatigue, and poor performance status. It is multifactorial in nature and associated with mechanical factors, changes in taste, cytokines, and psychological factors.

[0005] Many of the treatments for cancer, including chemotherapy and radiation therapy, can decrease a patient's appetite. Patients need to maintain their nutrition to allow normal tissue repair after aggressive cancer treatments. There can be significant problems with healing if the patient has poor nutrition. Poor nutrition can alter the ability of a patient to tolerate a specific treatment and cause adjustments in the dose of chemotherapy and radiation treatments. This could ultimately decrease the effectiveness of a particular therapy.

[0006] While the isoflavones, and genistein in particular, protect normal tissues from acute lethal doses of ionizing radiation (Landauer et al, WO 01/95937, published December 20, 2001), it has been demonstrated that they do not protect the tumor and have been shown to enhance the efficacy of both radiotherapy and chemotherapy. For example, genistein improved radiotherapy of prostate tumors (Hillman et al, 2004, Cancer Ther. 3:1271-9) chemotherapy of breast cancer cells (Satoh et al, 2003, Res Commun MoI Pathol Pharmacol. 2003; 113 & 149) and chemo-radiotherapy of lung carcinoma cells (McDonnell et al, 2004, J. Surg Res. 116:19).

[0007] Attempts have been made to create agents for administration before, at the time, or after chemotherapy, radiation therapy and alike to patients with cancer or exposed to radiation. However, problems have arisen with the potency, route of administration, selectivity of protecting normal tissues and organs without diminishing of the radioprotective or chemotherapeutic agent's desired effect. There is a need to to ameliorate weight loss and facilitate weight gain during recovery from chemotherapy, radiation therapy (or both chemotherapy and radiation therapy) treatment regimens.

SUMMARY

[0008] It has now been discovered that isoflavone compounds, in particular genistein, are ideal radioprotectants and chemoprotectants that possess desirable and important properties for treating and preventing radiation-induced and chemotherapy-induced weight loss in a patient. More specifically, as disclosed herein, are methods to ameliorate weight loss and facilitate weight gain during recovery from chemotherapy, radiation therapy (or both chemotherapy and radiation therapy) treatment regimens.

[0009] Accordingly, in one aspect, the invention provides methods of treating or preventing radiation-induced weight loss in a subject exposed to radiation, the method comprising administering to the subject a therapeutically effective amount of an isoflavone. In a related aspect, the radiation is an acute sub-lethal dose of ionizing radiation, a chronic low-dose of ionizing radiation, an acute sub-lethal dose of non-ionizing radiation, or a chronic low-dose of non-ionizing radiation. In a related aspect, the radiation is selected from the group consisting of diagnostic X-rays, radiation therapy in cancer treatment, CAT-scans, mammograms, radionuclide scans, interventional radiological procedures under CT or fluoroscopy guidance, tissue-incorporated radionuclides from ingestion of contaminated food or water, and uncontrolled exposure to ionizing radiation from nuclear weapons, radioactive spills, and/or cosmic radiation. In a further related aspect, the isoflavone is selected from the group consisting of genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O- desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof. In a related aspect, the isoflavone is administered orally, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, or rectally. In a further related aspect, the isoflavone is administered orally in the form of a capsule, a tablet, an inhaler, a troche, or a food supplement in the form of a food or beverage. In a related aspect, the isoflavone is administered chronically. In some aspects, the isoflavone is administered within 2 weeks prior to exposure to radiation, during radiation exposure, and/or within 2 weeks following radiation exposure. In other aspects, the isoflavone is administered within 4 days prior to radiation exposure, during radiation exposure, and/or within 4 days following radiation exposure.

[0010] In another aspect, the present invention provides methods for treating or preventing radiation-induced weight loss in a subject exposed to a dose of radiation, the method comprising administering to the subject before, during and/or after the dose of radiation a therapeutically effective amount of a compound of the formula:

wherein Rl, R2, R3, R4 are independently selected from the group consisting of hydrogen, hydroxyl and alkoxy. In a related aspect, the invention provides methods for treating or preventing radiation-induced weight loss in a subject exposed to a dose of radiation wherein the compound is genistein. In a related aspect, the invention provides methods for treating or preventing radiation-induced weight loss in a subject exposed to a dose of radiation, wherein the compound is administered to the subject during the time period of approximately 4 days prior to radiation exposure to approximately 4 days subsequent to the lethal dose of irradiation.

[0011] In another aspect, the invention provides methods for preventing or reducing weight loss in an individual undergoing treatment with a therapeutic agent, comprising administering to the patient an effective amount of an isoflavone. In a related aspect, the agent is a chemotherapeutic agent or a radiotherapeutic agent. In a further related aspect, the therapeutic agent is a chemotherapeutic agent and a radiotherapeutic agent. In a related aspect, the isoflavone is administered simultaneously with administration of the therapeutic agent. In a further related aspect, the isoflavone is administered prior to administration of the therapeutic agent. In a related aspect, the isoflavone is administered subsequent to administration of the therapeutic agent. In a further related aspect, the effective amount of the isoflavone is administered to the patient in a time period from about one month before administration of the chemotherapeutic agent to about one month after administration of the chemotherapeutic agent. In a related aspect, the effective amount of the isoflavone is administered to the patient in a time period from about 7 days before administration of the chemotherapeutic agent to about 7 days after administration of the chemotherapeutic agent. In a related aspect, the effective amount of the isoflavone is administered to the patient in a time period from about 4 days before administration of the chemotherapeutic agent to about 4 days after administration of the chemotherapeutic agent. In a related aspect, the isoflavone is genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, 0-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof. In a related aspect, the isoflavone is genistein. In a further related aspect, the isoflavone has the formula:

wherein Rl, KZ, JK3 and K4 are independently selected from the group consisting of hydrogen, hydroxy! and alkoxy. In a related aspect, the effective amount of the isoflavone is in the range of from about 0.1 mg/kg hody weight to about 1000 mg/kg body weight. In a related aspect, the effective amount of the isoflavone is in the range of from about 1 mg/kg body weight to about 500 mg/kg body weight. In a related aspect, the effective amount of the isoflavone is in the range of from about 20 mg/kg body weight to about 400 mg/kg body weight, hi a related aspect, the isoflavone is administered orally, hi a further related aspect, the isoflavone is administered in an effective amount in the range of from about 1 mg/kg body weight to about 50 mg/kg body weight, hi a further related aspect, the isoflavone is administered in an effective amount in the range of from about 2 mg/kg body weight to about 25 mg/kg body weight. In a further related aspect, the isoflavone is administered in an effective amount in the range of from about 4 mg/kg body weight to about 12 mg/kg body weight. In a related aspect, the isoflavone is administered once per day. In a related aspect, the isoflavone is administered twice per day. hi a further related aspect, the isoflavone is administered three or more times per day. In a further related aspect, the isoflavone is administered parenterally. hi a further related aspect, the parenteral administration is by slow intravenous infusion, hi a further related aspect, the isoflavone is administered parenterally by injection, hi a related aspect, the isoflavone is administered in an effective amount in the range of from about 0.1 mg/kg body weight to about 1000 mg/kg body weight, hi a further related aspect, the isoflavone is administered in an effective amount in the range of from about 1 mg/kg body weight to about 500 mg/kg body weight, hi a further related aspect, the isoflavone is administered in an effective amount in the range of from about 2 mg/kg body weight to about 10 mg/kg body weight, hi a further related aspect, wherein the isoflavone is orally administered beginning about 90 days prior to exposure to a sublethal dose of radiation. In a further related aspect, the isoflavone is orally administered beginning about 7 days prior to exposure to a sublethal dose of radiation, hi a further related aspect, the isoflavone is orally administered beginning about 90 days prior to and about 90 days after exposure to a sublethal dose of radiation, hi a further related aspect, the isoflavone is orally administered beginning about 30 days prior to and about 30 days after exposure to a sublethal dose of radiation, hi a further related aspect, wherein the isoflavone is administered 14 days prior to and about 14 days after exposure to a sublethal dose of radiation, hi a further related aspect, wherein the isoflavone is administered 7 days prior to and about 7 days after exposure to a sublethal dose of radiation, hi a further related aspect, wherein the isoflavone is administered a 1 day prior to and about 1 day after exposure to a sublethal dose of radiation, hi a related aspect, the isoflavone is administered as a food supplement. In a related aspect, the chemotherapeutic agent is administered for treatment of cancer. In a further related aspect, the cancer is breast cancer, lung cancer (small and/or non-small cell), acute lymphocytic leukemia, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, chronic myelocytic leukemia, hairy cell leukemia, ovary cancer, ovary (germ cell) cancer, melanoma, skin cancer, cervix cancer, ACTH-producing tumors, cancer of the adrenal cortex, bladder cancer, brain cancer, colorectal cancer, cutaneous T-cell lymphoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, gallbladder cancer, head & neck cancer, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, liver cancer, malignant peritoneal effusion, malignant pleural effusion, mesothelioma, multiple myeloma, neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma, pancreatic cancer, penis cancer, prostate cancer, retinoblastoma, soft-tissue sarcoma, squamous cell carcinomas, stomach cancer, testicular cancer, thyroid cancer, trophoblastic neoplasms, cancer of the uterus, vaginal cancer, cancer of the vulva, and Wihn's tumor.

[0012] In another aspect, the invention provides methods of preventing or reducing weight loss in a mammal, the method comprising: selecting a mammal having or at risk of having a condition characterized by weight loss; and administering to the mammal a composition comprising at least one isoflavonoid, wherein the administration prevents or reduces weight loss in the mammal, hi a related aspect, the weight loss is induced by radiation. In a further related aspect, the weight loss is induced by chemotherapy. In a further related aspect, the isoflavonoid is selected from the group consisting of genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof. In a further related aspect, the isoflavonoid is from a source selected from the group consisting of soy, soy products and clover.

BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1. Chemical structures of genistein and daidzein.

