IE20060312A1 - Radiation protection and immune recovery and improvement methods - Google Patents

Abstract

The invention relates to the use of compounds and compositions to ameliorate or treat conditions such as neutropenia, thrombocytopenia and anemia and other blood cell deficiencies or other exemplified conditions such as Reactive oxidative stress due to radiation exposure. Exemplary cytokinin compounds that can be used included N6 isopentyl adenosine, N6 benzyl adenosine or its analogues whose structure is herein designated as Formula 1 compounds alone or in combination with Oltipraz, or its analogues whose structure is herein designated as Formula 2 compounds. Compounds of Formula 1 are themselves radio protective and cause remission of neutropenia, thrombocytopenia and anemia as demonstrated in patient studies. Formula 2 compounds have demonstrated in irradiated mouse studies their ability to prevent neutropenia, thrombocytopenia and to counteract and neutralize the oxidative damage radiation generates in living tissue. Formula 1 and formula 2 compounds administered to patients per the terms of this invention will bring to radioprotection a unique synergy of benefits.

Description

The invention relates to methods to use compounds, such as isopentyl adenosine, and or Oltipraz, in combination or alone to treat, ameliorate or prevent neutropenia or related conditions or other pathological or disease conditions or their symptom^ BACKGROUND Immune dysregulation can be a component of many pathological diseases or conditions.

Such dysregulation may be a factor that favors the establishment, maintenance or progression of these diseases or conditions. Deficient immune responses or immune suppression can enhance a mammal's susceptibility to infection or to the development of cancer. Conversely, excessive or inappropriate immune responses can play a role in the establishment or progression of unwanted inflammation or autoimmune conditions. It would thus be advantageous to utilize agents that can modulate immune responses and to at least partially reverse-immune dysregulation when such dysregulation is a component of a given pathological condition. Some agents are known that can ameliorate some aspects of immune dysregulation, but typically such agents have their own unwanted effects on either the host's immune system or other organs or tissues. Agents such as glucocorticoid steroids have been used to reduce unwanted inflammation in a number of clinical conditions, but such compounds often have serious limitations, such as inducing immune suppression or causing unwanted mood changes.

Human and mammalian immune responses to infections or other conditions are often characterized by responses mediated by different immune effector cell populations. In some situations, helper T cells designated Thl in the murine system, facilitate immune effector functions that are typically dominated by cell-mediated responses. In other cases, helper T cells designated Th2 cells facilitate immune effector functions that are typically dominated by humoral responses. A vigorous Thl response is usually desirable to help clear infections or to slow the progression of an infection. When a subject's immune response is biased to, or dominated by, a Th2-type response, the cytokines associated with the Th2 response tend to suppress the immune system's capacity to mount a vigorous Thi response at the same time. The converse is also generally true. When mammalian immune responses begin to result in an increasing Thi response, Th2 responses tend to weaken. Insufficient Thi responses may be associated with progression of some infections or other conditions, see, e.g., M. Clerici and G. M. Shearer, Immunol. Today 14; 107-111,1993; M. Clerici and G. M. Shearer, Immunol. Today 15 15:575-581, 1994.

Some proteins such as interleukin-6 (IL-6), erythropoietin (EPO) and thrombopoietin (TPO) have been examined for their capacity to enhance various aspects hematopoiesis, e.g., Hematology-Basic Principles and Practice, 3.sup.rd edition, R. Hoffman, E. J. Benz Jr. et al., editors, Churchill Livingstone, New York, 2000 (see, e.g., Chapter 14 at pages 154202), O. J. Borge et al., Blood 1996 88:2859-2870, M. Cremer et al., Ann. Hematol. 1999 78:401-407, Y. Sasaki et al., Blood 1999 94:1952-1960, U.S. Pat. 5,879,673. Recombinant IL-6 was shown in model systems to affect platelet counts in peripheral circulation, e.g., Stahl et al., Blood 1991 78:1467-1475, although significanttoxicities are associated with its administration to humans, e.g., Andus et al., FEBS Lett. 1987 221:18, J. Gauldie et al., P.N.A.S. U.S.A. 1987 84:7251-7255, T. Geiger et al., Eur. J. Immunol. 1988 18:717-721. The IL-6 molecule has been described in detail, e.g., publication no. WO 88/00206. Administration of proteins is typically expensive, given factors such as the complexity of producing pharmaceutical grade material.

There is a current need for cost-effective pharmaceutical agents and treatment methods for treating various immune dysregulation conditions. The invention provides non steroid compounds that can be used in such treatments to treat or ameliorate one or more aspects of immune dysregulation conditions. Such agents can be used to treat autoimmune or inflammation conditions, immune suppression conditions, infections, blood cell deficiencies and other described conditions. The agents and methods are useful to ameliorate these conditions or one or more symptoms associated with any of these conditions. The use of these agents can be combined with one or more conventional treatments for these disorders.

DESCRIPTION OF THE INVENTION Summary of invention embodiments. In principal embodiments the invention provides therapeutic treatment methods.

The methods include a method to prevent, treat, ameliorate or slow the progression of blood cell deficiency disorder, neutropenia or thrombocytopenia in a subject, or to treat a symptom of, blood cell deficiency disorder, neutropenia or thrombocytopenia, comprising administering to a subject, or delivering to the subject's tissues, an effective amount of a Formula 1 compound, alone or in combination with a Formula 2 compound, Formula 1 compounds having the structure : FORMULA 1 COMPOUNDS are listed at the end of the text Other embodiments include (1) certain new fonnula 1 compounds, which are new chemical entities, (2) compositions that comprise a fonnula 1 compound and another compound or an excipient, (3) formulations that comprise a formula 1 compound and 1, 2, 3, 4, 5, 6 or more excipients. The formulations can be designed for human pharmaceutical use or they can be suitable for veterinary use. Therapeutic use embodiments include (1) use of a formula 1 compound for the preparation of a medicament and (2) use of a formula 1 compound for the preparation of a medicament for the prophylaxis or treatment of a condition or symptom disclosed herein.

The methods include a method to prevent, treat, ameliorate or slow the progression of blood cell deficency disorder, neutropenia or thrombocytopenia in a subject, or to treat a symptom of blood cell deficency disorder, neutropenia or thrombocytopenia, comprising administering to a subject, or delivering to the subject's tissues, an effective amount of a fonnula 2 compound having the structure FORMULA 2 COMPOUNDS are listed at the end of the text J 12 In another embodiment, the invention provides a method to determine if a mammal has a bone marrow or blood cell disorder or related disorder or the propensity to develop such a disorder, comprising: (a) obtaining a circulatory fluid sample from the mammal; (b) splitting the circulatory fluid sample into two, three or more suitable aliquots; (c) determining the hydrogen peroxide level in a first aliquot; (d) contacting a second aliquot with a sufficient amount of a one, two or more D-amino acids; (e) incubating the second aliquot for sufficient time and under conditions suitable to allow detectable metabolism of the one, two or more D-amino acids to determine the level of glutathione in the second aliquot; (f) determining the glutathione level of second first aliquot; and (g) comparing the glutathione level obtained from step (c) and step (f) and the, whereby a lower level of glutathione level indicates the presence of a bone marrow disorder or related disorder or the propensity to develop such a disorder.

Related embodiments provide a method to make the Formula 2 compound ,Oltipraz comprising (1) contacting pyrazine-2-carboxylic acid with methanol in the presence of an acid to form methyl-pyrazine-2-carboxylate; (2) condensing the methyl-pyrazine-2carboxylate with methyl propionate in the presence of a base to form methyl-2-methyl-3(pyrazin-2-yl)-3-oxopropionate; and (3) treating said methyl-2-methyl-3-(pyrazin-2-yl)-3oxopropionate with phosphorus pentasulfide to form Oltipraz.

Other embodiments include (1) certain new formula 2 compounds, which are new chemical entities, (2) compositions tliat comprise a Formula 1 and Formula 2 compound and another compound or an excipient, (3) formulations that comprise a Formula 1 compound an/or a Formula 2 and 1, 2, 3, 4, 5, 6 or more excipients. The formulations can be designed for human pharmaceutical use or they can be suitable for veterinary use. Therapeutic use embodiments include (1) use of formula 2 compounds for the preparation of a medicament -< and (2) use of a formula 2 compounds for the preparation of a medicament for the prophylaxis or treatment of a condition or symptom disclosed herein.

The methods include a method to prevent, treat, ameliorate or slow the progression of blood cell deficency disorder, neutropenia or thrombocytopoenia in a subject, or to treat a symptom of blood cell deficency disorder, neutropenia or thrombocytopenia, comprising administering to a subject, or delivering to the subject's tissues, an effective amount of a fonnula 2 compound having the structure or a metabolic precursor or a metabolite and oxides, derivatives and metabolites thereof, wherein the structure is DNA or RNA multi linked polymer of any Formula 1 compound, an example of which is set within the structures outlined in the definition of Fonnula 1 compounds.

Other phannaceutical compositions embodiments include (1) a composition of a Fonnula 1 and a Formula 2 compound (2) a composition of a Formula 1 compound (6) a composition of a Fonnula 2 compound. The compositions can be designed for human phannaceutical use or they can be suitable for veterinary use. Therapeutic use pharmaceutical compositions embodiments include (1) use of any composition compound for the preparation of a medicament and (2) use of any composition compound for the preparation of a medicament for the prophylaxis or treatment of a condition or symptom disclosed herein.

Other embodiments are as described elsewhere in the specification including the claims.

Definitions. As used herein and unless otherwise stated or implied by context, terms that are used herein have the meanings defined below. Unless otherwise contraindicated or implied, e.g., by including mutually exclusive elements or options, in these definitions and throughout this specification, the terms a and an mean one or more and the term or means and/or.

An invention formulation, formulation, pharmaceutical formulation or the like means a composition that one can administer to a subject, e.g., human, mammal or other animal, without further manipulations that change the ingredients or the ingredient proportions that are present. Formulations will typically comprise a single fonnula 1 compound and one or more excipients. Fonnulations are suitable for human or veterinary applications and would typically have expected characteristics for the formulation, e.g., parenteral fonnulations for y60 3 12 ιε 9 21 22 human use would usually be sterile and stored in a suitable closed container.

When referring to mixtures that contain a formula 1 compound and/or a Formula 2 compound, an invention composition, composition or the like means a composition, that is a formulation or that can be an intermediate one can use, e.g., to make a formulation or a formula 1 compound and/or formula 2 compound.. Compositions also include other types of mixtures, e.g., (1) reagents for assays or cells that are optionally contacted with a formula 1 compound or mixtures of Formula 1 and Formula 2 compounds with other compounds and (2) compounds used to a formula 1 compound or by-products of formula 1 compound synthesis, a formula 2 compound or by-products of formula 2 compound synthesis.

Invention compositions include compositions where further processing may be required before it is a formulation, e.g., mixing or addition of a desired amount of an excipient.

Phrases such as administration of a compound of formula 1, treatment with a formula 1 compound, use of a formula 1 compound or similar terms mean that the compound(s) is administered to, contacted with or delivered to, the subject or to the subject's cells or tissues in vitro or in vivo by one or more suitable methods, e.g., in vivo delivery can be by an oral, topical, subcutaneous, parenteral, buccal or sublingual route.

Phrases such as administration of a compound of formula 2, treatment with a formula 2 compound, use of a fonnula 2 compound or similar terms mean that the compound(s) is administered to, contacted with or delivered to, the subject or to the subject's cells or tissues in vitro or in vivo by one or more suitable methods, e.g., in vivo delivery can be by an oral, topical, subcutaneous, parenteral, buccal or sublingual route.

Expressions such as a Formula 1,Formula 2 compound(s), a formula 1 compound a formula 2 compound and the like mean invention compositions or formulations where one or more than one formula 1, and/or Formula 2 compound(s) is present, e.g., in a composition, or is used in the disclosed method, typically 1,2, 3 or 4, usually 1. Any reference to a formula 1 compound, one or more compounds of formula 1 or the like 060312 means that the formula 1 compound can have a different chemical formula structure or orientation or any other structure disclosed herein that is within the definition of formula 1 compounds.

Any reference to a formula 2 compound, one or more compounds of formula 2 or the like means that the formula 2 compound can have a different chemical formula structure or orientation or any other structure disclosed herein that is within the definition of formula 2 compounds.

The phrase formula 3 compound or formula 3 compound(s) is sometimes abbreviated as F3C or F3C(s) and formula 3 compounds may be abbreviated as F3Cs is a reference to any formula or substance to be administered per the embodiments of this inventions which shall comprise any single component or combination of the compounds described as Formula 1 and/or Formula 2 compounds within this patent specification. Note the individual components of any F3C formula may be not less than one or greater than thirty formula structures belonging to any combination of Formula 1, and/or Formula 2 compounds. Any reference to an F3C salt shall mean a salt or other similar chemical moiety of any single or multiple components of F3C. F3C formula compound components can be administer independently, simultaneously or concurrently or at various time intervals. An F3C compound need only have one component of either a Formula 1 or a Formula 2 compound.

Reference to subject matter as disclosed herein such as a therapeutic treatment or agent as disclosed herein, a dosing protocol as disclosed herein or a clinical condition or symptom as disclosed herein or the like means a treatment, agent, protocol, condition, symptom or the like that is described herein or in any reference that is cited herein.

An excipient, carrier, pharmaceutically acceptable excipient, pharmaceutically acceptable carrier or similar terms mean one or more component(s) or ingredient(s) that is acceptable in the sense of being compatible with the other ingredients of invention compositions or formulations and not overly deleterious to the patient, animal, tissues or cells to which the F3C, composition or formulation is to be administered.

IE 06 0 3 12 A 'subject means a human or animal. Usually the animal is a mammal or vertebrate such as a primate, rodent, lagomorphs, domestic animal or game animal. Primates include chimpanzees, cynomologous monkeys, spider monkeys, and macaques, e.g., Rhesus or Pan. Rodents and lagomorphs include mice, rats, woodchucks, ferrets, rabbits and hamsters. Domestic and game animals include cows, horses, pigs, sheep, camel, deer, bison, buffalo, mink, felines, e.g., domestic cat, canines, e.g., dog, wolf and fox, avian species, e.g., chicken, turkey, emus and ostrich, and fish, e.g., trout, catfish and salmon. Subject includes any subset of the foregoing, e.g., all of the above, but excluding one or more groups or species such as humans, primates or rodents. Other subsets of subjects include subjects of a given species or group of species of varying ages, e.g., young humans, e.g., about 1 week of age to about 9 years of age, adolescent humans, e.g., about 10-19 years of age, adult humans, e.g., about 20-100 years of age, and mature adult or elderly humans, e.g., at least about 55 years of age, at least about 60 years of age, at least about 65 years of age or a range of ages such as about 60-100 years of age. Thus, as used herein, prevention or treatment of a disease, condition or symptom may include or exclude any subset of subjects that are grouped by age.

The terms effective amount, effective dose or the like with reference to a F3C(s) mean an amount of the F3C(s) that is sufficient to elicit a desired response, e.g., detectable restoration of normal immune responsiveness in an immunodeficiency subject to which it is administered, e.g., a human, or to detectable modulation or amelioration of an immune or cellular parameter or a clinical condition or symptom. An effective amount, e.g., for human therapeutic use, may be a single dose or two or more sub doses of a F3C administered in one day, or it may be administered as multiple doses over a period of time, e.g., over 1. 2, 3,4 or about 7 days to about 1 year.

Terms such as use, treat, treatment, address or the like in the context of using the F3Cs in the treatment methods or other methods disclosed herein mean that a F3C is administered to a subject, delivered to the subject's tissues or contacted with tissues, cells or IE ΰ cell free systems in vivo or in vitro, e.g., as described herein or a reference cited herein. Typically such use or treatment results in, e.g., (1) detectable improvement in or amelioration of the condition or symptom being treated, (2) detectable modulation in the activity, level or numbers of a relevant biomolecule, therapeutic immune cell population or a pathological cell population, (3) slowing of the progression of a condition or delaying its onset, or reduction of the severity of a symptom(s) of the condition or (4) another detectable response as described herein. Any such amelioration may be transient, e.g., lasting for at least a few, e.g., about 1 to 24, hours or days, e.g., about 1,2, 3,4, 5, 6 or 7 days, or amelioration may be prolonged, e.g., lasting about 8, 9, 10,11,12, 13, 14,15, 16, 17, 18, 19, 20,21,22, 24,26,28, 35, 42,49, 56 to about 60 days or more, or amelioration may be pennanent. A treatment may slow the progression of a disease or symptom or it may reduce the severity thereof, e.g., onset of a disease or a symptom may be delayed in at least some subjects for about 1-24 hours, about 2-10 days, about 2-30 days or for about 1-5 years compared to subjects who are not treated with sufficient amounts of the F3C. Thus, a F3C use or treatment typically results in detectable modulation in a relevant immune parameter such as modulation of the level, activity or relative amount of a target effectors or suppressor immune cell population, interleukin, cytokine, chemokine, immunoglobulin compared to a suitable control, e.g., untreated. A F3C treatment can also cause modulation of the level or activity of a relevant transcription factor, enzyme, cell biological activity or level or activity of the etiological agent of the disease such as a pathogen, tumor cell or auto reactive immune cell subset. A treatment with a F3C may be used to delay or prevent the onset of a disease, symptom or complication or to ameliorate or slow the progression of a preexisting disease, condition, symptom or complication, or to facilitate elimination of a disease, condition, symptom or complication.

Ameliorate, amelioration, improvement or the like means a detectable improvement or ·< a detectable change consistent with improvement occurs in a subject or in at least a minority of subjects, e.g., in at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 100% or in a range about between any two of these values. Such improvement or change may be observed in treated subjects as compared to IE 0 5 o J J 2 subjects not treated with a F3C, where the untreated subjects have, or are subject to developing, the same or similar disease, condition, symptom or the like. Amelioration of a disease, condition, symptom or assay parameter may be determined subjectively or objectively, e.g., self assessment by a subject(s), by a clinician's assessment or by conducting an appropriate assay or measurement, including, e.g., a quality of life assessment, a slowed progression of a disease(s) or condition(s),' a reduced severity of a disease(s) or condition(s), or a suitable assay(s) for the level or activity(ies) of a biomolecule(s), cell(s) or by detection of cell migration within a subject. Amelioration may be transient, prolonged or permanent or it may be variable at relevant times during or after a F3C is administered to a subject or is used in an assay or other method described herein or a cited reference, e.g., within about 1 hour of the administration or use of a F3C to about 3, 6, 9 months or more after a subject(s) has received a F3C.

The modulation of, e.g., a symptom, level or biological activity of a molecule, replication of a pathogen, cellular response, cellular activity or the like, means that the cell, level or activity, or the like is detectably increased or decreased. Such increase or decrease may be observed in treated subjects as compared to subjects not treated with a F3C, where the untreated subjects have, or are subject to developing, the same or similar disease, condition, symptom or the like. Such increases or decreases may be at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 1000% or more or about within any range about between any two of these values. Modulation may be determined subjectively or objectively, e.g., by the subject’s self assessment, by a clinician's assessment or by conducting an appropriate assay or measurement, including, e.g., quality of life assessments or suitable assays for the level or activity of molecules, cells or cell migration within a subject. Modulation may be transient, prolonged or permanent or it may be variable at relevant times during or after a F3C is administered to a subject or is used in an assay or other method described herein or a cited reference, e.g., within about 1 hour of the administration or use of a F3C to about 3, 6, 9 months or more after a subject(s) has received a F3C. °60 3 ig Terms such as antigen, immunogenic, antigenic fragment or the like mean a molecule that comprises one or more epitopes that are capable of stimulating a subject's immune system to make, e.g., a secretory, humoral or cellular antigen-specific response against the antigen, immunogen or fragment. Antigenic fragments are synthetic or natural derivatives of natural or intact antigens or immunogens that retain at least a detectable capacity, e.g., at least about 10%, 20%, 30%, 40%, 50% or more of the native antigen's antigenic capacity, to stimulate a subject’s immune system in a desired manner.

Vaccine composition, vaccine or similar terms mean an agent suitable for stimulating a subject's immune system to ameliorate a current condition or to protect against or to reduce present or future harm or infection, e.g., reduced tumor cell proliferation or survival, reduced pathogen replication or spread in a subject or a detectably reduced unwanted symptom(s) associated with a condition. Vaccines may modulate, typically detectably enhance, humoral, cell mediated or innate immune responses.

Immunization means the process of inducing a detectable and continuing moderate or high level of antibody or cellular immune response that is directed against an antigen to which the subject has been exposed. Such responses are typically detectably maintained for at least about 3-48 months or more.

At various locations in the present disclosure, e.g., in any disclosed embodiments or in the claims, reference is made to compounds, compositions, formulations, or methods that comprise one or more specified components, elements or steps. Invention embodiments also specifically include those compounds, compositions, formulations or methods that are or that consist of or that consist essentially of those specified components, elements or steps. The terms comprising, consist of' and consist essentially of’ have their normally ·» accepted meanings under U.S. patent law. For example, disclosed compositions or methods that comprise a component or step are open and they include or read on those compositions or methods plus an additional components) or step(s). Similarly, disclosed compositions or methods that consist of' a component or step are closed and they would °60ji2 not include or read on those compositions or methods having appreciable amounts of an additional component(s) or an additional step(s).

Nucleoside includes 3TC, AZT, D4T, ddl, ddC, G, A, U, C, T, dG, dA, dT and dC.

Polymer includes biocompatible organic polymers, e.g., polyethylene-glycols (PEGs) and polyhydroxyalkyl polymers. PEG means an ethylene glycol polymer that contains about 2, 3,4, 5,6,7, 8, 9,10,11,12 or more linked monomers, e.g., about 50-1000 linked monomers. Average molecular weights typically are about 80, 100, 200, 300,400 or 500, and mixtures thereof may are included, e.g., PEG100 and PEG200, PEG200 and PEG300, PEG 100 and PEG300 or PEG200 and PEG400.

As used herein, innate immunity refers to one or more components typically associated with nonspecific immune defense mechanisms in a subject. These components include the alternate complement pathway, e.g., Factor B, Factor D and properdin; NK cells, phagocytes (monocytes, macrophages), neutrophils, eosinophils, dendritic cells, fibrocytes; antimicrobial chemicals, e.g., one or more of defensins; physical barriers-skin, mucosal epithelium; or certain interleukins, chemokines, cytokines, lung or alveolar macrophage respiratory burst activity or a lung surfactant protein such as surfactant protein A or surfactant protein D. Innate immunity plays a role in resistance to intracellular parasite infections, e.g., white blood cell infection, a liver infection, and other infections, e.g., lymph node infections. Detectable enhancement of innate immunity mechanism by F3Cs or method described herein can also enhance phagolysosome fusion or movement, which some pathogens, e.g., intracellular bacteria such as mycobacteria, or Listeria inhibit.

Tenns such as immune dysregulation, immune dysregulation condition, unwanted immune response and the like mean that a subject has or is subject to developing an immune response that is not desirable or is suboptimal for the subject's condition. Such dysregulation or unwanted responses can arise from various clinical conditions or diseases or as a result of treatment of such conditions or diseases, e.g., inflammation, autoimmunity, °δθj }2 organ or tissue transplant rejection (e.g., allograft, xenograft), infections, cancers, chemotherapy treatments, trauma, allergy' conditions or in conditions where a subject mounts a Thi, Tel, Th2 or Tc2 immune response that is considered to be pathogenic, ineffective, insufficient or suboptimal. Immune dysregulation conditions are as described herein or in the cited references.

Tenns such as cellular response, cellular activity, biological response, biological activity and the like mean a response or activity that is detectably modulated in response to the presence of a F3C. Such responses or activities can be direct effects or indirect effects on one or more cellular activities or on the expression or level of one or more molecules that the affected cell(s) bind, sequester, synthesize or respond to. Such responses or activities include a detectable change in the synthesis or level of one or more cytokines, growth factors, transcription factors (including receptors and their cofactors), enzymes, Thi - or Th2-associated antibody subtype responses or the like. Typically, the cytokines, growth factors, transcription factors, enzymes or antibodies that are modulated are involved in the amelioration of a pathological condition or in the establishment, maintenance, severity or progression of a pathological condition.

As used herein, references to CD molecules, specific immune cell subsets, immune responses and the like, generally use nomenclature that applies to molecules, cells or the like that are found in humans. Analogs or counterparts of such molecules, cells or the like in other species may have a differing nomenclature, but are included in this invention. A description of the nomenclature and function of various CD molecules and immune cell subsets are as found in the scientific literature. References to ThO, Thi or Th2 cells and references to Thi or Th2 immune responses in the context of human patients refers to the human counterparts of the murine ThO, Thi or Th2 immune cells or responses.

Immunosuppressive molecule means molecules such as cyclosporin, cyclohexaniide, mitomycin C, Adriamycin, taxol and amphotericin B and steroids. These molecules tend to have toxicities toward the immune system and are directly or indirectly immunosuppressive, ,E6<>312 e.g., they are toxic to dividing cells, they inhibit proliferation of immune cell precursors or they can down regulate an otherwise desired or improved immune response or condition.

An agonist or an antagonist is a compound or composition, usually containing F3C, that respectively, either detectably increases or decreases the activity of a receptor, an enzyme or another biological molecule, which can lead to increased or decreased transcription or mRNA levels of a regulated gene or to another measurable effect such as altered level of activity of the gene product or protein. The increase or decrease in a receptor's or enzyme's activity will be an increase or a decrease of at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or a range about between any two of these values, for one or more measurable activities. Receptors, their accessory factors and associated transcription factors can modulate transcription of their target gene(s) by detectably increasing or decreasing transcription or mRNA levels. Biological activities of receptors may also include modulating biological responses such as signal transduction within a cell or ion flux, e.g., sodium, potassium or calcium, across cell or organelle membranes, e.g., across mitochondrial matrix..

Terms such as biologically active metabolite and the like mean derivatives of the F3Cs that retain a detectable level, e.g., at least about 10%, at least about 20%, at least about 30% or at least about 50%, of at least one desired activity of the parent compound, e.g., stimulation of a desired immune response. Determination of a desired activity is accomplished essentially as described herein. Such metabolites can be generated in the gastrointestinal track in blood or in one or more subject tissues. Such metabolites are detected using standard analytical methods, e.g., GC-MS analysis of an optionally radio labeled F3C and its metabolites, in blood, urine or other biological samples after it is administered to a subject by one or more routes as disclosed herein. Terms such as metabolic precursor of F3Cs and the like can include compounds that generate a detectable level of the F3C or a detectable level, e.g., at least about 10%, at least about 20%, at least about 30% or at least about 50%, of at least one desired activity of the F3C.

Determination of a desired activity is accomplished essentially as described herein. ΙΕ ν* δ ο 3 12 Conversion of metabolic precursors can occur in the gastrointestinal tract, in blood or in one or more subject tissues.

Salts of F3Cs. Invention embodiments include salts and complexes of F3Cs, including pharmaceutically acceptable or salts that are relatively non-toxic. Some of the F3Cs have one or more moieties that carry at least a partial positive or negative charge in aqueous solutions, typically at a pH of about 4-10, that can participate in forming a salt, a complex, a composition with partial salt and partial complex properties or other non-covalent interactions, all of which we refer to as a salt(s). Salts are usually biologically compatible or pharmaceutically acceptable or non-toxic, particularly for mammalian cells. Salts that are biologically toxic are optionally used with synthetic intermediates of F3Cs. When a water12 soluble composition is desired, mono-valent salts are usually used.

Metal salts typically are prepared by reacting the metal hydroxide with a compound of this invention. Examples of metal salts that are optionally prepared in this way are salts containing Li.sup.+, Na.sup.+, and K.sup.+. A less soluble metal salt can be precipitated from the solution of a more soluble salt by adding a suitable metal compound. Invention salts may be formed from acid addition of certain organic acids, such as organic carboxylic acids, and inorganic acids, such as alkylsulfonic acids or hydrogen halide acids, to acidic or basic centers on F3Cs.. Metal salts include ones containing Na.sup.+, Li.sup.+, K.sup.+, Ca.sup.++ or Mg.sup.+. Other metal salts may contain aluminum, barium, strontium, cadmium, bismuth, arsenic or zinc ion.

Salt(s) of F3Cs may comprise a combination of appropriate cations, to one many or all of its components, such as alkali and alkaline earth metal ions or ammonium and quaternary ammonium ions with the acid anion moiety of the phosphoric acid or phosphoric acid group, which may be present in polymers or monomers.

Salts are produced by standard methods, including dissolving free base in an aqueous, aqueous-alcohol or aqueous organic solution containing the selected acid, optionally IE 0 6 0 j j 2 followed by evaporating the solution. The free base is reacted in an organic solution containing the acid, in which case the salt usually separates directly or one can concentrate the solution.

Suitable amine salts include amines having sufficient basicity to form a stable salt, usually amines of low toxicity including trialkyl amines (tripropylamine, triethylamine, trimethylamine), procaine, dibenzylamine, N-benzyl-betaphenethylamine, ephenamine, Ν,Ν'-dibenzylethylenediamine, N-ethylpiperidine, benzylamine and dicyclohexylamine.

Salts include organic sulfonic acid or organic carboxylic acid salts, made for example by addition of the acids to basic centers, typically amines. Exemplary sulfonic acids include C.sub.6-16 aryl sulfonic acids, C.sub.6-16 heteroaryl sulfonic acids and C.sub.1-16 alkyl sulfonic acids such as phenyl sulfonic acid, a-naphthalene sulfonic acid, .beta.-naphthalene sulfonic acid, (S)-camphorsulfonic acid, methyl (CH.sub.3SO.sub.3H), ethyl (C.sub.2H.sub.5SO.sub.3H), n-propyl, i-propyl, n-butyl, s-butyl, ί-butyl, t-butyl, pentyl and hexyl sulfonic acids. Exemplary organic carboxylic and other acids include C.sub.1-16 alkyl, C.sub.6-16 aryl carboxylic acids and C.sub.4-16 heteroaryl carboxylic acids such as acetic, glycolic, lactic, pyruvic, malonic, glutaric, tartaric, citric, fumaric, succinic, malic, maleic, oxalic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, salicylic, nicotinic, 2-phenoxybenzoic, methanesulfonic, pamoic, propionic, toluenesulfonic and trifluoroacetic acids.

Invention salts include those made from inorganic acids, e.g., HF, HCl, HBr, HI, H.sub.2SO.sub.4, H,sub.3PO.sub.4, Na.sub.2CO.sub,3, K.sub.2CO.sub.3, CaCO.sub.3, MgCO.sub.3 and NaClO.sub.3. Suitable anions, which are optionally present with a cation such a Ca.sup.-Η-, Mg.sup.++, Li.sup.+, Na.sup.+ or K.sup.+, include arsenate, arsenite formate, sorbate, chlorate, perchlorate, periodate, dichromate, glycodeoxycholate, cholate, deoxycholate, desoxycholate, taurocholate, taurodeoxycholate, taurolithocholate, tetraborate, nitrate, nitrite, sulfite, sulfamate, hyposulfite, bisulfite, metabisulfite, thiosulfate, thiocyanate, silicate, metasilicate, CN.sup.-, gluconate, gulcuronate, hippurate, picrate, hydrosulfite, hexafluorophosphate, hypochlorite, hypochlorate, borate, metaborate, tungstate «60312 and urate.

Salts also include the F3C salts with one or more amino acids. Many amino acids are suitable, especially the naturally-occurring amino acids found as protein components, although the amino acid typically is one bearing a side chain with a basic or acidic group, e.g., lysine, arginine, histidine or glutamic acid, or a neutral group such as glycine, serine, threonine, alanine, isoleucine, or leucine.

The invention compositions include F3Cs, their hydrates and the compounds in their ionized, un-ionized, as well as zwitterionic form. Thus, for any F3Cs or compounds described herein with any substituent that contains a moiety that is partially or completely ionizable, e.g., a carboxyl group, the ionizable atom, usually hydrogen, may be replaced with one or more suitable counter ions such as a monovalent metal, a multivalent metal, an alkaline metal, or an ionizable organic moiety, e.g., Li.sup.+, Na.sup.+, K.sup.+, Ca.sup.+2, Mg.sup.+2, SO.sub.4.sup.-2, PO.sub.4.sup.-2, CH.sub.3C(O)O.sup.-, CF.sub.3C(O)O.sup.-, F.sup.-, Cl.sup.-, Br.sup.-, I.sup.-, NH.sub.4.sup.+, N.sup.+(CH.sub.3).sub.4, N.sup.+(C.sub.2H.sub.5).sub.4, HN.sup.+(C.sub.2H.sub.5).sub.3, H.sub.2N.sup.+(C.sub.2H.sub.5).sub.2, .beta.-hydroxyethyltrimethylammoniu- m, piperazinium, pyridinium, N-methylpyridinium, morpholimium, N,Ndimethylmorpholinium, p-toluidinium or another ionizable moiety described herein. When a F3C is under conditions, e.g., in a solution, where such moieties can partially or completely ionize, the ionizable moiety may be partially or completely charged, e.g., --C(O)—O.sup.-, -NH.sub.3.sup.+, ~C(O)~NH.sub.3.sup.+ or -O-S(O)(O)-O- may be partially for fully ionized.

F3C compositions can be comprised of any Formula 1 Compound and/or a Fonnula 2 compound, or a Fonnula 2 compound alone ,or a metabolic precursor, a metabolite or salt thereof of its constituent components, wherein such components can be an ester, a thioester, a thionoester, a phosphoester, a phosphothioester, a phosphonoester, a phosphiniester, a IE ^60312 sulfite ester, a sulfate ester, a sulfoxide, a sulfamate, a sulfonate, a sulfamide, a sulfinamide, a sulfurous diamide, an amide, an amino acid, a peptide, an ether, a thioether, an acyl group, a thioacyl group, a carbonate, a carbamate, a halogen, an acetal, a thioacetal a spiro ring, an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted aryl moiety, an optionally substituted heteroaryl moiety, an optionally substituted heterocycle, an optionally substituted monosaccharide, an optionally substituted oligosaccharide, a nucleoside, a nucleotide, an oligonucleotide, a polymer, or, one or more of two adjacent R.sup.l-R.sup.4, R.sup. 10, R.sup.lOA, R.sup.lOB, R.sup.lOC and R.sup,10D are an independently selected epoxide or cyclopropyl ring of any compound belonging to Formula 1 and/or Formula 2 compounds.

Invention embodiments include a composition comprising a F3C and 1,2, 3, 4 or more non13 aqueous liquid excipients.

Embodiments of F3Cs include or exclude any subset of compounds within the definition of formula 1 and/or Formula 2, provided that at least one component remains.

Specific F3Cs that can be used in the clinical treatments and other methods described herein.

Metabolites. The invention includes the therapeutic or other uses disclosed herein of metabolites of F3C, which include biologically active metabolites. Metabolites can arise from in vivo or in vitro metabolism. Metabolites of F3C include products that are new.

Metabolites may result for example from the oxidation, reduction, hydrolysis, amidation, esterification, glycosidation and the like of the administered formula 1 compound And/or Formula 2 compound due to enzymatic or chemical processes. A F3C compound can have multiple co-existing metabolites. Metabolites may be generated in vivo in a subject or they ·< may arise ex vivo from cells or tissues, e.g., from a mammal such as a human, rodent or a primate. Accordingly, the invention includes new F3Cs produced by a process comprising contacting an F3C with a subject or a subject's cells or tissue for a period of time sufficient to yield detectable amounts of a metabolic product thereof. Such products typically are 0 6 θ 3 ί 2 identified by preparing a radio labeled or mass labeled F3C that comprises, e.g., 1, 2, 3 or more .sup.l3C, .sup.l4C, .sup.3H, .sup.2H, .sup.1311, .sup.32P, ,sup.35S or .sup.99Tc atoms bonded to the compound component, and administering it as a trace labeled compound along with the unlabeled compound. The labeled and unlabeled compounds are administered by any suitable route (by, e.g., a buccal, sublingual, parenteral, topical or oral route) in a detectable dose (e.g. greater than about 0.1 .mu.g/kg, or at least about 10 .mu.g/kg or at least about 0.5 mg/kg of the labeled compound) to a subject, e.g., an animal or mammal such as rat, mouse, guinea pig, primate, or to a human. After administration sufficient time is allowed for metabolism to occur (typically about 30 seconds to 30 hours) and conversion products are isolated from one or more of the urine, blood, plasma, feces or other suitable biological sources. The amount of labeled formula 1 compound that is administered to a subject will vary with the specific activity of the labeled compound. Exemplary metabolic conversions of formula 1 compounds include modification of hydrogen atoms or other moieties that are bonded to, e.g., one or more ofthe 1, 2, 3, 4, 6, 7, 11, 15, 16 or 17 positions. Exemplary conversions at these one or more of positions include hydroxylation of ring atoms, e.g., ring carbon atoms, conjugation of hydroxyl groups that are bonded to one or more of those positions with moieties such as sulfate, phosphate or a monosaccharide or disaccharide such as glucuronic acid and hydrolysis of moieties such as esters or alkoxy groups.

Analytical characterization and reference standards. Individual F3Cs described or disclosed herein are suitable for use as standards for determining chemical or physical properties using one, two or more analytical methods, to determine the chemical structure or properties on any of its chemical components, e.g., for use in HPLC, reverse phase HPLC, MS (mass spectrometry), quadrupole MS, GC-MS, LC-MS, NMR (nuclear magnetic resonance spectrometry), .sup.2H-NMR, .sup.3H-NMR, .sup.l3C-NMR, .sup.l4C-NMR, infrared spectrometry (IR), Fourier transform-IR, optical rotary dispersion, loss on drying for water and solvent measurement, Karl Fisher titration for water determination, differential scanning calorimetry, melting point, density, refractive index, solubility characteristics in organic solvents, aqueous systems or aqueous-organic solvent mixtures, the partition coefficient in IE o J 12 immiscible solvent systems, e.g., octanohwater partition coefficient, heat stability or epimerization rate or characteristics of a given enantiomer. These analytical or chemical properties of each F3C component are collectively referred to as analytical characteristics.