[0014] Figure 2. Recovery of body weight of CD2F1 male mice following sublethal irradiation (7 Gy). Mice received a single subcutaneous injection of the isoflavone genistein (top panel) or daidzein (bottom panel), 24 hr prior to irradiation. * p < 0.05 from vehicle-treated irradiated group.

DETAILED DESCRIPTION

1. INTRODUCTION

[0015] The invention provides a number of methods, reagents, and compounds that can be used either for the treatment of radiation-induced weight loss and chemotherapy-induced weight loss, the development of treatments for radiation-induced weight loss and chemotherapy- induced weight loss, the practice of the other inventive methods described herein, or for a variety of other purposes.

[0016] It is to be understood that this invention is not limited to particular methods, reagents, compounds, compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. As used in this specification and the appended claims, the singular forms "a", "an" and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a cell" includes a combination of two or more cells, and the like.

[0017] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, the preferred materials and methods are described herein, hi describing and claiming the present invention, the following terminology will be used.

[0018] "Patient", "subject" or "mammal" are used interchangeably and refer to mammals such as human patients and non-human primates, as well as experimental animals such as rabbits, rats, and mice, and other animals. Animals include all vertebrates, e.g., mammals and non-mammals, such as sheep, dogs, cows, chickens, amphibians, and reptiles.

[0019] "Treating" or "treatment" includes the administration of the compositions, compounds or agents of the present invention to prevent or delay the onset of the symptoms, complications, or biochemical indicia of a disease, alleviating or ameliorating the symptoms or arresting or inhibiting further development of the disease, condition, or disorder (e.g., radiation- induced or chemotherapy-induced weight loss). "Treating" further refers to any indicia of success in the treatment or amelioration or prevention of the disease, condition, or disorder (e.g., radiation-induced or chemotherapy-induced weight loss), including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the disease condition more tolerable to the patient; slowing in the rate of degeneration or decline; or making the final point of degeneration less debilitating. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of an examination by a physician. Accordingly, the term "treating" includes the administration of the compounds or agents of the present invention to prevent or delay, to alleviate, or to arrest or inhibit development of the symptoms or conditions associated with ocular disease. The term "therapeutic effect" refers to the reduction, elimination, or prevention of the disease, symptoms of the disease, or side effects of the disease in the subject. "Treating" or "treatment" using the methods of the present invention includes preventing the onset of symptoms in a subject that can be at increased risk of radiation- or chemotherapy-induced weight loss but does not yet experience or exhibit symptoms, inhibiting the symptoms of radiation or chemotherapy-induced weight loss (slowing or arresting its development), providing relief from the symptoms or side- effects of radiation or chemotherapy-induced weight loss (including palliative treatment), and relieving the symptoms of radiation or chemotherapy-induced weight loss (causing regression). Treatment can be prophylactic (to prevent or delay the onset of the disease, or to prevent the manifestation of clinical or subclinical symptoms thereof) or therapeutic suppression or alleviation of symptoms after the manifestation of the disease or condition.

[0020] "Cancer" or "malignancy" are used as synonymous terms and refer to any of a number of diseases that are characterized by uncontrolled, abnormal proliferation of cells, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other parts of the body (i.e., metastasize) as well as any of a number of characteristic structural and/or molecular features. A "cancerous" or "malignant cell" is understood as a cell having ^" specific structural properties, lacking differentiation and being capable of invasion and metastasis. Examples of cancers are, breast, lung, brain, bone, liver, kidney, colon, and prostate cancer, (see DeVita et ah, Eds., Cancer Principles and Practice of Oncology, 6th. Ed., Lippincott Williams & Wilkins, Philadelphia, PA, 2001; this reference is herein incorporated by reference in its entirety for all purposes).

[0021] "Cancer-associated" refers to the relationship of a nucleic acid and its expression, or lack thereof, or a protein and its level or activity, or lack thereof, to the onset of malignancy in a subject cell. For example, cancer can be associated with expression of a particular gene that is not expressed, or is expressed at a lower level, in a normal healthy cell. Conversely, a cancer-associated gene can be one that is not expressed in a malignant cell (or in a cell undergoing transformation), or is expressed at a lower level in the malignant cell than it is expressed in a normal healthy cell.

[0022] In the context of the cancer, the term "transformation" refers to the change that a normal cell undergoes as it becomes malignant. In eukaryotes, the term "transformation" can be used to describe the conversion of normal cells to malignant cells in cell culture.

[0023] "Proliferating cells" are those which are actively undergoing cell division and growing exponentially. "Loss of cell proliferation control" refers to the property of cells that have lost the cell cycle controls that normally ensure appropriate restriction of cell division. Cells that nave lost such controls proliferate at a faster than normal rate, without stimulatory signals, and do not respond to inhibitory signals.

[0024] "Advanced cancer" means cancer that is no longer localized to the primary tumor site, or a cancer that is Stage III or IV according to the American Joint Committee on Cancer (AJCC).

[0025] "Well tolerated" refers to the absence of adverse changes in health status that occur as a result of the treatment and would affect treatment decisions.

[0026] "Metastatic" refers to tumor cells, e.g., human breast cancer cells, that are able to establish secondary tumor lesions in the lungs, liver, bone or brain of immune deficient mice upon injection into the mammary fat pad and/or the circulation of the immune deficient mouse.

[0027] "Non-metastatic" refers to tumor cells, e.g., human breast cancer cells, that are unable to establish secondary tumor lesions in the lungs, liver, bone or brain or other target organs of breast cancer metastasis in immune deficient mice upon injection into the mammary fat pad and/or the circulation. The human tumor cells used herein and addressed herein as non- metastatic are able to establish primary tumors upon injection into the mammary fat pad of the immune deficient mouse, but they are unable to disseminate from those primary tumors.

[0028] Radiation is administered in accordance with well known standard techniques using standard equipment manufactured for this purpose, such as AECL Theratron and Varian Clinac. The dose of radiation depends on numerous factors as is well known in the art. Such factors include the organ being treated, the healthy organs in the path of the radiation that might inadvertently be adversely affected, the tolerance of the patient for radiation therapy, and the area of the body in need of treatment. The dose will typically be between 1 and 100 Gy, and more particularly between 2 and 80 Gy. Some doses that have been reported include 35 Gy to the spinal cord, 15 Gy to the kidneys, 20 Gy to the liver, and 65-80 Gy to the prostate. It should be emphasized, however, that the invention is not limited to any particular dose. The dose will be determined by the treating physician in accordance with the particular factors in a given situation, including the factors mentioned above. Sources of radiation include, but are not limited to, diagnostic X-rays, radiation therapy in cancer treatment, CAT-scans, mammograms, radionuclide scans, interventional radiological procedures under CT or fluoroscopy guidance, tissue-incorporated radionuclides from ingestion of contaminated food or water, and uncontrolled exposure to ionizing radiation from nuclear weapons, radioactive spills, and/or cosmic radiation.

[0029] A "radioprotector" or "radioprotectant" is a synthetic or naturally occurring chemical agent that is capable of ameliorating at least one adverse effect of exposure to sublethal amounts oi radiation, in one preferred embodiment, one adverse effect of sublethal amounts of radiation (also referred to as "radiotherapy") is radiation-induced weight loss. In one preferred embodiment, a radioprotectant of the present invention is an isoflavone. In other preferred embodiments, a preferred isoflavone is genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof.

[0030] As used herein, the term "radioprotectant composition" refers to a composition that includes at least one radioprotectant, and can include more than one radioprotectants. Radioprotectant compositions can also include, in addition to one or more radioprotectant(s), pharmaceutically acceptable carriers that facilitate administration of a radioprotectant composition to a subject.

[0031] As used herein, the term "ameliorating at least one adverse effect of radiotherapy" includes: (a) reducing the magnitude and/or duration of at least one adverse effect of radiotherapy; and/or (b) completely eliminating at least one adverse effect of radiotherapy; and/or (c) preventing the onset of one or more adverse effect(s) of radiotherapy that would occur without administration of a radioprotectant composition of the invention.

[0032] The term "chemotherapeutic agents" include all chemical compounds that are effective in inhibiting cancer.

[0033] A "chemoprotector" or "chemoprotectant" is a synthetic or naturally occurring chemical agent that is capable of ameliorating at least one adverse effect of chemotherapy. In one preferred embodiment, one adverse effect of chemotherapy is chemotherapy-induced weight loss, hi one preferred embodiment, a chemoprotectant of the present invention is an isoflavone. In other preferred embodiments, a preferred isoflavone is genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof.

[0034] As used herein, the term "chemoprotectant composition" refers to a composition that includes at least one chemoprotectant, and can include more than one chemoprotectants. Chemoprotectant compositions can also include, in addition to one or more chemoprotectant(s), pharmaceutically acceptable carriers that facilitate administration of a chemoprotectant composition to a subject.

[0035] As used herein, the term "ameliorating at least one adverse effect of chemotherapy" includes: (a) reducing the magnitude and/or duration of at least one adverse effect of chemotherapy; and/or (b) completely eliminating at least one adverse effect of chemotherapy; and/or (c) preventing the onset of one or more adverse effect(s) of chemotherapy that would occur without administration of a chemoprotectant composition of the invention. In one preferred embodiment, one adverse effect of chemotherapy is chemotherapy-induced weight loss.

[0036] As used herein, the term "chemotherapeutic agent" is an agent that is administered to a mammalian subject to destroy, or otherwise adversely affect, cancer cells.

[0037] The administration of chemotherapeutic agents can be accomplished in a variety of ways including systemically by the parenteral and enteral routes. Examples of chemotherapeutic agents include alkylating agents, for example, nitrogen mustards, ethyleneiniine compounds and alkyl sulphonates; antimetabolites, for example, folic acid, purine or pyrimidine antagonists, mitotic inhibitors, for example, vinca alkaloids and derivatives of podophyllotoxin; cytotoxic antibiotics; compounds that damage or interfere with DNA expression; and growth factor receptor antagonists.