Thus, to aid in the determination of, e.g., the structure of a metabolite of a F3C component or a structurally related compound, the parent compound or another structurally related F3C could be used as a standard. Metabolism of F3Cs will often include one or more of oxidation, reduction, hydroxylation or conjugation, e.g., oxidation or reduction to a -OH or .dbd.0 moiety, or conjugation with a moiety such as sulfate, phosphate, amino acid, dipeptide or a monosaccharide such as glucuronic acid at, e.g., the 2, 3, 6, 7,11, 15,16,17 or other positions on the steroid nucleus. In these embodiments, the appropriate use of a F3C of known structure as a standard can aid in or verily the identification of metabolites that are projected to have closely related structures. Information regarding the identification can be useful or sometimes is necessary for, e.g., obtaining regulatory approval to market a therapeutic agent such as a F3C or understanding the potential biological role that a F3C or its metabolite can play in one of the applications disclosed herein or in a cited reference.

F3Cs and their metabolites are isolated or characterized using radio labeled or mass labeled atoms. F3Cs are also optionally isolated by the use of antibodies capable of binding to epitopes in F3Cs or in metabolites.

In general, analysis of F3C metabolites is accomplished in the same way as conventional drug metabolism studies, which are known to those skilled in the art. The conversion products, especially when they are not otherwise found in vivo, are useful in diagnostic assays for therapeutic dosing of the formula 1 compounds and/or formula 2 compounds, even if they possess only limited therapeutic activity of their own.

Embodiments include a method (the characterization method) to characterize or at least partially characterize a formula 1 compound or a formula 2 compound, that is at least partially uncharacterized for one or more given chemical or analytical properties, e.g., a known or potential metabolite of a parent formula 1 compound, or a formula 2 compound, comprising (a) providing a formula 1 compound ,or a formula 2 compound, having one, two Μ 6 ΰ 3 j 2 or more known characteristics, e.g., a known or at least partially known or characterized chemical structure, XRD spectrum or melting point (a CF3C1 j, and a formula 1 compound or a formula 2 compound, that is unknown or at least partially uncharacterized, i.e., is uncharacerized for at least one of the same analytical characteristics (a UCF3C j, (b) obtaining one, two or more analytical characteristics of the UCF3C, and (c) comparing the 1, 2 or more analytical characteristics of the CF3C with the analytical characteristics of the UCF3C. The steps in this method may be conducted in any suitable order, e.g., analytical or chemical data for the CF3C will usually be obtained before or at about the same time as one obtains the analytical or chemical data for the UCF3C. Usually the CF3C will be more completely characterized than the UCF3C, particularly with regard to its chemical structure or its relative degree of purity or with regard to the analytical or chemical data that is being sought. This method allows further characterization of the UCF3C, e.g., by confirming the UCF3C's chemical structure or by determining the UCF3C's stability under various storage or temperature conditions or in various formulations or by determining other analytical or chemical properties of interest. In this method, the CF3C itself may not be completely characterized, however, for the one, two or more analytical characteristics of interest, the CF3C will usually have a known or confirmed property or properties, while the UCF3C is unknown or at least unconfirmed for the same property or properties.

In some embodiments the characterization method is conducted by comparing dissimilar analytical characteristics. For example, the CF3C may be well characterized by GC-MS or by NMR, while an insufficient amount of the UCF3C is available for analysis with the same technique. In these cases, one can then, e.g., compare the GC-MS ofthe CF3C with the NMR of the UCF3C to obtain the same or essentially the same information for the UCF3C. Other examples of where this can be done is where DSC data is available for the CF3C, and only melting point data is available for the UCF3C. In this .case, the CF3C DSC data is compared to the UCF3C's melting point data. Also, in conducting the characterization method, one can optionally derivative or chemically modify the CF3C and/or the UCF3C to facilitate analysis ofthe compound(s). For example, in conducting MS, GC-MS or NMR analysis, one or more free hydroxyl or ketone moieties on the CF3C and/or the F2C can be IE Ο 6 0 312 silylated using, e.g., trimethylsilyl chloride, t-butyl-dimethylsilyl chloride or other suitable silylating agents. Similarly, the UCF3C may be treated or incubated with a cell line or tissue or with a glucuronidase, sulfatase, phosphatase, esterase, lipase, oxidoreductase or other enzyme and then characterized. This treatment may in some cases convert the UCF3C into the CF3C, but this conversion would usually be confirmed by one, two or more suitable analytical methods. Such treatments will usually generate additional data about the structure, properties or origin of the UCF3C.

Embodiments include modifications of the characterization method that use a CF3C and a second formula 1 compound or a formula 2 compound, that is believed or known to have a related structure or empirical formula. In these modifications, the CF3C is used as described and a second formula 1 compound or a formula 2 compound, or a UCF3C that is believed or known to be, e.g., an epimer or a salt, of the CF3C is compared to the CF3C. Invention embodiments include other modifications of the characterization method such as (1) comparing analytical or chemical data from a single CF3C with 2, 3, 4 or more UCF3C, (2) comparing analytical or chemical data from 2, 3,4 or more CF3C with a single UCF3C and (3) comparing analytical or chemical data from 2, 3,4 or more CF3C with 2, 3, 4 or more UCF3C. In these modifications, the CF3C or UCF3C are used essentially as described for the characterization method, except that data is obtained for the added formula 1 compound or a formula 2 compound.

Typically, when the 1,2 or more analytical characteristics of a CF3C or a UCF3C are obtained, which may be for use in the characterization method or for other purposes, each compound is analyzed under the same or essentially the same analytical conditions using the same or essentially the same analytical technique or instrument. Variations in an analytical technique may be used where the properties of a CF3C or a UCF3C require slightly different handling or specimen preparation. An example of a variation in analytical conditions is the comparison of a property of a CF3C, e.g., its stability to heat, humidity or prolonged storage at a given temperature, with the same property of the CF3C in a composition containing an excipient(s) or in a formulation (where the CF3C in a composition is then considered the UCF3C for the characterization method). This allows the determination of the stability of the CF3C as a pure compound compared to its stability in any desired composition.

As is apparent from the present disclosure, the F3C may be prepared synthetically and typical embodiments will utilize purified a F3C. Purified F3C can be free, essentially free or partially free, of other F3C or other compounds such as excipients. Thus, any given purified F3C can be present as a solid that contains, e.g., less than about 15% w/w or less than about 10% w/w or less than about 8% w/w or less than about 5% w/w or less than about 3% w/w or less than about 1% w/w of one, two or more other F3Cs, excipients, synthetic byproducts, decomposition products or synthesis or purification reactants or reagents.

Similarly, the F3C can be present in a solution or suspension that contains at least about 90% w/w or at least about 95% w/w or at least about 97% w/w of the F3C and one or more excipients and less than about 10% or 8% or 5% or 3% w/w or 1% w/w of one, two or more other F3Cs, excipients, synthetic by-products, decomposition products or synthesis or purification reactants or reagents.

Protecting groups are removed by known procedures, although it will be understood that the protected intermediates fall within the scope of this invention. The removal of the protecting group may be arduous or straightforward, depending upon the economics and nature of the conversions involved. In general, one will use a protecting group with exocyclic amines or with carboxyl groups during synthesis of a F3C. Note it is not necessary to place a protecting group on all components of an F3C. For most therapeutic applications amine groups should be deprotected. Protecting groups commonly are employed to protect against covalent modification of a sensitive group in reactions such as alkylation or acylation. Ordinarily, protecting groups are removed by, e.g. hydrolysis, elimination or aminolysis. Thus, simple functional considerations will suffice to guide the selection of a reversible or an irreversible protecting group at a given locus on the F3Cs.

Groups capable of biological cleavage typically include prodrugs. Some exemplary groups are described in Design of Prodrugs, Hans Bundgaard (Elsevier, N.Y., 1985, ISBN 0-444IE ^60312 80675-X) (Bundgaard) and will not be detailed here. In particular, Bundgaard, at pages 1-92, describes prodrugs and their biological cleavage reactions for a number of functional group types. Prodrugs for carboxyl and hydroxyl groups are detailed in Bundgaard at pages 3 to , for amides, imides and other NH-acidic compounds at pages 10 to 27, amines at pages 27 to 43, and cyclic prodrugs at pages 62 to 70. These moieties are optionally bonded to the steroid at one, two or more of the variable groups that are bonded to the rings in tlie F3Cs, e.g., one or more R.sup.l-R.sup.6, R.sup.10, R.sup.15, R.sup.17 and R.sup.18.

In some embodiments one or more F3Cs or groups of F3Cs may be excluded from one or more of the uses disclosed herein.

Dosages of F3C and dosing protocols or methods. In treating any of the conditions or symptoms disclosed herein, one can continuously or intermittently administer the F3C(s) to a subject having or susceptible to developing the condition or symptom. In treating a condition such as an infection, a hyper proliferation condition, an inflammation condition or another condition disclosed herein with a F3C using an intermittent dosing can avoid or ameliorate some of the undesired aspects normally associated with discontinuous dosing.

Such undesired aspects include development of resistance of a pathogen such as a pathogen disclosed herein, e.g., a virus or bacterium such as HIV or Staphylococcus aureus or a parasite such as a Plasmodium parasite, to the therapeutic agent, failure of the patient or subject to adhere to a daily dosing regimen or reduction of the dosages of other therapeutic agents and/or their associated unwanted side effects or toxicities, e.g., reduction or a toxic effect of a chemotherapy or radiation exposure. In any of the continuous or intermittent dosing protocols described herein, other appropriate treatments can be applied as the subject's clinical situation dictates. Suitable other appropriate treatments or therapeutic agents are described elsewhere herein and in the cited references.

In any of the continuous or in any step(s) in the intermittent dosing protocols described herein, or in treating any of the diseases, conditions or symptoms described herein, the F3C(s) can be administered by one or more suitable routes, e.g., oral, buccal, sublingual, ie intramuscular (i.m.), subcutaneous (s.c.), intravenous (i.v.), intradermal, another parenteral route or by an aerosol. The effective daily dose in such methods will typically comprise about 0.05 mg/kg/day to about 200 mg/kg/day, or about 0.1 to about 100 mg/kg/day, including about 0.2 mg/kg/day, 0.5 mg/kg/day, about 1 mg/kg/day, about 2 mg/kg/day, about 4 mg/kg/day, about 6 mg/kg/day, about 10 mg/kg/day, about 20 mg/kg/day, about 40 mg/kg/day or about 100 mg/kg/day. Higher dosages, e.g., about 250 mg/kg/day, about 300 mg/kg/day or about 350 mg/kg/day can also be utilized, e.g., in veterinary applications. One can administer the F3C(s) orally using about 4 to about 60 mg/kg/day, usually about 6-30 mg/kg/day. In some embodiments, the intermittent dosing methods exclude dosing protocols that are commonly used to deliver contraceptive steroids to, e.g., human females, such as daily dosing for 21 days, followed by no dosing for 7 days. For humans, dosing is generally about 0.005 mg/kg/day to about 30 mg/kg/day, typically about 0.5-5 mg/kg/day. Low dosages for humans such as about 0.005 mg/kg/day to about 0.2 mg/kg/day or about 0.25-10 mg/day, can be used with, e.g., local, topical, transmucosal or intravenous administration and higher dosages such as about 0.1 mg/kg/day to about 20 mg/kg/day or about 5-200 mg/day, can be used, e.g., for oral, subcutaneous or other systemic or local administration route. For non-human subjects, e.g., mammals such as rodents or primates, the effective daily dosage may comprise about 0.05 mg/kg/day to about 350 mg/kg/day. F3C formulation dosages or daily doses or unit doses or subdoses for subjects such as humans and mammals include, e.g., about 1, 5,10, 15,20, 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 400 or 450 mg of the F3C.

An effective dosage or an effective amount of a F3C(s) is one that is sufficient to result in, e.g., a detectable change in a symptom or an immune parameter such as one described herein. An effective dosage (or daily dosage) may be administered to a subject over a period of time, e.g., at least about 1-14 days before a symptom change or an immune parameter detectably changes. Effective amounts of a F3C can be delivered using the dosages and dosing protocols described herein. in some embodiments, F3C are used to treat, ameliorate, prevent or slow the progression of ϋ 6 0 J Μ a condition or disease described herein by continuous daily dosing of the F3C for 1 day to 1, 2, 3 years or more. In related embodiments, F3C are used to treat, ameliorate, prevent or slow the progression of a condition or disease described herein by continuous dosing the F3C every other day or dosing every third, fourth, fifth, sixth, seventh or 14.sup.th day over a time period of 3 days to 3, 2, 3 years or more, e.g., dosing for about 2, 3,4, 5, 6 or 7 days or about 1, 2, 3, 4, 6, 8, 10, 12, 16, 20,24 or more weeks. Daily doses in any of these dosing regimens or protocols may be subdivided into 2 or 3 sub doses.

Intermittent dosing protocols include administration of a F3C, e.g., orally, topically or parenterally as follows: (1) daily dosing or dosing every other day or dosing every third day or dosing every fourth day or dosing every fifth day or dosing every seventh day for about 2, 3, 4, 5, 6, 7, 8, 9, 10,11,12, 13,14,15,16,17,18,19, 20, 21 or 28 days to about 190 days or more, e.g., 1 or 2 years, (2) no dosing of the F3C for 1 to about 190 consecutive days (e.g., about 2, 3,4, 5, 6, 7, 8, 9,10,11 or 12 days to about 20 days), (3) daily dosing for about 3 to about 190 days (e.g., about 3 to about 20 days), and (4) optionally repeating step (2) or a variation of step (2) and (5) optionally repeating the steps (1), (2), (3) and (4) 1, 2, 3, 4, 5, 6, 10, 15, 20, 30 or more times. In some embodiments, the dosing of steps (1) and (3) are the same, while in others, step (1) dosing is for a longer time than step (3). Less frequently, step (1) dosing will be for a shorter time. In some embodiments, steps (1)-(4) or (1)-(5) of the dosing protocol described above where step (4) is included, is repeated at least one time, e.g., at least 2, 3,4, 5 or 6 times. For conditions that tend to remain chronic, e.g., HIV infection or other chronic conditions described herein, any of these intermittent dosing protocols can be maintained over a relatively long time period, e.g., for at least about 4 months or 6 months to about 5 or more years.

In some embodiments, the number of days of dosing in steps (1) and (3) is the same in each round of treatment, i.e., each time period in step (1) and (3) is the same in the initial and subsequent rounds of the method. In other embodiments they differ. Thus, in some embodiments, step (1) may comprise dosing of about 1 mg/day to about 1500 mg/day of a F3C for 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more consecutive days. Then, step (2) may comprise °6 03 12 not administering any F3C for at least about 2, 3, 4, 5, 6, 7, 14, 21, 28, 42, 56, 84, 98, 112 or more consecutive days. Step (3) could comprise dosing of a F3C for 1, 2, 3, 4, 5, 6, 7, 8, 9, or more consecutive days. When step (4) is included it is typically about 1 day to about 3 months, usually 3 days to about 6 weeks. On days when the F3C is administered to the subject, it may be delivered in a single dose or in two, three or more subdoses at, e.g., about 12 hour or about 8 hour time intervals.

Exemplary embodiments comprise (1) administering a F3C(s) once every (as a single dose or as 2 or 3 daily subdoses) 2 days, every 3 days or every 4 days or once per week for about 3, 5, 7, 9, 11, 13, 14, 15, 21,28 or more days, followed by (2) no dosing for about 2, 3,4, 5, 6, 10, 14, 15, 21, 20, 25, 28, 30, 35, 40, 42, 45, 49, 56, 60, 70, 77, 84, 98, 112 or more days and then (3) administering the F3C(s) at least once more on one day, e.g., administering the F3C(s) as described in step (1), (4) not dosing for 2, 3,4, 5, 7 or more days, e.g., as described in step (2) for 1, 2, 3,4, 5, 6, 7 or 8 weeks, and (5) optionally repeating steps (1), (2), (3) and (4) 1, 2, 3, 4, 5 or 6 times or more.

Other embodiments comprise (1) administering a F3C(s) once every day (as a single dose or as 2 or 3 daily subdoses) for 3-15 or about 8-12 days, followed by (2) no dosing for 1, 2, 3, 4, 5, 6, 10,15, 20, 25, 30, 35,40, 45, 50, 56, 70, 84, 98, 112 or more days and then (3) administering the F3C(s) at least once more on one day, e.g., administering the F3C(s) once per day for about 3-15 or about 8-12 consecutive days essentially as described in step (1) and (4) optionally repeating steps (1), (2) and (3) 1,2, 3,4, 5 or 6 times or more. In a subset of these embodiments (1) comprises administering a F3C(s) once every day for about 5,6, 7, 8, 9 or 10 days, followed by (2) no dosing for about 10-40 days, (3) administering the F3C(s) at least once more on one day, e.g., administering the F3C(s) once per day for about 10 days (4) repeating step (2) or a variation, e.g., no dosing for about 5-45 days, and (5) optionally repeating steps (1), (2), (3) and (4) or a variation thereof those steps 1, 2, 3, 4, 5 or 6 times or more.

One aspect of invention intermittent dosing is monitoring the subject's response to a °60jf2 particular dosing regimen or schedule, e.g., to any intermittent administration method disclosed herein. Once a response is observed dosing can be continued for one, two or three additional days, followed by discontinuing the dosing for at least one day (at least 24 hours), usually for at least about 2, 3,4, 5, 6, 7, 14, 21, 28, 42, 56, 70, 84, 98, 112 or more days. Once the subject's response shows signs of remission (e.g., a symptom begins to intensify, viral serum DNA or RNA begins to increase or an immune parameter, e.g., as described herein, begins to deteriorate), dosing can be resumed for another course. An aspect of the subject's response to F3C(s) is that the subject may show a measurable response within a short time, usually about 5-10 days, which allows straightforward tracking of the subject's response, e.g., by monitoring peripheral white blood cells (PBMC), by measuring the blood or by measuring a white blood cell population(s) or expression of a cytokine or interleukin by e.g., white blood cells or a subset(s) thereof. One may monitor one or more immune cell subsets, e.g., NK, LAK, dendritic cells or cells that mediate ADCC immune responses, during and after intermittent dosing to monitor the subject's response and to determine when further administration of the F3C is indicated. These cell subsets are monitored as described herein, e.g., by flow cytometery.

For any of the treatments or methods described herein, prolonged beneficial effects or a sustained immune response by a subject may result from a single administration or short course, e.g, about 1-5 days or about 8 days to about 4 months, of continuous or intermittent administration of a F3C. A single administration means that a F3C is administered to the subject in one, two, three or more doses within a 24 hour period and no further administration of any F3C to the subject occurs for at least about 4-90 days, e.g., about for at least about 30 days to about 2 months, or for about 1.5,2, 3,4, 5, 6 or more months. Prolonged beneficial effects or immune responses may also persist after a short course of treatment has been completed (e.g., daily dosing for 2, 3, 4, 5 or 6 days) and the subject is no longer receiving any F3C, or, in some cases, any other therapeutic treatment to treat the primary cause of the subject's pathological condition. Such beneficial effects can persist for more than about 5-30 days, e.g., for at least about 21, 28,42, 56,70, 84, 98,112 or more days. Thus, administration of a F3C provides a method to help protect a subject against progression of an infection or against adverse consequences of unwanted immune reactions, e.g., inflammation or immunosuppression or as disclosed herein, without any dosing of the compound for at least 1,2 or 3 months after an initial dosing protocol.

Other intermittent dosing embodiments comprise administering to a subject having or susceptible to a condition as described herein an effective amount of a F3C using an initial induction or high dosing regimen. The high dosing regimen may comprise, e.g., 1, 2, 3, 4, 5, 6, 7 or more daily doses of about 4 to about 40 mg/kg that are administered daily, every other day, every 3.sup.rd day, every 4.sup.th day or every 5.sup.th day. Then, the subject is not dosed with a F3C for a period, e.g., of about 5, 7,14, 21,28, 42, 56, 70, 84, 98,112 or more consecutive days. Then a lower daily dosing regimen is administered to the subject, e.g., about 0.2 mg/kg to about 4 or about 6 mg/kg, essentially as described for the high dosing regimen. Alternatively, this low dosing regimen may comprise 1, 2, 3, 6 or more rounds of a low to moderate initial level, e.g., about 2 to about 10 mg/kg/day, optionally followed by subsequent rounds of daily dosing that decrease the initial low to moderate level by about 10%, 20%, 30%, 40% or more in each subsequent round of treatment, which is continued until administration is discontinued. These embodiments can be used with any of the dosing protocols described herein.

Dosages of the F3C, continuous or intermittent dose protocols, routes of administration and the use of combination therapies with other standard therapeutic agents or treatments could be applied essentially as described above for any of the diseases or conditions that are disclosed herein. Thus, the F3Cs may be administered prophylactically or therapeutically in chronic or acute conditions. In acute conditions, the F3Cs may also be administered at the time of occurrence or relatively soon after an acute event such as the onset of surgery, a migraine or the occurrence of trauma, e.g., a central nervous system injury, a cerebral stroke or myocardial infarction. For acute events, a F3C may thus be administered concurrently, e.g., within about 15 minutes or about 30 minutes or about 45 minutes of the onset or occurrence of the acute event, or at a later time, e.g., at about 1,2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15,16, 18, 20, 22, 24,26, 28, 30, 36,42,48, 54, 60, 72, 84, 96,108 or 120 hours after the onset or occurrence of the acute event or at any range of times defined by any two of these later times. The F3Cs may thus be administered at about 4-120 hours, about 6-120 hours, about 848 hours, 8-24 hours, 8-12 hours, 10-12 hours, 10-14 hours, 10-16 hours, about 10-24 hours, 12-14 hours or about 12-16 hours after an acute event starts, occurs or is believed to have begun, e.g., after a surgical procedure has been completed or after a radiation treatment has ended or after a cytotoxic chemotherapy or a myelosuppressive cancer chemotherapy has been administered to the subject.

Alternatively, the F3Cs may be administered before, e.g., within about 15 minutes, about 30 minutes or about 45 minutes before the onset or occurrence of a planned or anticipated acute event, or at an earlier time, e.g., at about 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 22, 24, 26, 28, 30, 36, 42,48, 54, 60, 72, 84, 96, 108 or 120 hours before the onset or occurrence of the acute event. The F3Cs may thus be administered at about 6-120 hours, about 8-48 hours, about 10-24 hours, about 10-16, or about 12-16 hours before the planned or anticipated acute event, e.g., before a planned surgery or a radiation treatment starts or occurs.

Formulations and compositions for preparing fonnulations. Invention embodiments include formulations described here and elsewhere in this disclosure. While it is possible for the F3C(s) to be administered to a subject or incubated with a subject's cells in vitro as the compound alone, it is usual to use F3C in a formulation or at least in a composition that contains 1,2,3,4, 5, 6 or more excipients. The fonnulations, which are useful for veterinary or human pharmaceutical use, comprise at least one F3C, together with 1, 2, 3, 4, 5, 6 or more excipients and optionally one or more additional therapeutic ingredients.

The invention includes compositions comprising one or more pharmaceutically acceptable excipients or carriers. The compositions are used to prepare formulations suitable for human or animal use. Formulations may be designed or intended for oral, rectal, nasal, topical or transmucosal (including buccal, sublingual, ocular, vaginal and rectal) and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal, intraocular and epidural) administration. In general, aqueous and non-aqueous liquid or cream formulations are delivered by a parenteral, oral or topical route. In other embodiments, the F3C(s) may be present as an aqueous or a non-aqueous liquid formulation or a solid formulation suitable for administration by any route, e.g., oral, topical, buccal, sublingual, parenteral, aerosol, a depot such as a subcutaneous depot or an intraperitoneal or intramuscular depot or a rectal or vaginal suppository. The preferred route may vary with, for example, the subject's pathological condition or weight or the subject's response to therapy with a F3C or other therapy that is used or that is appropriate to the circumstances. The F3C formulations can also be administered by two or more routes, e.g., subcutaneous injection and buccal or sublingual, where these delivery methods are essentially simultaneous or they may be essentially sequential with little or no temporal overlap in the times at which the compound is administered to the subject.

The formulations include those suitable for the foregoing administration routes. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods known in the art of pharmacy. Techniques, excipients and dosage forms are found in, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1985,17.sup.th edition; Nema et al., PDA J. Pharm. Sci. Tech. 1997 51:166-171; Pharmaceutical Coating Technology, 1995, G. Cole, et al., editors, Taylor & Francis, ISBN 0 136628915; Pharmaceutical Dosage Forms, 1992 2.sup.nd revised edition, volumes 1 and 2, H. A. Lieberman, et al., editors, Marcel Dekker, ISBN 0824793870; Pharmaceutical Preformulation, 1998, pages 1-306, J. T. Carstensen, Technomic Publishing Co. ISBN 1566766907; and Encyclopedia of Pharmaceutical Technology, volumes 1,2 and 3, 2.sup.nd edition, 2002, J. Swarbrick and J. C Boylan, editors, Marcel Dekker, Inc., New York, N.Y.

Methods to make invention formulations include the step of bringing into association or contacting a F3C(s) with one or more excipient, such as one described herein or in the cited references. In general the formulations are prepared by uniformly and intimately bringing into association the F3C(s) with liquid excipients or finely divided solid excipients or both, and then, if appropriate, shaping the product.

/£ Formulations suitable for oral administration are prepared as discrete units such as capsules, soft gelatin capsules (softgels), cachets, tablets or caplets each containing a predetermined amount of the F3C(s). F3C formulations can also be present as a powder or granules or as a solution or a suspension, colloid or gel in an aqueous liquid or base or in a non-aqueous liquid or base; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The F3C formulations may also be a bolus, electuary or paste. Suspension formulations will typically contain about 0.5% w/w or about 1% w/w to about 5%, 10%, 15% or 20% w/w of the F3C, which can be for parenteral use or for other routes of administration, e.g., oral softgels.

A tablet is made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the F3C(s) in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluents, preservative, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered or granulated F3C and one or more excipients, which are optionally moistened, with an inert liquid diluent or excipient. The tablets may optionally be coated or scored and optionally are formulated so as to provide slow or controlled release of the F3C(s) therefrom. An exemplary tablet or caplet formulation suitable for buccal or sublingual delivery of a F3C to a subject's tissues comprises about 25 or 50 mg of a F3C comprising per 25 mg of the F3C about 6.2 mg povidone, about 0.62 mg magnesium stearate, about 45 mg mannitol and about 48 mg of compressible sucrose.

For infections of the eye or other external tissues e.g., the mouth or skin, the formulations are typically applied as a topical ointment or cream containing the F3C(s) in an amount of, for example, about 0.075 to about 20% w/w (including F3C(s) in a range between about 0.1% and 20% in increments of 0.1% w/w such as about 0.6% w/w, about 0.7% w/w, about 1 % w/w, about 1.5% w/w, about 2% w/w, about 2.5 w/w, about 3% w/w, about 5% w/w, about 7% w/w, about 10% w/w etc.), including about 0.2 to 15% w/w and about 0.5 to 10% w/w. When formulated in an ointment, the F3C(s) may be employed with either a paraffinic υόθ3ΐ2 or a water-miscible ointment base. Alternatively, they may be formulated in a cream with an oil-in-water cream base.

If desired, the aqueous phase of the cream base may include, for example, at least 30% w/w of a polyhydric alcohol, i.e. an alcohol having two or more hydroxyl groups such as propylene glycol, butane 1,3-diol, butane 1,4-diol, mannitol, sorbitol, glycerol and a polyethylene glycol (including, e.g., PEG 300 and PEG 400) and mixtures thereof. The topical formulations may include a compound that enhances absorption or penetration of the F3C(s) through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulphoxide and related analogs.

The oily phase of the emulsion formulations may be constituted from known excipients in a known manner. While the phase may comprise an emulsifier or emulgent, it typically comprises a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. A hydrophilic emulsifier may be included together with a lipophilic emulsifier, which acts as a stabilizer. Some embodiments include both an oil and a fat Together, the emulsifier(s) with or without stabilizers) make up the so-called emulsifying wax, and the wax together with the oil and fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

Emuigents and emulsion stabilizers suitable for use in the formulations include Tween6.TM., Span80.TM., cetostearyl alcohol, benzyl alcohol, myrisfyl alcohol, glyceryl mono-stearate and sodium lauryl sulfate. Other excipients include emulsifying wax, propyl gallate, citric acid, lactic acid, polysorbate 80, sodium chloride, isopropyl palmitate, glycerin and white petrolatum.

The choice of suitable oils or fats for the formulation is based on achieving the desired cosmetic properties. Creams are generally a non-greasy, non-staining and washable products with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, °6o312 propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils are used.

Formulations suitable for topical administration to the eye include eye drops, which are usually sterile, wherein the F3C(s) is dissolved or suspended in a suitable excipient(s), including an aqueous solvent for a F3C(s) that comprise at least about 0.5, one, two or more charges at pH values near neutrality, e.g., about pH 6-8. The F3C(s) is typically present in such formulations in a concentration of about 0.5-20% w/w, about 1-10% w/w or about 25% w/w.

Formulations suitable for topical administration to oral mucosa include lozenges or tablets comprising the F3C(s) in a flavored basis or a monosaccharide or disaccharide such as sucrose, lactose or glucose and acacia or tragacanth; pastilles comprising the F3C(s) in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the F3C(s) in a suitable liquid excipient(s). In some embodiments, the lozenges or tablets optionally comprise the property of rapid dissolution or disintegration, e.g., disintegration within about 15 seconds to about 2 minutes, while in others, the lozenges or tablets comprise the property of slower dissolution or disintegration, e.g., disintegration within about 2 minutes to about 10 minutes or more.

Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.

Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the F3C(s) such excipients as are known in the art to be appropriate. /£ 9 21 22 Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats, salts (e.g., NaCl, potassium or sodium carbonate or bicarbonate or potassium or sodium phosphates) and solutes which render the formulation isotonic with the blood of the intended subject; and aqueous and non-aqueous sterile suspensions which may include suspending agents or thickening agents. In general, the F3C that is present in liquid compositions or formulations is completely dissolved in aqueous or non-aqueous excipients. However, in some embodiments, e.g., transient compositions or some formulations, the F3C is partially dissolved while the remaining portion is present as a solid, which can be a suspension or a colloid.

Formulations suitable for parenteral delivery of F3Cs to subjects such as humans or animals typically comprise 1, 2, 3, 4, 5, 6 or more excipients. Exemplary embodiments include (1) any two, three or four of propylene glycol, PEG200, PEG300, ethanol, benzyl alcohol and benzyl benzoate and (2) any two, three or four of propylene glycol, PEG 100, PEG200, PEG300, PEG400, benzyl alcohol and benzyl benzoate. Typically such fonnulations will contain both propylene glycol and one or more PEGs, e.g., PEG 100, PEG200, PEG300 or PEG400, which enhance the solubility of the F3C by a co-solvent effect.

Fonnulations, or compositions disclosed herein for use to make formulations suitable for administration by the routes disclosed herein optionally comprise an average particle size in the range of about 0.01 to about 500 microns, about 0.1 to about 100 microns or about 0.5 to about 75 microns. Average particle sizes include a range between 0.01 and 500 microns in 0.05 micron or in 0.1 micron or other increments, e.g., an average particle size of about 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.7, 1, 1.5,2.0,2.5, 3.0, 3.5,4.0,4.5, 5.0, 5.5,6, 7, 8, 9, 10,15, 20, 25, 30, 35, 40, 50, 60, 75, 85, 100, 120, 150, etc. microns). The F3C itself that is used to make a formulation can have one, two or more of these average particle sizes. When F3Cs or compositions that comprise a F3C are used as intermediates to make a formulation, they may comprise one, two, three or more of these average particle sizes, or size ranges. In preparing any of the compositions or formulations that are disclosed herein and that ΰ 6 0 J ί 2 comprise a F3C (and optionally one or more excipients), one may optionally mill, sieve or otherwise granulate the compound or composition to obtain a desired particle size, e.g., as described above.

Milling or micronisation by any other method may occur before or after the F3C is contacted with one or more excipients. For example, one may mill a F3C to obtain an average particle size (or diameter) of about 0.05-50 .mu.M or about 0.5-10 .mu.M (e.g., about 0.02, 0.04, 0.05, 0.07, 0.1, 0.5, 1, 1.5, 2, 2.5, 5, 10, 15, 20, 30, 40, 50, 60, 80, 100 or 120 .mu.M average particle size or diameter) before contacting the milled F3C with a liquid or solid excipient.

In some cases the F3C is milled or sieved to obtain an average particle size of about 5 .mu.m or about 10 .mu.m before it is contacted with a solid or liquid excipient(s) to obtain a solution or suspension or a powder suitable for making a tablet, capsule or other dosage fonn as described herein or in the cited references. Micronized compound may be prepared using any suitable process for obtaining small particles, e.g., controlled precipitation from a solution, micronizing or milling, a number of which are known in the art. The micronized particles may include a percentage of particles that are less than or equal to about 0.1-20 .mu.m in diameter. Ranges of average particle sizes include F3Cs of about 0.04-0.6 .mu.m, about 0.04-1.0 .mu.m, about 0.05-0.6 .mu.m, about 0.05-1.0 .mu.m, about 0.1-0.4 .mu.m, about 0.5-1 .mu.m, about 1-20 ,mu.m or about 2-50 .mu.m.

As used herein, reference to an average particle size or an average particle diameter means that the material, e.g., a F3C(s), an excipient(s) or a composition that comprises both, is ground, milled, sieved or otherwise treated so as to comprise the specified average size. It is to be understood that some particles may be larger or smaller, but the composition or the F3C(s) will comprise a significant proportion of the material with the specified size or within an acceptable range of the specified size, e.g., at least about 70% or about 80% of the particles within about 30% to about 50% of the average size or diameter. Micronization methods include milling by ball mills, pin mills, jet mills (e.g., fluid energy jet mills) and grinding, sieving and precipitation of a compound(s) from a solution, see, e.g., U.S. Pat.

Nos. 4,919,341, 5,202,129, 5,271,944, 5,424,077 and 5,455,049. Average particle size is determined by known methods, e.g., transmission electron microscopy, scanning electron l£ yt>0 3 microscopy, light microscopy, X-ray diffractometry, light scattering methods or Coulter counter analysis.

Thus, the F3Cs may comprise a powder that consists of one, two or more of these average particle sizes and the powder may be contacted with a solid excipient(s), suitably mixed and optionally compressed or formed into a desired shape. Alternatively, such a F3C(s) is contacted with a liquid excipient(s) to prepare a liquid formulation or a liquid composition that is incorporated into a solid formulation. Suitable micronized formulations thus include aqueous or oily solutions or suspensions of the F3C(s).

Fonnulations suitable for aerosol administration typically will comprise a fine powder, e.g., having an average particle size of about 0.1 to about 20 microns or any one, two or more of the average particle sizes within this range that are described above. The powder is typically delivered by rapid inhalation through the nasal passage or by inhalation through the mouth so as to reach the bronchioles or alveolar sacs of the lungs.

Fonnulations suitable for aerosol, dry powder or tablet administration may be prepared according to conventional methods and may be delivered with other therapeutic agents such as compounds heretofore used in the treatment or prophylaxis of infections as described herein. Such formulations may be administered, e.g., orally, parenterally (e.g., intravenous, intramuscular, subcutaneous, intradermal, intrathecal), topically, sublingually or by a buccal or sublingual route.

Micronized F3C is useful, e.g., to facilitate mixing, dissolution or uniform suspension of the F3C in one or more liquid or solid excipients, e.g., a PEG such as PEG 300 or PEG 400, propylene glycol, benzyl benzoate, a complexing agent, such as a cyclodextrin (e.g., an .alpha.-, .beta,- or .gamma.-cyclodextrin such as hydroxypropyl-.beta.-cy- clodextrin). Micronized F3C is also useful to facilitate uniformly distributing drug substance when the micronized compound is contacted with one or more solid excipients (e.g., a filler, a binder, a disintegrant, complexing agent (e.g., a cyclodextrin such as hydroxypropyl-.beta.IE Ogo 312 cyclodextrin), a preservative, a buffer or a lubricant).

In related embodiments, suitable compositions or fonnulations comprise a F3C that is present in two or more physical forms. For example, a liquid composition or formulation may comprise a F3C that is present in solution and as undissolved particles, which may be milled as described herein. Alternatively, a solid composition or formulation may comprise a F3C that is present as an amorphous fonn and as a crystal or in an encapsulated granule.

Such encapsulated granules may comprise a slow release type formulation and the F3C that is present may be in one or more physical forms, e.g., liquids or solids as described herein, but usually as a solid in tablets or other solid fonnulations.

The fonnulations are presented in unit-dose or multi-dose containers, for example sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example water for injection, immediately prior to use. In general, solid, liquid or other fonnulations or compositions that comprise a F3C, e.g., unit dosages for solid or liquid fonnulations, are stored in a sealed container, which may optionally be opaque or nearly opaque (e.g., amber or blue glass or brown plastic) to reduce the amount of light that reaches the formulation or composition. Such containers are also optionally sealed, e.g., hermetically sealed, to prevent or limit exchange of air, water or other gases between the container's contents and air. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets as described above. Unit dosage fonnulations are those containing a daily dose or unit daily-sub-dose, as recited herein, or an appropriate fraction thereof, of the F3C(s).

It should be understood that in addition to the ingredients particularly mentioned above the formulations of this invention may include other agents or .excipients conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents. Excipients include liquids, such as benzyl benzoate, cottonseed oil, Ν,Ν-dimethylacetamide, a C.sub.2-12 alcohol (e.g., ethanol), glycerol, peanut oil, vitamin E, poppyseed oil, safflower oil, sesame oil, soybean oil and If 06 0 312 vegetable oil. Excipients may optionally exclude one or more excipient, e.g., chloroform, dioxane, vegetable oil, DMSO, other excipients or any combination of these. Other excipients are components typically used in the pharmaceutical formulation arts, e.g., one, two or more of fillers, binders, disintegrants, dispersants, preservatives, glidants and lubricants, e.g., povidone, crospovidone, com starch, carboxymethyl cellulose, hydroxypropyl methylcellulose, microcrystallme cellulose, gum arabic, polysorbate 80, butylparaben, propylparaben, methylparaben, BHA, EDTA, sodium lauryl sulfate, sodium chloride, potassium chloride, titanium dioxide, magnesium stearate, castor oil, olive oil, vegetable oil, buffering agents such as sodium hydroxide, monobasic sodium phosphate, dibasic sodium phosphate, potassium hydroxide, monobasic potassium phosphate, dibasic potassium phosphate, tribasic potassium phosphate, potassium carbonate, potassium bicarbonate, ammonium hydroxide, ammonium chloride, saccharides such as mannitol, glucose, fructose, sucrose or lactose any of which may be compressible or any of which may be spray dried, milled, micronized or otherwise treated to obtained desired characteristics.