[0038] Additionally, chemotherapeutic agents include antibodies, biological molecules and small molecules, as described above.

[0039] Particular examples of chemotherapeutic agents or chemotherapy combined with the compositions, compounds or agents of the present invention, include but are not limited to aldesleukin, altretamine, aniifostine, asparaginase, bleomycin, busulfan, capecitabine, carboplatin, carmustine (BCNU), cladribine, cisapride, cisplatin, cyclophosphamide, cytarabine, dacarbazine (DTIC), dactinomycin, docetaxel (taxotere), doxorubicin (adriamycin), dronabinol, duocarmycin, epoetin alpha, etoposide, filgrastim, floxuridine, fludarabine, fluorouracil, gemcitabine, granisetron, hydroxyurea, idarubicin, ifosfamide, interferon alpha, irinotecan, lansoprazole, leuprolide, levamisole, leucovorin, lomustine (CCNU), megestrol, melphalan, mercaptopurine, mesna, methotrexate, metoclopramide, mitomycin, mitotane, mitoxantrone, omeprazole, ondansetron, paclitaxel (Taxol™), plicamycin, procarbazine, pilocarpine, prochlorperazine, rituximab, saproin, streptozocin, tamoxifen, testolactone, thioguanine, thiotepa, topotecan hydrochloride, trastuzumab, uracil mustard, vinblastine, vincristine and vinorelbine tartrate.

[0040] "Dosage unit" refers to physically discrete units suited as unitary dosages for the particular individual to be treated. Each unit can contain a predetermined quantity of active compound(s) calculated to produce the desired therapeutic effect(s) in association with the required pharmaceutical carrier. The specification for the dosage unit forms can be dictated by (a) the unique characteristics of the active compound(s) and the particular therapeutic effect(s) to be achieved, and (b) the limitations inherent in the art of compounding such active compound(s). [0041] Pharmaceutically acceptable excipient "means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients can be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous.

[0042] "Pharmaceutically acceptable salts and esters" means salts and esters that are pharmaceutically acceptable and have the desired pharmacological properties. Such salts include salts that can be formed where acidic protons present in the compounds are capable of reacting with inorganic or organic bases. Suitable inorganic salts include those formed with the alkali metals, e.g., sodium and potassium, magnesium, calcium, and aluminum. Suitable organic salts include those formed with organic bases such as the amine bases, e.g., ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. Such salts also include acid addition salts formed with inorganic acids (e.g., hydrochloric and hydrobromic acids) and organic acids (e.g., acetic acid, citric acid, maleic acid, and the alkane- and arene- sulfonic acids such as methanesulfonic acid and benzenesulfonic acid). Pharmaceutically acceptable esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the compounds, e.g., C_1-6 alkyl esters. When there are two acidic groups present, a pharmaceutically acceptable salt or ester can be a mono-acid-mono-salt or ester or a di-salt or ester; and similarly where there are more than two acidic groups present, some or all of such groups can be salified or esterified. Compounds named in this invention can be present in unsalified or unesterified form, or in salified and/or esterified form, and the naming of such compounds is intended to include both the original (unsalified and unesterified) compound and its pharmaceutically acceptable salts and esters. Also, certain compounds named in this invention can be present in more than one stereoisomeric form, and the naming of such compounds is intended to include all single stereoisomers and all mixtures (whether racemic or otherwise) of such stereoisomers.

[0043] The terms "pharmaceutically acceptable", "physiologically tolerable" and grammatical variations thereof, as they refer to compositions, carriers, diluents and reagents, are used interchangeably and represent that the materials are capable of administration to or upon a human without the production of undesirable physiological effects to a degree that would prohibit administration of the composition.

[0044] A "therapeutically effective amount" means the amount that, when administered to a subject for treating a disease, is sufficient to effect treatment for that disease. [0045] Except when noted, the terms "subject" or "patient" are used interchangeably and refer to mammals such as human patients and non-human primates, as well as experimental animals such as rabbits, rats, and mice, and other animals. Accordingly, the term "subject" or "patient" as used herein means any mammalian patient or subject to which the compositions of the invention can be administered. In some embodiments of the present invention, the patient will be suffering from weight loss induced by radiation or chemotherapy.

[0046] "Concomitant administration" of a known cancer therapeutic drug, a radiotherapeutic drug, or both, with a pharmaceutical composition of the present invention means administration of the drug and the isoflavone composition at such time that both the known drug and the composition of the present invention will have a therapeutic effect. Such concomitant administration can involve concurrent {i.e., at the same time), prior, or subsequent administration of the antimicrobial drug with respect to the administration of a compound of the present invention. A person of ordinary skill in the art, would have no difficulty determining the appropriate timing, sequence and dosages of administration for particular drugs and compositions of the present invention.

[0047] "In combination with", "combination therapy" and "combination products" refer, in certain embodiments, to the concurrent administration to a patient of a first therapeutic and the compounds as used herein. When administered in combination, each component can be administered at the same time or sequentially in any order at different points in time. Thus, each component can be administered separately but sufficiently closely in time so as to provide the desired therapeutic effect.

2. OVERVIEW

[0048] The present invention is directed to a method for preventing, reducing and/or treating weight loss in a patient undergoing treatment with a chemotherapeutic agent, a radiotherapeutic agent, or exposed to sublethal radiation by administering an effective amount of an isoflavone to the patient. The isoflavone chemoprotectant compound of the invention is given to the mammals prior to, during, or immediately after the radiotherapy, chemotherapy (or combinations thereof), or exposure to sublethal radiation. Isoflavone compounds particularly useful in the present invention include compounds having the general formula:

wherein R₁, R₂, R₃ and R₄ are independently selected from the group consisting of hydrogen, hydroxy! and alkoxy.

[0049] Isoflavone compounds of interest include genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethyangolensin, equol and the like, their glucosides and derivatives, and/or mixtures thereof. Of particular importance is genistein, also known as 5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-l benzopyran-4-one or 4', 5,7- trihydroxyisoflavone.

[0050] The isoflavone compounds can be derived from any suitable source such as soy, legumes, clover and the like using any of the techniques well known to one of ordinary skill in the art. Preferably the isoflavone compounds used in the present invention are naturally occurring substances which can be found in plants such as legumes, clover, and the root of the kudzu vine (pueraria root). Common legume sources of these isoflavone compounds include soy beans, chick peas, and various other types of beans and peas. Clover sources of these isoflavone compounds include red clover and subterranean clover. Soy beans are a particularly preferred source of the isoflavone compounds (except biochanin A which is not present in soy).

[0051] The isoflavone compounds can be isolated from the plant sources in which they naturally occur, or can be synthetically prepared by processes known in the art. For example, daidzein canbe isolated from red clover as disclosed by Wong, 1962, J. Sd. FoodAgr., 13:304) or canbe isolated from the mold Micromonospora halophytica as provided by Ganguly and Sarre (Chem. & Ind. (London), 1970, pg. 201), both references of which are incorporated by reference herein. Daidzein can be synthetically prepared by the methods provided by Baker et al (J Chem. Soc, p. 27), Wesley et al, 1033, 66:685), Mahal et al, 1934, J. Chem. Soc, p. 1769), Baker et al, 1953, J. Chem. Soc, p. 1852), or Farkas, 1957, Ber. 90:2940), each reference of which is incorporated herein by reference. The isoflavone glucoside daidzin can be synthetically prepared by the method of Farkas et al (1959, Ber. 92:819), incorporated herein by reference. The daidzein isoflavone glucoside conjugates 6'-0-MaI daidzin and 6'-0-Ac daidzin can be prepared by a conventional saponification of daidzin with a malonyl or an acetyl anhydride, respectively. [0052] Genistein can be synthetically prepared by the methods provided by Baker et al , 1928, J Chem. Soc, p. 3115); Narasimhachari et al, 1953, J Sd. Ind. Res. 12:287); Yoder et al, 1954, Proc. Iowa Acad. Sd. 61:271); and Zemplen et α/., 1959, Acta. CHm. Acad. Set Hung. 19:277), each reference of which is incorporated herein by reference. The isoflavone glucoside genistin can be synthetically prepared by the method of Zemplen et al, 1943, Ber. VoI 76B:1110), incorporated herein by reference. The isoflavone glucoside conjugates of genistein, 6'-0-MaI genistin and 6'-0-Ac genistin, can be prepared by a conventional saponification of genistin with a malonyl or an acetyl anhydride, respectively. Biochanin A can be synthetically prepared by the method provided by Baker et al, 1952, Nature 169:706), incorporated herein by reference. Biochanin A can also be separated from red clover by the method provided by Pope et al, 1953, Chem. & Ind. (London) p.1092), incorporated herein by reference. Formononetin can be synthetically prepared by the methods disclosed by Wessely et al 1933, Ber. 66:685) and Kagel et al, 1962, Tetrahedron Letters, p. 593), both references of which are incorporated herein by reference. Formononetin can be isolated from soybean meal by the method of WaIz, 1931, Ann. 489 : 118) or can be isolated from clover species by the method of Bradbury et al , 1951 , J Chem. Soc. p. 3447), both references of which are incorporated herein by reference.

[0053] The isoflavones useful in the present invention can be extracted from the plant materials in which they naturally occur by any suitable means known to one of skill in the art. Many of the isoflavone compounds are available commercially. For example, genistein, daidzein, and glycitein are commercially available and can be purchased, for example, from Indofine Chemical Company Inc., P.O. Box 473, Somerville, NJ. No. 08876, and biochanin A is available from Aldrich Chemical Company, Inc., 940 West Saint Paul Avenue, Milwaukee, Wis. 53233.

[0054] The administration of this compound to patients suffering from radiation- induced weight loss or chemotherapy-induced weight loss provides selective protection or recovery from weight loss induced by radiation or chemotherapy.

[0055] The isoflavone radioprotectant and chemoprotectant compositions of the present invention can be administered in conjunction with a therapy regimen used for the treatment of cancer. Such cancers can include, but are not limited to, types of cancer including breast cancer, lung cancer (small and/or non-small cell), acute lymphocytic leukemia, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, chronic myelocytic leukemia, hairy cell leukemia, ovary cancer, ovary (germ cell) cancer, melanoma, skin cancer, cervix cancer, ACTH-producing tumors, cancer of the adrenal cortex, bladder cancer, brain cancer, colorectal cancer, cutaneous T-cell lymphoma, endometrial cancer, esophageal cancer, Swing's sarcoma, gallbladder cancer, head & neck cancer, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, liver cancer, malignant peritoneal effusion, malignant pleural effusion, mesothelioma, multiple myeloma, neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma, pancreatic cancer, penis cancer, prostate cancer, retinoblastoma, soft-tissue sarcoma, squamous cell carcinomas, stomach cancer, testicular cancer, thyroid cancer, trophoblastic neoplasms, cancer of the uterus, vaginal cancer, cancer of the vulva, and Wihn's tumor. The administration of these isoflavone radioprotectant and chemoprotectant compositions to patients with different types of cancer and other diseases treatable with radiotherapeutic and chemotherapeutic agents will provide a selective protection or recovery from weight loss induced by radiation or chemotherapy.

[0056] Radiotherapeutic and chemotherapeutic agents can also be used in some diseases not related to cancer. For example, chemotherapeutic agents can be administered as part of a treatment regimen for a collagen- vascular disease, such as lupus erythematosus, rhematoid arthritis, or the like, inflammatory bowel disease, such as Crohn's disease, and temporal arthritis. In such cases the chemotherapeutic agent of choice is azathioprine, which after undergoing cellular metabolism transforms into active but toxic 6-thioquanine (6-TG).

[0057] Chemotherapeutic agents are also used in organ transplantation procedures requiring immunosuppression with chemodrugs to prevent the rejection of the transplant. Transplant organs can include the kidney, heart, liver, bone marrow, lung, skin, and the like. In such cases, the chemotherapeutic agent of choice is usually either cyclosporin or methotrexate.

[0058] In all of these cases, the use of the isoflavone radioprotectant and chemoprotectant compositions of the present invention confer a protective effect to a subject and thus prevent, reduce and/or treat various toxic side effects associated with or resulting from use of sublethal exposure to radiation or chemotherapeutic agents.

[0059] According to the present invention the isoflavone compound of the invention can be administered to a mammal or patient prior to, during, and/or following the radiation exposure or chemotherapy. The isoflavone is administered for a time period sufficient to confer a protective effect to the subject from the radiation-induced or chemotherapy-induced weight loss, or to allow a reduction in the severity of the effects, and/or to treat the side effects from the sublethal exposure to radiation or chemotherapy. One of ordinary skill in the art would be able to readily determine a suitable time period taking into account the dosage to be administered, the route of administration, the periodic intervals for administration, the age, weight and health of the patient, the cancer or other disease to be treated, and the like. [0060] The isoflavone compositions of the present invention can be administered by any conventional route known to one of skill in the art, either in a typical pharmaceutical preparation form or as a food supplement (discussed in further detail below).

[0061] These isoflavone compounds can be administered alone, or in combination with the other drug compounds discussed herein, in the form of the water-soluble acid, free base, or as physiologically acceptable salts, including acid addition salts formed with organic and inorganic acids, for example, hydrochlorides, hydrobromides, sulfates, phosphates, citrates, fumarates, and lnaleates, and cations such as sodium, potassium, and the like These compounds can be formulated for administration to humans and animals with pharmaceutically acceptable carriers, excipients, and diluents, such as sterile distilled water, Ringer's solution, normal saline, 5% glucose, dextrose, fructose, sucrose, and the like, and mixtures thereof, as is well known in the art. Antimicrobial agents, preservatives, and the like, can also be included. Compositions for oral administration can include coloring and flavoring agents. Additional methods of formulating compounds of the present invention for administration in the methods described herein can be found, for example, in Remington 's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA 18^th ed., 1990, incorporated herein by reference).

[0062] For these purposes, the compounds of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir in dosage formulations containing conventional non-toxic pharmaceutically- acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraperitoneally, intrathecally, intraventricularly, and other routes acceptable to one of skill in the art for drug delivery.

[0063] In addition, administration can be by a single dose, it can be repeated at intervals or it can be by continuous infusion. Where continuous infusion is preferred, pump means often will be particularly preferred for administration.

[0064] Unlike known prior art radioprotectants or chemoprotectants, the isoflavone radioprotectant and chemoprotectant compositions of the present invention are highly suited for administration to the patient as a food supplement. Suitable food supplements include any ingestible preparation containing therapeutically effective amounts of the isoflavone compositions of the present invention which are capable of imparting the desired radioprotectant and chemoprotectant effects to the mammal ingesting the food supplement. The food supplement can be freshly prepared, dried, cooked, boiled, lyophilized or baked. Breads, teas, soups, cereals, pills and tablets, are among the vast number of different food products contemplated. [0065] The isoflavone radioprotectant and chemoprotectant compositions of the present invention can be effectively used in methods for treating human and animal patients undergoing treatment with radiotherapeutic agents, cheniotherapeutic agents, or patients exposed to sublethal amounts of radiation, to prevent or reduce side effects associated therewith and to prolong survival. These methods comprise administering to the patient an appropriate effective amount of an isoflavone radioprotective and/or chemoprotective agent prior to, simultaneously with, or subsequent to administration of a radiotherapeutic, chemotherapeutic agent, and/or exposure to sublethal amounts of radiation. Combinations of these time periods can also be employed.

3. PHARMACEUTICAL COMPOSITIONS AND METHODS OF ADMINISTRATION A. TREATMENT REGIMES

[0066] The invention provides pharmaceutical compositions comprising one or a combination of isoflavones, e.g., genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, formulated together with a pharmaceutically acceptable carrier. Some compositions include a combination of multiple (e.g., two or more) protective agents (i.e., radioprotectants or chemoprotectants).

[0067] In prophylactic applications, pharmaceutical compositions or medicaments are administered to a patient susceptible to, or otherwise at risk of a disease or condition (i.e., radiation-induced weight loss or chemotherapy-induced weight loss) in an amount sufficient to eliminate or reduce the risk, lessen the severity, or delay the outset of the disease or condition, including biochemical, histologic and/or behavioral symptoms of the disease, its complications and intermediate pathological phenotypes presenting during development of the disease. In therapeutic applications, compositions or medicants are administered to a patient suspected of, or already suffering from such a disease or condition in an amount sufficient to cure, or at least partially arrest, the symptoms of the disease or condition (biochemical, histologic and/or behavioral), including its complications and intermediate pathological phenotypes in development of the disease or condition. An amount adequate to accomplish therapeutic or prophylactic treatment is defined as a therapeutically- or prophylactically-effective dose, hi both prophylactic and therapeutic regimes, agents are usually administered in several dosages until a sufficient response has been achieved. Typically, the response is monitored and repeated dosages are given if the response starts to wane.

[0068] In a preferred embodiment of the invention, the isoflavone can be administered chronically to the patient for a suitable time period of from about 90 days before administration of the chemotherapeutic agent or radiotherapeutic agent (or both chemotherapeutic and radiotherapeutic agents) to about 30 days or longer after administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic and radiotherapeutic agents). Preferably the isoflavone is administered about 90 days preceding the administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic and radiotherapeutic agents). More preferably, the isoflavone is administered about 30 days preceding the administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic agent and radiotherapeutic agent). Most preferably, the isoflavone is administered about 7 to 14 days preceding the administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic agent and radiotherapeutic agent). However, protective effects can still be conferred by administration 1 or 2 days, or even hours or less before administration of the chemotherapeutic agent or radiotherapeutic agents. The isoflavone can also be administered concurrently with the administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic agent and radiotherapeutic agent). Less preferred, but still conferring a protective effect is the administration of the isoflavone after the administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic agent and radiotherapeutic agent), preferably for a time period from about 7 to 14 days after administration of the chemotherapeutic agent or radiotherapeutic agent (or after administration of both chemotherapeutic agent and radiotherapeutic agent). In other preferred embodiments, the time period can be two weeks, one month or more, or years after administration of the radiotherapeutic agent, chemotherapeutic agent (or both radiotherapeutic agent and chemotherapeutic agent).

[0069] The pharmaceutical compositions comprising one or a combination of isoflavones, e.g., genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, 0-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, useful in the present compositions and methods can be administered to treat a human patient per se, in the form of a stereoisomer, prodrug, pharmaceutically acceptable salt, hydrate, solvate, acid salt hydrate, N-oxide or isomorphic crystalline form thereof, or in the form of a pharmaceutical composition where the compound is mixed with suitable carriers or excipient(s) in a therapeutically effective amount.

[0070] Pharmaceutically acceptable carriers are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of pharmaceutical compositions for administering tήe antibody compositions (see, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA 18^th ed., 1990, incorporated herein by reference). The pharmaceutical compositions are generally formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.

[0071] The isoflavone compositions of the present invention can be administered by any conventional route known to one of skill in the art, either in a typical pharmaceutical preparation form or as a food supplement.

[0072] These isoflavone compounds can be administered alone, or in combination with the other drug compounds discussed herein, in the form of the water-soluble acid, free base, or as physiologically acceptable salts, including acid addition salts formed with organic and inorganic acids, for example, hydrochlorides, hydrobromides, sulfates, phosphates, citrates, fumarates, and maleates, and cations such as sodium, potassium, and the like. These compounds can be formulated for administration to humans and animals with pharmaceutically acceptable carriers, excipients, and diluents, such as sterile distilled water, Ringer's solution, normal saline, 5% glucose, dextrose, fructose, sucrose, and the like, and mixtures thereof, as is well known in the art. Antimicrobial agents, preservatives, and the like, can also be included. Compositions for oral administration can include coloring and flavoring agents. Additional methods of formulating compounds of the present invention for administration in the methods described herein can be found, for example, in Remington^'' s Pharmaceutical Sciences, Mack Publishing Co., Easton, PA 18^th ed., 1990, incorporated herein by reference).