Formulations made from or comprising a F3C are optionally stored under conditions that limit the amount of light or water that reaches the formulation, e.g., in a sealed container that holds a formulation or unit dosage form and optionally contains silica gel or activated carbon. Water permeation characteristics of containers have been described, e.g., Containers-Permeation, Chapter, USP 23, 1995, U.S. Pharmacopeial Convention, Inc., Rockville, Md., p. 1787. Storage of BrEA hemihydrate or formulations that contain it is typically at about 4-30.degree. C.

The invention further provides veterinary compositions comprising at least one F3C together with a veterinary excipient(s) thereof. Veterinary excipients are materials useful for the purpose of administering the composition and may be solid, liquid or gaseous materials that '' are otherwise inert or acceptable in the veterinary art and are compatible with the F3C(s).

These veterinary compositions may be administered orally, parenterally or by any other desired route. ΙΕ θ 6 0 3 12 Invention formulations include controlled release or slow release formulations containing a F3C(s) in which the release of the F3C(s) is controlled or regulated to allow less frequency dosing or to improve the pharmacokinetic or toxicity profile of a given F3C(s). Polymers and other materials that is suitable to prepare controlled release formulations that comprise a F3C are known.

Formulations may thus contain microcapsules, granules or other shaped forms and may comprise a F3C and a slow release polymer or polymer matrix that comprises or consists of one or more of ethylene dimethacrylate, diethylene glycol dimethacrylate, diethylene glycol diacrylate, triethylene glycol dimethacrylate, triethylene glycol diacrylate, tetrathylene glycol dimethacrylate, tetraethylene glycol diacrylate, polyethylene glycol dimethacrylate, polyethylene glycol diacrylate, diethylaminoethyl dimethacrylate, glycidyl methacrylate, epoxy acrylate, glycidyl acrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, hydroxypropyl methacrylate, hydroxypropyl acrylate, hydroxybutyl methacrylate, hydroxybutyl acrylate, hydroxyhexyl methacrylate, hydroxyhexyl acrylate, butanediol dimethacrylate, butanediol diacrylate, propanediol dimethacrylate, propanediol diacrylate, pentanediol dimethacrylate, pentanediol diacrylate, hexanediol dimethacrylate, hexanediol diacrylate, neopentyl glycol dimethacrylate, neopentyl glycol diacrylate, trimethylopropane triacrylate, trimethylolpropane trimethacrylate, trimethyloethane triacrylate, trimethylolethane trimethacrylate, polypropyleneglycol diacrylate, and polypropylene glycol dimethacrylate.

Formulations may comprise a liposome or lipid complex that comprises or contains a F3C(s). Such formulations are prepared according to known methods, e.g., U.S. Pat. Nos. 4,427,649, 5,043,165, 5,714,163, 5,744,158, 5,783,211, 5,795,589, 5,795,987, 5,798,348, 5,811,118, 5,820,848, 5,834,016 and 5,882,678. The liposomes optionally contain an additional therapeutic agent(s), e.g., amphotericin B. cis-platin, adriamycin, a protease inhibitor, a nucleoside or a nucleotide analog, such as one of those mentioned herein. Formulations that comprise liposomes can be delivered to a subject by any standard route, e.g., oral, aerosol or parenteral (e.g., s.c., i.v. or i.m.).

IE 0 so 312 Liposome formulations can be used to enhance delivery of the F3C(s) to certain cell types such as tumor cells (see e.g., U.S. Pat. No. 5,714,163) or to cells of the reticuloendothelial system (RES). The RES includes macrophages, mononuclear phagocytic cells, Kupfer cells, cells lining the sinusoids of the spleen, lymph nodes, and bone marrow, and the fibroblastic reticular cells of hematopoietic tissues. In general, RES cells are phagocytic and they are targets for targeted delivery of a F3C(s) in vitro or in vivo using liposomes, or other compositions or fonnulations. Thus, one can deliver F3C to a neoplasm that is derived from reticuloendothelial tissue (reticuloendothelioma). The liposomes may optionally comprise a peptide from an infectious agent such as a malaria parasite, a virus or a tumor associated antigen. The peptides may facilitate the generation of a MHC class II and B cell response. In other cases a liposomal F3C formulation is useful to obtain suspension F3C fonnulations.

Invention embodiments include the product made by a process of combining, mixing or otherwise contacting a F3C and one, two or more excipients. Such products are produced by routine methods of contacting the ingredients. Such products optionally contain a diluent, a disintegrant, a lubricant, a binder, or other excipients described herein or in references cited herein.

Other embodiments include compositions that transiently occur when a method step or operation is performed. For example, when a F3C, containing less than about 3% w/w water is contacted with an excipient, e.g., a PEG, an alcohol, propylene glycol benzyl alcohol or benzyl benzoate, the composition before addition of one ingredient with another is a nonhomogenous mixture. As the ingredients are contacted, the mixture's homogeneity increases and the proportion of ingredients relative to each other approaches a desired value. Thus, invention compositions, which contain less than about 3% w/w or less than about 2% w/w or less than about 1% w/w or less than about 0.5% w/w water can comprise about 0.0001-99% w/w of a F3C and 1, 2, 3 or more excipients. These transient compositions are intennediates that necessarily arise when one makes an invention composition or formulation and they are included in invention embodiments.

When a F3C and an exeipient(s) is contacted or mixed, the final composition may comprise a homogenous mixture or it may comprise a mixture that is not homogenous for one or more of the compounds that are present in the composition. Compositions and formulations that are either homogenous or non-homogenous are included in the scope of the invention. Nonhomogenous compositions can be used, e.g., to make controlled release formulations.

Invention embodiments include compositions and formulations that comprise less than about 3% water, a F3C and a compound that is not generally considered suitable for human use but is useflil to make an invention formulation for veterinary use. Veterinary formulations are compositions useful for the purpose of administering invention compositions to primates, cats, dogs, horses, cows, rabbits and other subjects and may contain excipients acceptable in the veterinary art and are compatible with F3Cs. These veterinary compositions may not always be suitable for human use because they contain an excipient that is not suitable for human use, e.g., an alcohol other than ethanol such as methanol, propanol or butanol. Typically such excipients will be present at relatively low levels, e.g., about 1-30%, usually about 1-5%.

Invention embodiments include non-aqueous compositions and formulations, e.g., unit dosage forms and sterile solutions or suspensions, that comprise about 1-25% w/w of a F3C, about 20-60% w/w propylene glycol, about 15-55% w/w of more or more PEGs, e.g., PEG100 or PEG200, about 0-5% w/w benzyl benzoate, about 0-5% w/w benzyl alcohol and optionally one or more additional excipients. These non-aqueous formulations will usually contain less than about 3%, 2%, 1%, 0.8%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% w/w of water. In formulations that contain non-aqueous excipients, the F3C will usually be relatively hydrophobic and will usually not contain any easily charged or ionizable moieties such as free carboxyl groups.

F3Cs may be administered to subjects by transmucosal dosing, e.g., by buccal or sublingual administration. The buccal area generally refers to the subject's mouth and pharynx, and the Ι£ ϋ § q j 12 buccal mucosa includes the mucosa of the mouth and pharynx. The sublingual area refers generally to the mucosa below and adjacent to the tongue. Formulations suitable for buccal or sublingual administration typically comprise about 1-100 mg of F3C per unit dose, often about 2-60 mg. Transmucosal dosages may comprise a slow release or a rapid release formulation or tablet that contains about 1, 5,10, 15, 20, 25, 30, 35, 40, 50 or 60 mg of a F3C. Slow release formulations will generally degrade or release the F3C from the dosage over a period of about 2 minutes to about 60 minutes or more. Rapid release formulations will generally release the F3C over a period of about 4 seconds to about 2 minutes, typically over about 0.1 to about 1 minute. Solid and liquid buccal or sublingual formulations optionally include one, two, three or more excipients such as fillers, binders, lubricants, antioxidants, preservatives, flavoring agents or disintegrants, e.g., lactose, sucrose, mannitol, Tween-80, magnesium stearate, butylated hydroxyanisole, butylated hydroxytoluene, cyclodextrins (e.g., a-cyclodextrins,.beta.-cyclodextrins, .gamma.-cyclodextrins, hydroxypropyl-.beta.-cyclodextrin, .beta.-cyclodextrin ether comprising one or more hydroxybutyl sulfonate moieties, cyclodextrins as described in U.S. Pat. No. 6,610,671 or U.S. Pat. No. 6,566,347), carbomers, hydrolyzed polyvinylalcohol, polyethylene oxide, polyacrylates, hydroxypropylmethylcellulose, hydroxypropylcellulose, and combinations thereof. Such formulations may be a unit solid such as a tablet or powder or a liquid. Buccal tablets may comprise a concave surface for contacting the buccal mucosa and adhering to it. A buccal or sublingual dosage may comprise a compressed tablet of a substantially unifonn mixture of a bioerodible polymeric carrier, which on sustained contact with the oral mucosa, substantially or completely erodes within a predetennined period in the range of about 10 minutes to about 24 hours. In some embodiments, the F3C is administered by a method for administering the compound to the subject, e.g., to a mammal or a human, comprising affixing a unit dosage or tablet to the subject's buccal mucosa in a region at or near the upper gum between the first bicuspid on the left and the first bicuspid on the right (or an alternative location for the dosage unit is the inner lip area opposing the this upper gum area) and optionally allowing the tablet to remain in place until erosion thereof is complete or nearly complete. Exemplary excipients may comprise a combination of polyethylene oxide and a carbomer, e.g., wherein the polyethylene oxide and the carbomer are in an ,Eu60jf2 approximately 1:5 to 5:1 ratio by weight.

Tablets or unit dosages for buccal or sublingual delivery may be about 5 mm in diameter and 2 mm in height, so that the unit dosage occupies about 40 mm.sup.3. Such dosages will typically weigh less than about 100 mg (e.g., about 5 to 60 mg), with a contact surface area of about 10-30 mm.sup.2, e.g., about 15-20 mm.sup.2. Such dosages will generally be about 4-10 mm in diameter and about 1-3 mm in height. When a polymer excipient is used, it optionally comprises a polymer having sufficient tack to ensure that the dosage unit adheres to the buccal mucosa for a sufficient time period, e.g., the time period during which drug is to be delivered to the buccal mucosa. The polymeric excipient is gradually bioerodible, and it hydrolyzes, dissolves, erodes or disintegrates (collectively erodes) at a predetermined rate upon contact with water or saliva. The polymeric carrier is generally sticky when moist, but not when dry, for convenience in handling. The average molecular weight of the polymer may be about 400 to 1,000,000, or about 1,000 to 100,000. Higher the molecular weight polymers generally erode more slowly.

For these buccal and sublingual dosages, a pharmaceutically acceptable polymer(s) can be used. Such polymers will provide a suitable degree of adhesion and the desired drug release profile, and are generally compatible with the drug to be administered and any other components that may be present in the buccal dosage unit. The polymeric carriers optionally comprise hydrophilic (water-soluble and water-swellable) polymers that adhere to the wet surface of the buccal mucosa. Examples of polymeric carriers that are useful herein include acrylic acid polymers, e.g., those known as carbomers (Carbopo.TM., which may be obtained from B. F. Goodrich, is one such polymer). Other suitable polymers include hydrolyzed polyvinylalcohol; polyethylene oxides (e.g., Sentry Polyox.TM. water soluble resins, available from Union Carbide); polyacrylates (e.g., Gantrez.TM., which may be obtained from GAF); vinyl polymers and copolymers; polyvinylpyrrolidone; dextran; guar gum; pectins; starches; and cellulosic polymers such as hydroxypropyl methylcellulose, (e.g., Methocel.TM., which may be obtained from the Dow Chemical Company), hydroxypropyl cellulose (e.g., Klucel.TM., which may be obtained from Dow), v 6 0 3 ί 2 hydroxypropyl cellulose ethers (see, e.g., U.S. Pat. No. 4,704,285 to Alderman), hydroxyethyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, cellulose acetate phthalate, cellulose acetate butyrate, and the like. The carrier may also comprise two or more suitable polymers in combination, for example, a carbomer combined in an approximately 1:5 to 5:1 ratio, by weight, with a polyethylene oxide.

Buccal dosages may contain only the F3C and the polymer(s). However, it may be desirable in some cases to include one or more additional excipients. For example, a lubricant may be included to facilitate the process of manufacturing the dosage units; lubricants may also optimize erosion rate and drug flux. If a lubricant is present, it may optionally represent about 0.01 wt. % to about 2 wt. %, or about 0.01 wt. % to 0.5 wt. %, of the dosage unit. Suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, sodium stearylfumarate, talc, hydrogenated vegetable oils and polyethylene glycol. However, modulating the particle size of the components in the dosage unit and/or the density ofthe unit can provide a similar effect, e.g., improved manufacturability, and optimization of erosion rate and drug flux without addition of a lubricant.

Other excipients are also optionally incorporated into buccal unit dosages. Such additional optional excipients include one or more disintegrants, diluents, binders, enhancers, or the like. Examples of disintegrants that may be used include, but are not limited to, cross-linked polyvinylpyrrolidones, such as crospovidone (e.g., Polyplasdone.TM. XL, which may be obtained from GAF), cross-linked carboxylic methylcelluloses, such as croscarmelose (e.g., Ac-di-sol.TM., which may be obtained from FMC), alginic acid, and sodium carboxymethyl starches (e.g., Explotab.TM., which may be obtained from Edward Medell Co., Inc.), methylcellulose, agar bentonite and alginic acid. Suitable diluents are those which are generally useful in pharmaceutical formulations prepared using compression techniques, e.g., dicalcium phosphate dihydrate (e.g., Di-Tab.TM., which may be obtained from Stauffer), sugars that have been processed by cocrystallization with dextrin (e.g., cocrystallized sucrose and dextrin such as Di-Pak.TM., which may be obtained from Amstar), u6 0 3 12 lactone, calcium phosphate, cellulose, kaolin, mannitol, sodium chloride, dry starch, powdered sugar and the like. Binders, if used, are those that enhance adhesion. Examples of such binders include, but are not limited to, starch, gelatin and sugars such as sucrose, dextrose, molasses, and lactose. Permeation enhancers may also be present in the novel dosage units in order to increase the rate at which the active agent passes through the buccal mucosa. Examples of permeation enhancers include, but are not limited to, polyethylene glycol monolaurate (PEGML), glycerol monolaurate, lecithin, the 1-substituted azacycloheptan-2-ones, particularly l-n-dodecylcyclaza-cycloheptan-2-one (available under the trademark Azone.TM. from Nelson Research & Development Co., Irvine, Calif.), lower alkanols (e.g., ethanol), SEPA.TM. (available from Macrochem Co., Lexington, Mass.), cholic acid, taurocholic acid, bile salt type enhancers, and surfactants such as Tergitol.TM., Nonoxynol-9.TM. and TWEEN-8.TM..

Flavorings are optionally included in buccal or sublingual formulations. Any suitable flavoring may be used, e.g., one or more of mannitol, sucrose, glucose, lactose, lemon, lemon lime, orange, menthol or artificial sweeteners such as aspartame, saccharin sodium, dipotassium glycyrrhizinate, stevia and thaumatin. Some sweeteners such as sucrose may also aid in dissolution or erosion of solid formulations. Coloring agents may also be added, e.g., any of the water soluble FD&C dyes or mixtures thereof, e.g., one or more of FD&C Yellow No. 5, FD&C RED No.2, FD&C Blue No.2, etc., food lakes or red iron oxide. In addition such formulations dosages may be formulated with one or more preservatives or bacteriostatic agents, e.g., methyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol, benzalkonium chloride, or the like.

Other embodiments include solid buccal or sublingual formulations comprising (i) a F3C formulation and (ii) erythritol, (iii) crystalline cellulose and (iv) a disintegrant, e.g., crospovidone. These formulations are capable of buccal disintegration or dissolution and may further comprise mannitol. These formulations may dissolve completely in solely saliva within about 1-10 minutes of administration to a subject. The erythritol is optionally contained in a proportion of about 5-90 parts by weight, based on 100 parts by weight of the /£uo03J2 solid buccal formulation. The crystalline cellulose is optionally contained in a proportion of about 3-50 parts by weight, based on 100 parts by weight of the formulation. The disintegrant is optionally contained in a proportion of 1-10 parts by weight. In any of the solid buccal or sublingual formulations the ingredients are generally uniformly mixed, although non-uniform mixtures may be used. An exemplary formulation comprises a solid capable of buccal disintegration or dissolution, which comprises (i) about 0.3-50 parts by weight of a F3C, (ii) about 50-80 parts by weight of erythritol, (iii) about 5-20 parts by weight of crystalline cellulose and (iv) about 3-7 parts by weight of a disintegrant, which optionally is one or more of crospovidone, croscannellose, croscarmellose sodium, carmellose calcium, carboxymethylstarch sodium, low substituted hydroxypropyl cellulose or com starch. Examples of the crystalline cellulose include products of various grade such as CEOLUS KG801, avicel PH101, avicel PH102, avicel PH301, avicel PH302, avicel RC13 591 (crystalline cellulose carmellose sodium) and so on. One crystalline cellulose may be used or two or more species may be used in combination. The disintegrant, e.g., crospovidone, may be used singly or in combination with other disintegrants. Crospovidone includes any cross-linked l-ethenyl-2-pyrrolidinone homopolymer, and may comprise a polymer of molecular weight of 1,000,000 or more. Examples of commercially available crospovidone include Cross-linked povidone, Kollidon CL, Polyplasdone XL, Polyplasdone XL-10, INF-10 (manufactured by ISP, Inc.), polyvinylpolypyrrolidone, PVPP and l-vinyl-220 pyrrolidinone homopolymer. The disintegrants are optionally incorporated in a proportion of about 1-15 parts by weight, or about 1-10 parts by weight, or about 3-7 parts by weight, based on 100 parts by weight of the solid formulation.

Some embodiments include a solid buccal or sublingual formulation containing a F3C where unit doses of the formulation substantially or completely disintegrates or erodes within about -120 seconds in water at 37.degree. C. or on insertion of the unit dose into the buccal area x or upon placement under the tongue. Such formulations may comprise a swellable hydrophilic excipient, a water-soluble or a water-dispersible excipient, e.g., one or more of partially hydrolyzed gelatin, hydrolyzed dextran, dextrin, mannitol, alginates, polyvinyl alcohol, polyvinyl pyrrolidine, water soluble cellulose derivatives, methylcellulose, ethyl “003,2 cellulose, carboxymethyl cellulose, hydroxymethylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, alginates, gelatin, guar gum, gum tragacanth, gum acacia, polyacrylic acid, polymethacrylic acid, polysilicic acid, polylactic acid, polymaleic acid, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, nonionic blocked polymers, carbomers, polycarbophils, a water soluble starch, dicalcium phosphate, calcium carbonate, silica or polyethyleneglycol, e.g., PEG 1000, PEG2000 or a polyethylene oxide (PEO'j, PE01000, ΡΕΘ100000 or PE05000000.

Other embodiments include the product obtained by storing invention compositions or formulations, e.g., unit dosage forms or compositions used to make formulations, at about 44O.degree. C. for at least about 30 days, e.g., storage at ambient temperature for about 1-24 months. Invention formulations will typically be stored in hermetically or induction sealed containers for these time periods. Compositions and formulations that comprise a F3C will typically be held in closed or sealed containers, particularly when the composition is a fonnulation for pharmaceutical or veterinary use.

Typical containers for storage of compositions and formulations that comprise a F3C will limit the amount of water that reaches the materials contained therein. Typically, formulations are packaged in hermetically or induction sealed containers. The containers are usually induction sealed. Water permeation characteristics of containers have been described, e.g., Containers—Permeation, chapter, USP 23 <671>, United States Pharmacopeial Convention, Inc., 12601 Twinbrook Parkway, Rockville, MD 20852, pp.: 1787 et seq. (1995).

Immune modulation. The F3Cs, or the biologically active substances produced from these compounds by hydrolysis or metabolism in vivo, have a number of clinical and non-clinical χ applications. The compounds are generally useful to correct immune dysregulation, e.g., imbalanced immune responses to disease conditions, pathogens or the like, suppression of an innate or acquired immune response(s) and inflammation conditions in vertebrate or mammalian subjects, e.g., as disclosed herein. Thus, while the compounds will generally IE Ο 6 ο 312 enhance a deficient immune response in a given clinical condition, they will generally reduce the same immune response when it is too active in a different clinical condition. For example, they can enhance insufficient or suboptimal Thl immune responses, reduce excess or undesirable Th2 immune responses, reduce excess or undesirable Th 1 immune responses or enhance insufficient or suboptimal Th2 immune responses or they can reduce excess or undesirable inflammation or one or more of its symptoms. The compounds will generally also modulate dysregulated Tel and Tc2 immune responses (associated with CD8.sup.+ T cells) in a similar manner, e.g., excessive. Tel or Tc2 responses will be detectably decreased and deficient or suboptimal Tel or Tc2 responses will generally be detectably enhanced.

Invention embodiments include a method to modulate a subject's innate immunity, Thl immune responses, Tel immune responses, Th2 immune responses or Tc2 immune responses comprising administering an effective amount of a F3C to a subject or delivering the F3C to the subject's tissues. Other methods include modulating an immune or cellular response in a subject in need thereof comprising administering to the subject, or delivering to the subject's tissues, an effective amount of a compound of formula 1. Immune and cellular response modulation includes enhancing Thl immune responses, reducing Th2 immune responses, reducing Thl immune responses, enhancing Th2 immune responses, reducing unwanted or pathological inflammation, enhancing hematopoiesis or modulating the synthesis, level or a biological activity of a biomolecule such as (1) a transcription factor such as a nuclear hormone receptor or an associated receptor factor, (2) a purine such as adenosine, (3) a nucleotide cofactor such as NADPH, (4) a cytokine or interleukin or a receptor for a cytokine or interleukin, or (5) another biomolecule as disclosed herein. Such enhancements, reductions, levels or activities are usually in an easily detectable range, e.g., a change compared to a suitable control of at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or a range that is between about any two of these values.

Typically the subject is in need of such treatment, e.g., by having a clinical condition disclosed herein or being subject to developing such a condition, e.g., having been exposed or potentially exposed to a pathogen or having a predisposing condition such as pre-cancer. 6 0 J 12 In modulating one or more activities of Thl, Th2, Tel or Tc2 cells or their function(s), the F3Cs will typically detectably modulate one, two, three or more factors, e.g., immune cell subsets or populations, cytokines, interleukins, surface antigens such as a CD molecule(s) and/or their receptors that affect the development, migration, numbers or biological function(s) of such cells. When a Thl or Tel cell or population is affected, the F3Cs will typically increase or decrease the synthesis or level of one, two or more of an associated effector factor, e.g., IFN.gamma., IL-2, IL-12, IL-18, T-bet, PPAR.alpha. and PPAR.gamma. or a cell surface molecule, e.g., as disclosed herein or in the cited references, that is associated with or needed for normal, optimal or enhanced Thl or Tel cells or cell function. Such molecules are generally associated with development or enhancement of Thl or Tel cells or their biological function(s). When a Th2 or Tc2 cell or population is affected, the F3Cs will typically increase or decrease the synthesis or level of one, two or more of an associated effector factor, e.g., IL4, IL-5, IL-6, IL-8, IL-10, IL-13, GATA-3, COX-2 or a cell surface molecule, e.g., as disclosed herein or in the cited references, that is associated with or needed for normal, optimal or enhanced Th2 or Tc2 cells or cell function(s). Such molecules are generally associated with development or enhancement of Th2 or Tc2 cells or their biological function(s).

Similarly, when a subject has or is subject to developing an unwanted or excessive inflammation, the F3Cs will generally detectably modulate one or more relevant effector factors for inflammation, e.g., a detectable decrease of one, two, three or more of IL1.alpha., IL-1.beta., TNF.alpha., TNF.beta., MIP-1.alpha., MIP-2, TGF-.beta.l, IP-10, LT,beta., .gamma.IFN, IL-6, IL-8, IL-10 and COX-2, lipoxygenase, or an increase of one or more suppressor factors or antagonists of inflammation. Such modulation can comprise increases or decreases of at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 100%, 200%, 300%, 500%, 1000%, 5000% or within a range between any two of these values, e.g., between about 5-95%, about 1090%, about 5-60% or about 40-95%. In general, such changes leads to a detectable amelioration of an inflammation-associated disease, condition, symptom or to the detectable slowing of the progression thereof or to a detectably reduced incidence or severity of or ΙΕ ο ό 0 3 12 susceptibility to developing an unwanted inflammatory response.

In conditions where an unwanted or excessive Thi, Tel, Th2 or Tc2 response is associated with or causes a disease(s), disease(s) progression, disease(s) state maintenance, condition(s) or symptom(s), the F3Cs will generally decrease the level or one or more biological activity of one, two or more of their respective associated effector molecules. In conditions where a deficient or suboptimal Thi, Tel, Th2 or Tc2 response is associated with or causes a disease(s), disease(s) progression, disease(s) state maintenance, condition(s) or symptom(s), the F3Cs will generally increase the level or one or more biological activity of one, two, three or more of their respective associated effector molecules. Such changes in the level or biological activity(ies) the associated effector molecules is generally detectable using standard methods and is typically an increase (when a response is insufficient) or a decrease (when a response is in excess) of at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or within a range between any two of these values, e.g., between about 5-95%, about 10-90%, about 5-60% or about 40-95%. In general, such changes leads to a detectable amelioration of a disease, condition, symptom or to the detectable slowing of the progression thereof or to a detectably reduced incidence or severity of or susceptibility to developing a disease(s) or the occurrence of a symptom(s) for a at least a portion of subjects that are treated with a F3C, e.g., at least about 5%, 10%, 20%, 40%, 60% or 80% of treated subjects. The F3Cs may facilitate the clinical cure of a disease(s), prolong remission of a disease(s) or eliminate or ameliorate a clinically detectable symptom(s).

The F3C will generally also affect the function of other immune cell subsets in a similar manner. Thus, when an insufficient macrophage, dendritic cell or neutrophil response is associated with the establishment, maintenance or progression of a disease, symptom or a condition, the F3Cs will generally enhance of the level or a biological activity(ies) of one or more effector molecule associated with or needed for an optimal or more normal response or immune function that is mediated by the macrophages, dendritic cells or neutrophils. Similarly, when the subject suffers from a excessive or pathological activity associated with 0603J2 macrophages, dendritic cells or neutrophils, which is associated with the establishment, maintenance or progression of a disease, symptom or a condition, the F3Cs will generally detectably reduce the level or a biological activity(ies) of one or more effector molecule associated with or needed for an optimal or more normal response or immune function that is mediated by the macrophages, dendritic cells or neutrophils. Such effector molecules are as described herein or in the cited references.

As used herein, reference to Thl or Th2 immune responses means such responses as observed in mammals generally and not as observed in the murine system, fom which the Thl and Th2 terminology originated. Thus, in humans, Thl cells are CD4.sup.+ T lymphocytes and they usually preferentially display chemokine receptors CXCR3 and CCR5, while Th2 cells are CD4.sup.+ T lymphocytes and usually preferentially express the CCR4, CCR8 and/or CXCR4 chemokine receptor molecule(s) and generally a smaller amount of CCR3, see, e.g., U. Syrbe et al., Springer Semin. Immunopathol. 1999 21:263285, S. Sebastiani et al., J. Immunol. 2001 166:996-1002. Tel and Tc2 immune responses are mediated by CD8.sup.+ lymphocytes and means to identify these cells and their associated lymphokines, cell specific antigens and biological activities have been described, see, e.g., Μ. B. Faries et al., Blood 2001 98:2489-2497, W. L. Chan et al., J. Immunol. 2001 167:1238-1244, C. Prezzi et al., Eur. J. Immunol. 2001 31:894-906, H. Ochi et al., J. Neuroimmunol. 2001 119:297-305, D. H. Fowler and R. E. Gress, Leukemia and Lymphoma 2000 38:221-234.

The F3Cs are useful in re-establishing normal immune function in various immune dysregulation or immune suppression conditions. For example, they are useful to treat, slow progression of or to ameliorate one or more symptoms associated with one or more of an autoimmune condition(s), a inflammation condition(s), an infection(s), a cancer(s), a precancer(s), a chemotherapy(ies), radiation therapy, a burn(s), a trauma(s), a surgery(ies), a pulmonary condition, a cardiovascular disease(s) and a neurological or neurodegenerative disease(s).

The F3Cs are useful to modulate delayed-type hypersensitivity (DTH) responses and anergic conditions in subjects having to subject to developing abnormal DHT responses or anergy. Means to measure such responses and conditions are known and can be used to characterize the effects of the F3Cs on these responses and conditions. See, e.g., A. E.

Brown, et al., J. Med. Assoc. Thailand 83:633-639 2000, R. A. Smith et al., J. Adolesc.

Health 27:384-390 2000, Ν. M. Ampel, Med. Mycology 37:245-250 1999. The compounds will generally detectably enhance or restore DTH in immune suppression conditions. They will also generally detectably reduce or eliminate anergy in subjects having significantly reduced or no immune response to, e.g., specific antigens or pathogens.

The invention provides a method to detectably enhance an antigen specific immune response, cell mediated immune response or a delayed-type hypersensitivity immune response in a subject having impaired or negligible antigen specific immune response, cell mediated immune response or delayed-type hypersensitivity immune response, comprising administering to the subject, or delivering to the subject's tissues, an effective amount of a F3C. The antigen specific immune response, cell-mediated immune response or delayedtype hypersensitivity immune response can be enhanced at least about 25%, at least about 40%, at least about 50%, at least about 60%, at least about 75% or at least about 90%. Some of the subjects may have an antigen specific immune response, cell mediated immune response or a delayed-type hypersensitivity immune response that is impaired or negligible, e.g., about 50% or less or about 30% or less or about 10% or less of the response that an otherwise normal subject would be expected to have. Such subjects may not detectably respond to at least 1 antigen out of 2, 3, 4 or 5 antigens that a normal subject would respond to. The responses to treatment with a F3C may be quantitated by, e.g,, mixed lymphocyte reaction, ELIspot analysis or flow cytometric analysis of, e.g., circulating blood cells such as ' CD4.sup.+ or CD8.sup.+ T cells or for levels of cytokines (e.g,, IL-2, TNF.alpha. or IFN.gamma.) in such cells. Such analyses have been described, e.g., V. P. Badovinac and J.

T. Hardy, J. Immunol. Methods 2000,238:107-117, N. Favre et al., J. Immunol. Methods 1997, 204:57-66, E. Hagiwara et al., Cytokine 1995, 7:815-822, N. W. Lukacs et al., Blood °SO312 1993, 82:3668-3674, M. Umemoto et al., Clin. Exp. Immunol. 1998, 112:459463, A. Fietta et al.. Gerontology 1994, 40:237-245, C. H. Orteu et al., J. Immunol. 1998,161:1619-1629.

Clinical indications that have an association with or have a symptom(s) that is consistent or associated with an excessive or unwanted Th2 immune response include, e.g., fatigue, pain, fever or an increased incidence of infection, schizophrenia, acute myelitis, tumor progression, progressive systemic sclerosis, Omenn's syndrome, atopic disease, atopy, allergen hypersensitivity, atopic asthma, atopic dermatitis, bums, trauma (e.g., bone fracture, hemorrhage, surgery), immune responses to xenotransplantation, chronic periodontitis, SLE (systemic lupus erythematosus), discoid lupus erythematosus, osteoporosis, myasthenia gravis, Graves disease, mite-associated ulcerative dermatitis, rheumatoid arthritis and osteoarthritis. Excessive Th2 immune responses are also associated with an unwanted symptom or pathology, e.g., fatigue, pain, fever or an increased incidence of infection, that is associated with aging, allergy and inflammation conditions, allergic respiratory disease, allergic rhinitis, atopic dermatitis, subepithelial fibrosis in airway hyperresponsiveness, chronic sinusitis, perennial allergic rhinitis, fibrosing alveolitis (lung fibrosis). This common underlying immune component is at least part of the pathology or symptoms of all of these conditions. This allows a F3C to be effectively used to prevent or treat the condition or to treat or ameliorate one or more symptoms that are associated with these conditions. Thus, in some embodiments, an unwanted or excessive Th2 response is present and amelioration of one or more symptoms associated with this condition is accomplished by administering an effective amount of a F3C according to the methods described herein, e.g., F3C is administered using a formulation and a route of administration essentially as described herein on an intermittent or a daily basis.

Typically, unwanted Th2 immune responses are associated with, or caused by, increased expression of one or more cytokines or interleukins such as one, two, three or more of cortisol, IL4, IL-5, IL-6, IL-10 and IL-13. Administration of a F3C will generally reduce the expression of one or more of the Th2-associated cytokines or interleukins. At the same time, the compounds generally enhance the expression of one or more cytokines or interleukins Ούθ5ΐ2 associated with Thl immune responses. Because of their capacity to modulate or to balance Thl and Th2 immune responses, the compounds are useful for a variety of clinical conditions, e.g., infection, immunosuppression or cancer, where an enhanced Thl immune response is desired. Effects of the F3Cs in treating, preventing or slowing the progression of the clinical conditions described herein can include one or more of (1) enhancing the Thl character of a subject's immune response or immune status, (2) increasing the intensity of a Thl or a Th2 immune response or both and (3) decreasing inflammation or a symptom thereof.

Exemplary conditions where an immune imbalance or an excessive Thl immune response is involved include autoimmune diseases such as multiple sclerosis, Crohn's disease (regional enteritis), ulcerative colitis, inflammatory bowel disease, rheumatoid arthritis, reactive arthritis, acute allograft rejection, sarcoidosis, type 1 diabetes mellitus, Helicobacter pylori associated peptic ulcer, graft versus host disease and Hashimotos' thyroiditis. Because these conditions are associated with a similar type immune dysfunction, a F3C can be effectively used to prevent or treat these conditions or to treat or ameliorate one or more symptoms associated therewith. Thus, in some embodiments, an unwanted or excessive Thl response is present and amelioration of one or more symptoms associated with this condition is accomplished by administering an effective amount of a F3C according to the methods described herein, e.g., F3C is administered using a formulation and a route of administration essentially as described herein on an intermittent or a daily basis. In other embodiments, an deficient Thl response is enhanced, which is optionally observed as a detectable increase in one or more of IFN.gamma., IL-2, IL-12 or IL-18 in Thl cells or in accessory cells such as a dendritic cell or macrophage. In all of the conditions where an insufficient or excess Thl, Th2, Tel or Tc2 response is present, amelioration of one or more symptoms associated with the condition is accomplished by administering an effective amount of a F3C according to v the methods described herein.

Aspects of the invention include the use or administration of compositions or formulations that comprise a carrier and an amount of at least one F3C effective to detectably modulate an immune parameter. For example, to enhance the relative proportion of a desired immune cell subset, e.g., CD4.sup.+ T cells, CD8.sup.+ T cells, NK cells, LAK cells, neutrophils, granulocytes, basophils, eosinophils or dendritic cells, or to modulate (detectably increase or decrease) one or more functions of immune cell subsets. The F3Cs can modulate the expression of CD molecules or alter the proportion of cell subsets, e.g., CD4.sup.+ or CD8.sup.+ T cells, or their relative numbers in a subject's blood or tissues. CD and related molecules participate in the function of various immune cell subsets and can be useful as markers for immune function in vivo. In some aspects, the F3Cs activate immune cells which generally alters (increases or decreases) expression of, or changes the numbers of cells that express one or more of, CD4, CD6, CD8, CD25, CD27, CD28, CD30, CD36, CD38, CD39, CD43, CD45RA, CD45RO, CD62L, CD69, CD71, CD90 or HLA-DR molecules. Often, the numbers of cells that express these molecules are increased, e.g., CD25, CD36, CD16 or CD69. Typically, such increases are observed as an increased proportion of circulating white blood cells that express one or more of these molecules or white blood cells, e.g., T cells or dendritic cells, that express CXCR3, CCR5, CCR4, CCR8 and/or CXCR4. In some cases the number of such molecules per cell is detectably altered.

Expression of one or more adhesion molecules CD2, CD5, CD8, CD11 a, CD1 lb, GDI lc, CD18, CD29, CD31, CD36, CD44, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD50, CD54, CD58, CD 103 or CD 104 are also detectably modulated after administration of the F3Cs to a subject. Often, the numbers of cells that express these molecules are increased, e.g., CD5 or CD56. The adhesion molecules function in various aspects of immune responses, such as binding to class IMHC molecules, transducing signals between cells or binding to molecules in the extracellular matrix associated with endothelial or other cell types. Administration of the F3Cs to a subject also affects the numbers of certain immune cell subsets, e.g., NK cells (e.g., CD8.sup.-, CD56.sup.+ or CD8.sup.+, CD56.sup.+) or lymphokine activated killer cells (LAK). Increased circulating NK or LAK cells are typically observed, which is reflected in increased numbers of cells that express one or more of CD16, CD38, CD56, CD57 or CD94. Also, increased numbers of circulating dendritic cell precursors are observed, as shown by increases in cells that express one or more of CD1 lc, CD80, CD83, CD106 or CD123. Although one can observe an increased proportion of circulating white blood cells that express one or more of these molecules, in some instances the number of such molecules per cell is detectably altered. Both the cell numbers and the density of CD molecule per cell can also be detectably modulated. Modulation of immune cell subsets typically occurs on intermittent dosing of a F3C, but will arise from any suitable dosing regimen, e.g., as described herein.