[0073] For these purposes, the compounds of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir in dosage formulations containing conventional non-toxic pharmaceutically- acceptable carriers, adjuvants and vehicles. In one preferred embodiment, the preferred vehicle is polyethylene glycol (PEG) {see Landauer et al., U.S. Application No. 20040238781; the disclosure of which is herein incorporated by reference for all purposes.). The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraperitoneally, intrathecally, intraventricularly, and other routes acceptable to one of skill in the art for drug delivery.

[0074] In addition, administration can be by a single dose, it can be repeated at intervals or it can be by continuous infusion (described below in further detail). Where continuous infusion is preferred, pump means often will be particularly preferred for administration.

[0075] The teachings presented herein permit the design of therapeutic regimens that can be employed to reduce the undesirable side effects of radiotherapeutic agents, chemotherapeutic agents, and;^' radiation exposure by increasing the dosing of such agents to obtain a higher cancer cure rate or treatment rate for non-cancer conditions, and perhaps include weaker patients in treatment protocols employing such radiotherapeutic agents, chemotherapeutic agents, from which they are currently excluded because they cannot withstand the toxicities associated therewith, In particular, the weight loss induced by these agents. The presently disclosed teachings also permit the design of therapeutic regimens useful for preventing or reducing the undesirable decreased patient survival due, in part, to weight loss.

B. EFFECTIVE DOSAGES

[0076] Pharmaceutical compositions suitable for use with the present invention include compositions wherein the active ingredient is contained in a therapeutically effective amount, i.e., an amount effective to achieve its intended purpose. Of course, the actual amount of active ingredient will depend on, among other things, its intended purpose. For example, when administered to cancer patients in conjunction with radiation or chemotherapy, such compositions will contain an amount of active ingredient effective to, inter alia, ameliorate the radiation-induced or chemotherapy-induced weight loss effects of ionizing radiation or chemotherapeutic agents to individuals. Determination of an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure herein.

[0077] For any compound described herein the therapeutically effective amount can be initially estimated from cell culture assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range of compound, and/or a metabolite thereof, that includes an effective concentration as determined in cell culture.

[0078] Therapeutically effective amounts for use in humans can also be estimated from animal models. For example, a dose for humans can be formulated to achieve a circulating concentration found to be effective in animals, as described herein.

[0079] A therapeutically effective dose can also be estimated from human pharmacokinetic data. While not intending to be bound by any particular theory, it is believed that efficacy is related to a subject's total exposure to an applied dose of administered drug, and/or an metabolite thereof, as determined by measuring the area under the blood concentration- time curve (AUC). Thus, a dose administered according to the methods of the invention that has an AUC of administered compound (and/or an metabolite thereof) within about 50% of the AUC of a dose known to be effective for the indication being treated is expected to be effective. A dose that has an AUC of administered compound (and/or an metabolite thereof) within about 70%, 80% or even 90% or more of the AUC of a known effective dose is preferred. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above, particularly on the blood concentration and duration of administered compound and/or its metabolites is well within the capabilities of the ordinarily skilled artisan.

[0080] For use as a protectant against the radiation-induced and/or chemotherapy- induced weight loss by radiation or chemotherapeutic agents, a circulating concentration of administered compound (and/or and metabolite thereof) of about 2 μM to 100 μM is expected to be effective, with about 5 μM to 50 μM being preferred. Alternatively, or in addition, a tissue concentration of administered compound (and/or a metabolite thereof) of about 4 μM to 700 μM is expected to be effective, with about 20 μM to 35 μM being preferred.

[0081] Generally, dosage levels for nonprimates (i.e., mice or rats) on the order of about 0.1 mg to about 1,000 mg per kilogram of body weight of the isoflavone active ingredient compound are useful in the treatment of the above conditions, with preferred levels of about 1 mg to about 500 mg per kilogram of body weight, and more preferred amounts of 10 mg to about 400 mg per kilogram of body weight. The amount of isoflavone active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.

[0082] It is understood, however, that a specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the severity of the particular disease being treated and form of administration. In some embodiments, dosage levels range from about 1-6000 mg/dose for a 70 kg patient. Although one dose per day can be sufficient, up to 4 doses per day can be given. For example, specific dosage and treatment regimens will depend on factors such as the patient's general health profile, the severity and course of the patient's disease, or disorder, or disposition thereto, and the judgment of the treating physician.

[0083] For a 70 kg patient oral doses for administration of the compounds (assuming average body weight for a human patient is 70 kg) of the invention usually range from about 50 mg/day to 6000 mg/day, commonly from about 100 mg/day to 4000 mg/day, and typically from about 200 mg/day to 3000 mg/day. Stated in terms of patient body weight, usual dosages range from about 0.7 to 86 mg/kg/day, commonly from about 1.4 to 57 mg/kg/day, and typically from about 2.9 to 50 mg/kg/day. Stated in terms of patient body surface areas, usual dosages range from about 23 to 4000 mg/m²/day, commonly from about 45 to 2666 mg/m²/day, and typically from about 90 to 2333 mg/m²/day.

[0084] For subcutaneous administration of the compounds patient dosages usually range from about 50 mg/day to 1500 mg/day, commonly from about 100 mg/day to 1000 mg/day and typically irom about 2UU mg/day to /50 mg/day. Stated in terms of body weight, usual dosages range from 0.7 mg/kg/day to 21 mg/kg/day, commonly from about 1.4 mg/kg/day to 14.2 mg/kg/day and typically from about 2.8 mg/kg/day to 10.7 mg/kg/day. Stated in terms of patient body surface areas, usual doses range from about 22 mg/m² 2/day to 1000 mg/ m²/day, commonly from about 45 mg/m²/day to 666 mg/m.sup.2/day and typically from about 133 mg/m²/day to 500 mg/ m²/day.

[0085] For use as a protectant against radiation-induced weight loss radiation, or as a protectant against chemotherapy-induced weight loss, the dose should be administered far enough in advance of exposure to radiation or chemotherapy to provide effect. For subcutaneous administration, the dose is preferably administered within at least about 3 hours prior to the administration of radiation therapy. Preferably the dose is subcutaneously administered at least about 6 hours prior to the administration of radiation therapy, more preferably at least about 8 hours, and most preferably about 12 to about 24 hours prior to the administration of radiation therapy or chemotherapy.

[0086] Administering isoflavones, e.g., genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, are particularly useful in treating radiation-induced weight loss, hi these cases subcutaneous administration is one preferred embodiment and is very effective at reducing radiation-induced weight loss. In other preferred embodiments, oral administration is preferred.

[0087] Administering isoflavones, e.g., genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, are particularly useful in treating chemotherapy-induced weight loss. In these cases subcutaneous administration is one preferred embodiment and is very effective at reducing chemotherapy-induced weight loss. In other preferred embodiments, oral administration is preferred.

[0088] In one preferred embodiment for radiation-induced and chemotherapy-induced weight loss, the dose is administered as at least two subcutaneous injections, hi other preferred embodiments, the dose is administered as one subcutaneous injection.

[0089] For other modes of administration, dosage amount and interval can be adjusted individually to provide effective plasma and/or tissue levels of the administered compound, and/or an metabolite thereof, according to the pharmacokinetic profiles described herein, as previously described. [υυ^yυj ine actual amount ot composition administered will, of course, be dependent on the subject being treated, the subject's weight, the severity of the affliction, the mode of administration and the judgment of the prescribing physician.

C. ROUTES OF ADMINISTRATION

[0091] Compounds of the invention, for example one or more isoflavones, their glucosides and derivatives, and mixtures thereof, or pharmaceutically acceptable addition salts or hydrates thereof, can be delivered to a patient so as to avoid or reduce undesirable side effects according to the invention using a wide variety of routes or modes of administration. Suitable routes of administration include, but are not limited to, inhalation, transdermal, oral, rectal, transmucosal, intestinal and parenteral administration, including intramuscular, subcutaneous and intravenous injections.

[0092] For any mode of administration, the actual amount of the compounds of the invention, as well as the dosing schedule necessary to treat or prevent radiation-induced or chemotherapy-induced weight loss, will depend, in part, on such factors as the bioavailability of compounds of the invention, the disorder being treated, the desired therapeutic dose, and other factors that will be apparent to those of skill in the art. The actual amount delivered and dosing schedule can be readily determined by those of skill without undue experimentation by monitoring the blood plasma levels of administered compounds of the invention, and adjusting the dosage or dosing schedule as necessary.

[0093] For example, for intravenous administration the advantageous profiles of the compounds of the invention can be obtained by utilizing a significantly slower rate of infusion than is conventionally used, or by using an ambulatory pump.

[0094] Compounds of the invention, for example one or more isoflavones, their glucosides and derivatives, and mixtures thereof, or pharmaceutically acceptable salts and/or hydrates thereof, can be administered singly, in combination with other compounds, and/or in combination with other therapeutic agents, including radiotherapeutic agents and cancer chemotherapeutic agents. The compounds of the invention, e.g., one or more isoflavones, their glucosides and derivatives, and mixtures thereof, can be administered alone or in the form of a pharmaceutical composition, wherein the compound or compounds are admixture with one or more pharmaceutically acceptable carriers, excipients or diluents. Pharmaceutical compositions for use in accordance with the present invention can be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the compounds of the invention into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. IU095J For injection, the agents of the invention can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0096] For oral administration, the compounds of the invention, for example, one or more isoflavones, their glucosides and derivatives, and mixtures thereof can be formulated readily by combining the compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the inventions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained from a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0097] When administered orally, the isoflavone radioprotective and chemoprotective agent(s) should be given in an amount that will result in a therapeutically effective blood serum level equivalent to that achieved by the parenterally administered doses. Such effective oral doses can easily be determined by one of ordinary skill in the art via conventional in vitro or in vivo assays and methods.

[0098] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments can be added to the tablets or dragee coatings for identification or to characterize different combinations of the compounds of the invention, for example, one or more isoflavones, their glucosides and derivatives, and mixtures thereof.

[0099] Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with πiier sucn as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers can be added. All formulations for oral administration should be in dosages suitable for such administration.

[0100] For buccal administration, the compositions can take the form of tablets or lozenges formulated in conventional manner.

[0101] For administration by inhalation, the compounds of the invention, for example, one or more isoflavones, their glucosides and derivatives, and mixtures thereof, according to the present invention, are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoro ethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0102] The compounds of the invention, for example, one or more isoflavones, their glucosides and derivatives, and mixtures thereof, can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. It is preferred that the compounds of the invention be administered by continuous infusion subcutaneously over a period of 15 minutes to 24 hours. Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compounds of the invention can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

[0103] Pharmaceutical formulations for parenteral administration include aqueous solutions of the compounds of the invention in water-soluble form. Additionally, suspensions of the compounds of the invention, for example, one or more isoflavones, their glucosides and derivatives, and mixtures thereof can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension can also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Luiutj Alternatively, me active ingredient can be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. The compounds of the invention, for example, one or more isoflavones, their glucosides and derivatives, and mixtures thereof can also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

[0105] In addition to the formulations described previously, the compounds of the invention can also be formulated as a depot preparation. Such long acting formulations can be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds can be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[0106] The pharmaceutical compositions also can comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

D. FORMULATION

[0107] The compositions of the present invention, e.g., the radioprotectant and chemoprotectant compositions, are often administered as pharmaceutical compositions comprising an active therapeutic agent, i.e., an isoflavone, such as genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, and a variety of other pharmaceutically acceptable components (see, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA 18^th ed., 1990).

[0108] The preferred form depends on the intended mode of administration and therapeutic application. The compositions can also include, depending on the formulation desired, pharmaceutically-acceptable, non-toxic carriers or diluents, which are defined as vehicles commonly used to formulate pharmaceutical compositions for animal or human administration. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological phosphate-buffered saline, Ringer's solutions, dextrose solution, and Hank's solution, hi addition, the pharmaceutical composition or formulation can also include other carriers, adjuvants, or nontoxic, nontherapeutic, nonimmunogenic stabilizers and the like.

[0109] Pharmaceutical compositions can also include large, slowly metabolized macromolecules such as proteins, polysaccharides such as chitosan, polylactic acids, poiygiycυiic awus ana copolymers ^such as latex functionalized Sepharose™, agarose, cellulose, and the like), polymeric amino acids, amino acid copolymers, and lipid aggregates (such as oil droplets or liposomes). Additionally, these carriers can function as immunostimulating agents (i.e., adjuvants).

[0110] For parenteral administration, compositions of the invention can be administered as injectable dosages of a solution or suspension of the substance in a physiologically acceptable diluent with a pharmaceutical carrier that can be a sterile liquid such as water oils, saline, glycerol, or ethanol. Additionally, auxiliary substances, such as wetting or emulsifying agents, surfactants, pH buffering substances and the like can be present in compositions. Other components of pharmaceutical compositions are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, and mineral oil. In general, glycols such as propylene glycol or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

[0111] Typically, compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection can also be prepared. The preparation also can be emulsified or encapsulated in liposomes or micro particles such as polylactide, polyglycolide, or copolymer for enhanced adjuvant effect, as discussed above. Langer, Science 249: 1527, 1990 and Hanes, Advanced Drug Delivery Reviews 28: 97-119, 1997. The agents of this invention can be administered in the form of a depot injection or implant preparation which can be formulated in such a manner as to permit a sustained or pulsatile release of the active ingredient.

[0112] Additional formulations suitable for other modes of administration include oral, intranasal, and pulmonary formulations, suppositories, and transdermal applications.

[0113] For suppositories, binders and carriers include, for example, polyalkylene glycols or triglycerides; such suppositories can be formed from mixtures containing the active ingredient in the range of 0.5% to 10%, preferably l%-2%. Oral formulations include excipients, such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, and magnesium carbonate. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders and contain 10%- 95% of active ingredient, preferably 25%-70%.

Topical application can result in transdermal or intradermal delivery. Topical administration can be facilitated by co-administration of the agent with cholera toxin or detoxified derivatives or subunits thereof or other similar bacterial toxins. Glenn et ah, 1998, nature sy i :^QΌ i . ^υ-aαmmistration can be achieved by using the components as a mixture or as linked molecules obtained by chemical crosslinking or expression as a fusion protein.

[0114] Alternatively, transdermal delivery can be achieved using a skin patch or using transferosomes. Paul et al, 1995, Eur. J. Immunol. 25:3521-24; Cevc et al, 1998, Biochem. Biophys. Acta 1368:201-15.

[0115] The pharmaceutical compositions are generally formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.

E. TOXICITY

[0116] Treatment dosages generally can be titrated to optimize safety and efficacy. Typically, dosage-effect relationships from in vitro initially can provide useful guidance on the proper doses for patient administration. Studies in animal models also generally can be used for guidance regarding effective dosages in accordance with the present invention, hi terms of treatment protocols, it should be appreciated that the dosage to be administered will depend on several factors, including the particular isoflavone that is administered, the route administered, the condition of the particular patient, and the like. Generally speaking, one will desire to administer an amount of the agent that is effective to achieve a serum level commensurate with the concentrations found to be effective in vitro. Determination of these parameters are well within the skill of the art.

[0117] Toxicity of the compounds and methods described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD₅₀ (the dose lethal to 50% of the population) or the LD₁₀₀ (the dose lethal to 100% of the population). The dose ratio between toxic and therapeutic effect is the therapeutic index. The data obtained from these cell culture assays and animal studies can be used in formulating a dosage range that is not toxic for use in human. The dosage of the proteins described herein lies preferably within a range of circulating concentrations that include the effective dose with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See, e.g., Fingl et al, 1975, In: The Pharmacological Basis of Therapeutics, Ch. 1).

F. PHARMACOGENOMICS

[0118] In conjunction with the therapeutic methods of the invention, pharmacogenomics {i.e., the study of the relationship between a subject's genotype and that subject's response to a toreign compound or drug) can be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Thus, a physician or clinician can consider applying knowledge obtained in relevant pharmacogenomics studies in determining whether to administer a radioprotectant agent or chemoprotectant agent which modulates radiation-induced, chemotherapy-induced weight loss (or both radiation-induced and chemotherapy-induced weight loss), as well as tailoring the dosage and/or therapeutic regimen of treatment with an agent which modulates radiation-induced, chemotherapy-induced weight loss (or both radiation-induced and chemotherapy-induced weight loss).

[0119] Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See, for example, Eichelbaum, M. et al, 1996, Clin. Exp. Pharmacol. Physiol. 23:983-985 and Linder, M. W. et al, 1997, Clin. Chem. 43:254-266. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare genetic defects or as naturally-occurring polymorphisms. For example, glucose-6-phosphate aminopeptidase deficiency (G6PD) is a common inherited enzymopathy in which the main clinical complication is haemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.

[0120] One pharmacogenomics approach to identifying genes that predict drug response, known as "a genome-wide association", relies primarily on a high-resolution map of the human genome consisting of already known gene-related markers {e.g. , a "bi-allelic" gene marker map which consists of 60,000-100,000 polymorphic or variable sites on the human genome, each of which has two variants). Such a high-resolution genetic map can be compared to a map of the genome of each of a statistically significant number of patients taking part in a Phase II/III drug trial to identify markers associated with a particular observed drug response or side effect. Alternatively, such a high resolution map can be generated from a combination of some ten million known single nucleotide polymorphisms (SNPs) in the human genome. As used herein, a "SNP" is a common alteration that occurs in a single nucleotide base in a stretch of DNA. For example, a SNP can occur once per every 1000 bases of DNA. A SNP can be involved in a disease process, however, the vast majority can not be disease-associated. Given a genetic map based on the occurrence of such SNPs, individuals can be grouped into genetic categories depending on a particular pattern of SNPs in their individual genome. Ih such a manner, treatment regimens can be tailored to groups of genetically similar individuals, taking into account traits that can be common among such genetically similar individuals.

4. KITS

[0121] Also within the scope of the invention are kits comprising the compositions (e.g., isoflavones such as genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof) of the invention and instructions for use. The kit can further contain a least one additional reagent, or one or more additional isoflavone. Kits typically include a label indicating the intended use of the contents of the kit. The term label includes any writing, or recorded material supplied on or with the kit, or which otherwise accompanies the kit.

[0122] The following Examples of specific embodiments for carrying out the present invention are offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way.

EXEMPLARY EMBODIMENTS

EXAMPLE 1 Materials and Methods

Animals

[0123] Young adult (12 weeks of age) male CD2F1 mice (Charles River Laboratories, NCI-FCRDC, Frederick, MD), still in their growth phase, weighing between 28 and 30 g at the beginning of the experiment were used. All mice were acclimated upon arrival and representative animals were screened for evidence of disease. Mice were housed in a facility accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care, International. Animal rooms were maintained at 21 ± 2 ⁰C with 50% ± 10% humidity on a 12-hr light/dark cycle. Commercial rodent ration (Harlan Teklad Rodent Diet 8604) was freely available as was acidified (pH = 2.5 - 2.8) water to control opportunistic bacterial infections (McPherson 1963). All animal handling procedures were performed in compliance with the guidelines from the National Research Council and were approved by the Institutional Animal Care and Use Committee of the Armed Forces Radiobiology Research Institute (AFRRI).

Isoflavones

[0124] Genistein and daidzein (Figure 1) were obtained from Sigma Chemical Company (St Louis, MO). The isoflavones were solubilized in a mixture of DMSO/PEG-400 in a 15:85 ratio, on the day of the experiment by 20 sec of sonication (Heat Systems-Ultrasonics Inc., Plainview, NY).