Expression of one or more homing or other receptors or receptor subunits such as CD62L, CLA-1, LFA1, CD44, ICAM, VCAM or ECAM may also be detectably affected after administration of the F3Cs to a subject. The numbers of cells that express these molecules, or the relative amounts per cell of, e.g., CD44 or CD62L, may be increased where a desired immune response is desired, e.g., migration of T cells to mucosal tissues or exposure of nave T cells to antigen in lymph nodes. Alternatively, numbers of cells that express these molecules, or the relative amounts per cell of, e.g., CLA-1, may be decreased where inhibition of an undesired immune response, such as an inflammatory response is desired. The subject's response to such enhanced expression includes migration of cells such as movement of nave T cells to peripheral lymph nodes in response to modulation of CD62L or other homing receptor expression. Thus, the F3Cs can also facilitate migration of various immune cell types, e.g., dendritic cells, NK cells, LAK cells, macrophages or lymphocytes, from one location to another within a subject. For example, the compounds can enhance dendritic cell or lymphocyte migration from areas such as the skin tissues to the gut associated lymphoid tissue (GALT), lymph nodes or spleen. Such migration may facilitate the function of those cell types by increasing their transit to tissues where their effector functions, e.g., antigen presentation by dendritic cells, normally occur. The migration period is often relatively transient (e.g., observable over about 1-7 days) or occasionally longer (e.g., occurring for about 8-40 days), depending on the dosing regimen and other factors. This migration can be observed by standard methods, e.g., by cell staining, by PCR analyses or by detennining the presence of a given cell type in circulation or determining a decrease in the number circulating cells. A decrease would generally reflect sequestration of an immune cell population(s) in a tissue(s) where the immune cell normally exercises its effector functions.

Thus, in some embodiments, the migration of one or more immune cell subsets such as CD1 lC.sup.+ cells from tissue such as skin or lung through the blood to immune tissue such as lymph nodes or GALT is seen as a transient increase in the level of circulating GDI lC.sup.+ cells in response to exposure of the subject's tissues to a suitable amount ofa F3C. Thus, the level of CD1 lC.sup.+ cells in the blood will generally detectably increase, e.g., a statistically significant increase, plateau and then decrease as migration of the cells to immune tissue subsides. In these embodiments, the proportion of the cells of the affected immune cell subset is typically relatively low in most physiological immune states, e.g., normal or abnormal immune conditions, compared to the total white blood cell population in circulation. In other embodiments, the migration of one or more immune cell subsets such as CD123.sup.+ cells from the circulation to immune tissue such as lymph nodes or GALT results in a decrease. In these embodiments, the decrease in the numbers of circulating immune cells reflects the migration of the immune cells from the blood to immune tissue such as lymph nodes or GALT. Such a decrease may be transient and followed by recovery of the affected immune cell subset(s) over about 2 to 24 weeks. In conducting these embodiments, administration of the F3C to the subject is accomplished using the fonnulations or the methods as described herein.

Thus, an aspect of the invention is a method to enhance the migration of one or more immune cell types in a subject from one location (e.g., bone marrow, circulating blood or a tissue such as the skin, liver, central nervous system or lung) to another (e.g., to the blood or to a lymphoid tissue such as a lymph node, spleen or a mucosal tissue such as GALT) by administration to a subject as described herein of an effective amount of a F3C essentially as described by any of the methods disclosed herein. A related aspect is the monitoring, e.g., by suitable blood counts or tissue biopsy, of the subject's response to determine the timing and extent of such immune cell migration.

Other CD molecules that are modulated by the presence of the F3Cs in a subject include cytokine receptor molecules such as one or more of CD115, CDW116, CD117, CD118, CDW119, CD120a, CD120b, CD121a, CD121b, CD122, CD123, CD124, CD125 CD126, CDW127, CDW128 or CDW130. Often, the numbers of receptor molecules per cell will be modulated. For example, receptors for cytokines that mediate or facilitate Thi immune responses or innate immune responses (e.g., one or more of IL-1.alpha., IL-1.beta., IL-2, IL4, IL-12, .gamma.IFN or .alpha.-interferon) will typically increase in or on cells that mediate Th 1 or innate immune responses. Modulation of these molecules may be by direct interactions with a receptor(s) in the cell that expresses the cytokine receptor or indirectly by modulation of cytokine synthesis in the affected cells or in other cells, typically immune cells that may interact with the cells whose receptor synthesis is being modulated. Thus, autocrine or paracrine mechanisms may underlie some of the effects associated with administration of a F3C(s) such as altered cytokine profiles in immune cells or altered immune cell populations. Endocrine cytokine mechanisms may also contribute to desired immune responses.

Treatment of a subject with a F3C can result in a change of at least about 20-80% or about 25-50% above or below (e.g., at least 30% or at least 40% above or below) the control or basal level of affected immune cell subsets. For example, increases of more than about 30% in the total numbers of activated CD8.sup.+ T cells, e.g., CD8.sup.+, CD69.sup.+, CD25.sup.+ T cells, CD8.sup.+, CD69.sup.+, CD25.sup.- T cells or CD8.sup.+, CD69.sup.-, CD25.sup.+ T cells, can occur by days after a single dose of a F3C to a subject. Such increases may be greater than 50%, 60% or 100% in the total numbers of activated CD8.sup.+ T cells or subsets of activated CD8.sup.+ T cells in individual subjects. Typically such increases are about in the total numbers of activated CD8.sup.+ T cells or subsets of activated CD8.sup.+ T cells averages about 30-40%, with individual subjects experiencing increases over 100% in the numbers of activated CD8.sup.+ T cells per unit blood volume compared to the basal level.

Administration of the F3Cs can affect other immune cell subsets. For example, the concentration of circulating CD4.sup.+, CD69.sup.+, CD25.sup.- (Thi helper cells) and νοϋj 12 CD8.sup.+, CD16.sup.+, CD38.sup.+ LAK cells or CD8.sup.-, CD16.sup.+, CD38.sup.+ LAK cells typically increases during or after the course of dosing a subject with a F3C. Also, CD8.sup.-, CD16.sup.+, CD38.sup.+ and CD8.sup.+, CD16.sup.+, CD38.sup.+ (ADCC effector cells) and low side scatter Lin.sup.-, DR.sup.+, CD123.sup.+ (dendritic precursors) or low side scatter Lin.sup.-, DR.sup.+, CD1 lc.sup.+ (dendritic cells or precursors) may show modest to significant increases.

In subjects that are immunosuppressed, e.g., from certain infections or from chemotherapy (e.g., a cancer chemotherapy or a radiation therapy), administration of the F3Cs to the subject results in a favorable shift in the balance of Thi or Th2 responses the subject can mount in the face of immunosuppression. When Thi responses are suboptimal or insufficient, treatment with a F3C results in enhancement of Thi responses or a reduction in Th2 responses. Conversely, when Th2 responses are suboptimal or insufficient, treatment with a F3C results in enhancement of Th2 responses, which may occur with a concomitant modulation (increase or decrease) in Thi responses. The F3Cs can thus be used to shift the nature of a subject's immune response to result in a more balanced immune response from immunosuppression. Alternatively, the compounds can selectively suppress inappropriate or unwanted immune responses. Enhanced Thi responses appears to be at least partly due to one or more of (i) a reduction in biological restraints, e.g., high levels of IL4 or IL-10, on Thi functions by preexisting primed Thi effector cells, (ii) enhanced differentiation of ThO cells to Thi cells or enhanced responses mediated by Thi cells, (iii) enhanced function of accessory cell function, e.g., antigen presentation by dendritic cells, dendritic precursor or progenitor cells or by macrophages or their precursors or progenitors, (iv) enhanced proliferation and differentiation of Thi precursor or progenitor cells, (v) enhanced IL-12 expression in dendritic cells or their precursors, which results in enhanced differentiation of Thi cells from ThO precursors, (vi) enhanced expression or activity of factors associated with Thi functions, e.g., IL-2, gamma interferon (.gamma.IFN or IFN.gamma.), IL-18 or lymphotoxin.

An aspect of the invention methods is an alteration in the expression of IL4 or IL-10 that IE 0 6 0312 occurs after administration of a F3C to a subject. A consistent observation is that extracellular IL-4 or IL-10 levels rapidly decrease to levels that are undetectable by ELISA.

Intracellular IL-10 levels are reduced to levels that are near or below the limits of detection by flow cytometry. The administration of a F3C to a subject thus provides a means to inhibit either or both of these interleukins. Such inhibition may be associated with enhancement of Thl immune responses relative to Th2 or ThO responses, e.g., in subjects where Thl responses are suppressed (e.g., from viral, bacterial or parasite infection (HIV, HCV, etc) or chemotherapy) or are otherwise suboptimal. In many subjects, levels of either IL-4 or IL-10, usually IL-10, before dosing with a F3C is low or undetectable. In these subjects, dosing with the F3C results in a rapid drop in the interleukin that is detectable, usually IL-4.

Clinical conditions are described in more detail below where the F3Cs are usefi.il for treating, preventing, slowing the progression of, or ameliorating one or more symptoms associated with the described conditions. In any these conditions, any F3C disclosed herein can be used according to one or more of the dosing methods that are disclosed herein. For these conditions, dosages for the F3Cs, formulations and routes of administration are as described herein. Additional information regarding these and other clinical conditions or symptoms that can be treated, prevented or ameliorated with the F3Cs are found at e.g., The Merck Manual, 17.sup.th edition, Μ. H. Beers and R. Berkow editors, 1999, Merck Research Laboratories, Whitehouse Station, N.J., ISBN 0911910-10-7, or in other references cited herein.

Responses to treatment of a subject having a condition disclosed herein with a F3C is optionally monitored by observing changes in one or more immune or other appropriate clinical parameters, e.g., as described herein or in D. S. Jacobs et al., editors, Laboratory Test Handbook, 4.sup.th edition, pages 11-686, Lexi-Comp Inc., Hudson, Ohio, ISBN 027 916589-36-6, or in any of the references cited herein, or by monitoring the progression or severity of the underlying condition according to known methods, e.g., J. B. Peter, editor, Use and Interpretation of Laboratory Tests in Infectious Disease, 5.sup.th Edition, pages 130 309, 1998, Specialty Laboratories, Santa Monica, Calif., ISBN 1-889342-13-0. ΙΕ ν ό ΰ 3 12 For any of the infections disclosed herein, a subject who has the infection, or is susceptible of developing the infection, e.g., by suspected or potential exposure to an infectious agent, is treated by administering an effective amount of a F3C to the subject. Such subjects may have, or be susceptible to developing another condition, e.g., an autoimmune condition, inflammation condition, cardiovascular condition or a cancer or pre-cancer as described herein, such as rheumatoid arthritis, systemic lupus erythematosis, Crohn's disease, ulcerative colitis, type 1 diabetes, type 2 diabetes, peptic ulcers, skin ulcers, oral cavity ulcers, asthma, multiple sclerosis, coronary artery disease, acute or chronic rheuinatuc heart disease, atherosclerosis, stroke or lung cancer, that can be related to or exacerbated by the infection. In these embodiments, the F3Cs can function by one or more mechanisms, including enhancing innate immune responses, modulating, e.g., detectably increase or decrease, the level or activity of one or more of the transcription factors, enzymes or other biomolecules described herein, e.g., IL-1.alpha., IL-l.beta.,TNF.alpha., TNF-.beta., IL-6, IL-8, IL-10, gro-.alpha., IFN-.gamma., IFN-.alpha., MCP-1.alpha., MIP-1.alpha., MIPl.beta., MIP-2, IP-10, LT-.beta., GM-CSF, RANTES or their isotypes or homologs or cortisol. For example, molecules such as ILl.alpha., TNF.alpha., MIP-Lalpha. or MCP-1 are generally decreased in infections where there is an overexpression of one or more of these molecules. A detectable decrease of one or more of these molecules often occurs.

During or after treatment, the patient is optionally monitored and the amelioration of one or more symptoms or a slowed disease progression is observed. Such symptoms can include one or more of a red-brown bump with swelling at the edges, blisters, formation of a black scab or eschar at the site of skin infection and edema. Symptoms of cutaneous anthrax that can be ameliorated include fever, headache, muscle ache, nausea, and vomiting. In treating ·< B. anthracis infections, the F3Cs will typically decrease tissue damage associated with inflammation, enhance innate immune responses, enhance immune responses, reduce TNF.alpha., IL-1.alpha, or IL-l.beta. levels or activity or enhance killing or phagocytosis of pathogen in the infected subject or the subject's immune cells, e.g., monocytes, neutrophils ΙΕ ο ρ q j 2 or macrophages.

For a pulmonary anthrax infection, amelioration of one or more of fever, bleeding and necrosis of lymph nodes near the lung, local chest infection, shock, coma or death can occur. Infection of the brain and meningoencephalitis may occur and is treated in a similar manner, although an increased dosage can be utilized, e.g., about 20-50 mg/kg/day of the F3C is administered by a parenteral, e.g., intravenous, sublingual or buccal route. In any of these skin, pulmonary or gastrointestinal infections, the subject is also optionally treated using one or more standard antibiotics and routes of administration, e.g., procaine penicillin G, of streptomycin, tetracycline, erythromycin, ciprofloxacin, doxycycline, levofloxacin, norfloxacin or oxofloxacin.

The use of the F3Cs will generally ameliorate the inflammation, sepsis or shock that can occur when antibiotics are administered to subjects having a systemic or pulmonary B. anthracis infection. A potential adverse effect of antibiotic use to treat a systemic or pulmonary B. anthracis infection is serious or potentially lethal inflammation, sepsis and/or shock that results from release of anthrax lethal toxin or factor or other inflammatory molecules on lysis of the bacteria. Release of bacterial lethal factor from lysed bacterial cells is associated with an intense inflammation, which is at least partially mediated by one or more inflammatory factors such as TNF.alpha., IL-1.beta., IL-1.alpha., IL-6, IL-8 or COX-2. The F3Cs detectably reduce the level and/or biological effects of such inflammatory factors and can also detectably maintain or facilitate macrophage viability or one or more desired macrophage function(s) at the same time.

The antiviral or antimicrobial agents or treatments in combination therapies with a F3C will be or are used essentially according to new or to known dosing and administration methods χ for those agents or treatments. Their use may precede, overlap or be coincident in time with or follow a treatment protocol with a F3C. In some embodiments, the other therapeutic agents or treatments will overlap and will thus be administered on one or more of the same days on which a F3C is administered to a subject having a viral infection, or subject to a IE 060 312 viral infection. In other embodiments, the other therapeutic agents or treatments will be administered to such a subject within about 1 day to about 180 days before or after a treatment protocol or a dosing period with a F3C begins or ends. In exemplary embodiments, the other suitable treatment or agent is administered within 1 day, 2 days, 3 days, 4 days, about 7 days, about 14 days, about 28 days or about 60 days before or after a treatment protocol or a dosing period with a F3C begins or ends.

Although the forgoing combination therapies have been described in the context of viral or other infections, the protocols and methods that employ a F3C can be used in conjunction with any suitable new or known therapeutic agent(s) or treatment protocol(s) for other any other clinical condition described herein. Any of these additional treatments can be coupled with the administration of any of the F3Cs in any of the embodiments described herein. Exemplary conditions include one or more of a non-viral pathogen infection(s), a cancer(s), a pre-cancer(s), an inflammation condition(s), an autoimmune condition(s), an immunosuppression condition(s), a neurological disorders), a cardiovascular disorder(s), a neurological disorder(s), diabetes, obesity, wasting, anorexia, anorexia nervosa, a cancer chemotherapy(ies) side-effect(s), a side-effect(s) of a chemotherapy(ies) or a radiation therapy(ies) of any other clinical condition disclosed herein or in the cited references, or the like. Thus, invention embodiments include the use of a F3C before, during or after a treatment that uses another suitable therapeutic agent(s) or therapeutic treatment(s) for any of the diseases or conditions disclosed herein, any of which diseases or conditions may be acute, chronic, severe, mild, moderate, stable or progressing.

Examples of such agents, treatments or chemotherapies include the use of one or more adrenergic agents, adrenocortical suppressants, aldosterone antagonists, anabolics, analeptics, analgesics, anesthesia, anthelmintics, antiacne agents, anti-adrenergics, antiallergies, anti-amebics, anti-androgens, antianginals, anti-anxiety agents, anti-arthritics, antiasthmatic agents, anti-atherosclerotic agents, antibacterials, anticholinergics, anticoagulants, anticonvulsants, antidepressants, antidiabetics, antidiarrheals, antidiuretics, anti-emetics, anti-epileptics, anti-estrogens, antifibrinolytics, antifungals, antihistamines.

IE tf 6 0 3 12 antihyperlipidemia agents, antihyperlipoproteinemic agents, antihypertensive agents, antihypotensives, anti-infectives, anti-inflammatory agents such as entanercept (a dimeric fusion coprising a portion ofthe human TNF receptor linked to the Fc protion of human IgGl containing the C.sub.H2 and C.sub.H3 domain and hinge regions of IgGl) or a COX-2 inhibitor such as celexicob (4-5[-(4-methylphenyl)-3-(trifluoromethyl)-lH-pyrazole-l-ylj benzenesulfonamide) or rofecoxib (4-[4-methylsulfonyl)phenyl]-3-phenyl-2(- 5H)furanone), antimaiarial agents, antimicrobials, antimigraine agents, antimycotic agents, antinausea agents, antineoplastic agents, antiparasitics, antiparkinsonian agents, antiproliferatives, antiprostatic hypertrophy agents, antiprotozoals, antipruritics, antipsychotics, antirheumatics, antischistosomals (e.g., praziquantel, artemisinin), blood glucose regulators, bone resorption inhibitors, bronchodilators, cardiac depressants, cardioprotectants, choleretics, depressants, diuretics, dopaminergic agents, enzyme inhibitors, free oxygen radical scavengers, glucocorticoids, peptide hormones, steroid hormones, hypocholesterolemics, hypoglycemics, hypolipidemics, hypotensives, immunomodulators, liver disorder treatments, mucosal protective agents, nasal decongestants, neuromuscular blocking agents, plasminogen activators, platelet activating factor antagonists, platelet aggregation inhibitors, post-stroke and post-head trauma treatments, progestins, psychotropics, radioactive agents, relaxants, sclerosing agents, sedatives, sedative-hypnotics, selective adenosine Al antagonists, serotonin antagonists, serotonin inhibitors, serotonin receptor antagonists, thyroid inhibitors, thyromimetics, tranquilizers, vasoconstrictors, vasodilators, wound healing agents, xanthine oxidase inhibitors or a treatment(s) or therapeutic agent(s) for amyotrophic lateral sclerosis, ischemia, e.g., cereberal ischemia, cardiac ischemia or cardiovascular ischemia, or unstable angina. The selection and use of these agents for a particular subject will typically use dosing methods, dosages and routes of administration essentially according to known methods, dosages and routes of administration. Such methods, dosages and routes of administration are described in detail at, e.g., Textbook of Autoimmune Diseases, R. G. Lahita, editor, Lippincott Williams & Wikins, Philadelphia, Pa., 2000, ISBN 0-7817-1505-9, pages 81-851, Holland.cndot.Frei Cancer Medicine .sup.e.5, 5.sup.th edition, R. C. Bast et al., editors, 2000, ISBN 1-55009-113-1, pages 168-2453, B. C. Becker Inc. Hamilton, IE Ontario, Canada, Hematology, Basic Principles and Practice, 3.sup.rd edition, R. Hoffman, et al., editors, 2000, ISBN 0-443-779544, pages 115-2519, Churchill Livingstone, Philadelphia, Pa., Rheumatology, 2.sup.nd edition, J. H. Klippel et al., editors, 1998, ISBN 0-7234-2405-5, volume 1, sections 1-5 and volume 2, sections 6-8, Mosby International, London, UK, Alzheimer's Disease and Related Disorders: Etiology, Pathogenesis and Therapeutics, K. Iqbal, et at, editors, 1999, ISBN 0-471986386,John Wiley & Son Ltd, and Cardiovascular Medicine, E. J. Topol, editor, Lippincott Williams & Wikins, Philadelphia, Pa., 1998, ISBN 0781716810.

In some infections, the F3C(s) effects an improvement of one or more of the symptoms associated with the infection or a symptom thereof. For example, treatment of subjects who are immune suppressed, e.g., cancer chemotherapy or other cause, generally show improvement of one or more associated symptoms, such as weight loss, fever, anemia, pain, fatigue or reduced infection symptoms that are associated with a secondary infection(s), e.g., HSV-1, HSV-2, papilloma, human cytomegalovirus (CMV), Pneumocystis (e.g., P. carinil) or Candida (C. albicans, C. krusei, C. tropicalis) infections.

In some embodiments, the F3C(s) is administered as a nonaqueous liquid formulation as described herein or the F3C(s) is administered according to any of the intermittent dosing protocols described herein using a solid or liquid formulation(s). In the case of a subject who has a retroviral infection, e.g., a human with an HIV infection, with symptoms that include one or more of, a relatively low CD4 count (e.g., about 10-200, or about 20-100 or about 2050), one or more additional pathogen infections (HSV-1, HSV-2, HHV-6, HHV-8, CMV, HCV, a HPV, P. carinii or Candida infection) and one or more of anemia, fatigue, Kaposi's sarcoma, fever or involuntary weight loss (at least about 5% of body weight), administration of about 0.1 to about 10 mg/kg/day (usually about 0.4 to about 5 mg/kg/day) of a F3C(s) to \ the subject typically results in noticeable improvement of one or more of the symptoms within about 1-4 weeks. In other embodiments, the F3C(s) is administered to a subject who has a condition that appears to be associated with a viral infection, e.g., pneumonia or retinitis associated with CMV, nasopharyngeal carcinoma or oral hairy leukoplakia It « 6 0 3 12 associated with Epstein-Barr virus, progressive pancephalitis or diabetes associated with Rubella virus or aplastic crisis in hemolytic anemia associated with Parvovirus 19.

One or more intermittent dosing protocols disclosed herein or one or more of the liquid nonaqueous formulations described herein can be applied by routine experimentation to any of the uses or applications described herein. For a F3C(s) that is a new compound per se, the compound(s) can be administered to a subject according to an invention intermittent dosing protocol(s) or by other protocols, e.g., continuous daily dosing of a single dose or two or more subdoses per day. In addition any of the F3Cs, e.g., one or more F3Cs that are new per se, can be present in any solid or liquid formulation described herein. These formulations and dosing protocols can be applied by routine methods to any of the uses or applications described herein.

Antibodies, vaccines and vaccine adjuvants. The F3Cs can be used to enhance cellular or humoral responses to vaccination against, e.g., infectious agents or malignant cells. F3Cs can also be used to make antibodies that bind to the F3Cs themselves or their metabolic products. Antibodies that bind to the F3Cs can be used, e.g., in diagnostic, quality control, or the like, methods or in assays for the F3Cs or their metabolites. In addition, the F3Cs are useful for raising antibodies against otherwise non-immunogenic polypeptides, in that the compounds may serve as haptenic sites stimulating an immune response against the polypeptide.

Immunogens that are used to make antibodies that bind to a F3C comprise a F3C that has 1 or more epitopes and optionally another immunogenic substance. The immunogenic substance can be covalently bonded to the F3C to form an immunogenic conjugate or it can be in a mixture of non-covalently bonded materials, or a combination of the above. Immunogenic substances include adjuvants such as Freund's adjuvant, immunogenic proteins such as viral, bacterial, yeast, plant and animal polypeptides, including keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin or soybean trypsin inhibitor, and immunogenic polysaccharides. Typically, the F3C having one, two or more epitopes is ΙΕ ΰ 6 0 3 12 covalently conjugated to an immunogenic polypeptide or polysaccharide by the use of a polyfunctional (ordinarily bifunctional) cross-linking agent. Methods for the manufacture of immunogens that comprise one or more haptens are conventional per se. Methods for conjugating haptens to immunogenic polypeptides or the like are used here. Such conjugates are prepared in conventional fashion. For example, the cross-linking agents Nhydroxysuccinimide, succinic anhydride or C.sub.2-8 alkyl-N.dbd.C.dbd.N--C.sub.2-8 alkyl are useful in preparing the conjugates. The conjugates comprise a F3C that is attached by a bond or a linking group of 1-100, typically, 1-25, more typically about 1-10 carbon atoms to the immunogenic substance. Typically a polypeptide, polysaccharide or other suitable immunogenic moiety is conjugated to a site on a F3C in a location that is distant from the epitope on the F3C to be recognized. The conjugates are separated from starting materials and by-products using chromatography or the like, and then are optionally sterile filtered, or otherwise sterilized, or are optionally vialed for storage. Synthetic methods to prepare hapten-carrier immunogens have been described, see e.g., G. T. Hermanson, Bioconjugate Techniques Academic Press, 1996, pages 419493.

Animals or mammals are typically immunized once, twice or more times against the immunogenic conjugates that comprise a F3C and an immunogen. Polyclonal antisera or monoclonal antibodies are prepared in conventional fashion. In some embodiments, about 0.0001 mg/kg to about 1 mg/kg, e.g., about 0.001 or about 0.01 or about 0.1 mg/kg, of immunogenic conjugate or derivative is used on one, two, three or more occasions to immunize the subject as described herein. The immunogenic conjugates are administered, orally, topically or parenterally as described herein, e.g., by i.m. or s.c. injection. Methods to prepare antibodies, including methods to obtain antibodies that bind to steroids have been described, see, e.g., R. O. Neri et al., Endocrinology 74:593-598 1964, M. Ferin et al.. Endocrinology 85:1070-1078 1969, J. Vaitukaitis et al., J. Clin. Endocr. Metab. 33:988-991 i 1971 and M, Ferin et al., Endocrinology 94:765-775 1974. Such methods can be used essentially as described to prepare antibodies or monoclonal antibodies that bind to a F3C. Embodiments include serum or other preparations that comprise any polyclonal or monoclonal antibodies that bind to a F3C(s), methods to make such antibodies and ΙΕ νού j U compounds or compositions that are used in conducting these methods.

In other embodiments the F3Cs are used as adjuvants to enhance a subject's immune response to antigens such as proteins, peptides, polysaccharides, glycoproteins or killed or attenuated viruses or cell preparations. In these methods, an effective amount of the F3C is administered at about the same time that the antigen is delivered to the subject, e.g., within about 1, 2, 3, 4, 5, 6, or 7 days of when the antigen is administered to the subject. In some embodiments, the F3C is administered 1, 2, 3,4 or more times (usually once or twice per day) at 1, 2, 3 or 4 days before or after the antigen is administered to the subject. In other embodiments, the F3C is administered on the same day that the antigen is administered to the subject, e.g., within about 1-4 hours. Such immunization methods may be repeated once, twice or more as needed. The F3C can be administered to the subject using any of the formulations or delivery methods described herein or in the references cited herein. Subjects suitable for these vaccinations include young and elderly mammals, including humans, e.g., humans about 3-36 months of age or older and humans about 60, 65, 70, 75 years of age or older. The amount of antigen used can be about 0.01 .mu.g/kg to about 20 mg/kg, typically about 1-100 .mu.g/kg. Dosages of the F3C used in these vaccinations is essentially as described herein, e.g., about 5 mg to about 1000 mg of a F3C is used per day on days when it is administered as part of the vaccination method.

Related embodiments include compositions or formulations that comprise a F3C, an antigen(s) or antigen(s) preparation and optionally one or more excipients. The antigen is essentially as disclosed herein or in a cited reference. Antigen preparations may comprise one or more of (1) lethally or sublethally radiated cells or pathogens, (2) disrupted cells or viruses or such as attenuated viruses, (3) a nucleic acid or DNA vaccine. (4) an antigenic protein, glycoprotein, polysaccharide or a fragment or derivative of any of these molecules, \ (5) chemically treated cells or pathogens, e.g., formalin or detergent treated cells, viruses or cell or virus extracts and (6) genetically engineered viral or bacterial vectors that express one or more antigens or antigen fragments. Pathogens include prions or the etiologic agents of, e.g., Creutzfelt-Jacob disease, bovine spongifonn encephalopathy and scrapie in sheep, IE 0 6 0 3 12 goats or mice. Where cells or disrupted are present in an antigen preparation, they may by genetically modified, e.g., to express one or more antigens or epitopes against which an immune response is desired. Antigens in these embodiments are moieties that can elicit a detectable immune response when it is administered to a subject. In some embodiments, the antigen is foreign to the subject. For foreign antigens, the subject to be vaccinated may not encode or express the antigen, while the antigen is usually part of or expressed by a pathogen or by a subject or mammal of a different species. In other embodiments, antigens are endogenous or non-foreign to the subject, e.g., they are usually encoded or expressed by the subject or another subject of the same species. Endogenous antigens are suitable for use in, e.g., tumor vaccination methods.

Exemplary tumors from which a suitable antigen(s) may be obtained are as described herein or in the cited references. A DNA vaccine as used here typically comprises a nucleic acid, usually DNA, that encodes one or more antigens or epitopes that a pathogen, e.g., a parasite, fungus, virus or bacterium, or a tumor encodes or can express. Tumor antigens that are suitable for use in vaccination methods that employ a F3C include tumor-associated antigens and tumor-specific antigens. These molecules typically comprise one or more protein, glycoprotein, carbohydrate or glycolipid. Vaccinations that employ a tumor antigen(s) may comprise autologous tumor cells or allogenic tumor cells, which are optionally disrupted and optionally used with a non-formula 1 adjuvant, such as bacillus Calmette-Guerin (BCG), purified protein derivative, Freund's complete adjuvant, Corynebacterium parvum, Mycobactedium vaccae, oligonucleotides that consist of or comprise an unmethylated CpG dimer or an alum precipitate. In some embodiments, tumor cells treated with neuraminidase comprise all or part of the tumor antigen source. The non-formula 1 adjuvants are also optionally used in any of the vaccination methods disclosed herein. As used here, tumor associated antigens, e.g., the carcinoembryonic antigen, .alpha.-fetoprotein or the prostate specific antigen, are molecules that are often associated with or detectably expressed by premalignant or malignant cells or cell populations and also with some normal tissues during at least part of the subject's life cycle.

IE 3 Other suitable antigens include STn, sialyl Tn-KLH, carbohydrate conjugates, carcinogenic embryonic antigen, MAGE-1, MUC-1, HER-2/neu, prostate specific antigen, p53, T/Tn, bacterial flagella antigens or capsular polysaccharide antigens (e.g., Staphylococcus aureus capsular polysaccharide antigens) and antigenic fragments or antigenic synthetic derivatives of any of these molecules, e.g., a fragment or derivative that retains at least about 20% or 30% of the antigenicity of the native or intact molecule. See, e.g., L. A. Holmberg et al.. Bone MaiTow Transplant. 2000 25:1233-1241, J. W. Hadden, Int. J. Immunopharmacology 1999 21:79-101, G. Ragupathi et al., Glycoconj. J. 1998 15:217-221, A. I. Fattom et al.. Infect. Immun. 199866:4588-4592, U.S. Pat. Nos. 5,770,208, 5,866,140 and 6,194,161 and citations elsewhere herein, including the preceding paragraph.

An antigenic protein, peptide or glycoprotein can be identified by standard methods, e.g., protein or nucleic acid sequencing, for any of the infectious agents or tumors that are described herein or in the cited references. Thus, in some embodiments, an effective amount of a F3C and an antigen are administered to a subject, or delivered to the subject's tissues, to stimulate an immune response against the antigen. The antigen may comprise one, two or more antigenic epitopes, which may come from one, two or more genes. In some embodiments, the subject is optionally monitored to follow or determine the immune, dendritic cell, B cell, T cell, antibody or cytokine response, such as one disclosed herein, e.g., modulation or increase in .gamma.IFN, IL-2 or IL-12 levels or measurement of the production of one or more immunoglobulin types or subtypes. The subject may also be monitored by in vitro cell assays, e.g., for activation of T cells or subsets of T cells or other relevant white blood cell types. Such assays include measuring T cell activation using chromium release assays, or mixed lymphocyte assays. The subject is optionally treated with one or more additional booster vaccinations, when this is called for under the circumstances.

Nucleic acid or DNA vaccines as used here will typically comprise a nucleic acid comprising an expressible region that encodes one, two or more suitable antigens or epitopes, e.g., all or an antigenic portion of a viral, bacterial, fungal or parasite protein or glycoprotein. The expressible region will usually comprise a transcription promoter and Κ 060 3 12 optionally other control sequences that are operatively linked to the antigen coding region where the promoter and control sequences are transcriptionally active in the intended subject or tissue. Suitable control sequences include enhancers, recognition sequences for transcription factors and termination sequences. Such expression vectors may optionally comprise one, two or more expressible genes or gene fragments, which may each comprise their attendant operatively linked expression sequences. Suitable methods and expression vectors to deliver nucleic acids for vaccine purposes have been described, e.g., U.S. Pat.

Nos. 5,223,263, 5,580,859, 5,703,055, 5,846,946 and 5,910,488.

Vaccinations that utilize a F3C and an antigen(s) are generally suitable for eliciting or enhancing desired immune responses in conjunction with exposure of a subject to an antigen(s), compared to vaccination without the compound. Antigen specific humoral antibody responses or antigen specific T cell responses may be enhanced or elicited.

Typically vaccination using a F3C and a suitable antigen is conducted to prevent a potential infection or to reduce the severity of a future infection. However, in some cases the vaccination is conducted in a subject that has an infection such as a chronic or a latent infection such as a parasite or a retrovirus or herpesvirus infection, which may be latent or in relapse. In other cases the subject may have a cancer or precancer. Thus, the subject may be exposed to, or contain, one or more of the antigens that are used in one of these vaccination procedures. Such vaccinations are included within the scope of the invention.

In related embodiments, the F3Cs are useful to facilitate preparation of hybridoma clones that express monoclonal antibodies. In these methods, a suitable amount of a F3C, e.g., about 100 .mu.g to about 2 mg for a small mammal, is administered to a subject, e.g., a mouse, to enhance the immune response to the desired antigen, which is also administered to the subject. After antigen challenge, suitable cells are recovered from the subject, e.g., anti- ι antigen immunoglobulin expressing HPRT.sup.+ spleen cells from a mouse. These cells are then fused with suitable immortal cells (e.g., mouse melanoma cells) using, e.g., PEG or Sendai virus, and selected in suitable selection growth medium, e.g., tissue culture medium that contains hypoxanthine, aminopterin and thymidine, to obtain a group or panel of IE Ο60 3 12 hybridomas that express anti-antigen monoclonal antibodies. The hybridoma panel is used to generate individual clones, which are optionally screened to determine the antibody specificity and antigen binding properties. About one, 100, 1000, 10,000, 100,000 or more individual clones are screened by standard methods. The monoclonal antibodies may be from any suitable source, e.g., murine, human, human-murine hybrid or the like. Methods to obtain human, human-murine hybrid or related monoclonal antibodies have been described, e.g., U.S. Pat. Nos. 5,562,903, 5,461,760, 5,705,154, 5,854,400, 5,858,728, 5,874,082, ,874,540, 5,877,293, 5,882,644, 5,886,152, 5,889,157, 5,891,996, 5,916,771, 5,939,598, ,985,615, 5,998,209, 6,013,256, 6,075,181, 6,901,001, 6,114,143, 6,114,598, 6,117,980.

The F3Cs can be used in any of the methods disclosed in these references to facilitate generation or recovery of hybridoma panels and clones that express monoclonal antibodies.

An aspect of these methods comprise a product, i.e., a hybridoma panel or a hybridoma clone, that is obtained by the process of contacting a subject (such as a mouse) with (1) a suitable amount of a F3C and (2) a suitable amount of an antigen, allowing sufficient time to generate an immune response in the subject against the antigen and then fusing suitable anti17 antigen immunoglobulin producing cells from the subject, e.g., the subject’s spleen cells, with a suitable immortal cell line (e.g., a HPRT.sup.+ mouse myeloma). The antigen or immunogen is as described above, e.g., a suitable protein, protein fragment or glycoprotein such as an interleukin, cytokine or antigen from an infectious agent. In these methods, a mouse is typically the subject, but other mammals, e.g., humans or other rodents, are also suitable according to known methods.

The amount of antigen for immunization used in preparing monoclonal antibodies in a human or a mammal will typically be about 1 .mu.g to about 1000 .mu.g, e.g., about 2 mu.g, 5 .mu.g, 10 .mu.g, 50 .mu.g or 100 .mu.g of antigen. The antigens are essentially as described in the vaccination methods described above, e.g., disrupted cell, a protein or glycoprotein, which is optionally combined with a suitable amount of an adjuvant such as Freund's complete adjuvant, alum precipitate, a bacterial lipopolysaccharide or BCG. ΙΕ “ 60 3 12 Related embodiments include a method comprising administering to a subject (e.g., a mammal such as a human or a primate), or delivering to the subject's tissues, an effective amount of a F3C and a specific antigen. Immune responses that are enhanced include a mucosal immune response to an antigen such as a protein, peptide, polysaccharide, microorganism, tumor cell extract or lethally radiated tumor or pathogen cells (e.g., antigens or cells from melanoma, renal cell carcinoma, breast cancer, prostate cancer, benign prostatic hyperplasia, virus or bacteria, or other tumor or pathogen as disclosed herein). Aspects of these embodiments include enhancement ofthe subject's immune response when an antigen or immunogen is administered intranasally or orally. In these aspects, the F3C is administered about simultaneously witli the antigen or within about 3 hours to about 6 days of antigen administration. The use of immune modulating agents to enhance immune responses to a vaccine has been described, e.g., U.S. Pat. No. 5,518,725.

Other uses for the F3C(s) include administering the compound(s) to a subject who suffers from a pathological condition(s). The treatment may treat or ameliorate the source of the condition(s) and/or symptoms associated with the pathological condition(s) such as infection with a pathogen(s) (viruses, bacteria, fungi), a malignancy, unwanted immune response, i.e., an immune response that causes pathology and/or symptoms, e.g., autoimmune conditions or allergy or conditions such as hypoproliferation conditions, e.g., normal or impaired tissue growth, or wound healing or bum healing, or in immunosuppression conditions, e.g., conditions characterized by an absence of a desired response and/or an inadequate degree of a desired response.