Genistein Daidzein

[0125] Figure 1. Chemical structures of genistein and daidzein. Experimental Design

[0126] Animals were randomly assigned to one of six groups, with 12 mice per group. Each mouse received a single subcutaneous injection of vehicle, genistein or daidzein, 24 hrs before sham-irradiation or a sublethal dose of total body irradiation (7 Gy at a dose rate of 0.6 Gy/min cobalt-60 irradiation). The genistein and daidzein were administered at a dose of 200 mg/kg (approximately 5 mg/mouse). This dose was previously demonstrated to be the optimal dose for protection against radiation-induced lethality and determined to be non-toxic (Landauer M.R., Srinivasan V., and Seed T.M., 2003, JAppl Toxicol 23: 379-85). Mice were housed in polycarbonate cages in groups of four and individually marked. To observe radiation-induced weight loss, mice were weighed to the nearest tenth of a gram over a 30-day period using a digital Sartorius balance.

[0127] The dose and time of isoflavone administration used in this experiment was based on previous experiments in our laboratory. Optimal radioprotection was observed after a single subcutaneous 200 mg/kg genistein or daidzein was administered 24 hr prior to irradiation. We previously demonstrated that a single administration of isoflavones at the dose used had no effect on the behavior, body weight, or pathology, thereby demonstrating that that 200 mg/kg is a safe dose in the mouse (Landauer M.R., Srinivasan V., and Seed T.M., 2003, JAppl Toxicol 23: 379-85). Studies in mice from our laboratory also demonstrated radiation protection with repeated oral doses of genistein at 100 mg/kg. hi these protocols, no clinical signs of toxicity were observed. Based on differences in surface area and the more rapid metabolism of isoflavones in mice, a conversion factor of 12 (Freireich, E.J., 1966, Cancer Chemother Rep 50: 219-44) to 20 (Cline J.M. et al., 2004, Toxicol Pathol 32: 91-9) is often used to estimate the human equivalent dose from the mouse dose. Using the more conservative allometric scaling number of 12, doses of 100 and 200 mg/kg genistein in the mouse would be approximately to 8 and 16 mg/kg in humans. Clinical trials have used doses as high as 16-mg/kg body weight of genistein in humans. These doses were reported to be well tolerated with no clinically related drug toxicity (Bergan R.C. et al, 2001, Urology 57: 77-80). Consequently, the experimental doses used in this experiment appear to be clinically feasible.

Radiation Procedure

[0128] Mice were irradiated in a bilateral gamma radiation field at the APRRI cobalt-60 facility (Carter R.E., Verrelli D.M., 1973, AFRRI cobalt whole-body irradiation Technical Report 73-3, Armed Forces Radiobiology Research Institute, Bethesda, MD). The midline tissue dose (MLT) to the animals was 7 Gy, with a dose-rate of 0.6 Gy/min. Control animals were sham irradiated. The dose-rate was established in an acrylic mouse phantom by use of a 0.5-ml tissue- equivalent ionization cnamber (calibration factor traceable to the National Institute of Standards and Technology). The field was uniform to within ± 3%. Dose measurements followed the American Association of Physicists in Medicine protocol (AAPM American Association of Physicists in Medicine 1983). The dose conversion factor (DCF) was 0.96. MLT doses were determined by applying the DCF to dose measurements made free in air (FIA). The DCF was determined by taking the ratio of two measurements. The first measurement was the MLT dose- rate taken at a well-defined point in the midpoint of the phantom. The second measurement was made by removing the phantom and taking an FIA tissue dose-rate measurement at a convenient point in the region occupied earlier by the phantom. Once determined for a particular experimental array, the DCF value can be applied to all future FIA measurements to obtain the MLT dose-rate using the same array.

Results and Discussion

[0129] The average body weight of each individual mouse was compared to its pre- injection baseline weight taken 4 days prior to irradiation. We observed that non-irradiated mice treated with vehicle, genistein, or daidzein exhibited normal weight gain over the 30 days after irradiation. In contrast, a significant reduction in body weight was observed for all groups of irradiated animals. Beginning 1-3 days after irradiation, mice in the genistein/irradiated and daidzein/irradiated groups exhibited significant amelioration of weight loss when compared with animals that did not receive the isoflavones (p < 0.05, t-test). Therefore, a single injection of genistein or daidzein protected mice against radiation-induced weight loss. By 20 days after irradiation, the body weight of the genistein and daidzein irradiated animals had returned to control levels, while untreated irradiated animals had not (Figure 2).

[0130] These results demonstrate that the isoflavones genistein and daidzein can facilitate the recovery of weight loss following a sublethal dose of irradiation. Isoflavones appear to have great utility when used in conjunction with radiotherapy or other situations where exposure to radiation can occur. The protection observed for normal tissue in previous studies as evidenced by prevention of radiation-induced lethality (Landauer M.R., Srinivasan V., and Seed T.M., 2003, JAppl Toxicol 23: 379-85), coupled with a reduction in weight loss following sublethal doses of irradiation, provides a novel use for isoflavones. The mechanism for the amelioration of body weight is unknown at this time, but can be involved in reversing the side effects associated with radiation-induced anorexia, dehydration, and intestinal malabsorption. The recovery of radiation-induced weight loss will likely be facilitated by multiple isoflavone treatments before, during, and/or after irradiation, hi addition, the isoflavones could be administered as a pharmaceutical or dietary supplement by parenteral injection, by suppository, buccal administration, or orally by gelatin capsule/tablet.

EXAMPLE 2

Chemotherapy-Induced Weight Loss Model

[0131] Sickness and loss of body weight is induced in mice with by injection of 10 mg/kg of the chemotherapeutic agent cisplatin. Genistein or other phytoestrogenic isoflavones (e.g daidzein, equol) are administered by injection 24 hr before cisplatin injection, while other groups of mice receive daily oral gavage of genistein for 2 weeks before, 2 weeks after, or 2 weeks before and 2 weeks after chemotherapy. AU administration schedules are effective in reducing cisplatin-induced weight loss, with the order of most effective to least effective being: 1) injection, gavage: 2) before and after cisplatin, 3) before only, and 4) after only. Similar effects are observed when bleomycin is used as an alternative chemotherapeutic agent, indicating that the effect is not cisplatin specific. Genistein treatments also ameliorate weight loss in radiation alone (7 Gy) or chemotherapy/radiation combination experiments when mice are injected with a single dose of cisplatin (7.5 mg/kg) followed by irradiation (6.5 Gy) one hour after chemotherapy administration. In addition, while recovery from the loss body weight induced by chemotherapy or chemo/radiation therapy are evidenced, genistein does not protect tumors from chemotherapy, radiotherapy or the combination of these treatment regimens. Genistein enhances the effects of chemotherapeutic agents and radiation therapy both alone and in combination, as evidenced by a reduction of in tumor size.

EXAMPLE 3

[0132] Isoflavones selected from the group consisting of genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, are administered to patients with malignant brain tumors at time periods starting one month, one week, 5 days, 2 days, and 1 day prior to the beginning of chemotherapy and continued concomitantly with the chemotherapy drugs regimen (or radiotherapy drugs regimen). The isoflavones are administered as food supplements and as pharmaceutical preparations with the daily estimated preferred dose of genistein varying from about 0.1 mg/kg to about 1,000 mg/kg of body weight per day, with a preferred dose from about 1 mg/kg to about 500 mg/kg of body weight per day, with the most preferred dose from about 10 mg/kg to about 400 mg/kg of body weight per day.

[0133] The use of the isoflavones such as genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and denvatives, and mixtures thereof, act to ameliorate weight loss and facilitates weight gain during recovery from chemotherapy, radiation therapy (or both chemotherapy and radiation therapy), in the treatment regimens in patients with brain tumors. Prolonged or chronic administration of isoflavones, particularly genistein, in a pharmaceutical formulation or in a form of food supplement, prevents the occurrence or delays development of weight loss condition through treatment with chemotherapeutic agents, radiotherapeutic agents (or both chemotherapeutic agents and radiotherapeutic agents).

EXAMPLE 4

[0134] Isoflavones selected from the group consisting of genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof, are administered to group of patients with prostate cancer. Administration of the isoflavones is commenced at different periods starting at one month, one week, 5 days, 2 days and 1 day prior to the beginning of chemotherapy and continued concomitantly with the chemotherapy drags regimen, radiotherapy drugs regimen, or both chemotherapy drugs regimen and radiotherapy drugs regimen. The isoflavones are administered as food supplements and as pharmaceutical preparations with the daily estimated preferred dose of genistein is varied from about 0.1 mg/kg to about 1,000 mg/kg of body weight per day, with a preferred dose from about 1 mg/kg to about 500 mg/kg of body weight per day, with the most preferred dose from about 10 mg/kg to about 400 mg/kg of body weight per day.

[0135] Patients with this tumor receive chemotherapy agents, radiotherapy agents, or both, that induce a weight loss condition through treatment with these agents. Administering genistein either in a form of pharmaceutical agent or as a food supplement, prevents the occurrence or delays development of a weight loss condition induced by treatment with chemotherapeutic agents, radiotherapeutic agents (or both chemotherapeutic agents and radiotherapeutic agents).

EXAMPLE 5

[0136] Isoflavones selected from the group consisting of genistein, genistin, daidzein, daidzin, glycditein, glycitin, biochannin A, formononetic, O-desmethylangolensin, and equol, their glucosids and derivatives, and mixtures thereof, are administered to a group of patients prior to, during, immediately following, or combinations thereof, commencement of the administration of immunosuppressant drags (chemodrags) prior to transplantation of organs, including kidney, heart, liver lung, skin and the like. Administration of the isoflavones is commenced at different periods starting at one month, one week, 5 days, 2 days and 1 day prior to tϊie beginning oi me administration of the chemodrugs, during the chemodrug regimen, or for different periods starting at one month, one week, 5 days, 2 days and 1 day prior to the beginning of the administration of the chemodrug. The chemodrugs can include cyclosporin and methotrexate. The isoflavones are administered as food supplements and as pharmaceutical preparations with the daily estimated preferred dose of genistein is varied from about 0.1 mg/kg to about 1,000 mg/kg of body weight per day, with a preferred dose from about lmg/kg to about 500 mg/kg of body weight per day, with the most preferred dose from about 10 mg/kg to about 400 mg/kg of body weight per day.

[0137] The isoflavones ameliorate weight loss in patients brought about by treatment with chemodrugs during the chemodrug regimen described above.

EXAMPLE 6

[0138] The procedures of Examples 4 and 5 are followed in conjunction with treatment of an inflammatory bowel disease, including Crohn's disease, by administration of a chemotherapeutic agent such as azathioprine, which after undergoing cellular metabolism transforms into active but toxic 6-thioquanine (6-TG).

EXAMPLE 7

[0139] The procedures of Examples 4 and 5 are followed in conjunction with treatment of temporal arthritis by administration of a chemotherapeutic agent such as azathioprine, which after undergoing cellular metabolism transforms into active but toxic 6-thioquanine (6-TG).

[0140] AU publications and patent applications cited in this specification are herein incorporated by reference in their entirety for all purposes as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference for all purposes.

[0141] Each recited range includes all combinations and sub-combinations of ranges, as well as specific numerals contained therein.

[0142] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to one of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Claims

What is Claimed:

1. A method of treating or preventing radiation-induced weight loss in a subject exposed to radiation, the method comprising administering to the subject a therapeutically effective amount of an isoflavone.

2. The method of claim 1, wherein the radiation is an acute sub-lethal dose of ionizing radiation, a chronic low-dose of ionizing radiation, an acute sub-lethal dose of non-ionizing radiation, or a chronic low-dose of non-ionizing radiation.

3. The method of claim 2, wherein the radiation is selected from the group consisting of diagnostic X-rays, radiation therapy in cancer treatment, CAT-scans, mammograms, radionuclide scans, interventional radiological procedures under CT or fluoroscopy guidance, tissue-incorporated radionuclides from ingestion of contaminated food or water, and uncontrolled exposure to ionizing radiation from nuclear weapons, radioactive spills, and/or cosmic radiation.

4. The method of claim 1, wherein the isoflavone is selected from the group consisting of genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O- desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof.

5. The method of claim 1, wherein the isoflavone is administered orally, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, or rectally.

6. The method of claim 5, where the isoflavone is administered orally in the form of a capsule, a tablet, an inhaler, a troche, or a food supplement in the form of a food or beverage.

7. The method of claim 1, wherein the isoflavone is administered chronically.

8. The method of claim 1, wherein the isoflavone is administered within 2 weeks prior to exposure to radiation, during radiation exposure, and/or within 2 weeks following radiation exposure.

9. The method of claim 8, wherein the isoflavone is administered within 4 days prior to radiation exposure, during radiation exposure, and/or within 4 days following radiation exposure.

10. A method lor treating or preventing radiation-induced weight loss in a subject exposed to a dose of radiation, the method comprising administering to the subject before, during and/or after the dose of radiation a therapeutically effective amount of a compound of the formula:

wherein Rl, R2, R3, R4 are independently selected from the group consisting of hydrogen, hydroxyl and alkoxy.

11. The method for treating or preventing radiation-induced weight loss in a subject exposed to a dose of radiation as defined in claim 10 wherein the compound is genistein.

12. A method for treating or preventing radiation-induced weight loss in a subject exposed to a dose of radiation as defined in claim 2, wherein the compound is administered to the subject during the time period of approximately 4 days prior to radiation exposure to approximately 4 days subsequent to the lethal dose of irradiation.

13. A method for preventing or reducing weight loss in an individual undergoing treatment with a therapeutic agent, comprising administering to the patient an effective amount of an isoflavone.

14. The method of claim 13, wherein the agent is a chemotherapeutic agent or a radiotherapeutic agent.

15. The method of claim 13, wherein the therapeutic agent is a chemotherapeutic agent and a radiotherapeutic agent.

16. The method of claim 13, wherein the isoflavone is administered simultaneously with administration of the therapeutic agent.

17. The method of claim 13, wherein the isoflavone is administered prior to administration of the therapeutic agent.

18. The method of claim 13, wherein the isoflavone is administered subsequent to administration of the therapeutic agent.

19. The method of claim 13, wherein the effective amount of the isoflavone is administered to the patient in a time period from about one month before administration of the chemotherapeutic agent to about one month after administration of the chemotherapeutic agent.

20. The method of claim 13, wherein the effective amount of the isoflavone is administered to the patient in a time period from about 7 days before administration of the chemotherapeutic agent to about 7 days after administration of the chemotherapeutic agent.

21. The method of claim 13, wherein the effective amount of the isoflavone is administered to the patient in a time period from about 4 days before administration of the chemotherapeutic agent to about 4 days after administration of the chemotherapeutic agent.

22. The method of claim 13, wherein the isoflavone is genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, 0-desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof.

23. The method of claim 13, wherein the isoflavone is genistein.

24. The method of claim 13, wherein the isoflavone has the formula:

wherein Rl, R2, R3 and R4 are independently selected from the group consisting of hydrogen, hydroxyl and alkoxy.

25. The method ot claim 15, wnerem the effective amount of the isoflavone is in the range of from about 0.1 mg/kg hody weight to about 1000 mg/kg body weight.

26. The method of claim 13, wherein the effective amount of the isoflavone is in the range of from about 1 mg/kg body weight to about 500 mg/kg body weight.

27. The method of claim 13, wherein the effective amount of the isoflavone is in the range of from about 20 mg/kg body weight to about 400 mg/kg body weight.

28. The method of claim 13, wherein the isoflavone is administered orally.

29. The method of claim 28, wherein the isoflavone is administered in an effective amount in the range of from about 1 mg/kg body weight to about 50 mg/kg body weight.

30. The method of claim 28, wherein the isoflavone is administered in an effective amount in the range of from about 2 mg/kg body weight to about 25 mg/kg body weight.

31. The method of claim 28, wherein the isoflavone is administered in an effective amount in the range of from about 4 mg/kg body weight to about 12 mg/kg body weight.

32. The method of claim 28, wherein the isoflavone is administered once per day.

33. The method of claim 28, wherein the isoflavone is administered twice per day.

34. The method of claim 28, wherein the isoflavone is administered three or more times per day.

35. The method of claim 13 , wherein the isoflavone is administered parenterally.

36. The method of claim 35, wherein the parenteral administration is by slow intravenous infusion.

37. The method of claim 35, wherein the isoflavone is administered parenterally by injection.

38. The method of claim 37, wherein the isoflavone is administered in an effective amount in the range of from about 0.1 mg/kg body weight to about 1000 mg/kg body weight.

39. The method of claim 37, wήerem the isoflavone is administered in an effective amount in the range of from about 1 mg/kg body weight to about 500 mg/kg body weight.

40. The method of claim 37, wherein the isoflavone is administered in an effective amount in the range of from about 2 mg/kg body weight to about 10 mg/kg body weight.

41. The method of claim 28, wherein the isoflavone is orally administered beginning about 90 days prior to exposure to a sublethal dose of radiation.

42. The method of claim 28, wherein the isoflavone is orally administered beginning about 7 days prior to exposure to a sublethal dose of radiation.

43. The method of claim 28, wherein the isoflavone is orally administered beginning about 90 days prior to and about 90 days after exposure to a sublethal dose of radiation.

44. The method of claim 28, wherein the isoflavone is orally administered beginning about 30 days prior to and about 30 days after exposure to a sublethal dose of radiation.

45. The method of claim 28, wherein the isoflavone is administered 14 days prior to and about 14 days after exposure to a sublethal dose of radiation.

46. The method of claim 28, wherein the isoflavone is administered 7 days prior to and about 7 days after exposure to a sublethal dose of radiation.

47. The method of claim 28, wherein the isoflavone is administered a 1 day prior to and about 1 day after exposure to a sublethal dose of radiation.

48. The method of claim 28, wherein the isoflavone is administered as a food supplement.

49. The method of claim 14, wherein the chemotherapeutic agent is administered for treatment of cancer.

50. The method of claim 46, wherein the cancer is breast cancer, lung cancer (small and/or non-small cell), acute lymphocytic leukemia, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, chronic myelocytic leukemia, hairy cell leukemia, ovary cancer, ovary (germ cell) cancer, melanoma, skin cancer, cervix cancer, ACTH-producing tumors, cancer of the adrenal cortex, bladder cancer, bram cancer, colorectal cancer, cutaneous T-cell lymphoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, gallbladder cancer, head & neck cancer, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, liver cancer, malignant peritoneal effusion, malignant pleural effusion, mesothelioma, multiple myeloma, neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma, pancreatic cancer, penis cancer, prostate cancer, retinoblastoma, soft-tissue sarcoma, squamous cell carcinomas, stomach cancer, testicular cancer, thyroid cancer, trophoblastic neoplasms, cancer of the uterus, vaginal cancer, cancer of the vulva, and Wihn's tumor.

51. A method of preventing or reducing weight loss in a mammal, the method comprising: selecting a mammal having or at risk of having a condition characterized by weight loss; and administering to the mammal a composition comprising at least one isoflavonoid, wherein the administration prevents or reduces weight loss in the mammal.

52. The method of claim 51 , wherein the weight loss is induced by radiation.

53. The method of claim 51 , wherein the weight loss is induced by chemotherapy.

54. The method of claim 51 , wherein the isoflavonoid is selected from the group consisting of genistein, genistin, daidzein, daidzin, glycitein, glycitin, biochannin A, formononetin, O- desmethylangolensin, and equol, their glucosides and derivatives, and mixtures thereof.

55. The method of claim 51, wherein the isoflavonoid is from a source selected from the group consisting of soy, soy products and clover.