Enhanced antibody responses include detectable enhancement of antibody titer or a shift in the antibody, e.g., an antibody response from a Th2 biased response to an increased Thl biased component of the response. In such antibody shifts, the Thl and Th2 character of the 3 response is determined by known methods. For example, a relatively low ratio of IgG 1 (or the analogous antibody subclass in humans and other subjects) to IgG2a (or the analogous antibody subclass in humans and other subjects), e.g., about 6:1 to about 12: 1, that is generated after exposure of a subject (a mouse for the IgGl and IgG2a subclasses) to an ΙΕ υόο3 12 antigen indicates a Thi biased antibody response. Conversely a higher ratio, e.g., about 20:1 to about 30:1 indicates a Thi biased antibody response. Generation of antigen-specific IgG 1 generation involves T-helper type 2 (Th2) cells, and for IgG2a, T-helper type 1 (Thi) cells.

The F3Cs can detectably increase the Thl character of an antibody response to an antigen or they can increase the magnitude of both the Thl and Th2 response.

Exemplary pathogens or cells that are suitable sources for antigens or a gene(s) that encode suitable antigens include pathogens described herein or in the cited references.

Cancer and hyper proliferation conditions. Many cancers, pre-cancers, malignancies or hyper-proliferation conditions are associated with an unwanted Th2 immune response, a deficient Thl response or unwanted inflammation. An insufficient Thl immune response may play a role in the capacity of malignant or pre-malignant cells to escape immune surveillance. Any of the F3Cs disclosed herein, may thus be used to treat, prevent or slow the progression of one or more cancers, pre-cancers or cell hyper-proliferation conditions or they may be used to ameliorate one or more symptoms thereof. In these conditions, the F3Cs are useful to enhance the subject's Th 1 responses or to reestablish a more nonnal Th 1 -Th2 balance in the subject's immune responses. The F3Cs may function at least in part by decreasing inflammation or inflammation associated markers such as IL-6 and/or by enhancing hematopoiesis in many of these conditions.

These conditions include cancers or pre-cancers comprising carcinomas, sarcomas, adenomas, blastoma, disseminated tumors and solid tumors such as one associated with or arising from prostate, lung, breast, ovary, skin, stomach, intestine, pancreas, neck, larynx, esophagus, throat, tongue, lip, oral cavity, oral mucosa, salivary gland, testes, liver, parotid, biliary tract, colon, rectum, cervix, uterus, vagina, pelvis, endometrium, kidney, bladder, central nervous system, glial cell, astrocyte, squamous cell, blood, bone marrow, muscle or thyroid cells or tissue. The F3Cs are thus useful to treat, prevent, slow the progression of, or ameliorate one or more symptoms of a precancer, cancer or related hyperproliferation condition such as myelodysplastic syndrome, actinic keratoses, endometriosis, Barrett's l£ esophagus, leiomyoma, fibromyoma, benign or precancerous intestinal or bowel polyps or benign prostatic hyperplasia. The compounds can also be used to treat, prevent, slow the progression of, slow the replication or growth of, or to ameliorate one or more symptoms of a primary tumor, a metastasis, an advanced malignancy, a blood bom malignancy, a leukemia or a lymphoma. Any of these conditions may be in an early or mild fonn or can be moderate or advanced in the existent or progression of the disease or a symptom.

In treating endometriosis, the use of an F3C will slow the rate of disease progression and decrease the severity or frequency of one or more symptoms such as irregular menstrual periods, infertility abdominal pain or cramping and pain in the lower back or pelvic area, which may precede menstruation or may accompany sexual intercourse or bowel movements. Beneficial effects from F3C treatment will be mediated in patients with endometriosis at least partially by increasing the patient's Thi immune responses and/or by decreasing anti-endometrial antibodies or abenent Th2 immune responses. Treatment of emdometriosis could be accompanied by other suitable treatments, e.g., treatment with one or more of estrogen, progesterone, danazol, follicle stimulating hormone antagonists, leutinizing hormone antagonists, gonadotropin-releasing hormone antagonists such as nafarelin acetate or analgesics such as codeine, tylenol or aspirin.

The F3Cs can be used to treat paraneoplastic syndromes or conditions such as ones associated with lung or breast cancers that secrete calcitonin or that enhance osteoclast activity. Such conditions include hypercalcemia, Cushing's syndrome, acromegaly and non23 islet cell tumor hypoglycemia. The compounds are used to decrease osteoclast activity or other symptoms associated with such conditions.

Hyper-proliferation conditions that can be treated include melanoma, Kaposi's sarcoma, leiomyosarcoma, non-small cell lung cancer, small cell lung cancer, bronchogenic carcinoma, renal cell cancer or carcinoma, glioma, glioblastoma, pancreatic or gastric adenocarcinoma, gastrointestinal adenocarcinoma, human papillomavirus associated cervical intraepithelial neoplasia, cervical carcinoma, hepatoma, hepatocellular carcinoma, IE U6O312 hepatocellular adenoma, cutaneous T-cell lymphoma (mycosis fungoides, Sezary syndrome), colorectal cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, ALL or follicular lymphoma, multiple myeloma, carcinomas with p53 mutations, colon cancer, cardiac tumors, adrenal tumors, pancreatic cancer, retinoblastoma, a small cell lung cancer, a non-small cell lung cancer, intestinal cancer, testicular cancer, stomach cancer, neuroblastoma, neuroma, myxoma, myoma, endothelioma, osteoblastoma, osteoclastoma, osteosarcoma, chondrosarcoma, adenoma, breast cancer, prostate cancer, Kaposi's sarcoma, ovarian cancer, squamous cell carcinoma of the gastrointestinal tract and non-myeloid tumors. Treating a subject with a F3C can ameliorate one or more side effects of chemotherapy or cancer symptoms such as alopecia, pain, fever, malaise, chronic fatigue and cachexia or weight loss. Other cancers, precancers or their symptoms that can be treated, prevented or ameliorated are described in, e.g., Holland.cndot.Frei Cancer Medicine .sup.e.5, S.sup.th edition, R. C. Bastet al., editors, 2000, ISBN 1-55009-113-1, pages 1682453, B. C. Becker Inc. Hamilton, Ontario, Canada or The Merck Manual, 17.sup.th edition, Μ. H. Beers and R. Berkow editors, 1999, Merck Research Laboratories, Whitehouse Station, N.J., ISBN 0911910-10-7.

In some of these embodimants, the subject's hyper-proliferation or malignant condition may be associated with or caused by one or more pathogens. Such conditions include hepatocellular carcinoma associated with HCV or HBV, Kaposi's sarcoma associated with HIV-1 or HIV-2, T cell leukemia associated with HTLV I, Burkitt’s lymphoma associated with Epstein-Barr virus or papillomas or carcinoma associated with papilloma viruses (e.g., human HPV 6, HPV 11, HPV 16, HPV 18, HPV 31, HPV 45) or gastric adenocarcinoma, gastric MALT lymphoma or gastric inflammation associated with Helicobacter pylori, lactobacillus, enterobacter, staphylococcus or propionibacteria infection.

In some of these embodiments, the F3Cs may be used to treat, prevent or slow the progression of or ameliorate one or more conditions in a subject having or subject to developing a hyperproliferation condition where angiogenesis contributes to the pathology. Abnormal or unwanted angiogenesis or neovascularization contributes to the development or ΙΕ ϋ 6 ο 3 12 progression of solid tumor growth and metastases, as well as to arthritis, some types of eye diseases such as diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, rubeosis, retinoblastoma, uvietis and pterygia or abnormal blood vessel growth of the eye, and psoriasis. See, e.g., Moses et al,, Biotech. 9:630-634 1991, Folkman et at, N. Engl. J. Med., 333:1757-1763 1995, and Auerbach et al., J. Microvasc. Res. 29:401-411 1985.

Dosages of the F3C, routes of administration and the use of combination therapies with other standard therapeutic agents or treatments could be applied essentially as described above for cancer or hyperproliferation conditions or other conditions as disclosed herein. Thus, in some embodiments, the use of the F3C is optionally combined with one, two or more additional therapies for a cancer or precancer(s), e.g., one, two or more of surgery and treatment with an antiandrogen or an antiestrogen as described herein or in the cited references, an antineoplastic agent such as an alkylating agent, a nitrogen mustard, a nitrosourea, an antimetabolite, a cytotoxic agent, a cytostatic agent, platinum agents, anthracyclines, taxanes or treatment with an analgesic such as propoxyphene napsylate, acetaminophen, morphine or codeine. Exemplary anticancer and adjunct agents include tamoxifen, paclitaxel, taxol, docetaxil, methotrexate, vincristine, vinblastine, 5-fluorouracil, thioguanine, mercaptopurine, adriamycin, chlorambucil, cyclophosphamide, cisplatin, carboplatin, transplatinum, irinotecan, procarbazine, hydroxyurea, erythropoietin, G-CSF, bicalutamide, anastrozole, fludarabine phosphate, doxorubicin and any suitable form of any of these agents, e.g., salts and solvates. Such therapies would be used essentially according to standard protocols and they would precede, be essentially concurrent with and/or follow treatment with a F3C. In some embodiments, such additional therapies will be administered at the same time that a F3C is being used or within about 1 day to about 16 weeks before or after at least one round of treatment with the F3C is completed. In other embodiments, a course of therapy is administered to the subject, e.g., treatment with a myelosuppressive amount of a myelosuppressive agent such as 5-fluorouracil, cyclophosphamide or a platinum compound such as cisplatin, followed within about 1, 2, 3, 4, 5 or 6 days by administration of one or more courses of treatment with a F3C. Other suitable exemplary therapeutic agents and their use have been described in detail, see, e.g., Physicians Desk Reference 54.sup.th edition, 2000, pages 303-3250, ISBN 1-56363-330-2, Medical Economics Co., Inc., Montvale, N.J. One or more of these exemplary agents can be used in combination with a F3C to ameliorate, slow the establishment or progression of, prevent or treat any of the appropriate cancers, precancers or related conditions described herein, or any of their symptoms. hr treating cancers or hyperproliferation conditions, the F3Cs may detectably modulate, e.g., decrease or increase, the expression or level or activity of one or more biomolecules associated with the prevention, establishment, maintenance or progression of the cancer or hyperproliferation condition. Such biomolecules include one or more of carcinoembryonic antigen, prostate specific antigen, her2/neu, Bcl-XL, bcl-2, p53, IL-1.alpha., IL-1.beta., IL-6, or INF.alpha., GATA-3, COX-2, NF.kappa.B, IkB, an IkB kinase, e.g., IkB kinase-.alpha., IkB kinase-.beta. or IkB kinase-.gamma., NF AT, calcineurin, calmodulin, a ras protein such as H-ras or K-ras, cyclin D, cyclin E, xanthine oxidase, or their isoforms, orthologs, homologs or mutant forms, which may be observed as either reduced or increased levels or biological activity(ies). Biomolecule levels or their activity(ies) that can be at least transiently detectably increased include one or more IL-2, IFN.gamma., IL-12, T-bet, 06methylguanine-DNA-methyltransferase, calcineurin, calmodulin, a superoxide dismutase (e.g., Mn, Zn or Cu), a tumor suppressor protein such as the retinoblastoma protein (Rb) or CDKN2A (p 16), BRCA1, BRCA2, MeCP2, MBD2, PTEN, NBR1, NBR2 or the isoforms, orthologs, homologs or mutant forms, which may have either attenuated or enhanced biological activity(ies), of any of these molecules. In treating a cancer described herein such as prostate cancer, one or more of ELAC2, 2',5'-oligoadenylate dependnet RNAse L (RNASEL), macrophage scavenger receptor 1 (MSR1), BRCA2 can be modulated or decreased.

The F3Cs can modulate the synthesis or a biological activity of one or more other gene products such as transcription factors, enzymes or steroid or other receptors that are associated with the establishment, progression or maintenance of a cancer or precancer or K *> 6 0 J 12 associated symptom. The compounds can inhibit AIB-1 coactivator or HER2/neu synthesis or activity in breast cancer cells or breast cancer conditions. They can enhance the synthesis or an activity of an estrogen receptor such as ER.alpha., ER.beta.l or ER.beta.2 or progesterone receptor in breast cancer or colon cancer cells or conditions. These effects can include modulation of the expression or one or more biological activities of proteins or enzymes that contribute to disease establishment or progression. Thus, the compounds can decrease IL4, IL-6 or IL-13 expression by stromal cells or immune cells that are in proximity to or adjacent to solid or diffuse tumor cells in a subject such as a human or another mammal. In the cancers or precancers described herein, the compounds can thus directly or indirectly modulate (e.g., decrease) the activity or expression of relevant enzymes such as STAT-6, neutral endopeptidase, a hydroxysteroid dehydrogenase, such as a 17.beta.12 hydroxysteroid dehydrogenase, 11 ,beta.-hydroxysteroid dehydrogenase, 7.beta.13 hydroxysteroid dehydrogenase or a 3 .beta.-hydroxysteroid dehydrogenase.

In some embodiments, the F3Cs are used to treat tumors or cancers wherein proliferation of the tumor or cancer cells is enhanced in response to sex steroids such as natural or synthetic androgens or estrogens. In other embodiments, the tumor or cancer cells are not responsive to such hormones or they are only slightly responsive to the presence of such compounds.

The F3Cs can be used to treat, prevent or ameliorate one or more symptoms of myocardial diseases or pathological myocardial or vascular conditions such as alcoholic cardiomyopathy, congestive cardiomyopathy, hypertrophic cardiomyopathy, aortic subvalvular stenosis, pulmonary subvalvular stenosis, restrictive cardiomyopathy, Chagas cardiomyopathy, endocardial fibroelastosis, myocardial fibrosis, endomyocardial fibrosis, Kearns Syndrome, myocardial reperfusion injury, myocarditis, cardiovascular or vascular diseases such as dissecting aneurysms, false aneurysms, infected aneurysms, ruptured aneurysms, aortic aneurysms, cerebral aneurysms, coronary aneurysms, heart aneurysms, and iliac aneurysms, angiodysplasia, angiomatosis, bacillary angiomatosis, Sturge-Weber Syndrome, angioneurotic edema, aortic diseases, Takayasu's Arteritis, aortitis, Leriche's Syndrome, arterial occlusive diseases, arteritis, enarteritis, polyarteritis nodosa. /£ osq Jf2 cerebrovascular diseases, disorders, and/or conditions, diabetic angiopathies, diabetic retinopathy, thrombosis, erythromelalgia, hemorrhoids, hepatic veno-occlusive disease, hypertension, hypotension, idiopathic pulmonary fibrosis, peripheral vascular diseases, phlebitis, pulmonary veno-occlusive disease, Raynaud's disease, CREST syndrome, retinal vein occlusion, Scimitar syndrome, superior vena cava syndrome, telangiectasia, atacia telangiectasia, hereditary hemorrhagic telangiectasia, varicocele, varicose veins, varicose ulcer, vasculitis, yenous insufficiency and arterial occlusive diseases such as arteriosclerosis, intermittent claudication, carotid stenosis, fibromuscular dysplasias, mesenteric vascular occlusion, Moyamoya disease retinal artery occlusion, thromboangiitis obliterans or atherosclerosis, any of which may be at an early stage or at a more advanced or late stage.

The F3Cs can also be used to treat, prevent or ameliorate one or more symptoms of cerebrovascular diseases, thrombosis, and/or conditions such as carotid artery diseases, cerebral amyloid angiopathy, cerebral aneurysm, cerebral anoxia, cerebral arteriosclerosis, cerebral arteriovenous malformation, cerebral artery diseases, cerebral embolism and thrombosis, carotid artery thrombosis, sinus thrombosis, Wallenberg's syndrome, cerebral hemorrhage, epidural hematoma, subdural hematoma, subarachnoid hemorrhage, cerebral infarction, cerebral ischemia (including transient), subclavian steal syndrome, periventricular leukomalacia, vascular headache, cluster headache, migraine, vertebrobasilar insufficiency, air embolisms, embolisms such as cholesterol embolisms, fat embolisms, pulmonary embolisms or amniotic fluid embolism, thromoboembolisms, thrombosis such as coronary thrombosis, hepatic vein thrombosis, retinal vein occlusion, carotid artery thrombosis, sinus thrombosis, Wallenberg's syndrome, and thrombophlebitis.

The F3Cs can also be used to treat, prevent or ameliorate one or more symptoms of vascular ischemia or myocardial ischemias, vasculitis and coronary diseases, including angina \ pectoris, coronary aneurysm, coronary arteriosclerosis, coronary thrombosis, coronary vasospasm, myocardial infarction and myocardial stunning, cerebral ischemia, ischemic colitis, compartment syndromes, anterior compartment syndrome, myocardial ischemia, reperfusion injuries, peripheral limb ischemia, aortitis, arteritis, Behcet's Syndrome, mucocutaneous lymph node syndrome, thromboangiitis obliterans, hypersensitivity vasculitis, Schoenlein-Henoch purpura, allergic cutaneous vasculitis, Wegener's granulomatosis or metabolic syndrome, which may be accompanied by one, two or more of obesity, insulin resistance, dyslipidemia, hypertension or other related symptoms or conditions.

Exemplary symptoms that the use of the F3Cs can ameliorate include one or more of pain such as arm, jaw or chest pain, edema or swelling, high blood pressure, shortness of breath or dyspnea, e.g., on exertion or while prone, fatigue or malaise and low cardiac injection fraction. In treating a cardiovascular condition in a subject or in improving one or more symptoms thereof, the F3Cs may accomplish one or more of increasing cardiac ejection volume or fraction, decreasing levels of IL-6, decreasing levels of C reactive protein, fibrinogen, cardiac creatinine kinase, increasing fatty acid metabolism or utlization by cardiac tissue, increasing camityl palmitoyl fatty acid transferase or other cardiac metabolic enzymes, activating potassium dependent calcium channels, vasodilating or enhancing oxygen delivery to ischemic tissues or decreasing levels of scarring or plaque formation that occurs, e.g., after vascular damage. Symptoms associated with a cardiovascular condition such as ischemia that can be ameliorated also include acidosis, expression of one or more immediate early genes in, e.g., glial cells, vascular smooth muscle cells or endothelial cells, neuronal membrane depolarization and increased neuronal extracellular calcium and glutamate concentration. Other biological effects associated with treatment using a F3C may also be monitored, e.g., and increase or decrease of a cell surface antigen, a cytokine or an interleukin as disclosed herein.

Useful biological effects of the F3Cs in cardiovascular indications such as myocardial ischemias also include preventing or reducing heart or vascular cell death and subsequent fibrosis. These effects are associated with a decreased oxidative capacity of heart cells or myocytes, which is associated with a decreased capacity of the cells to metabolize fatty acids efficiently. The compounds enhance fatty acid metabolism and ameliorate the deleterious effects of a limited oxidative capacity. j 12 The F3Cs also can limit inflammation or cell injury that is associated with ischemia or oxygen reperfusion after ischemia. Ischemia, which is a detrimental decrease in oxygenated blood delivery to affected cells or tissues, may arise from a cardiovascular condition or event such as an infarction, or from thermal injury or bums. Ischemia may also arise from accidental or surgical trauma. Reperftision after cells have become hypoxic for a sufficient period of time can lead to tissue or cell injury that varies from slight to lethal. The compounds can reduce cell or tissue injury or death associated with ischemia and reperfusion, by, e.g., reducing inflammation or the level of a molecule associated with inflammation. Thus, levels of a proinflammatory cytokine or molecule such as leukotriene B4, platelet activating factor or levels of extracellular P-selectin may result from administration of a F3C to a subject who may experience reperftision injury. Thus, the compounds can reduce injury or death of, e.g., neuron, cardiac, vascular endothelium, myocardial, pulmonary, hepatic or renal cells or tissues. Without wishing to be bound by any theory, the compounds may act in part by reducing one or more of neutrophil activation, platelet activation, platelet aggregation, endothelial cell activation and neutrophil adherence or adhesion to endothelial cells in these conditions.

The F3C-s are useful to treat autoimmune or metabolic conditions or disorders, or their symptoms, in subjects such as mammals or humans, that relate to impaired insulin synthesis or use or that relate to abnormal or pathological lipid or cholesterol metabolism or levels.

Such conditions and symptoms include polycystic ovarian syndrome, Type 1 diabetes (including Immune-Mediated Diabetes Mellitus and Idiopathic Diabetes Mellitus), Type 2 diabetes (including forms with (1) predominant or profound insulin resistance, (2) predominant insulin deficiency and some insulin resistance, (3) forms intermediate between these), obesity, hyperglycemia and dyslipidemia, unwanted hyperlipidemia conditions such λ as hypertriglyceridemia and hypercholesterolemias such as hyper-LDL cholesterolemia, (4) unwanted hypolipidemias, e.g., hypo-HDL cholesterolemia or low HDL cholesterol levels and (5) angina pectoris. In diabetes, the compounds are useful to (1) enhance .beta.-cell function in the islets of Langerhans (e.g., increase insulin secretion), (2) reduce the rate of 312 islet cell damage, (3) increase insulin receptor levels or activity to increase cell sensitivity to insulin and/or (4) modulate glucocorticoid receptor activity to decrease insulin resistance in cells that are insulin resistant. The compounds are thus useful to treat, prevent, ameliorate or slow the progression of a metabolic or cardiovascular condition such as diabetes or hyperglycemia, or a related symptom or condition such as a dyslipidemia in a subject such as a human or a mammal.

Beneficial effects that can the F3Cs can exert on such related symptoms or conditions include improved glucose tolerance, improved glucose utilization, decreased severity or slowed progression of vascular disease (e.g., microvascular or macrovascular disease, including nephropathy, neuropathy, retinopathy, hypertension, cerebrovascular disease and coronary heart disease) or a decreased severity or slowed progression of atherosclerosis, an arteriosclerosis condition (e.g., coronary arteriosclerosis, hyperplastic arteriosclerosis, peripheral arteriosclerosis or hypertensive arteriosclerosis), decreased severity or slowed progression of diabetic osteoarthropathy, skin lesions, rhabdomyolysis, ketosis, detectably decreased generation of islet cell autoantibodies, decreased levels or activity of inflammatory macrophages (foam cells) in atherosclerotic plaques, or detectably decreased expression or levels of one or more of human (or mammalian) angiopoietin-like 3 gene product, apolipoprotein C-l, inducible or constitutive nitric oxide synthase, e.g., in endothelial cells, macrophages or the like, pyruvate dehydrogenase kinase 4, carboxyl ester lipase, cholesteryl ester transfer protein, endothelial lipase, vascular wall lipoprotein lipase, anti-lipoprotein lipase autoantibodies, triglyceride-rich lipoproteins, LDL cholesterol, Creactive protein, high sensitivity C-reactive protein, fibrinogen, plasma homocysteine, VCAM-1, IL-1 (e.g., IL-1.beta.), IL-6, a TNF (e.g., TNF.alpha.), AP-1, NF-.kappa.B, and IFN-.gamma.. In these any of these diseases or conditions, the F3Cs can also modulate, e.g., detectably increase, the activity or level of one, two or more of human (or mammalian) LOX-1, apolipoprotein A-l, apolipoprotein A-2, LPDL lipase, hormone sensitive lipase, paraoxonase, brain natriuretic peptide, a brain natriuretic peptide receptor, e.g., Nprl or Npr3, hepatic lipase, LDL receptor, HDL apoliporpotein E, HDL apoliporpotein J, HDL cholesterol, VLDL receptor, ATP-binding casette transporter 1, leukemia inhibitor)' factor. °60 312 CD36, LXR.alpha., LXR.beta., CAR.beta., RXR, PPAR.alpha., PPAR.beta., PPAR.gamma. or a lipoprotein lipase, e.g., marophage lipoprotein lipase. As used herein, obesity includes a human with a body mass index of at least about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36 or greater. Obese humans that are treated with a F3C may have one or more of the conditions descrbed here.

The F3Cs are useful in treating insulin resistance and associated symptoms and conditions. Insulin resistance is typically observed as a diminished ability of insulin to exert its biological action across a broad range of concentrations. This leads to less than the expected biologic effect for a given level of insulin. Insulin resistant subjects or human have a diminished ability to properly metabolize glucose or fatty acids and respond poorly, if at all, to insulin therapy. Manifestations of insulin resistance include insufficient insulin activation of glucose uptake, oxidation and storage in muscle and inadequate insulin repression of lipolysis in adipose tissue and of glucose production and secretion in liver. Insulin resistance can cause or contribute to polycystic ovarian syndrome, impaired glucose tolerance, gestational diabetes, hypertension, obesity, atherosclerosis and a variety of other disorders. Insulin resistant individuals can progress to a diabetic state. The compounds can also be used in the treatment or amelioration of one or more condition associated with insulin resistance or glucose intolerance including an increase in plasma triglycerides and a decrease in high-density lipoprotein cholesterol, high blood pressure, hyperuricemia, smaller denser low-density lipoprotein particles, and higher circulating levels of plasminogen activator inhibitor-1. Such diseases and symptoms have been described, see, e.g., G. M. Reaven, J. Basic Clin. Phys. Pharm. 1998,9: 387-406, G. M. Reaven, Physiol. Rev. 1995, 75: 473-486 and J. Flier, J. Ann. Rev. Med. 1983, 34:145-60.

The compounds can thus be used in diabetes, obesity, hyperlipidemia or hypercholesterolemia conditions to reduce body fat mass, increase muscle mass or to lower one or more of serum or blood low density lipoprotein, triglyceride, cholesterol, apolipoprotein B, free fatty acid or very low density lipoprotein compared to a subject that would otherwise be considered normal for one or more of these characteristics. These ^60312 beneficial effects are typically obtained with little or no effect on serum or blood high density lipoprotein levels. The F3Cs are useful to reduce or slow the rate of myocardial tissue or myocyte damage, e.g., fibrosis, or to enhance cardiac fatty acid metabolism in conditions, such as inflammation, where fatty acid metabolism is depressed or decreased. Elevated cholesterol levels are often associated with a number of other disease states, including coronary artery disease, angina pectoris, carotid artery disease, strokes, cerebral arteriosclerosis, and xanthoma, which the F3Cs can ameliorate or slow the progression or severity of. Abnormal lipid and cholesterol conditions that can be treated include exogenous hypertriglyceridemia, familial hypercholesterolemia, polygenic hypercholesterolemia, biliary cirrhosis, familial combined hyperlipidemia, dysbetalipoproteinemia, endogenous hypertriglyceridemia, mixed hypertriglyceridemia and hyperlipidemia or hypertriglycidemia secondary to alcohol consumption, diabetic lipemia, nephrosis or drug treatments, e.g., corticosteroid, estrogen, colestipol, cholestyramine or retinoid treatments. Dosages, routes of administration and dosing protocols for the F3Cs are essentially as described herein. Where the condition is chronic, the F3Cs will generally be administered to a subject such as a human for a relatively long time period, e.g., for about 3 months to about 10 years or more. Dosages, routes of administration and dosing protocols for the F3Cs are essentially as described herein. Dosing of the compound can be daily or intermittent using a dosing protocol using dosages as described herein, e.g., about 0.01 to about 20 mg/kg of a F3C administered to a subject once or twice per day daily or intermittently. The use of the F3Cs can be combined with one, two or more other suitable treatments, e.g., treatment for cessation of smoking, diet control, e.g., caloric restriction or reduced fat intake, or treatment with fibrates, non-steroidal anti-inflammatory drugs, angiotensin-converting enzyme inhibitors or HMG-CoA reductase inhibitors such as aspirin, clofibrate, fenofibrate, ciprofibrate, gemfibrozil, Simvastatin.TM., Pravastatin.TM., Mevastatin.TM. or Lovastatin.TM.

The use of any F3C or species in any genus of F3Cs disclosed herein to treat, prevent or ameliorate any of these cardiovascular or metabolic disorders or symptoms will generally use one or more of the routes of administration, dosages and dosing protocols as disclosed herein. Thus, in exemplary embodiments, about 0.5 to about 100 rng/kg or about 1 to about 25 rng/kg, of the F3C will be administered per day by an oral, buccal, sublingual or parenteral route. Such administration can be, e.g., daily for about 5 to about 60 days in acute conditions or it can be intermittent for about 3 months to about 2 years or more for chronic conditions. Alternatively, intermittent dosing can be used essentially as described herein for acute cardiovascular conditions. In some embodiments, for conditions such as ischemia or trauma, administration of the F3C is provided before or as soon after the ischemic or traumatic event as possible, e.g., within about 6 hours of an ischemic or traumatic event or about 12-24 hours before an anticiapted ischemic or traumatic event. In other embodiments, administration of the F3C can be delayed for, e.g., about 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 18,19, 20, 21, 24, 28, 32, 36,40, 48 or more hours after an ischemic or traumatic event has occurred and a course of daily or intermittent dosing is initiated at one of these times, or in a range between any of these times after the event. Thus, administration of the F3C can begin at about 10-14 hours, at about 11-13 hours or at about 8-16 hours after the ischemic or traumatic event.

In another aspect of the invention, the F3Cs can be used to prevent, treat or to reduce the severity of vascular or microvascular occlusions in human blood cell deficency disorders (SCD). SCD is heterogenous and includes subgroups with high transcranial velocities, which is a group with an increased risk of infarctive stroke or cereberal thrombosis. SCD types also include sickle cell-.beta..sup.+ thalassemia, sickle cell-.beta..sup..smallcircle. thalassemia, sickle cell-.delta..beta..sup.- .smallcircle. thalassemia and sickle cell-HPFH (hereditary' of persistent fetal hemoglobin). Another subgroup of SCD patients is characterized by the presence of a Plasmodium parasite infection. SCD is usually accompanied by acute vasoocclusive episodes such as microvascular occlusions, ischemia and infarctions that arise from adhesion of sickle cells and other blood cell types, e.g., platelets or leukocytes, to vascular endothelial cells. Reduced sickle cell adhesion in response to treatment with a F3C and related responses is facilitated at least in part by decreased production or activity of one or more biological response mediators such as one, two, three or more of thrombospondin, von Willebrand factor, epinephrine, C reactive protein, cAMP, basal cell adhesion molecule/Lutheran (BCAM/Lu), P-selectin, L-sclectin, E-selectin, VCAM-1, ICAM-1, fibronectin, annexin V, placenta growth factor, superoxide, CD1 la, CD1 lb, CD1 lc, CD15, CD18, CD31, CD36, TNF.alpha., NF-.kappa.B, IL-1 .beta, or IL-6 by endothelial cells or one or more immune cell types as described herein. In treating blood cell deficency disorder, the F3Cs will also increase the activity or levels of one, two or more desired response mediators including fetal hemoglobin, erythropoietin, heme oxygenase, nitric oxide, PPAR.alpha., PPAR.gamma. or GM-CSF. The F3Cs will thus ameliorate one or more symptoms of blood cell deficency disorder such as anemia, stroke, pain, e.g., chest or abdominal pain, skin ulcers, dyspnea, organ damage, retinopathy or the level of infected red cells in Plasmodium-infected subjects. Treatment of acute SCD episodes or of chronic SCD with F3Cs can be combined with other suitable therapies, e.g., inhaled nitric oxide, hydroxyurea treatment, anti-adhesion molecule antibody treatment or analgesic use such as morphine, oxycodone, or codeine. The F3Cs can also be used to reduce cellular damage from reactive oxygen species associated with hydroxyurea treatment, since the F3Cs will enhance cellular antioxidant capacity.

As is apparent from the foregoing, the use of the F3C is optionally combined with one or more additional therapies for cardiovascular or related disorders, e.g., insulin therapy, vascular surgery, cardiac surgery, angioplasty, or treatment with andrenergic blockers, coronary vasodilators, calcium channel blockers, nitrates, angiotensin converting enzyme inhibitors, anti-hypertensives, anti-inflammatory agents, diuretics, anti-arrhythmia agents, thrombolytic agents, enzyme inhibitors such as hydroxymethylglutaryl CoA reductase inhibitors or xanthine oxidase inhibitors. Exemplary hydroxymethylglutaryl CoA reductase inhibitors include statins such as mevastatin, lovastatin, pravastatin, simvastatin or compounds described in U.S. Pat. Nos. 4,346,227, 4,448,979,4,739,073, 5,169,857, 5,006,530 or 5,401,746. Other therapies that can be applied include diet control, dietary calorie restriction or diet modification for subjects who are or who are susceptible to developing a cardiovascular or related condition such as pulmonary hypertension, diabetes, a dyslipidemia or obesity, e.g., humans having a body mass index of 27, 28, 29, 30, 31, 32, 33, 34, 35, 36 or greater. Diet modifications include limiting or restricting salt, alcohol, caffeine, ΙΕ ο ό ϋ 3 12 cigarette, drugs, e.g., opiate, hallucinogen, sedative, narcotic or amphetamine, sugar, refined sugar and/or fat or cholesterol intake, use or abuse. Additional therapies include treatment with one or more of digoxin, nitroglycerin, doxazosin mesylate, nifedipine, enalapril maleate, indomethicin, tissue plasminogin activator, urokinase, acetylsalicylic acid or the like. Any of such additional therapies would be used essentially according to standard protocols and such therapies would precede, be concurrent with or follow treatment with a F3C. In some embodiments, such additional therapies will be administered at the same time that a F3C is being used or within about 1 day to about 16 weeks before or after at least one round of treatment with the F3C is completed. Other exemplary therapeutic agents and their use have been described in detail, see, e.g., Physicians Desk Reference 54.sup.th edition, 2000, pages 303-3251, ISBN 1-56363-330-2, Medical Economics Co., Inc., Montvale, N.J.; Harrison's Principles of internal Medicine, 15.sup.th edition, 2001, E. Braunwald, et al., editors, McGraw-Hill, New York, N.Y., ISBN 0-07-007272-8, especially chapters 231, 241 248 and 258-265 at pages 1309-1318, 1377-1442 and 1491-1526. One or more of these exemplary agents or treatments can be used in combination with a F3C to treat any of the appropriate cardiovascular and related disorders described herein and in the references cited herein.

Respiratory and Pulmonary conditions. F3Cs can be used to treat, ameliorate, prevent or slow the progression of a number of pulmonary conditions or their symptoms such as 1, 2, 3 or more of cystic fibrosis, bronchiectasis, cor pulmonale, pneumonia, lung abcess, acute bronchitis, chronic bronchitis, chronic obstructive pulmonary diseases, emphysema, pneumonitis, e.g., hypersensitivity pneumonitis or pneumonitis associated with radiation exposure, alveolar lung diseases and interstitial lung diseases, e.g., associated with asbestos, fumes or gas exposure, aspiration pneumonia, pulmonary hemorrhage syndromes, amyloidosis, connective tissue diseases, systemic sclerosis, ankylosing spondylitis, allergic granulomatosis, granulomatous vasculitides, asthma, e.g., acute asthma, chronic asthma, atopic asthma, allergic asthma or idiosyncratic asthma, cystic fibrosis and associated conditions, e.g., allergic bronchopulmonary aspergillosis, chronic sinusitis, pancreatic insufficiency, inflammation or Haemophilus influenzae, S. aureus or Pseudomonas 060312 aeruginosa infection. In some of these conditions where inflammation plays a role in the pathology of the condition, the F3Cs can ameliorate or slow the progression of the condition by reducing damage from inflammation. In other cases, the F3Cs act to limit pathogen replication or pathogen-associated lung tissue damage.

For these conditions, the severity of the disease or the type or severity of associated symptoms can vary. For example, in humans having pediatric, e.g., infants or children of about 1 month or about 4 months of age to about 16 or 17 years of age, or adult cystic fibrosis (CF), the disease may be associated with the presence of one or more symptoms, syndromes, genetic mutations or the like. Symptoms or syndromes that can be observed in human CF patients include 1, 2, 3, 4 or more of Staphylococcus (e.g., S. aureus), Haemophilus influenzae, Pseudomonas or Burkholderia respiratory tract or lung infection or propensity to develop detectable infection or colonization, coughing, wheezing, cyanosis, bronchiolitis, bronchospasm, pneumothorax, hemoptysis, pancreatic exocrine insufficiency, bronchiectatic lung disease, atelectasis-consolidation, pulmonary edema, increased lung vascular hydrostatic pressure, increased lung vascular permeability, sinusitis, respiratory insufficiency, bronchial wall or interlobular septa thickening, reduction of forced expiratory volume in 1 second, dyspnea, impaired male fertility, elevated sweat chloride (e.g., >60 mmol/L), mucous plugging, tree-in-bud sign, mosaic perfusion pattern, glucose intolerance or abnormal elevation of one or more of IL-4, IL-8, RANTES, neutrophil elastase, eosinophils, macrophages, neutrophils, eosinophil cationic protein or cysteinyl leukotrienes. Any of these symptoms or syndromes can be acute, intermittent or chronic and/or mild, moderate or severe. Relevant mutations include, e.g., a homozygous or heterozygous, dominant or recessive deletion, insertion and/or point mutation in (1) the cationic trypsinogen gene or (2) the cystic fibrosis transmembrane conductance regulator (CFTR) gene, such as one, two or more of, a CFTR F508del deletion mutation or CFTR lacking phe508, 3272-26A>G/F508deI, 3659deIC, 394deITT, S1455X or .DELTA.26,I1234V, 2183AA>G, 2043deIG, 548A>T, I148T, R334W, SI 196X, 4041 C>G, 1161 deIC, 1756G>T or 312O+1G>A mutation.

IE 0 60 312 The use of a F3C to treat, ameliorate or slow the progression of conditions such as CF can be optionally combined with other suitable treatments. For CF, this includes, e.g., one, two or more of oral or aerosol corticosteroid treatment, ibuprofen treatment, DNAse or IL-10 treatment, diet control, e.g., vitamin E supplementation, vaccination against pathogens, e.g., Haemophilus influenzae, or chest physical therapy, e.g., chest drainage or percussion.

Humans or other subjects who have one or more of these conditions can be treated with other suitable therapeutics. Pulmonary conditions that can be treated with the F3Cs and other therapeutic methods and agents that can be used in conjunction with the F3Cs have been described in detail, see, e.g., Harrison's Principles of Internal Medicine, 15.sup.th edition, 2001, E. Braunwald, et al., editors, McGraw-Hill, New York, N.Y., ISBN 0-0712 007272-8, especially chapters 252-265 at pages 1456-1526; Physicians Desk Reference 54.sup.th edition, 2000, pages 303-3251, ISBN 1-56363-330-2, Medical Economics Co., Inc., Montvale, N.J. One or more of these exemplary agents or treatments can be used in combination with a F3C to treat any of the appropriate cardiovascular and related disorders described herein and in the references cited herein. Treatment of any of these respiratory and pulmonary conditions using a F3C is accomplished using the treatment regimens described herein. For chronic conditions, intermittent dosing of the F3C can be used to reduce the frequency of treatment. Intermittent dosing protocols are as described herein.

Applications in autoimmunity, allergy, inflammation and related conditions. As mentioned above, the F3Cs may be used to treat, prevent or slow the progression of one or more autoimmune allergic or inflammatory diseases, disorders, or conditions, or to ameliorate one or more symptoms thereof in a subject. These diseases and conditions include Addison's Disease, autoimmune hemolytic anemia, autoimmune sensorineural hearing loss, antiphospholipid syndrome, acute or chronic rheumatoid arthritis and other synovial disorders, an osteoarthritis including post-traumatic osteoarthritis and hypertrophic pulmonary osteoarthropathy, psoriatic arthritis, polyarthritis, epichondylitis, type I diabetes, type II diabetes, rheumatic carditis, bursitis, ankylosing spondylitis, multiple sclerosis, a dermatitis such as contact dermatitis, atopic dermatitis, exfoliative dermatitis or seborrheic ΙΕ ν 6 0 3 12 100 dermatitis, mycosis fungoides, allergic encephalomyelitis, autoimmune glomerulonephritis, Goodpasture's Syndrome, Graves’ Disease, Hashimoto's Thyroiditis, multiple sclerosis, myasthenia gravis, neuritis, bullous pemphigoid, pemphigus, polyendocrinopathies, purpura, Reiter's Disease, autoimmune thyroiditis, systemic lupus erythematosus, scleroderma, fibromyalgia, chronic fatigue syndrome, autoimmune pulmonary inflammation, GuillainBarre Syndrome, type 1 or insulin dependent diabetes mellitus, autoimmune inflammatory eye disease, hepatitis C virus associated autoimmunity, postinfectious autoimmunity associated with, e.g., virus or bacterial infection such as a parvovirus such as human parvovirus B19 or with rubella virus, autoimmune skin and muscle conditions such as pemphigus vulgaris, pemphigus foliaceus, systemic dermatomyositis or polymyositis or another inflammatory myopathy, myocarditis, asthma such as allergic asthma, allergic encephalomyelitis, allergic rhinitis, a vasculitis condition such as polyarteritis nodosa, giant cell arteritis or systemic necrotizing vasculitis, chronic and an acute or chronic inflammation condition such as chronic prostatitis, granulomatous prostatitis and malacoplakia, ischemiareperfusion injury, endotoxin exposure, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine induced lung injury, cachexia, sarcoidosis, inflammatory bowel disease, regional enteritis, ulcerative colitis, Crohn's disease, inflammatory bowel disease or inflammation associated with an infection, e.g., septic shock, sepsis, or systemic inflammatory response syndrome. Any of these diseases or conditions or their symptoms may be acute, chronic, mild, moderate, severe, stable or progressing before, during or after the time administration of the F3C to a subject such as a human, is initiated. In general, a detectable improvement is observed in the subject within a period of about 3 days to about 12 months after initiation of a dosing protocol, e.g., the severity of the disease or condition will detectably decrease, the rate of progression will detectably slow or the severity of a symptom(s) will detectably decrease.

As used herein, acute inflammation conditions are characterized as an inflammation that typically has a fairly rapid onset, quickly becomes moderate or severe and usually lasts for only a few days or for a few weeks. Chronic inflammation conditions as used herein are characterized as an inflammation that may begin with a relatively rapid onset or in a slow, or 101 even unnoticed manner, tends to persist for at least several weeks, e.g., about 3-6 weeks, months, or years and may have a vague or indefinite termination. Chronic inflammation may result when the injuring agent (or products resulting from its presence) persists in the lesion, and the subject's tissues respond in a manner (or to a degree) that is not sufficient to overcome completely the continuing effects of the injuring agent. Other exemplary conditions are described in, e.g., Textbook of Autoimmune Diseases, R. G. Lahita, editor, Lippincott Williams & Wikins, Philadelphia, Pa., 2000, ISBN 0-7817-1505-9, pages 175851 and Rheumatology, 2,sup.nd edition, J. H. Klippel et al., editors, 1998, ISBN 0-72342405-5, volume 1, sections 1-5 and volume 2, sections 6-8, Mosby International, London, UK.

A F3C can be used to inhibit or ameliorate one or more inappropriate immune responses or their symptoms in autoimmunity, inflammation, allergy or related conditions. The effects of the F3Cs include detectably ameliorating one or more of (1) the proliferation, differentiation or chemotaxis of T cells, (2) reducing unwanted cytotoxic T cell responses, (3) reducing unwanted autoantibody or other antibody synthesis, e.g., an unwanted IgA, IgE, IgG or IgM, in allergy, asthma or another autoimmune or inflammation condition, (4) inhibiting the development, proliferation or unwanted activity of autoreactive T or B cells, (5) altering the expression of one or more cytokines, interleukins or cell surface antigens, e.g., a cytokine, interleukin or cell surface antigen described herein (decreasing IL-8 in an autoimmune condition, decreasing the level of acute phase proteins such as C reactive protein or fibrinogen in inflammation conditions, (6) decreasing eosinophilia in allergy conditions, (7) detectably decreasing the level or activity of one or more of ICAM-1, IL-l.alpha., IL-l.beta., TNF.alpha., IL-6 or IL-8 in, e.g., inflammation conditions or in autoimmune conditions such as an arthritis or a myocarditis condition such as osteoarthritis, rheumatoid arthritis, toxic myocarditis, indurative myocarditis or idiopathic myocarditis, (8) decreasing the level or biological activity of one or more of anti-islet antibody, TNF, IFN-.gamma., IL-1, an arthritis symptom(s), nephritis, skin rash, photosensitivity, headache frequency or pain, migraine frequency or pain, abdominal pain, nausea or anti-DNA antibodies in, e.g., insulin dependent diabetes mellitus or an autoimmune or inflammation condition such as systemic /£ νού3 t2 102 lupus erythematosus, rheumatoid arthritis or Crohn's disease, (9) reducing induction of arachidonic acid metabolism or reducing eicosanoid metabolites such as thromboxanes or prostaglandins in, e.g., inflammation, asthma or allergy, (10) reducing IL-4, IL-8 or IL-10 synthesis, levels or activity in, e.g., allergy or inflammation such as idiopathic pulmonary fibrosis or allergic asthma or (11) reducing or interfering with neutrophil chemotaxis by, e.g., reducing thioredoxin release from affected cells in conditions such as cancer, infections, inflammation or autoimmunity.

Exemplary symptoms that the use of the F3Cs can ameliorate in these autoimmune, inflammatory and allergy conditions include one or more of pain such as shoulder, hip, joint, abdominal or spine pain, joint stiffness or gelling, bursitis, tendonitis, edema or swelling, fatigue or malaise, headache, dyspnea, skin rash, fever, night sweats, anorexia, weight loss, skin or intestine ulceration, muscle weakness, pericarditis, coronary occlusion, neuropathy and diarrhea. In treating one of these conditions in a subject or in improving one or more symptoms thereof, the F3Cs may accomplish one or more of decreasing levels of one or more of IL-1, IL-4, IL-6 or TNF.alpha., decreasing levels ofC reactive protein, fibrinogen or creatinine kinase. Other biological effects associated with treatment using a F3C may also be monitored or observed, e.g., an increase or decrease of a cell surface antigen, a cytokine or an interleukin as disclosed herein.

In another aspect of the invention, the F3Cs can be used to treat or to reduce the severity of chronic allergies or atopic diseases such as allergic rhinitis, psoriasis, eczema, gastrointestinal allergies, atopic dermatitis conditions, allergic asthma, food allergies and hay fever. These conditions are typically characterized by the presence of elevated levels of allergen specific antibodies of the IgE isotype. In treating or ameliorating these conditions, the F3Cs reduce the generation of IgE by isotype switching, which is increasing allergenspecific IgA production and/or decreasing IgE production from preexisting allergen-primed cells. Allergen specific IgG may also be increased from new cells that might otherwise have responded to allergen exposure by generating unwanted IgE.

Ogo 312 103 The IgA and IgG are allergen specific, which will enhance clearance of allergen from inucosa or other tissue and reduction of chronic or late phase allergic responses. The F3C’ can thus also be used to increase the biological clearance of allergens from tissue and mucosa. Reduced generation of the levels or activity of IgE by B cells in response to treatment with a F3C and related responses is facilitated at least in part by decreased production of one or more biological response mediators, e.g., cytokines or response mediators such as protein kinase A inhibitors, substance P neuropeptide, thymus- and activation-regulated chemokine, e.g., by airway smooth muscle cells, proteinase activated receptor-2 by neurons, intracellular signal-transducing protein-6 (STAT6), Janus kinase 1, Janus kinase 6, CD40, CD86 and/or NF-kB by B cells, CD1 54 in T cells, and suppressor of cytokine signalling-3, phosphodiesterase 4, TNF-.alpha., MCP-1, RANTES, CXCL10, CXCL8 (IL-8), prostaglandin E2 receptor, IL-l.beta., IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, and IL-23, by one or more other cell types such as immune cells as described herein, airway smooth muscle cells, mucosal cells or keratinocytes. In these treatments, the F3Cs will also increase the activity or levels of one or more desired response mediators including soluble CD23, cathepsin E, epidermal growth factor receptor, IFN.gamma., IL-2, IL-12 or IL-1 8. In treating these conditions, Treatment can be combined with other suitable therapies, e.g., corticosteroids such as fluticasone propionate. The F3Cs can also be used to reduce rebound phenomena following withdrawal of corticosteroid therapies, since the F3Cs have an antiinflammatory effect without having immunosuppressive side effects. Use ofthe F3Cs to generate any of these biological responses or treatments can be by daily or intermittent administration of the F3C to the subject.

In a related embodiment, the F3Cs are used in allergen vaccination protocols to enhance levels or activity of allergen specific IgA or IgG, which contributes to reducing IgE responses to allergen exposure. Such protocols are used to decrease a subject's sensitivity to allergen exposure. Typically such allergies are chronic or atopic. In these applications, the vaccination protocol typically uses the allergen(s) or an active fragment(s) of the allergen that is associated with the allergy or atopic condition. In these methods, a F3C is administered to a subject who has an IgE mediated allergy or atopy condition in conjunction 104 ΙΕ ο 6 0 3 12 with administration of the allergen. Allergens typically used include dermatophagoides, house dust, cat allergen and pollen. In any of these inethods isotype switching or vaccination methods, the F3C is typically administered as described herein, e.g., by administering the F3C about 1, 2, 3, 4, 5, 6, 7, 8, or more days before the allergen is administered to the subject. The subject may receive about 1-20 mg/kg of a F3C at 2, 3 and 4 days before the allergen is administered or injected. The F3C treatment increases allergen specific IgA or IgG responses or levels relative to untreated controls. The use of F3C with allergens will reduce the total number of anti-allergic vaccinations that are needed, increase the quality or length of an effective response and/or increase the proportion of subjects in which allergy shots are effective. An effective response is seen in about 55%, 60%, 65%, 70%, 75%, 80% or more of vaccinated patients who also receive the F3C compared to about 40-50% of vaccinated patients who do not receive the F3C.

In treating inflammation or any condition described herein where inflammation contributes to the condition, the F3Cs may detectably modulate, e.g., decrease or increase, the expression or level or activity of one or more biomolecules associated with the prevention, establishment, maintenance or progression of the inflammation condition. Such biomolecules include one or more of carcinoembryonic antigen, prostate specific antigen, her2/neu, Bcl-XL, bcl-2, p53, IL-1.alpha., IL-1.beta., IL-6, or TNF.alpha., GATA-3, COX-2, NF.kappa.B, IkB, an IkB kinase, e.g., IkB kinase-.alpha,, IkB kinase-.beta. or IkB kinase.gamma., NF AT, a ras protein such as H-ras or K-ras, cyclin D, cyclin E xanthine oxidase, or their isoforms, orthologs, homologs or mutant forms, which may have either reduced or enhanced biological activity(ies), and which may be detectably decreased. Biomolecules that can be detectably increased include IL-2, IFN.gamma., IL-12, T-bet, O6-methylguanineDNA-methyltransferase, calcineurin, calmodulin, a superoxide dismutase (e.g., Mn, Zn or Cu), a tumor suppressor protein such as the retinoblastoma protein (Rb) or CDKN2A (pi6), BRCA1, BRCA2, MeCP2, MBO2, PTEN, NBR1, NBR2 dr the isoforms, orthologs, homologs or mutant forms, which may have either attenuated or enhanced biological activity(ies), of any of these molecules. One or more of these biomolecules may be modulated in any inflammation condition described herein. ί£ 0 δ G j ΐ £ 105 The use of any F3C or species in any genus of F3Cs disclosed herein to treat, prevent or ameliorate any of these autoimmune, inflammatory or allergy conditions or symptoms will generally use one or more of the routes of administration, dosages and dosing protocols as disclosed herein. Thus, in exemplary embodiments, about 0.5 to about 100 mg/kg or about 1 mg/kg to about 15 mg/kg, of the F3C will be administered per day by, e.g., an oral, buccal, sublingual, topical or parenteral route. Such administration can be, e.g., daily for about 5 to about 60 days in acute conditions or it can be intermittent for about 3 months to about 2 years or more for chronic conditions. Alternatively, intermittent dosing can be used essentially as described herein for acute autoimmune, inflammatory and allergy conditions.

In another aspect of the invention, the F3Cs can be used to treat or to reduce the severity of chronic allergies or atopic diseases such as allergic rhinitis, psoriasis, eczema, gastrointestinal allergies, atopic dermatitis conditions, allergic asthma, food allergies and hay fever. These conditions are typically characterized by the presence of elevated levels of the IgE isotype and of B cells that generate IgE. In treating or ameliorating these conditions, the F3Cs reduce the generation of IgE by facilitating an isotype switch from B cells that produce IgE to cells that produce antigen-specific IgA and/or IgG4. The IgA and IgG4 are allergen specific, which will facilitate clearance of allergen from mucosa or other tissue and reduction of chronic or late phase allergic responses. The F3C can thus be used to increase the biological clearance of allergens from tissue. Reduced generation of the levels or activity of IgE by B cells in response to treatment with a F3C and related responses is facilitated at least in part by decreased production of one or more biological response mediators, e.g.. cytokines or response mediators such as protein kinase A inhibitors, substance P neuropeptide, thymus- and activation-regulated chemokine, e.g., by airway smooth muscle cells, proteinase activated receptor-2 by neurons, intracellular signal-transducing protein-6 (STAT6), Janus kinase 1, Janus kinase 6, CD40, CD86 and/or NF-.kappa.B by B cells, CD 154 in T cells, and suppressor of cytokine signalling-3, phosphodiesterase 4, TNF.alpha., MCP-1, RANTES, CXCL10, CXCL8 (IL-8), prostaglandin E.sub.2 receptor, ILl.beta., IL4, IL-5, IL-6, IL-10, IL-13 and IL-18 by one or more other cell types such as 106 IE 0 g q j 12 immune cells as described herein, airway smooth muscle cells, mucosal cells or keratinocytes. In these treatments, the F3Cs will also increase the activity or levels of one or more desired response mediators including cathepsin E, epidermal growth factor receptor, IFN.gamma., IL-2, IL-12. In treating these conditions, Treatment can be combined with other suitable therapies, e.g., corticosteroids such as fluticasone propionate. The F3Cs can also be used to reduce rebound phenomena following withdrawal of corticosteroid therapies, since the F3Cs have an anti-inflammatory effect without having immunosuppressive sideeffects. Use of the F3Cs to effect any of these biological responses or treatments can be by daily or intermittent administration of the F3C to the subject.

In a related embodiment, the F3Cs are used to enhance isotype switching from IgE to IgG in these chronic allergies or atopic diseases in vaccination protocols that use the allergen(s) or an active fragment(s) of the allergen that is associated with the allergy or atopic condition. In these methods, a F3C is administered to a subject who has an elevated IgE allergy or atopy condition in conjunction with administration of the allergen. The subject's response to the allergen is an enhanced proportion of B cells that produce IgG compared to B cells that generate IgE. Allergens typically used include dermatophagoides, house dust, cat allergen and pollen. In these methods, the F3C is typically administered about 1, 2, 3, 4, 5, 6, 7, 8, or more days before the allergen is administered to the subject, e.g., the subject receives about 1-20 mg/kg of a F3C at 2, 3 and 4 days before the allergen is administered or injected. The F3C facilitates isotype switching to IgG. The use of F3C with allergens will reduce the total number of anti-allergic vaccinations that are needed, increase the quality or length of an effective response and/or increase the proportion of subjects in which allergy shots are effective, e.g., an effective response is seen in about 55%, 60%, 65%, 70%, 75%, 80% or more of vaccinated patients who also receive the F3C compared to about 40-50% of vaccinated patients who do not receive the F3C.

The F3Cs are suitable for enhancing immune responses in aging in subjects such as humans or primates. In humans at about 50 to 60 years of age and later, one or more aspects of immune responses will typically decrease by a detectable amount compared to typical IE 0 β Q 312 107 immune responses at younger ages, e.g., at about 18-50 years of age. The F3Cs can be used on an intermittent basis or continuously in aged subjects. Intennittent administration of a F3C can occur as described herein, e.g., daily dosing or dosing every other day or every third day for about 1, 2, 3,4, 5, 6, 7, 8 or 9 days, followed by about 2, 3,4, 5, 6, 7, 8, 9 or 10 weeks of no dosing, optionally followed by about 1, 2, 3, 4, 5, 6, 7 or 8 days of daily dosing or dosing every other day or every third day and then followed by about 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks of no dosing. Such dosing cycles can be repeated indefinitely or as needed.

Such treatments can be used prophylactically or therapeutically. In prophylaxis the F3C are administered, e.g., before or during influenza outbreaks, or in aged patients in hospitals or in aged patients in long term living or care facilities such as retirement communities or nursing homes. In therapeutic applications, the F3C are used to treat trauma, e.g., bone fractures or active infections. The F3C treatments in these embodiments will result in enhanced immune responses, including increased innate and specific responses to, e.g., infectious agents. These treataients will typically also have other beneficial effects including enhancing bone marow production of blood cells or blood components such as neutrophils or improving levels of dysregulated immune response mediators, e.g., decreasing elevated cortisol, IL-6, IL-10, COX-2 or C reactive protein levels or increasing low IL-2 or IL-12 levels.

In related embodiments, the use of the F3C is optionally combined with one or more additional known or experimental therapies for an autoimmune, inflammatory or allergy disorder(s), e.g., one or more of surgery and treatment with a corticosteroid or glucocorticoid such as hydrocortisone, hydrocortisone acetate, fludrocortisone, prednisone, prednisolone, prednisolone acetate, methylprednisolone, dexamethasone, dexamethasone acetate or triamcinolone acetonide, leflunomide, an antibody, e.g., a human or humanized monoclonal antibody, that decreases the activity or level of C5 complement, TNF.alpha. or TNF.alpha, receptor, an antirheumatic drug such as methorexate, D-penicillamine, sodium aurothiomalate, sulfasalazine or hydroxychloroquine, immunosuppressive agents such as 6thioguanylic acid, chlorambucil, cyclophosphamide or cyclosporin, a non-steroidal antiinflammatory agent such as celecoxib, ibuprofin, piroxicam or naproxin, an antihistamine such as loratidine or promethazine hydrochloride, an analgesic such as IE a 6 o j 12 108 propoxyphene napsylate, acetaminophen or codeine or administration of vitamins (e.g., multivitamins, individual vitamins), antioxidants or other agents (e.g., vitamin E, folinic acid, carnitine, a C2-8 alkanoyl carnitine such as acetyl or propionyl L-camitine) or nutritional supplements (e.g., liquid protein or carbohydrate preparations). Such therapies would be used essentially according to standard protocols and such they would precede, be concurrent with or follow treatment with a F3C. In some embodiments, such additional therapies will be administered at the same time that a F3C is being used or within about 1 day to about 16 weeks before or after at least one round of treatment with the F3C is completed. Other exemplary therapeutic agents and their use have been described in detail, see, e.g., Physicians Desk Reference 54.sup.th edition, 2000, pages 303-3267, ISBN 156363-330-2, Medical Economics Co., Inc., Montvale, N.J. One or more of these exemplary agents can be used in combination with a F3C to ameliorate, prevent or treat any of the appropriate autoimmune, inflammatory or allergy conditions or disorders described herein or any of their symptoms.

Where a natural or synthetic antiinflammatory glucocorticoid is used to treat one more of the conditions disclosed herein or wherein endogenous levels of glucocorticoid such as cortisol are elevated to an unwanted level in a subject, the use of a F3C will ameliorate unwanted side-effects of such glucocorticoid use or excess. Typically the F3C will be administered during, before and/or after glucocorticoid levels are elevated or during, before and/or after a therapeutic glucocorticoid is administered to the subject, e.g., within about 1, 2, 3, 4, 5, 6 or 7 days or within about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16,20 or 24 weeks before or after glucocorticoid use or elevated glucocorticoid levels exist. Typically, the use of the F3C to counteract unwanted side-effects of therapeutic glucocorticoid use In these embodiments, will reduce or ameliorate the onset, severity or progression of one or more unwanted sideeffects of glucocorticoid therapy such as a detectable immune suppression, an increased occurrence or incidence of infection, an undesirable alteration of mood (e.g., increased anxiety, depression or schizophrenia) or a detectable loss or alteration of memory.

Regeneration and wound healing. The F3Cs can be used to facilitate cell differentiation.

IE 060 3 12 109 proliferation or repair where regeneration of tissues is desired. The regeneration of tissues could be used to repair, replace, protect or limit the effects of tissue damaged by congenital defects, trauma (wounds, bums, incisions, or ulcers), age, disease (e.g. osteoporosis, osteoarthritis, periodontal disease, liver failure), surgery, including cosmetic plastic surgery, fibrosis, reperfusion injury, toxin exposure or systemic cytokine damage. Ulcers or skin lesions can arise from ionizing radiation exposure, cytotoxic chemotherapy or pressure, e.g., a pressure or decubitis ulcer or vascular insufficiency, e.g., associated with diabetes or vascular occlusion. Tissues for which regeneration may be enhanced include organs (e.g., pancreas, liver, lung, intestine, kidney, skin, endothelium, oral mucosa, gut or intestinal mucosa), muscle (e.g., smooth, skeletal or cardiac), vasculature (including vascular and lymphatics), central or peripheral nervous tissue, hemnatopoietic tissue, and skeletal tissue (e.g., bone, cartilage, tendon, and ligament). Decreased scarring or an increased rate or quality of healing may accompany these effects.

[0506] The F3Cs are thus useful to enhance healing or tissue repair in a subject having a bone ffacture(s), e.g., a simple or compound skull, spine, hip, arm or leg bone fracture. Similarly, nerve or brain tissue treatment using a F3C allows treating, slowing the progression of, ameliorating or preventing diseases such as central and peripheral nervous system diseases, neuropathies, or mechanical and traumatic diseases, disorders, and/or conditions (e.g., spinal cord disorders, head trauma, cerebrovascular disease, and stoke). The compounds are useful to treat diseases associated with peripheral nerve injuries, peripheral neuropathy (e.g., resulting from chemotherapy, radiation exposure or therapy or other medical therapies), localized neuropathies, and central nervous system diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis. The subjects undergoing treatment in these conditions may be elderly, e.g., a human at least about 55, 60, 65 or 70 years of age. Where the condition is acute, e.g., a bone fracture or a bum, the treatment may comprise administration of a F3C to the subject on a daily or intermittent basis for about 3 days to about 12 months, e.g., administration for about 2-12 weeks beginning after the subject sustains an injury.

IE a60 3 12 110 An aspect of the F3Cs is their capacity to facilitate wound or trauma healing by increasing the proliferation or self-renewal of stem cells and pluripotent derivatives of stem cells and/or the rate of differentiation of stem cells or their pluripotent derivatives to mature cell types. Thus, the F3Cs can increase the numbers, rate of differentiation or activity of stem cells in, e.g., skin, central or nervous system tissue, blood vessels, heart tissue, lung, liver, pancreas, kidney, thymus, spleen, oral mucosa, intestine or bone marrow, some of which is discussed elsewhere herein. Increased numbers of mature cell types typically is observed beginning at about 2-28 days after treatment with a F3C is started, usually after about 2-21 days. Thus, the F3Cs can enhance the numbers, activities or differentiation of, e.g., crypt cells in intestinal mucosa, skin stem cells in the oral mucosa or cardiac precursor cells after damage to those cells or tissues. Such damage can arise, e.g., from trauma, infection, ionizing radiation exposure, toxin exposure and/or cytotoxic chemotherapy. Optimal modulation of stem cell survival, self-renewal and differentiation in these embodiments is usually obtained by dosing the F3C at a time period near the time that the subject is exposed to an agent, event or treatment that can cause significant tissue damage. Typically this time period is about 1, 2, 3, 4 or 5 days before, on the same day as or within 1, 2, 3, 4 or 5 days after the damaging event or exposure occurs. For chronic toxin exposure, e.g., alcohol, chronic continuous or intermittent administration of the F3C can be used. Dosages of the F3Cs, routes of administration and dosing protocols for these embodiments are as described herein.

As noted above, the F3Cs are useful to enhance healing in a subject who has experienced or who is expected to experience one or more traumas or acute injuries such as a wound, bum, bone fracture, nervous system tissue trauma, gastrointestinal damage or intestinal cell damage or other traumatic events. In some embodiments, such subjects have experienced a trauma and who are immune suppressed or are anticipated to become immune suppressed. The immune suppression may arise from, e.g., a myelosuppressive cancer therapy, a glucocorticoid therapy or from radiation exposure. Thus, in some cases a subject such as a human or a primate who has experienced a trauma, e.g., a bone fracture, a chemical or thermal burn, a cut or a laceration, is also exposed to, e.g., an ionizing radiation as described herein such as .gamma.-radiation, .beta.-radiation, X-radiation or neutron radiation in an 312 111 immune suppressive amount or dose, e.g., about 0.3 Gy (gray) to about 30 Gy, typically about 0.5 Gy to about 12 Gy or about 0.7 Gy to about 8 Gy. The subject's radiation exposure can be localized or whole body and can occur rapidly, e.g., over a period of up to about 20 minutes, or more slowly, e.g., over a period of about 5-25 minutes to about 5-72 hours or more. A Gy of radiation is 1 joule per kg of absorbed ionizing radiation. The trauma event and the radiation exposure event may occur at about the same time, e.g., on the same day, or within a time period of about 1, 2, 3,4, 5 or 6 days to about 1, 2, 3 or 4 weeks, when detectable clinical effects of both events are present. Treatment with the F3C will use the dosing protocols, dosages and routes of F3C administration as described herein, e.g., dosing daily or every other day for about 1-12 days using dosages of about 0.1 mg/kg to about 30 mg/kg, depending on the route of administration and the subject's condition Dosing of the F3C will usually commence within a few days of the radiation exposure event, e.g., within 0, 1, 2, 3 or 4 days. Similarly, such healing or repair of traumas in subjects who are or are expected to become immune suppressed, e.g., from an immunosuppressive chemotherapy, cancer, stress, infection or from aging, can be treated in the same manner.

Neurological conditions. Nervous system diseases, disorders, conditions, or their symptoms (collectively 'neurological conditions') that can be ameliorated, treated or prevented with any of the F3Cs disclosed herein include, but are not limited to, nervous system trauma or injury, and neurological conditions that result in an unwanted pathology or symptom, e.g., demyelination, pain, impairment of cognitive function, discernable memory loss, depression, anxiety, a disconnection of axons, a diminution of neuron, astrocyte or glia function or degeneration or death of nervous system cells or tissues such as one or more of those described herein.

Neurological conditions, including nervous system lesions that may be treated, prevented, or ameliorated in a subject include but are not limited to, the following lesions of either the central ( including spinal cord, brain) or peripheral nervous systems. Exemplary neurological conditions include (1) ischemic lesions, in which a lack of oxygen in a portion of the nervous system results in neuronal injury or death, including cerebral infarction, ischemia or ιε ο β ο 112 312 stroke, or spinal cord infarction or ischemia, (2) traumatic lesions, including lesions caused by physical injury or associated with surgery, for example, lesions which sever a portion of the nervous system, or compression injuries, (3) malignant lesions, in which a portion of the nervous system is destroyed or injured by malignant tissue which is either a nervous system associated malignancy or a malignancy derived from non-nervous system tissue, (4) infectious lesions, in which a portion of the nervous system is destroyed or injured as a result of infection, for example, by an abscess or associated with infection by human immunodeficiency virus, herpes zoster, or herpes simplex virus or with Lyme disease, tuberculosis or syphilis, (5) degenerative lesions or conditions, in which a portion of the nervous system is destroyed or injured as a result of a degenerative process including but not limited to degeneration associated with Parkinson's disease, Alzheimer's disease, Huntington's chorea, AIDS associated dementia, epileptic dementia, presenile dementia, senile dementia, vascular dementia, post stroke dementia, post traumatic dementia or amyotrophic lateral sclerosis (ALS), (6) lesions associated with nutritional diseases, disorders, and/or conditions, in which a portion of the nervous system is destroyed or injured by a nutritional disorder or disorder of metabolism including but not limited to, vitamin B 12 deficiency, folic acid deficiency, Wernicke disease, tobacco-alcohol amblyopia, Marchiafava-Bignami disease (primary degeneration of the corpus callosum), and alcoholic cerebellar degeneration, (7) neurological lesions associated with systemic diseases including, but not limited to, diabetes (diabetic neuropathy, Bell's palsy), systemic lupus erythematosus, carcinoma, or sarcoidosis, (8) lesions caused by toxic substances including alcohol, lead, or neurotoxins, (9) demyelinated lesions in which a portion of the nervous system is destroyed or injured by a demyelinating disease including, but not limited to, multiple sclerosis, human immunodeficiency virus-associated myelopathy, progressive multifocal leukoencephalopathy, and central pontine myelinolysis or a myelopathy, e.g., diabetic meylopathy or a transverse myelopathy, (10) neurological conditions such as insomnia (e.g., transient or chronic), epilepsy, schizophrenia, psychosis, delusion, a unipolar mood disorder, a bipolar mood disorder, psychomotor dysfunction, depression, anxiety, addiction to or abuse of a drug substance such as tobacco, nicotine, caffeine, alcohol, a barbiturate, a tranquilizer, a narcotic such as hydromorphone HCl, propoxyphene napsylate, <£ °60jj2 113 meperidine HCl, valium, codeine, cocaine, morphine, heroin or methadone, (11) cognitive dysfunction conditions or diseases such as one or more of impaired long-term or short-term memory, impaired concentration, impaired attention or impaired learning, where the cognitive dysfunction condition or disease is optionally associated with chemotherapy, radiation therapy or exposure, aging, trauma, e.g., CNS trauma, or neurodegeneration and (12) genetic disorders with a neurological pathology or component such as Down's syndrome or Tay Sach’s disease.

The F3Cs are useful to ameliorate, treat or prevent the onset, severity or length of other neurological diseases or conditions such as headache or a migraine condition or symptom such as classic migraine, cluster headache, abdominal migraine, common migraine, hemiplegic migraine, ocular migraine, fulminating migraine, complicated migraine or a symptom of any of these such as head pain, vertigo, nausea, vomiting or potophobia.

In some embodiments, the F3C is used to protect neural cells from the damaging effects of cerebral hypoxia, cerebral ischemia or neural cell injury associated with cerebral infarction, heart attack, stroke or elevated levels of glucocorticoids such as cortisol. The compounds that are also useful for treating or preventing a nervous system disorder may be selected, e.g., by assaying their biological activity in promoting the survival or differentiation of neurons. For example, and not by way of limitation, the F3Cs can be used to elicit any of the following useful effects: (1) increased survival time of neurons in culture, (2) increased sprouting of neurons in culture or in vivo, (3) increased production of a neuron-associated molecule in culture or in vivo, e.g., dopamine or choline acetyltransferase or acetylcholinesterase with respect to motor neurons or (4) decreased symptoms of neuron dysfunction in vivo. Such effects may be measured by any method known in the art.

Increased survival of neurons may be measured using known methods, such as, for example, the method set forth in Arakawa et al. (J. Neurosct. 10:3507-3515 1990); increased sprouting of neurons may be detected by methods known in the art, such as the methods set forth in Pestronk et al. (Exp. Neurol. 70:65-82 1980) or Brown et al. (Ann. Rev. Neurosci. 4:1742 1981). Increased production of neuron-associated molecules may be measured by, 114 IE V6O3 12 e.g., bioassay, enzymatic assay, antibody binding or Northern blot assay, using techniques known in the art and depending on the molecule to be measured. Motor neuron dysfunction may be measured by assessing the physical manifestation of motor neuron disorder, e.g., weakness, motor neuron conduction velocity, or functional disability. Motor neuron conditions may arise from infarction, cancer, infection, exposure to toxin, trauma, surgical damage or a degenerative disease that affects motor neurons as well as other components of the nervous system.

Other neurological conditions that can be treated using F3Cs include conditions that selectively affect neurons or adjacent tissues such as amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, primary lateral sclerosis, infantile and juvenile muscular atrophy, poliomyelitis and the post polio syndrome, hereditary motorsensory neuropathy, spinal cord compression and a myelitis such as necrotizing myelitis, transverse myelitis, ascending myelitis, bulbar myelitis, concussion myelitis, demyelinated myelitis, postinfectious myelitis, systemic myelitis or transverse myelitis.

In some neurological conditions such as mood changes, depression, anxiety, memory loss or motor function impairment, the F3Cs can modulate one or more biological activities of a transcription factor or a nuclear hormone receptor such as ER.alpha. in tissue such as the hypothalamus or amygdala or ER.beta. in tissue such as the hippocampus, thalamus or entorhinal cortex.

In neurological conditions or other conditions where loss or damage to nervous system cells or tissue is typically present, e.g., multiple sclerosis, cerebral infarction, cerebral trauma, elevated glucocorticoid levels or Alzheimer's disease, use of the F3Cs can lead to detectable ·<. repair of damaged cells or replacement of at least some killed cells. Elevated glucocorticoids can result from endogenous production of natural glucocorticoids, e.g., cortisol or hydrocortisone, or from administration of synthetic glucocorticoids, e.g., dexamethasone, triamcinolone, betamethasone or other synthetic agents disclosed herein or in the cited 1£ ο § ο j j £ 115 references. Repair or replacement can occur for cell types that are present in nervous system tissues, e.g., neurons, Schwann cells, glial cells, astrocytes, oligodendrocytes, macroglia cells, endothelial cells, or stem or progenitor cells of any of tliese cell types. The cells may reside in discrete regions of nervous organs, e.g., hippocampus, cerebrum or cerebellum, or they may reside in multiple regions. Any of the neurological conditions that cen be treated with the F3Cs may be acute, subacute or chronic and they may be subclinical (having few or no overt symptoms), mild, moderate or severe.

In treating neurological conditions, the F3Cs will generally enhance function, self renewal and/or differentiation of stem or progenitor cells and/or they will reduce the severity of cell damage or impairment compared to similar subjects that are not treated with the F3Cs. In cases where myelin damage or nerve death occurs, the F3Cs can reduce the rate at which damage or death occurs or they can detectably reverse damage or enhance replacement of killed cells, particularly where the extent of such damage or killing is mild or moderate. Without wishing to be bound to any theory, the F3Cs may exert these properties (1) by directly acting as a hormone, growth factor or modulator of a biomolecule disclosed herein such as an enzyme, a glucocorticoid receptor, PPAR.alpha., a neural stem cell helix-loophelix transcription factor such as HES1 or an estrogen receptor to enhance replication, synaptogenesis or other repair or maintenance functions, (2) by enhancing recruitment and/or differentiation of cells involved in cell or tissue repair, e.g., enhanced recruitment and differentiation of oligodendrocyte cells to a demyelinated lesion in multiple sclerosis and/or (3) indirectly by modulating the level or activity of autocrine, paracrine or endocrine factors such as one or more inflammatory cytokines or markers as disclosed herein that can modulate disease progression, e.g., cortisol, IL-1.alpha., IL-1.beta., TNF-.alpha., IL-6, a thromboxane, a prostaglandin or a neuregulin.

In treating chronic or progressive disorders such as multiple sclerosis or Alzheimer's disease, the F3Cs will typically slow the rate of progression of the disease. The F3Cs act at least in part by decreasing the activity or levels of chemokines and/or pro-inflammatory cytokines, e.g., one, two or more of MCP-1, MIP-1, ICAM, V-CAM, E-selectin, RANTES, IL-1.alpha., ® 6 0 3 f 2 116 IL-1.beta., IL-6, IL-8 and TNF-.alpha.. This reduction can be accompanied by a reduced rate of deposition of amyloid-.beta. (AJ3) protein, which results in slowed disease progression and in reduced severity and/or frequency of one or more symptoms such as short term memory loss, impaired concentration, impaired judgement, episodes of disorientation or confusion and periods of mood or behavior changes such as irritability, anxiety or aggression. Treatment of chronic or progressive disorders such as Alzheimer's disease with a F3C is optionally accompanied by other suitable treatments, e.g., treatment with one or more non-steroidal anti-inflammatory drugs or other palliative measures.

Factors such as increased levels of cortisol or thromboxane, that are associated with increased cell or tissue damage or with inhibition of cell growth or differentiation are generally decreased or reregulated to express in a normal manner by the appropriate cells such as neurons, astrocytes, glial cells or their stem or precursor cells. Factors that facilitate normal differentiation or repair, e.g., basic fibroblast growth factor 2 or neuregulin, are generally increased or reregulated to express in a normal manner by the appropriate cells such as neurons, astrocytes, glial cells or their stem or precursor cells.

Because of these properties, the F3Cs can be used in various protocols or methods to enhance differentiation or proliferation of these cell types in vivo or in vitro. Typically, the concentration of the F3Cs will exert one or more of these beneficial effects at extracellular concentrations of about 1.times. lO.sup.-12 M to about 5.times.l0.sup.-6 M, e.g., about 1 .times. 10.sup.-l 1 M to about 5.times.l0.sup.-7 M or about Ltimes.10.sup.-10 M to about 1.times. 10.sup.-7 M. Such concentrations can suitably be established transiently, e.g., for about 10 minutes to about 6 hours or about 12 hours once or twice per day on one, two or more days. Alternatively, such concentrations may be maintained more or less constantly, e.g., within these ranges for at least about 12 hours per day for one, two or more days, particularly for in vitro use to enhance cell or tissue growth, differentiation or viability in tissue culture. Methods to administer the F3Cs for in vivo use are essentially as described herein.

IE 0 6o 312 117 For any of these neurological conditions or their associated symptoms, the presence of the condition or its pathological manifestation, e.g., cell or tissue damage, or symptom may be determined by suitable objective or subjective means, e.g., assays to detect tissue damage, levels of diagnostic markers or an etiological agent, performance of histopathological examination of cells or tissues, patient questionnaires or behavior performance tests, measurement of a diagnostic marker(s), e.g., an enzyme, hormone, cytokine or drug substance in blood or tissue, electroencephalography, imaging methods such as X-ray, MRI scan or CAT scan, observation and diagnosis of clinical features or symptoms or biopsy of affected tissue or cells, e.g., aspiration biopsy, needle biopsy, incision biopsy or punch biopsy of tissue or cells. Neurological conditions, diseases and symptoms, which the F3Cs can be used to treat or ameliorate and methods to diagnose and characterize such conditions or diseases have been described. See, e.g., Ph. Demaerel, A. L. Baert et al., eds. Recent Advances in Diagnostic Neuroradiology (Medical Radiology: Diagnostic Imaging) 2001 Springer Verlag, ISBN: 3504657231, W. G. Bradley et al., Neurology in Clinical Practice: Principles of Diagnosis and Management 1995, see, e.g., vol. 1 Ch. 1-55 and vol. 2. Ch. 166, Butterworth-Heinemann Medical, ISBN 0750694777, H. J. M. Barnett et al., eds. Stroke: Pathophysiology, Diagnosis and Management 3.sup.rd edition, 1998, see, e.g., pages 101450, Churchill Livingstone, ISBN 0443075514, P. J. Vinken et al., eds. Neurodystrophies and Neurolipidoses 2.sup.nd ed. 1996, see, e.g., pages 8-780, Elsevier Science, ISBN 0444812857, P. L. Peterson and J. W. Phillis eds. Novel Therapies for CNS Injuries: Rationales and Results 1995, see, e.g., pages 8-380, CRC Press, ISBN 0849376521, D. Schiffer, Brain Tumors: Pathology and Its Biological Correlates 2.sup.nd ed. 1997, see, e.g.. pages 5-450, Springer Verlag, ISBN 3540616225 and E. Niedermeyer and F. Lopes Da Silva, eds. Electroencephalography: Basic Principles, Clinical Applications and Related Fields 4.sup.th ed. 1999 see, e.g., pages 13-1238, Lippincott, Williams & Wilkins, ISBN 0683302841.

The use of the F3Cs in these conditions is optionally combined with one or more of the therapeutic treatments that are described in these references. The F3C may be administered before, during or after another treatment is employed to prevent, treat or ameliorate a given 118 0 3 12 - o (/ 3 / 2 neurological condition or symptom thereof. Any of these neurological conditions or symptoms may be mild or at an early stage, moderate or severe or advanced.

Dosages of the F3C, routes of administration and the use of combination therapies with other standard therapeutic agents or treatments could be applied essentially as described above for cardiovascular conditions or as disclosed elsewhere herein. Thus, the F3Cs may be administered prophylactically or therapeutically in chronic conditions or they may be administered at the time of or relatively soon before or after an acute event such as an epileptic seizure, onset of a migraine or occurrence of trauma, before, during or after surgery, accidental head or central nervous system injuiy or a cerebral stroke or infarction.

For acute events, the formula 1 compounds may thus be administered concurrently, e.g., within about 15 minutes or about 30 minutes of the onset or occurrence of the acute event, or at a later time, e.g., at about 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,18, 20, 22, 24, 26, 28, 30, 36, 42, 48, 54, 60, 72, 84, 96, 108 or 120 hours after the onset or occurrence of the acute event. The F3Cs may thus be administered at about 6-120 hours, or about 8-48 hours, about 10-24 hours or about 12-16 hours after an acute event starts or occurs. In other embodiments, the F3C can be administered before an expected acute event such as a planned surgery. In these cases, the F3Cs may be administered before, e.g., within about 15 minutes or about 30 minutes of the onset or occurrence of the acute event, or at an earlier time, e.g., at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18,20, 22, 24, 26, 28, 30, 36, 42, 48, 54, 60, 72, 84, 96, 108 or 120 hours before the onset or occurrence of the acute event.

Skin treatments. The affect of the F3Cs on immune function permits their use to improve the function of organs or organ systems that rely on the optimal functioning of one or more immune responses. Thus, the F3Cs can be administered to a subject to prevent, treat, ameliorate, slow the progression of or enhance the healing of certain skin conditions such as '<· skin inflammation, lesions, atrophy or rash. Conditions that can give rise to skin pathology or an unwanted skin condition include autoimmune diseases, inflammation, allergy, age, exposure to sunlight, cancer, infection or the like.

IE 0 6 Ο 3 12 119 As used here, skin includes external skin and internal skin or surfaces such as oral, intestinal and rectal mucosa. These conditions include lesions, rashes or inflammation associated with, e.g., bums, infections and the thinning or general degradation of the dermis often characterized by a decrease in collagen or elastin as well as decreased number, size and doubling potential of fibroblast cells. Such skin conditions include keratoses such as actinic keratosis, psoriasis, eczema, warts such as papillomavirus-induced warts, ulcers or lesions such as herpesvirus-induced ulcers or lesions or diabetes associated ulcers or lesions, discoid lupus erythematosus, erythema nodosum, erythema multiform, cutaneous T cell lymphoma, atopic dermatitis, inflammatory vasculitis, relapsing polychondritis, exfoliative dermatitis, sarcoidosis, bums, melanoma, rash or irritation from poison oak, poison ivy or poison Sumac, blemished or hyperpigmented skin, hyperkeratotic skin, dry skin, dandruff, acne, inflammatory dermatoses, scarring such as from a chemical or thermal bum and age-related skin changes. In these embodiments, treatment with the F3Cs is optionally combined with other appropriate treatments or therapies essentially as described herein, e.g., one or more of a corticosteroid such as hydrocortisone or cortisol, prednisone, or prednisolone, an .alpha.16 hydroxybenzoic acid or an ,alpha.-hydroxycarboxylic acid(s) is coadministered with a F3C to treat, prevent or ameliorate a skin condition such as atrophy or a lesion, .alpha.18 Hydroxybenzoic acids and .alpha.-hydroxycarboxylic acids suitable for use in these embodiments are described in, e.g., U.S. Pat. Nos. 5,262,407, 5,254,343, 4,246,261, 4,234,599 and 3,984,566. The F3C can be used to minimize cutaneous atrophy caused by corticosteroids, a side effect of their application to the skin.

In these embodiments that address skin conditions, dosages, routes of administration and dosing protocols for the F3Cs are essentially as described herein. In some embodiments, the F3C is administered to the subject in the form of a topical cream, ointment, spray, foam or gel. These topical formulations will optionally comprise about 0.1 % w/w to about 20% w/w, or about 0.2% w/w to about 10% w/w of a F3C in a composition that comprises one or more excipients that are suitable for such topical formulations, including, e.g., one or more agents that enhance penetration or delivery of the F3C into the skin. Such topical formulations can be administered, e.g., once, twice or three times per day using about 0.1 g to about 8 g or IE 0 6 0 3 12 120 about 0.2 g to about 5 g of the topical formulation on each occasion. Administration may be daily for about 1 to about 28 days, or it may be intermittent and used as needed. The amount of a topical formulation that can be administered may be higher, e.g., about 15 g or about 20 g, if the size of the area to be treated is relatively large, e.g., at least about 30 cm.sup.2 to about 100 cm.sup.2 or more. Alternatively, systemic administration of the F3C such as oral, parenteral, sublingual or buccal delivery may be used, particularly when the area of the skin to be treated is relatively large. In some cases, both topical and systemic administration of a F3C can be used. Excipients that topical or other formulations may contain include those described herein, or agents that enhance permeation or solubilization of the F3C, e.g., DMSO or an alkylalkanol, such as a 2-alkylalkanol or a 3-alkyloctanol that comprises about 8-36 carbon atoms (e.g., 8, 9, 10, 11,12,13, 14, 15, 16,17,18,19 or 20 carbon atoms) such as 2-ethyloctanol, 2-propyloctanol, 2-octyldodecanol, 2-butyloctanol, 2-hexyldecanol, 2pentylnonanol, 3-ethyloctanol, 3-propyloctanol, 3-octyidodecanol, 3-butyloctanol, 3hexyidecanol, 3-pentylnonanol, isostearyl alcohol, isocetyl alcohol, or mixtures thereof.

Such alkylalkanol moieties include those having the structure HO--CH.sub.2(CH.sub.2).sub.0-4—CH(Cl-10 alkyl)-(CH.sub.2).sub.0-6-CH.- sub.3, any of which are optionally substituted at the alkanol or the alkyl moiety with one, two, three or more independently selected substituents as described herein, e.g., with one, two, three or more independently selected --0--, -F, -OH, --CN or -CH.dbd.CH-- moieties. Such formulations can be used in therapeutic applications described herein or in cosmetic applications.

Enhancement of hematopoiesis. The invention includes methods to modulate hematopoiesis by administering a F3C to a subject, which can be used to treat or prevent various blood cell deficiencies such as thrombocytopenia (TP) or neutropenia (NP). Hematopoiesis or hemopoiesis is the formation and development of the various types of blood cells and their < progenitor cells. Mature cells are found in circulation or tissues such as the lymph nodes, spleen or the thymus. Many of the stem cells that give rise to mature forms reside in the bone marrow, although some may circulate in the blood for some time. Clinical blood cell deficiencies such as thrombocytopenia, neutropenia or erythropenia can arise from causes IE 060 3 12 121 such as impaired hematopoiesis or abnormal loss or destruction of mature or immature blood cells.

Without being bound to any theory, the treatment methods at least in part result in enhanced hematopoiesis, enhanced movement of blood cells into the circulation and/or in reduced loss of blood cells such as platelets or neutrophils. The F3Cs can enhance self-renewal or numbers of hematopoietic stem cells, precursor cells, mature blood cells and/or they can enhance or accelerate differentiation of stem or any progenitor cell that can give rise to a mature blood cell. The stem or progenitor cells include early lineage cells showing little or no characteristics of fully differentiated blood cells and/or they can be partially differentiated. Increased platelet or neutrophil production, enhanced survival or reduced loss is typically observed as increased circulating blood cell counts. Increases in blood cells appear to arise from enhanced proliferation of precursor cells and/or from enhanced or accelerated differentiation of precursor cells. Increased cell numbers, e.g., platelets or neutrophils, can also arise from from reduced loss or death of such cells, increased demargination of cells such as neutrophils from the vasculature into circulating blood or other tissues and/or shorter transit time of mature or precursor cells from the bone marrow into blood.

Thus, invention embodiments comprise a method to treat or prevent a blood cell deficiency such as TP or NP in a subject in need thereof, comprising administering to the subject, or delivering to the subject's tissues, an effective amount of a F3C. Related embodiments include a method to increase self-renewal of hematopoietic stem cells or hematopoietic progenitor cells or to increase the commitment of such cells to transition to a more differentiated blood precursor cell or mature blood cell. In other embodiments, the invention provides a method for stimulating the proliferation or differentiation of neutrophil precursors < or to increase demargination of neutrophils or to reduce transit time from bone marrow to blood in a subject having or susceptible to developing NP comprising administering an effective amount of a F3C to the subject in need thereof. The F3C treatment will stimulate the activity of, e.g., neutrophils, or enhance their production from progenitor cells, enhance ΙΕ Ο6Ο312 122 their survival and/or limit their loss. Hematopoietic stem cells, e.g., GEMM cells, are pluripotent and can give rise to more than one type of mature blood cell, while hematopoietic progenitor cells are usually not pluripotent, but are bipotent or monopotent. Hematopoietic progenitor cells reside primarily in bone marrow, but can also be found in blood, spleen or lymph tissue or fluids.

Normal ranges of various white blood cells or blood components in adult (about 18-49 years of age) human blood are as follows. Total adult white blood cell counts average about 7500/mm.sup.3, with an approximate normal range of about 4.511.0.times.l0.sup.3/mm.sup.3. The normal basophil level is about 35/mm.sup,3, with a nonnal range of about 10/mm.sup.3 to about 100/mm.sup.3. The normal adult neutrophil level is about 4400/mm.sup.3, with a normal range of about 2000-7700/mm.sup.3. The nonnal eosinophil level is about 275/mm.sup.3, with a normal range of about 150300/mm.sup.3. The normal monocyte level is about 540/mm.sup.3, with a nonnal range of about 300-600/mm.sup.3. The nonnal adult platelet level is about 2.5. times.l0.sup.5/mm.sup.3, with a nonnal range of about 2.1.times.l0.sup.52.9.times. 10.sup.5/mm.sup.3. The nonnal human adult red cell mass corresponds to about 4.6. times. 10.sup.12 red cells/L in females and about 5.2.times.l0.sup.l2 red cells/L in males.

A human patient in need of treatment will typically have, or be subject to developing, a cell count below these values. For example, the subject may have a cell count that is about 2% to about 90% below the lower or upper values of these ranges, e.g., about 5%, about 10%, about 20%, about 30%, about 50% or about 70% below any of these values. As used herein, neutropenia means generally a circulating neutrophil count of less than about 2000/mm.sup.3, typically less than about 1500/mm.sup.3 or usually less than about 1300/mm.sup.3. Under the common terminology criteria for adverse events, version 3.0, published at http://ctep.cancer.gov, grade 1 neutropenia in humans is the lower limit of nonnal to 1500 neutrophils/mm.sup.3, less than 1500 to 1000 neutrophils/mm.sup.3 is grade 2 neutropenia, about 1000-500 neutrophils/mm.sup.3 is grade 3 neutropenia and less than ΙΕ θ 6 0 3 12 123 about 500 neutrophils/mm.sup.3 is considered to be grade 4 neutropenia. Febrile NP is NP accompanied by a fever, e.g., about 39.5.degree. C. to about 43.degree. C. or more, that is at least transient, e.g., lasting about 2 or more hours.

Thrombocytopenia generally means a circulating platelet count of less than the normal circuating range, e.g., less than about about 1.6.times.l0.sup.5/mm.sup.3, less than about 1.5. times.l0.sup.5/mm.sup.3, less than about 1.3.times.l0.sup.5/mm.sup.3 or less than about 1.0.times.l0.sup.5/mm.sup.3. Under common terminology criteria for adverse events, version 3.0, grade 1 thrombocytopenia is the lower normal limit to 75,000 platelets/mm.sup.3, grade 2 thrombocytopenia is <75,000-50,000 platelets/mm.sup.3, grade 3 thrombocytopenia is <50,000-25,000 platelets/mm.sup.3 and grade 4 is <25,000 platelets/mm.sup.3. Anemia generally means a red cell mass corresponding to less than about 4.0.times.l0.sup.l2 red cells/L in adult females and less than about 4.5. times.l0.sup.l2 red cells/L in adult males (a hemoglobin level of less than about 12.0 g/dL in adult females and less than about 13.5 g/dL in adult males).

In some cases, the diagnosis of a deficiency may cover a cell count that falls outside these ranges, due, e.g., to individual variations in a subject's age, sex, race, animal strain or nonnal blood cell status for the individual. Such variations are identified by known means such as by identification of a change from the subject's nonnal status or by multiple cell measurements over time that reveal a deficiency. See, e.g., Hematology-Basic Principles and Practice, 2.sup.nd edition, R. Hoffman, E. J. Benz Jr. et al., editors, Churchill Livingstone, New York, 1995. Subjects with an identified or identifiable deficiency outside these standard ranges are included in the definition of a blood cell deficiency or a subject in need of treatment, as used herein.

In exemplary embodiments, use of the F3Cs for treating subjects including primates or humans who are subject to developing a NP condition will typically result in a decreased in the severity and/or duration of NP. Typically, the F3C treatment will comprise treating the subject daily, every other day or every third day for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 or 124 IE 060 3 12 days with about 0.1 mg/kg to about 5 mg/kg, usually about 0.5 mg/kg to about 4 mg/kg. For these dosages, the F3C is typically administered by parenteral, e.g., intravenous, subcutaneous or intramuscular, or transmucosal delivery. Oral administration will generally use dosages that are about 3-25 mg/kg higher, e.g,, about 4-30 mg/kg of the F3C. Human unit dosages will typically comprise about 1-1500 mg, usually about 10-150 mg, which can be subdivided, e.g., into two or three subdoses. Treatment of subjects who may develop a NP condition from a chronic or slow onset condition will generally begin when reduced neutrophil counts are observed, e.g., when the subject has grade 1 or 2 NP. In situations where NP can arise over a short time period, e.g., from an inducing event such as a chemotherapy, an acute infection or radiation exposure, treatment with the F3C will generally begin at about the time of the inducing event. Thus, for subjects who will be subjected a chemotherapy or radiation therapy, dosing with the F3C can begin about 1, 2, 3 or 4 days before, during (essentially simultaneous with or on the same day as) or about 1,2, or 4 after the inducing event. Typically dosing the F3C begins in a period from 2 days before to 2 days after the subject is exposed to the NP inducing event.

Treatment with a F3C will reduce the severity of NP, e.g., by preventing the development of grade 3 or 4 NP or febrile NP in subjects who would otherwise be expected to develop or susceptible to developing grade 3 or 4 NP. The F3C will also typically reduce the duration of, e.g., grade 3 or 4 NP, in subjects who would otherwise be expected to develop or susceptible to developing such NP. The reduction in the duration of NP, grade 3 or 4 NP, can range from 100% to a detectable level, e.g., a reduction of at least about 10%. Typically, the reduction of the period during which a subject has grade 3 or 4 NP or febrile NP is about 25% to about 85%, e.g., about 30%, 40%, 50%, 60%, 70%, 80% or more.

Individual responses can vary depending on factors such as the subject's initial neutrophil status, when dosing with the F3C is initiated, dosage of the F3C and the route of administration of the F3C. NP in subjects susceptible to developing NP can arise from conditions or treatments as described herein, e.g., autoimmune conditions, cancer, cancer chemotherapy, an infection, antimicrobial chemotherapy, bone marrow transplantion, an IE 060 3 12 125 immunosuppressive therapy, bone marrow damage or exposure to or treatment with an ionizing radiation such as one or more of .gamma.-radiation, X-rays, fast neutrons, .beta.radiation or .alpha.-radiation.

TP, abnormally low platelet counts, can arise from impaired platelet production, sequestration of platelets in the spleen or abnormal loss of circulating platelets. Impaired production can result from causes such as chemotherapy, radiation exposure, e.g., a radiation therapy, or an from autoimmune condition. Abnormal loss of circulating platelets is often associated with autoreactive antibodies that bind to platelets and reduce their life span.

These underlying causes give rise to the various clinical forms of TP, such as autoimmune neonatal TP, immune thrombocytopenic purpura, radiation induced TP, chemotherapy induced TP and amegakaryocytic TP.

Other conditions that are amenable to prophylaxis or treatment by the invention methods include the acquired blood cell deficiencies. Exemplary deficiencies or groups of deficiencies that can be treated are neonatal alloimmune TP, immune TP, immune thrombocytopenic purpura, thrombotic thrombocytopenic purpura, post-transfusion purpura, radiation associated TP, chemotherapy associated TP (e.g., an anticancer, antiviral, antibacterial, antifungal or antiparasite therapy, NSAID treatments such as with indomethicin, ibuprofen, naproxen, phenylbutazone, piroxicam or zompirac, or .beta.-lactam antibiotic treatments such as with ampicillin, carbenicillin, penicillin G, ticarcillin, or cephalosporin treatments such as with cefazolin, cefoxitin or cephalothin, anticoagulant treatments such as heparin, hirudin, lepirudin or aspirin, treatment with plasma expanders or psychotropic drugs), amegakaryocitic TP, radiation associated TP, TP associated with solid organ allograft or xenograft rejection or immune suppression therapy in solid organ or other tissue transplants (e.g., liver, lung, kidney, heart, bone marrow, hematopoietic stem cell or endothelial cell transplant, implant or transfusion), cardiopulmonary bypass surgery, cardiovascular disease or therapy associated TP (e.g., congenital cyanotic heart disease, valvular heart disease, pulmonary embolism, pulmonary hypertension disorders or diltiazem, nifedipine, nitroglycerin or nitroprusside therapy), TP associated with chronic or acute renal IE 0 60 3 12 126 failure or treatment for these conditions (e.g., dialysis), TP associated with infection such as a virus or bacterial infection. NP conditions that can be treated include postinfectious NP, autoimmune NP, chronic idiopathic NP, basophilic leukopenia, eosinophilic leukopenia, monocytic leukopenia, neutrophilic leukopenia, cyclic NP, periodic NP, chemotherapy associated NP, radiation associated NP, NP associated with solid organ allograft or xenograft rejection or immune suppression therapy in solid organ or other tissue transplants (e.g., liver, lung, kidney, heart, bone marrow, hematopoietic stem cell or endothelial cell transplant, implant or transfusion), chemotherapy associated leukopenia, radiation associated leukopenia, leukopenia associated with solid organ allograft or xenograft rejection or immune suppression therapy in solid organ or other tissue transplants (e.g., liver, lung, kidney, heart, bone marrow, hematopoietic stem cell or endothelial cell transplant, implant or transfusion), immune hemolytic anemias, anemia associated with chronic or acute renal failure or treatment for these conditions (e.g., dialysis), anemia associated with chemotherapy (e.g., isoniazid, prednisone) or anemia associated with radiation exposure.

The F3Cs are thus useful to facilitate or speed up immune system recovery in autologous bone marrow transplant or stem cell transplant situations. In many cases it would be medically sound to continue the treatment associated with causing or exacerbating the blood cell deficiency. Thus, in some embodiments a F3C treatment is conducted with subjects who are undergoing another therapy at the same time or near the same time, e.g., within about 1, 2, 3, 4 or several days to within about 1-6 months. Such subjects typically will have an identified blood cell deficiency such as a NP or a TP, e.g., as disclosed herein. However, the F3Cs can be generally suitable for preventing the onset or reducing the severity of such deficiencies, and they can thus be used prophylactically in these indications, e.g., by administering a F3C beginning at about 1-60 days before administering another therapy that could lead to a cytopenia condition such as TP or NP.

In conditions such as NP, the F3Cs will typically function at least in part by modulating, e.g., increasing, the level or activity of biomolecules such as IL-1.beta., G-CSF, GM-CSF or one or more of their receptors, that can enhance generation or survival of a desired cell type IE 0 60 3 12 127 such as neutrophils. In this regard, the F3Cs can act as inducers of endogenous growth or differentiation factors that facilitate increased production or survival of neutrophils or other blood cell types. This aspect of the F3Cs allows one to eliminate or reduce the use of such molecules in treating conditions such as NP.

Use of a F3C in treating cytopenia conditions is thus optionally combined with the use of an effective amount of one or more growth factors or cytokines as a means to further enhance the effect of the F3Cs for their intended uses or to modulate, e.g., enhance, their effects or efficacy. Suitable growth factors and cytokines are as described herein or in the cited references. For example, when one administers the F3C to enhance generation of platelets in humans or other subjects, or their precursor cells such as CFU-blast cells, multipotent thymic precursor cells (CD34.sup.+, CD38.sup.+, CD7.sup.+, CD44.sup.+, CD33.sup.+, CD2.sup.-, CD5.sup.-, CDla.sup.-), Pro-DC2 cells, immature DC2 cells, immature NK cells, CFU-GEMM, BFU-Mk, CFU-Mk, CFU-G, CFU-GM, immature megakaryocytes or mature postmitotic megakaryocytes, one can also administer one or more of G-CSF, GMCSF, SCF, Steel factor (SF), leukemia inhibitory factor (LIF), interkeukin-1.alpha., (IL-1.alpha.), IL-3, IL-6, IL-11, TPO, EPO, their isoforms, their derivatives (e.g., linked to a PEG or fusions such as PIXY321) or their isofonns, orthologs or homologs for other species. Similarly, administration of the F3C to enhance the generation or function of myelomonocytic cells such as neutrophils, basophils or monocytes in humans or other subjects, can also be combined with administration of one or more of G-CSF, GM-CSF, MCSF, LIF, TPO, SF, interleukin-1 (IL-1), IL-2, IL-3, IL4, interleukin-5 (IL-5'j, IL-6, IL11, interleukin-12 (IL-12), interleukin-13 (IL-13'j, FLT3 ligand, their isoforms, orthologs, homologs or derivatives (e.g., linked to a PEG or fusions such as PIXY321) or their isoforms, orthologs or homologs for other species. To enhance generation of red cells or their precursor cells such as CFU-GEMM, BFU-E or CFU-E in humans being treated with a F3C, one can co-administer one or more of G-CSF, GM-CSF, IL-1, IL-3, IL-6, TPO, EPO, transforming growth factor-.sym.l, their isoforms, their derivatives (e.g., linked to a PEG or fusions such as PIXY321) or their isofonns, orthologs or homologs for other species. See, e.g., Hematology--Basic Principles and Practice, 3.sup.rd edition, R. Hoffman, >!ΐί: IE 06 0 3 12 128 E. J. Benz Jr. et al., editors, Churchill Livingstone, New York, 2000 (see, e.g., Chapters ΜΙ 7 at pages 154-260). The co-administration of such factors in these methods is intended to enhance the efficacy ofthe F3C treatment, which is optionally measured by taking suitable blood or tissue, e.g., bone marrow, samples at one or more times before and after the compounds have been administered. Such co-administration will generally be compatible with a subject's condition and other therapeutic treatments. Co-administration of such factors can precede, be simultaneous with, or follow the times of administration of the F3C(s) to the subject. Dosages of such growth factors would generally be similar to those previously described, e.g., typically an initial course of treatment comprises administering about 1.0 to about 20 .mu.g/kg/d for about 1-10 days, or as described in, e.g., Hematology—Basic Principles and Practice, 3.sup.rd edition, R. Hoffman, E. J. Benz Jr. et al., editors, Churchill Livingstone, New York, 2000 (see, e.g., Chapter 51 at pages 939-979 and the references cited therein).

In cases where a subject's blood cell deficiency is caused by, or associated with another therapy, the invention contemplates that the other therapy will continue, if this is reasonable under the circumstances. The timing of other therapies can precede, be simultaneous with, or follow the times of administration of the F3C(s) to the subject. For example, chemotherapy for some malignancies is accompanied by myelosuppression or a deficiency in one or more blood cell types, e.g., TP or NP. Continued treatment would be called for in some cases, and then the invention methods would be employed to deliver to the subject an effective amount of a F3C. Thus, alkylating agents, antimicrotubule agents, antimetabolites, vinca alkaloids, topoisomerase I or II inhibitors, or platinum compounds such as one or more of mechlorethamine, vincristine, vinblastine, bleomycin, doxorubicin, epirubicin, tamoxifen, cyclophosphamide, etoposide, methotrexate, ifosfamide, melphalan, chlorambucil, busulfan, carmustine, lomustine, streptozocin, dacarbazine, vinorelbine, paclitaxel (taxol), docetaxel, cytosine arabinoside, hydroxyurea, fludarabine, 2'-chlorodeoxyadenosine, 2’deoxycofonnycin, 6-thioguanine, 6-mercaptopurine, 5-azacytidine, gemeitabine, arabinofuranosylguanine, daunorubicin, mitoxantrone, amsacrine, topotecan, irinotecan, cisplatin, carboplatin, pilcamycin, procarbazine, aspariginase, aminoglutethimide, IE 0 6 0 3 12 129 actinomycin D, azathioprine and gallium nitrate may be administered in conjunction with administration of any F3C(s) that is disclosed herein. Treatments with other therapeutic agents such as heparin or nucleoside analogs such as 3-thiacytosine, azidothymidine or dideoxycytosine, or other antimicrobials such as cephalosporin, quinine, quinidine, gold salts (e.g., aurothioglucose), a fluoroquinolone (e.g., ciprofloxacin), clarithromycin, flucomazole, fusidic acid, gentamycin, nalidixic acid, penicillins, pentamidine, rifampicin, sulfa antibiotics, suramin or vancomycin may result in a blood cell deficiency(s) and they can thus be combined with administration of a F3C to treat the deficiency, or to ameliorate a symptom thereof. Similarly, anti-inflammatory drugs (e.g., salicylates, entanercept (a dimeric fusion comprising a portion of the human TNF receptor linked to the Fc portion of human IgGl containing the C.sub.H2 and C.sub.H3 domain and hinge regions of IgG 1) or a COX-2 inhibitor such as celexicob (4-5[-(4-methylphenyl)-3-(trifluoromethyl)-lH-pyrazole1 -yl] benzenesulfonamide) or rofecoxib (4-[4-methylsulfonyl)phenyl]-3-phenyl-2(- 5H)furanone) or an IL-1 receptor antagonist such as anakinra), cardiac drugs (e.g., digitoxin), .beta.-blockers or antihypertensive drugs (e.g., oxprenolol or captopril), diuretics (e.g., spironolactone), benzodiazepines, (e.g., diazepam) or antidepressants (e.g., amitriptyline, doxepin). Any of these methods also optionally include co-administration of one or more of the growth factors described above, e.g., IL-3, G-CSF, GM-CSF or TPO.

Other therapies for treating a blood cytopenia such as TP or NP also include administering one or more of glucocorticoid steroids (e.g., prednisone, prednisolone), human IgG antibodies, anti-Rh(D).sup.+ antibodies for Rh(D).sup.+ patients, an androgen such as danazol, vinca alkaloids (e.g., vincristine, vinblastine), thrombopoietin and immunosuppresants (e.g., azathioprine, cyclophosphamide, FK506 or cyclosporin). Splenectomy may also be indicated, for example when first line treatments fail. The goal of treatment for TP in humans is typically to increase platelet counts to at least about 20,000/mm.sup.3 or more typically to at least about 50,000/mm.sup.3 and to maintain these levels.

Although the treatment options to increase platelet levels are generally known, they usually *E 0 6 0 3 j 2 130 have a number of drawbacks. For example, infusion of IgG antibodies is not always effective and the treatment is relatively expensive. Other treatments, such as prednisone are also not always effective and they typically are discontinued or tapered off after several weeks due to toxicity or unwanted side effects. Splenectomy, which is relatively expensive and invasive, is also not always effective. The sources of thrombocytopenia and treatment options have been described. See, e.g., Hematology-Basic Principles and Practice, 3.sup.rd edition, R. Hoffman, E. J. Benz Jr. et al., editors, Churchill Livingstone, New York, 2000 (see, e.g., Chapters 126-129 and 131 at pages 2096-2154 and 2172-2186), PCT publication WO 200035466.

Neutropenia (NP), is considered to exist clinically when neutrophils drop to below a level considered normal. NP can arise from impaired production of neutrophil precursors or mature neutrophils, movement of neutrophils from the circulation to tissue, abnormal circulating neutrophil loss or a combination of these causes. Impaired neutrophil production can be acquired from, e.g., treatment with a cytotoxic or cytostatic drug, chemotherapy, radiation therapy or an autoimmune response as described herein. The abnormal loss of circulating neutrophils in autoimmunity is typically associated with autoreactive antibodies that bind to the cells and reduce their life span. These underlying causes give rise to the various clinical forms of NP, such as postinfectious NP, drug-induced NP, autoimmune NP, or chronic idiopathic NP. The sources of NP and treatment options have been described.

See, e.g., Hematology—Basic Principles and Practice, 3.sup.rd edition, R. Hoffman, E. J. Benz Jr. et al., editors, Churchill Livingstone, New York, 2000 (see, e.g., Chapters 19, 41, 51, 79, 134 and 137 at pages 297-331, 720-762, 939-979, 1443-1500, 2220-2248 and 22572263).

In some embodiments, the F3Cs that are used to enhance hematopoiesis or to treat associated conditions such as a TP or a NP disease or condition as disclosed herein, are characterized by having a lack of appreciable androgenicity. In these embodiments, the F3Cs are characterized by having about 15% or less, about 10% or less, about 5% or less, about 2% or less, about 1 % or less or about 0.5% or less of the androgenicity of a reference IE <>6o 312 131 androgen such as testosterone, testosterone proprionate, dihydrotestosterone or dihydrotestosterone proprionate as measured in a suitable assay using suitable positive and/or negative controls. F3Cs having, e.g., a substitution at the 6- or 7-position or having no double bond at the 4-5 or 5-6 positions, will generally have relatively low levels of androgen activity. Suitable assays for androgenicity of various compounds have been described, e.g., J. R. Brooks, et al., Prostate 1991, 18:215-227, M. Gerrity et al., Int. J. Androl. 1981 4:494-504, S. S. Rao etal., Indian J. Exp. Biol. 1969 7:20-22, O. Sunami et al., J. Toxicol. Sci. 2000 25:403-415, G. H. Deckers et al., J. Steroid Biochem. Mol. Biol. 2000 74:83-92. The androgenicity of the F3Cs are optionally determined as described or essentially as described in one or more of these assays or any other assay. Thus, one such embodiment comprises a method to enhance hematopoiesis or to treat TP or NP comprising administering to a subject in need thereof an effective amount of a F3C, or delivering to the subject's tissues an effective amount of a F3C, wherein the F3C has about 30% or less, about 20% or less, about 10% or less or about 5% or less of the androgenicity of an androgen such as testosterone, testosterone proprionate, dihydrotestosterone or dihydrotestosterone proprionate as measured in a suitable assay, e.g., as described in the citations above. In conducting such methods, the subjects, e.g., rodents, humans or primates, are optionally monitored for e.g., amelioration, prevention or a reduced severity of a disease, condition or symptom. Such monitoring can optionally include measuring one or more of cytokines (e.g., TNF.alpha., IL-l.beta.), WBCs, platelets, granulocytes, neutrophils, RBCs, NK cells, macrophages or other immune cell types, e.g., as described herein or in the cited references, in circulation at suitable times, e.g., at baseline before treatment is started and at various times during or after treatment with a F3C, e.g., at about 245 days after treatment with a F3C has ended.

In conducting any of these methods, one can monitor the subject's clinical condition at any relevant time before, during or after administration of the F3Cs, which treatments are optionally combined with any of the other agents or treatments described above. The subject's blood can be drawn on one, two or more occasions in advance of treatment to, e.g., obtain a baseline or initial level of white or red blood cells, to verify a presumptive diagnosis •Ε 0 6 0 3 12 132 of a blood cell deficiency or to determine a blood parameter such as circulating myelomonocyte counts, circulating neutrophil counts or circulating platelet counts. Then, during the course of treatment or thereafter the subject's blood can be drawn on one, two or more occasions to follow the subject's response, e.g., once treatment with a F3C has ended.

Invention embodiments include methods that comprise administering to a subject in need thereof an effective amount of a F3C and an effective amount of at least one fonn of interferon, such as .gamma.-interferon or a growth factor or interleukin such as G-CSF or IL-6. Interferons can enhance the biological activity of the white cells that arise from increased hematopoiesis. This can be particularly useful when the subject's circulating blood cell deficiency is associated with, e.g., an infection or a chemotherapy that suppresses hematopoiesis. Administration of a growth factor or an interleukin such as IL-6 can facilitate hematopoiesis by stimulating quiescent stem cells or other progenitors that give rise to deficient cell types. Related embodiments replace growth factor or interferon administration partially or completely by increasing endogenous production in the subject using conventional methods, e.g., administering double stranded RNA to stimulate .gamma.-IFN.

In these embodiments, the subject may have thrombocytopenia or neutropenia or the subject's circulating platelets, red cells, mature myelomonocytic cells, or their precursor cells, in circulation or in tissue may be detectably increased. In some cases the subject has renal failure. These methods may further comprise the steps of obtaining blood from the subject before administration of the F3C and measuring the subject's white or red cell counts and optionally, on one, two, three or more occasions, measuring the subject's circulating white cell or red cell counts after administration of the F3C, e.g., within about 12 weeks after an initial administration of a F3C or during or within about 12 weeks after a course of treatment as described herein.

Delayed radiation effects. Invention embodiments include a method to prevent, treat or ameliorate a symptom or condition associated with one or more delayed adverse effect, symptom or condition from ionizing radiation exposure in a subject in need thereof ΙΕ ο 6 Ο 3 12 133 comprising administering to the subject, or delivering to the subject's tissues, an effective amount of a F3C. In these embodiments, administration of the F3C commences at least 2 weeks after the subject has been exposed to a dose or subdose of radiation that could give rise to a delayed radiation effect. Dosing with the F3C can thus begin at 14 days to about 2 years or more after ionizing radiation exposure. Typically dosing will begin at about 2 weeks, 3 weeks, or 1,2, 3 or 4 months after exposure of the subject to sufficient ionizing radiation to potentially cause delayed effects. Radiation exposure may arise from a radiation therapy where exposure is intentional, or it may arise from an accidental exposure.

Radiation therapy (RT) can generate a number of late delayed-onset conditions or symptoms. Delayed radiation effects are conditions or symptoms that generally arise or become detectable to the subject or to a health care provider at least about 1 month after exposure to radiation. Thus the conditions or symptoms may be detectable at about 2 months, about 3 months, about 4 months, about 5 months, about 1 year, about 20 years or more after radiation exposure. For example, transient nervous system symptoms may develop early after RT, but progressive, permanent, often disabling nervous system damage may appear months or years later. The total radiation dose, size of the fractions, duration of RT, and volume of tissue irradiated influence the probability of the injury and its severity. Individual patient and tissue susceptibility to delayed injuries is variable, which factors into the selection of safe and effective radiation doses for RT. Total radiation doses that a subject may receive may comprise single doses or 2, 3, 4, or more doses within a range of about 1 to about 400 Gy, e.g., about 1, 1.4, 1.6, 1.8, 2, 2.5, 3, 5, 10,20,40, 50, 80, 100, 130, 150, 180, 200, 250, 300, 400 Gy. Typical doses are about 1-12 Gy or about 1-8 Gy. Such doses in a given course of treatment may be the same or different and can occur over a period of time, e.g., over 1 day to about 1 or 2 years.

In some embodiments, the total radiation dose occurs on a single exposure that occurs in a relatively short time period, e.g., about 1-20 minutes to about 12 hours. In other embodiments, the total dose is delivered to the subject in multiple doses or over a longer time, e.g., over about 2 days to about 12 months or more in multiple doses in, e.g., 2, 3, 4, 6, 134 ΙΕ ο 6 Ο 3 12 8, 10 or more individual doses. Ameliorating a side effect may comprise detectably slowing the progression of a symptom or condition or detectably reducing the ultimate expected severity of a symptom or condition. The affected condition or symptom may be detectably reduced as determined by the subject or the health care provider. Thus, after administration of a F3C, the target symptom or condition may be moderately reduced, slightly reduced, essentially nonexistent or subclinical, e.g., present at a low level that is not deemed significant by the subject or the health care provider. Amelioration of one or more conditions or symptoms that can be suitably quantified may be observed as a decrease of about 5% or more, e.g., at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 70%, at least about 80% or at least about 90% in the relative expected or potential severity or extent of the condition or symptom.

For example, in lung pneumonitis, administration of a F3C can lead to detectably increased oxygen saturation in the subject's blood by about 5% or by about 10% or more, e.g., oxygen saturation can rise from about 83% to about 88%, which would typically be detectable by the subject and the health care provider. Such decreased severity of a condition or symptom may be objectively measured in some instances, e.g., by detennining the number or activity of circulating platelets or neutrophils or by evaluation of fever, severity or frequency of diarrhea or blood oxygen saturation levels. For other symptoms or conditions, prevention may be subjectively observed by a significant or detectable improvement in a relevant score, e.g., decreased fever or pain or a decreased need for treatment of fever, pain or inflammation.

Symptoms or conditions of radiation exposure that can be treated also include encephalopathy, myelopathy, nausea, diarrhea, acute inflammation, chronic inflammation, edema, pain, fever, headache, depression, malaise, weakness, hair loss, skin atrophy, skin ulceration, skin lesion, keratosis, telangiectasia, infection, e.g., bacterial, viral, fungal or yeast infection, hypoplasia, atrophy, marrow hypoplasia, hemorrhage, fibrosis, e.g., lung fibrosis, pneumonitis, bone marrow hypoplasia, hemorrhage or cytopenia, e.g., anemia, leukopenia or thrombocytopenia, edema, fibrosis or hemorrhage or the need for edema, ΙΕ 0 3 12 135 fibrosis or hemorrhage treatment. Such symptoms or conditions may arise from one or more radiation-damaged tissues or cells, including lymphoid cells, bowel or intestinal epithelium or tissue, bone marrow, testicles, ovaries, brain tissue, spinal cord tissue or skin epithelium.

Exemplary symptoms or conditions associated with late effects of radiation exposure include (1) acute or chronic radiation-induced enteritis or diarrhea, e.g., in patients receiving pelvic radiotherapy, (2) pseudomembranous inflammation, (3) perivascular fibrosis, (4) endothelial cell damage or death, e.g., associated with vascular radiation therapy, (5) cardiac tissue inflammation or damage or pericardial disease, e.g., in pediatric or adult patients receiving radiation therapy for a leukemia, thoracic neoplasm or other malignancy, (6) pulmonary tissue inflammation or damage, (7) hematopoietic or marrow cell inflammation or damage, e.g., in wide field radiation therapy, (8) endocrine or thyroid dysfunction, e.g., in thalamic or hypothalamic tumors in pediatric or other patients, (9) decreased growth or decreased bone development or density, e.g., in pediatric patients receiving radiation therapy for a childhood leukemia or other malignancy, (10) central nervous system inflammation or damage, e.g., in pediatric or adult patients receiving radiation therapy for a leukemia (e.g., CNS acute lymphocytic leukemia) or other malignancy, (11) connective tissue damage after radiation therapy, (12) incidence or severity of a secondary leukemia such as acute myelogenous leukemia or myelodysplasia and (13) gastric ulceration, bleeding, small bowel obstruction or fistula formation in, e.g., patients receiving radiation therapy to the gastrointestinal tract. These symptoms or conditions are treated or ameliorated using the F3Cs essentially as disclosed herein.

In treating such symptoms or conditions, slowing the progression of a symptom, condition or side effect will detectably reduce the rate at which the condition, symptom or side effect worsens or intensifies. In some embodiments, pronounced slowing of the rate of progression -< is, e.g., the time needed to progress to an expected or a measurable point, which may be increased by a period of about 1, 2, 3, 4, 5,10, 20, 30 or more days to a period of about 1, 2, 3, 4, 6, 8, 10, 12, 18, 24, 36, 48, 72 or more months. ιε oso 312 136 Radiation-associated brain damage can give rise to acute encephalopathy with symptoms such as headache, nausea, vomiting, somnolence, depression, disorientation, and worsening neurologic signs. The encephalopathy may arise from the first, second or a subsequent radiation fraction, e.g., when high intracranial pressure has not been treated with, e.g., corticosteroids. Late-delayed radiation damage to the brain or nervous system can arise at about 2, 3,4, 5, 6, 7, 8, 9, or 10 months to 1, 2, 3 or more years after leukemia prophylaxis in children or after brain tumor prophylaxis or treatment in adults. Symptoms often include pain or headache and progressive dementia without focal signs and adults typically also develop an unsteady gait. Cerebral atrophy appears on CT scans in some cases. Late-delayed damage can arise at about 1 week, about 2 weeks about 2 months or about 1-2 years after irradiation of extracranial tumors or high-dose irradiation of intracranial tumors, e.g., brachytherapy or radiosurgery, although the symptoms are generally more focal. The invention method would be used during the time period when such symptoms would be expected to arise, e.g., commencing at about 1-5 days or about 7-60 days after radiation exposure and ending at about 0.5, 1, 2, 3, 4, 5 or more years later. Exemplary brachytherapies and unsealed source therapies include prostate .sup. 1251 seed implants in prostate conditions such as prostate cancer, .sup.90Yt conjugated to monoclonal antibodies or in endovascular brachial radiotherapy.

Early-delayed radiation spinal cord myelopathy follows radiation therapy to the spinal cord, neck, upper thorax or lumbar region or and it is often characterized by Lhermitte's sign, i.e., an electric shock-like sensation radiating down the back and into the legs on neck flexion. Late-delayed radiation myelopathy can arise months or years after therapy for extraspinal tumors, e.g., Hodgkin's disease. Other symptoms can include progressive weakness and sensory loss, such as a Brown-Sequard type, i.e., a proprioceptive sensory loss and weakness on one side of the body and loss of temperature and pain sensation on the other side. Progression times vary, but many human patients suffering from late-delayed radiation spinal cord myelopathy become paraplegic. Late-delayed radiation neuropathy may produce brachial neuropathy, e.g., after treatment for breast or lung cancer. Radiation can also give rise to gliomas, meningiomas, or peripheral nerve sheath tumors at about 1, 2, 3, 4, 5 or If Ο 6 ο 3 1 £ 137 more years after therapy. The F3Cs will generally be administered at about the time period when these symptoms would be expected to arise, e.g., commencing at about 1-5 days, or about 7-60 days or about 6 or 12 months after radiation exposure and ending at about 3, 4, 6 months later or about 1, 2, 3, 4, 5, 6 or more years later. In some embodiments, the F3C is administered to the subject on the same day that a planned or accidental radiation exposure occurs and dosing is continued for about 1, 2, 3,4, 8, 12 or more weeks to about 2, 3, 4, 5, 6 or more years, or for a time as disclosed elsewhere herein.

Early-delayed encephalopathy often arises or is detectable at about 2, 3 or 4 months after radiation therapy. This encephalopathy in adults, is distinguished from worsening or recurrent brain tumor by, e.g., computed tomography (CT) or magnetic resonance imaging (MRI). The condition in children can occur as a somnolence syndrome, e.g., after wholebrain irradiation for leukemia. The condition in children typically improves spontaneously over several days to weeks. Such encephalopathies can be prevented, delayed in onset, recede more rapidly and/or be less severe when a F3C is administered to the subject throughout the period when encephalopathy can arise, e.g., beginning about a week, two weeks or a month before the expected onset of a symptom or condition and ending about a week or month or two months after it would be expected to arise or to resolve.

In some embodiments, a radiation late effect is a symptom or condition that may arise months or years after radiation exposure, treatment with the F3C can commence shortly, e.g., about 0.5, 1, 2, 3, 4, 5, 10, 14, 21 or 28 days, after the radiation exposure or after initiation of a radiation treatment. In other embodiments, the invention treatment method can commence after radiation exposure has terminated, e.g., about 1-30 days or about 1-72 months or more after radiation exposure. In these embodiments, the treatment method can be administered over a period of months or years, e.g., about 0.5,1,2, 3, 4, 5, 6, 12, 18, 24, 36, 48, 72, 96 or more months. In some embodiments, dosing of the subject will occur for a period of about 2-12 months or for a period of about 4-6 months. Occasionally, treatment for radiation late effects will commence on the day of or before initiation of a planned radiation treatment, e.g., at about 1, 2, 3, 4, 5, 7, 14, 21, 28 or more days before a planned exposure to IE 0 Bq 312 138 radiation of a sufficient dose to a subject that will potentially generate, or is likely to generate, one or more radiation late effects, symptoms or conditions in the subject, e.g., any radiation-associated symptom or condition disclosed herein. In any of these embodiments, dosing of the subject with the F3C can be on a daily dosing basis or on an intermittent basis, e.g., using a treatment protocol essentially as described herein or in the cited references.

The F3Cs can be used to prevent, ameliorate, slow the progression and/or reduce the ultimate severity of marrow hypoplasia, hemorrhage, e.g., brainstem hemorrhage, cerebral hemorrhage or gastric hemorrhage or cytopenia, e.g., a blood cell count about 4-25% or more below the low end of a normal range for the subject, e.g., one or more of anemia (e.g., less than about 4.0.times.l0.sup.l2 red cells/L for adult human females and less than about 4.5.times.l0.sup.l2 red cells/L in adult human males or a hemoglobin level of less than about 12.0 g/dL in adult human females and less than about 13.5 g/dL in adult human males), late effect leukopenia (e.g., adult human white blood cell counts less than about 3,800, 4,000 or 4,300 mm.sup.-3; adult human basophil counts less than about 10 or 15 mm.sup.-3; adult human neutrophil counts less than about 1,600, 1,800 or 2,000 mm.sup.-3; human eosinophil level less than about 100, 120 or 150 mm.sup.-3; monocyte level less than about 260 or 300 mm.sup.-3) or late effect thrombocytopenia (e.g., human platelet counts less than about 15,000, 18,000 or 20,000 mm.sup.-3).

In some embodiments, an effective amount of a F3C is administered to a subject, or delivered to the subject's tissues, wherein the subject has received or has been exposed to a total radiation dose of at least about 0.5 Gy to about 100 Gy or more. The radiation dosage may comprise a single dose or two, three, four, five, six, 10 or more divided doses or subdoses. Thus, in exemplary embodiments, the subject may have received a total radiation dose in ranges of about 0.2-300 Gy, about 0.2-100 Gy, about 0.2-80 Gy 0.2-60 Gy, about 0.240 Gy, about 0.2-20 Gy, about 0.2-12 Gy, about 0.2-10 Gy, about 0.2-8 Gy, about 0.2-6 Gy or about 0.2-4 Gy. Subdivided doses may be administered on 1, 2, 3, 4, 5, 6, 7, 8, 9,10 or more occasions and such doses may be, e.g., about 0.05, 0.1, 0.3., 0.5, 0.8,1, 2, 3, 4, 5, 6 or more Gy per subdose. The subject may be exposed to radiation subdoses over a period of 139 about one day or over several days, e.g., about 2, 3, 4, 5, 6, 8, 10, 20 or 25 days, or over a period of months, e.g., about 1, 2, 3, 4, 5, 6, 8, 10,12,15, 18, 24, 36, 48 or more months.

When a subject is exposed to a full dose or a subdose of radiation, the exposure will occur over a period of about 1 minute to about 48 hours, typically about 2-120 minutes or about 460 minutes. Radiation doses or subdoses may be, e.g., about 0.01, 0.05, 0.1, 0.2, 0.5, 0.8, 1, 1.5, 2, 2.5, 3, 4, 5, 6 or 8 Gy per dose or subdose.

Administration of the F3C will typically commence at about 1 day to about 6 months after a subject has received a total radiation exposure, e.g., any dose or dose range disclosed herein. Typically, the F3C is used in the invention method commencing at about 2-120 days after radiation exposure or at about the time that radiation delayed effects become apparent to the subject or the subject's health care provider, e.g., within about 1-30 days after a condition or symptom is detected. Administration of the F3C may continue for a period of about 5 days to about 60 days for conditions or symptoms that tend to resolve over a relatively short time period. In other embodiments, the F3C is administered for a period of 2, 3, 4, 5, 6, 8, 10, 12, , 18, 24, 36, 48, 60 or more months for conditions or symptoms that tend to be chronic (e.g., neurological damage or inflammation), arise over a long time period (e.g., secondary cancers or neurological damage) or to progress over a relatively long time, e.g., about 1-5 years or more (e.g., cancers or neurological damage).

In any of the radiation exposure embodiments or dosing protocols disclosed herein, the F3C can be administered to the subject daily or on an intermittent basis, e.g., on about 1-5 days/week or about 2-10 days/month. In daily dosing embodiments, the F3C is administered to the subject daily for about 3 days to about 5 years or longer. Exemplary daily dosing embodiments include daily administration of a F3C for about 14, 30, 60, 90, 120, 180, 360 or more days. Daily doses may be administered in a single dose or as divided subdoses that \ are given, e.g., twice, three times, four times or more per day. In intermittent dosing embodiments, the F3C can be administered to the subject on 1, 2, 3, 4 or 5 days within a one week period, followed by a period of about 1, 2, 3,4, 5, 6, 7, 8,9, 10,12,16,20,24, 28 or 32 weeks without administration of the F3C, followed by administration of the F3C to the tS°603l2 140 subject on 1, 2, 3, 4 or 5 days within a one week period. In other intermittent dosing embodiments, the F3C is administered to the subject every other day, every two days, every three days, every 4 days or every seven days.

For any radiation exposure situation where delayed radiation effects may arise, e.g., a radiation exposure as disclosed herein, daily administration may comprise administering about 0.01 mg/kg to about 500 mg/kg of the F3C to a subject per day. Exemplary dosages are about 0.1-100 mg/kg/day and about 0.2-30 mg/kg/day. Exemplary unit doses comprise about 1, 5, 10,15, 20, 25,30,40, 50,75,100,200,300 or 500 mg of a F3C in a suitable formulation. Exemplary unit dosages for humans or other subjects disclosed herein comprise a formulation that comprises about 1-1000 mg of a F3C or about 5-400 mg or about 10-300 mg, e.g., about 5, 10, 20,25, 30, 40, 50, 60 75, 100, 150, 200,250, 300, 400 or 500 mg. Larger unit or daily dosages, e.g., about 5-400 mg, will generally be used with larger subjects such as humans, while smaller subjects such as rodents or dogs will generally utilize lower unit or daily dosages, e.g., about 0.3-25 mg.

Modulation of transcription factors, receptors and gene expression. In treating any of the diseases, conditions or symptoms disclosed herein, the F3Cs can modulate, i.e., detectably enhance or increase or detectably inhibit or decrease, the expression or a biological activity(ies) of one or more transcription factors or receptors. This can lead to detectable modulation of target gene activity or expression as part of the treatment or amelioration of the disease, condition or symptom. Such modulation can arise from changes in the capacity of a transcription factor or receptor to bind to or form a complex with other natural ligands such as a target DNA sequence(s), another transcription factor(s), a transcription cofactor, a receptor such as a nuclear hormone receptor or cell membrane receptor (e.g., a lipid, peptide, protein or glycoprotein receptor such as an interleukin receptor or a growth factor receptor), a receptor cofaetor or an enzyme such as a polymerase, kinase, phosphatase or transferase. The effects of F3Cs on these biomolecules can be exerted in immune cells or in non-immune tissue, e.g., cells or tissue adjacent to diseased tissue such as infected or malignant cells. The F3Cs may directly or indirectly modulate the capacity of any of these molecules to transduce signals that are part of normal signal trandsuction processes.

ME 6 o j 12 141 In many of the clinical conditions described herein, e.g., in cancers, infections, acute inflammation, chronic inflammation, trauma, neurological conditions or autoimmunity, the F3Cs can modulate, e.g., detectably decrease or increase, a biological activity(ies), protein or molecule level or RNA level of 1, 2, 3, 4, 5, 6 or more biomolecules that are involved in establishment, maintenance or progression of a disease, condition or symptom. Such biomolecules include 1, 2, 3, 4, 5, 6 or more of AP-1, a cyclooxygenase such as mammalian or human cyclooxygenase-1 (COX-1) or cyclooxygenase-2 (COX-2), a mammalian or human lipoxygenase, e.g., 5-lipoxygenase, TNF.alpha., TNF.alpha. receptor 1, TNF.alpha. receptor 2, TNF receptor-associated factor, TNF.beta., TNF.beta. receptor, MIP-1.alpha., monocyte chemoattractant-1 (MCP-1), interferon gamma (IFN.gamma. or .gamma.IFN), IL1.alpha., IL-1.beta., IL-1 .alpha, receptor, IL-1.beta, receptor, IL-2, IL-3, IL-4, IL-4 receptor (IL4R), IL-5, IL-6, IL-6 receptor (IL-6R), IL-8, IL-8 receptor (IL-8R), IL-10, IL-10 receptor (IL-10R), IL-12, an IL-12 receptor (e.g., IL-12R.beta.2), IL-13, IL-15, IL-17, IL-18, nuclear factor kappa B (NF.kappa.B), AP-1, c-maf, v-maf, rnafB, Nrl, mafK, mafG, the maf family protein pi8, reactive oxygen species, e.g., peroxynitrite, hydrogen peroxide or superoxide ion (collectively ROS), a 17.beta.-hydroxysteroid dehydrogenase (17.beta.-HSD) or an 11 .beta.-hydroxysteroid dehydrogenase (1 l.beta.-HSD), e.g., 11.beta.-HSD type 11 .beta.HSD type 2, 17.beta.-HSD type 1,17.beta.-HSD type 2 or 17.beta.-HSD type 5, a steroid aromatase, e.g., cytochrome P450 aromatase, steroid 5.alpha.-reductase, serum or blood cortisol, cytosolic phospholipase A2 (cPLA2), calcium-independent phospholipase A2 (iPLA2), a prostaglandin, e.g., prostaglandin E2 (PGE2) or prostaglandin D2 (PGD2), a leukotriene. e.g., leukotriene B4, inducible nitric oxide synthetase (iNOS), nitric oxide (NO), GM-CSF, RANTES (regulated on activation, nonnal T cells expressed and secreted), eotaxin, GATA-3, CCR1, CCR3, CCR4, CCR5, CXCR4, in, e.g., a subject's cell(s) or tissue(s) or in enzyme, tissue or cell-based assays. In these subjects, the levels of other biomolecules, their RNAs or the level of their activity can be detectably modulated include IFN.alpha., INF.alpha. receptor, PPAR.alpha., PPAR.gamma., PPAR.delta. or a transcription factor such as T-bet is detectably increased. Other biomolecules or their isofonns, polymorphs, orthologs, or homologs that the F3Cs directly or indirectly modulate ΙΕ ί 6 0 3 12 142 include one or more of, e.g., Janus kinase 1 (JAK1), Janus kinase 2 (JAK2), Janus kinase 3 (JAK3), signal transducer and activator of transcription 1 (STAT1), signal transducer and activator of transcription 2 (STAT2) and signal transducer and activator of transcription 3 (STAT3). The F3Cs can modulate the other biologically active analogs of any these enzymes, chemokines, cytokines, their receptors or ligands, including their isoforms, polymorphs, orthologs or homologs. In some cells or tissues, one or more of these biomolecules may be detectably increased, while in other cells or tissues, the same biomolecule may be detectably decreased. Thus, the biomolecules that the F3Cs can modulate, e.g., detectably increase or decrease, include the intracellular or extracellular level or biological activity of one or more enzyme, cytokine, cytokine receptor, chemokine and/or chemokine receptor. Exemplary chemokine receptors include one, two or more of CCR-1, CCR-2, CCR-2A, CCR-2B, CCR-3, CCR4, CCR-5, CXCR-3 and CXCR4.

The F3Cs can modulate the activity of certain biomolecules that mediate various biological responses that affect establishment or progression of a disease or that enhance or inhibit specific immune responses. Thus, in conditions where unwanted inflammation is present, the F3Cs can reduce inflammation, while enhancing Thi or Tel immune responses at the same time. The biomolecules that the F3Cs can modulate include, e.g., transcription factors or receptors, including orphan nuclear receptors, and the homologs, isoforms, orthologs and co-factors (e.g., co-repressors, co-activators, transcription factors, gene promoter regions, sequences or messenger moieties such as calcium ions, potassium ions or cAMP) of any of these molecules and related molecules that participate in their function. The compounds can directly or indirectly from complexes with such molecules or they can modulate (detectably increase or decrease) the synthesis, level or one or more biological activities of those molecules. These complexes include receptors or transcription factor complexes, which can comprise heterodimers, homodimers and trimer, tetramer, pentamer or higher homo or hetero complexes. A number of the orphan receptors or their isoforms, orthologs or homologs, e.g., PPAR.alpha., PPAR.beta., PPAR.gamma., PPAR.gamma.l, PPAR.gamma.2, PPAR.gamma.3, LXR.alpha., LXR.beta., SXR, PXR, CAR.alpha. and CAR.beta., can form heterodimers with one or more of RXR.alpha., RX.beta., and 143 RXR.gamma.. Exemplary mammalian, human or other biomolecules include steroidogenic factor-1 (SF-1), steroidogenic acute regulatory protein (StAR), a chicken ovalbumin upstream promoter-transcription factor (COUP-TF), chicken ovalbumin upstream promoter4 transcription factor (COUP-TFI) and its mammalian isoforms, orthologs and homologs, silencing mediator for retinoid and thyroid hormone receptor (SMRT) and its mammalian isoforms, orthologs and homologs, sterol regulatory element binding protein (SREBP) 1 a (SREBP-la), SREBP-lc, SREPB-2, NF-E3, FKHR-L1, COUP-TFII and its mammalian isoforms, orthologs and homologs, and the isoforms, orthologs and homologs of I.kappa.B, I.kappa.BA, AML-3, PEBP2.alpha.Al, Os£2, Cbfal, RUNX2, activating transcription factor 2 (ATF2), c-Jun, c-Fos, a mitogen activated kinase (MAP) such as p38 or JNK, a mitogen activated kinase kinase (MKK), a pl60 or steroid receptor coactivator-1 family (SRC-1, SRC-l/serum response factor), SRC-2, SRC-3, SET, nerve growth factor inducible protein B, StF-IT, NFAT, NF AT interacting protein 45 (NIP45), IkB, an IkB kinase, NFATp, NFAT4, an AP-1 family protein, a p300 protein, CREB, CREB-binding protein (CPB), p300/CBP, p300/CPB-associated factor, S WI/SNF and their human and other homologs, BRG-1, OCT-1/OAF, AP-1, AF-2, Ets, androgen receptor associated protein 54 (ARA54), androgen receptor associated protein 55 (ARA55), androgen receptor associated protein 70 (ARA70), androgen receptor-interacting protein 3 (ARIP3), ARIP3/PIASx .alpha, complex, PLASx .alpha., Mizl, Mizl/PIASx .beta, complex, PIASx .beta., PIAS1, PIAS3, GBP, GBP/PIAS1 complex, RAC3/ACTR complex, SRC-1.alpha., receptor interacting protein21 140 (RIP-140), transcription factor activator protein-1, activation function-2, glucocorticoid receptor-interacting protein-1 (GRIP-1), receptor interacting protein-160 (RIP-160), suppressor of gal4D lesions (SUG-1), transcription intermediary factor-1 (TIF-l), transcription intermediary factor-2 (TIF-2), SMRT, N-CoR, N-CoA-1, p/CIP, p65 (RelA), the 120 KD rel-related transcription factor, heat shock proteins (HSP) such as HSP90, HSP70 and HSP72, heat shock factor-1, Vpr encoded by the human immunodeficiency virus and its isoforms and homologs thereof, testicular orphan receptor 2 (TR2), testicular orphan receptor 4 (TR4), a thyroid hormone receptor .alpha., thyroid hormone receptor .alpha. 1 (TR.alpha. 1), thyroid hormone receptor .alpha.2 (TR.alpha.2), thyroid hormone receptor .beta. (TR.beta.), retinoid X receptor .alpha. (RXR.alpha.), retinoid X receptor .beta. 060312 144 (RXR.beta.), retinoid X receptor .gamma. (RXR.gamma.), TR .alpha. 1/RXR .alpha, heterodimer, direct repeat4 thyroid hormone response element (DR4-TRE), an estrogen receptor (ER) such as ER.alpha.or ER.beta., estrogen receptor related receptor .alpha. (ERR.alpha. or EER1), estrogen receptor related receptor .beta. (ERR.beta. orEER2), estrogen receptor related receptor .gamma. (ERR.gamma. or EER3), steroid xenobiotic receptor (SXR), a hepatocyte nuclear factor 4 (HNF4), hepatocyte nuclear factor 4.gamma. (HNF-4.gamma.), hepatocyte nuclear factor 3 (HNF-3), liver X receptors (LXRs), LXR.alpha., LXR.beta., estrogen receptor .alpha. (ER.alpha.), constitutive androstane receptor-.alpha. (CAR-.alpha.), constitutive androstane receptor-.beta. (CAR-.beta.), RXR/CAR-.beta. heterodimer, short heterodimer partner (SHP; NROB2), SHP/ER. alpha, heterodimer, estrogen receptor .beta., SHP/ER.beta. heterodimer, testicular orphan receptor TR4, TR2/TR4 heterodimer, pregnane X receptor (PXR) and isoforms, cytochrome P-450 monooxygenase 3A4, including its gene promoter region and isoforms thereof, HNF4/cytochrome P-450 monooxygenase 3A4 gene promoter region and isoforms complex, HIV-1 long terminal repeat (LTR), HIV-2 LTR, TR2/HIV-1 LTR complex, TR4/HIV-1 LTR complex, TR4/HIV-1 LTR complex, TR al/TR4/HIV-l LTR complex, TR2 isoforms (TR2-5, TR7, TR9, TRI 1), DAX-1 (NROB1), DAX-1/steroidogenic acute regulatory protein gene promoter region, RevErb, Rev-erbA .alpha., Rev-erb .beta., steroid receptor coactivator amplified in breast cancer (AIB 1), p300/CREB binding protein-interacting protein (p/CIP), thyroid hormone receptor (TR, T3R), thyroid hormone response elements (T3REs), retinoblastoma protien (Rb), tumor suppressor factor p53, transcription factor E2F, mammalian acute phase response factor (APRF), constitutive androstane receptor (CAR), Xehopus xSRC-3 and mammalian (e.g., human) isoforms, orthologs and homologs, TAK1, TAK1/peroxisome proliferator-activated receptor .alpha. (PPAR.alpha.) complex, PPAR.alpha./RXR.alpha. complex, peroxisome proliferator-activated receptor .beta. (PPAR.beta.), peroxisome proliferator-activated receptor .gamma. (PPAR.gamma.), peroxisome proliferator-activated receptor .delta. (PPAR.delta.), famesoid X receptor, retina X receptor, TAK-1/RIP-140 complex, a retinoic acid receptor (RAR), retinoic acid receptor.beta. (RAR.beta.), retinoic acid receptor-.gamma. (RAR.gamma.), TR4/RXRE complex, SF-l/steroid hydroxylase gene promoter region, SF-1/oxytocin, including its gene promoter J £0 g q j j £ 145 region, a bile acid receptor (FXR), nuclear receptor corepressor (NcoR), liver receptor homologous protein-1 (LRH-1; NR5A2), SF-l/ACTH receptor gene promoter region, rat Ear-2 and mammalian homologs, human TR3 orphan receptor (TR3), RLD-1, OR-1, androgen receptor, glucocorticoid receptor, estrogen receptor, progesterone receptor, mineralcorticoid receptor, aldosterone receptor, E6-associated protein (E6-AP), ORI, ORl/RXR.alpha. complex, TIF-1, CBP/P300 complex, TRIP1/SUG-1 complex, RIP-140, steroid receptor coactivator 1 (SRC1), SRCl.alpha./P160 complex and TIF-2/GRIP-1 complex, RAR/N-CoR/RIP13 complex, RAR/SMRT/TRAC-2 complex and protein X of hepatitis B virus. The homologs, orthologs and isoforms of these transcription factors, receptors and other molecules are included among the molecules that the F3Cs can modulate the synthesis or one or more biological activities of. Such factors are biologically active or function in one or more of a number of cell types such as T cells, B cells, macrophages, dendritic cells, platelets, monocytes, neutrophils, neurons, epithelial cells, endothelial cells, cartilage cells, osteoblasts, osteoclasts, splenocytes, thymocytes and GALT associated cells. Methods to identify these molecules and their biological activities have been described, e.g., U.S. Pat. Nos. 6,248,781, 6,242,253, 6,180,681, 6,174,676, 6,090,561, 6,090,542, 6,074,850, 6,063,583, 6,051,373, 6,024,940, 5,989,810, 5,958,671, 5,925,657, 5,958,671, 5,844,082, 5,837,840, 5,770,581, 5,756,673, and PCT publication Nos. WO 00/24245, WO 0073453 and WO 97/39721.

In one aspect, the compounds are used to treat, prevent or to ameliorate conditions or symptoms that are associated with unwanted or expression or activity of one or more of these molecules in conditions such as, e.g., acute inflammation, chronic inflammation or their symptoms, acute allergy, chronic allergy or their symptoms, e.g., allergic rhinitis or acute or chronic asthma, psoriatic arthritis, osteoporosis, osteoarthritis, rheumatoid arthritis, neurological dysfunction or their symptoms, e.g., dementias such as Alzheimer's Disease, Parkinson's Disease, or memory loss conditions, in osteoporosis or in cancer such as breast cancer. The compounds can prevent NF.kappa.B from translocating from the cytoplasm into the nucleus and thus can increase the ratio of cytoplasmic NF.kappa.B to nuclear NF.kappa.B. The F3Cs may inhibit activation of NF.kappa.B-mediated transcription while 512 146 NF.kappa.B is bound to target DNA sequences in the nucleus. Alternatively, the F3Cs can activate or enhance the expression of or one or more activity of a transcription factor such as T-bet in, e.g., a subject's cell(s) or tissue(s) or in enzyme or cell-based assays. In this aspect the compounds are used to treat, prevent or to ameliorate conditions or symptoms that are associated with deficient expression or activity of T-bet in conditions such as immune dysfunction in an immunosuppression condition, aging, an infection, a cancer or precancer as described herein or in the cited references.

The invention provides methods to identify compounds to regulate immune or other biological responses in a context-sensitive manner. Such compounds modulate differential expression in a cell of the level of or an activity of, eg., 4, 5, 6, 7, 8 or more genes or gene products, comprising administering an effective amount of a F3C. The genes or gene products are USF1, c-Fos, EGR1, Cull, RIPK2, Lkappa.B.alpha., I.kappa.BKb, NF.kappa.B, NF-.kappa.B2, NF-.kappa.Bl p50, Fnl4 (fibroblast growth factor-inducible 14), TWEAK (TNF-like weak inducer of apoptosis), NEMO (NF-.kappa.B essential modifier), FCAR, c-Fos/C/EBP.beta., RANTES, ICAM1, TSG (TNFAIP6), IL-2 receptor .alpha., GRO2, GRO3, HOI, Jun B, c-Fos/JunB complex, JunB/ATF3 complex, c-Jun, c-Fos/c-Jun complex, ATF-3, MMP1, TSG-6 (TNFAIP3), AP-1, EGR1, TGF.beta., ATF-3/c-Jun complex, c-Fos, MMP3, IL-8, STAT5A, STAT5B, CDKN1A, IFN.gamma. receptor 2 (IFN.gamma.R2), T-bet, C reactive protein, immunoglobulin E, an AP-1 family protein, GATA-3, Jak2, Tyk2, statl, stat3, stat4, stat5, MIP-l.alpha., MIP-2, IP-10, MCP-1, TNF.alpha., TNF-.beta., LT-.beta., IFN-.alpha., IFN-.beta., TGF-.beta.l, NF-.kappa.B, ILl.alpha., IL-l.beta., IL-4, L-6, IL-10, IL-12 receptor .beta.l, IL-12p35, IL-12p40, IL-23, IL23 receptor or another gene or gene product disclosed herein, including in Table 1. The compounds identified by the screening methods modulate the expression of dysregulated genes and restore or enhance normal immune responses in conditions where unwanted dysregulation contributes to the establishment or progression a pathological condition such as an infection, an autoimmune disorder, a cardiovascular condition or a neurological condition. ί o j f 2 147 Thus, in some embodiments, the level or a biological activity of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more of COX-2, IL-l.beta., TNF.alpha., TNF.alpha. receptor 1, TNF.alpha. receptor 2, TNF receptor-associated factor, MIP-l.alpha., MCP-1, IFN.gamma., IL4, IL4R, IL-6, IL-6R, IL-8, IL-8R, IL-10, IL-10R, IL-12, IL-12R, IL-18, IL-18RNF.kappa.B, IkB.alpha., AP-1, GATA-3, ll.beta.HSDl, cPLA2, iPLA2, cortisol, ROS, PGE2, PGD2, leukotriene A4, leukotriene B4, leukotriene C4, iNOS or GM-CSF are optionally measured and they are generally detectably reduced, e.g., RNA or protein levels are reduced by about 10-95% or about 20-95% or more compared to suitable untreated controls. In these embodiments, the level or a biological activity of 4, 5, 6 or more of IFN.alpha., INF.alpha. receptor, IL-12, an IL-12 receptor, (e.g., IL-12R.beta.2), PPAR.alpha., PPAR.gamma., and T-bet are optionally measured and they are generally detectably increased. In a chronic infection condition, e.g., HIV in humans, autoimmunity, a chronic fungal or parasite infection or in a precancer or cancer condition, e.g., benign prostatic hyperplasia, the progression of the condition may be slowed over a period of 1, 2, 3, 4, 5 or more years. In these embodiments, the subject's condition becomes more manageable with a reduced incidence or severity of side effects, e.g., a detectable halt, slowing, reversal or decreased incidence of wasting, dementia, CD4 cell count decreases or viral load increases, which tend to occur over time in HIV infected humans or a halt, slowing or reversal of pathogen or precancer or cancer cell replication.

These effects are typically observed after administration of an effective amount of a F3C using, e.g., a method or dose essentially as disclosed herein. The simultaneous reduction of multiple biomolecules provides a method to modulate immune responses by modulating multiple pathways that lead to a common condition such as inflammation. This provides a method to treat or ameliorate, e.g., acute or chronic inflammation, a cancer, an infection or a symptom associated therewith, or to slow the progression of or reduce the severity of these i conditions or their symptoms.

Previously described methods can be used to measure the amount, activity or cellular location of various biomolecules such as cytokines or transcription factors. See, e.g., U.S. u6oji2 148 Pat. Nos. 6,107,034, 5,925,657, 5,658,744, 4,016,043 and U.S. Pat. No. 3,850,752, S. Szabo et al., Cell 2000 100:655-669, Y. Nakamura et al., J. Allergy Clin. Immunol. 1999 103(2 pt. 1):215-222., R. V. Hoch et al., Int J. Cancer 1999 84:122-128. These methods can be used to measure the effects of the F3Cs on transcription factors or receptors in cells or tissues that have been exposed to the compounds.

Without wishing to be bound to any theory, the F3Cs may modulate multiple biomolecules in a microenvironment sensitive manner or context. The effects of the compounds can provide a decrease in a particular molecule such as IFN.gamma. and a decrease in inflammation associated with elevated IFN.gamma. levels or activity without eliminating beneficial effects of the molecule. This effect arises from decreasing the level or activity of a biomolecule such as IFN.gamma. in cells that are dysregulated, while allowing normal immune cells to produce sufficient amounts of the same molecule to perform normal immune functions. In locations where the biomolecule is needed for activity, e.g., in lymph nodes or spleen cells, sufficient amounts of the modulated molecule are present to elicit a desired response, while the level of the molecule in cells in circulation decreases. The compounds can increase IL-13, IL-15, IL-17 or IL-18 in conditions where a subject has a deficient Thl immune response, e.g., in infection or cancer. Conversely, the compounds can decrease IL-13, IL-15 or IL-18 in conditions such as allergy or autoimmune conditions, e.g., multiple sclerosis, where an excess Thl immune status may prevail.

In general, the F3Cs will detectably decrease the synthesis or one or more biological activity of one or more of these molecules (or other transcription factors or receptors disclosed herein) when such synthesis or activities is associated with the establishment, maintenance, progression or enhanced severity of a clinical condition or symptom disclosed herein. Conversely, the F3Cs will generally detectably increase the synthesis or one or more biological activities of one or more of these molecules (or other transcription factors or receptors disclosed herein) when such synthesis or activity is associated with the treatment, prevention, cure or amelioration of a clinical condition or symptom disclosed herein. ιεοβο 312 149 These decreases or increases compared to suitable controls can be relatively small, including changes near the lower limits of delectability for such molecules using known or new assays, e.g., a decrease or increase in the synthesis or biological activity of at least about 2%, about 5%, about 10% or about 20%. Such changes can be modest or relatively large, e.g., at least about a 50% change, at least about a 90% change, or at least about a 200% change, up to about a 5-fold, about a 10-fold, about a 100-fold or greater decrease or increase in the synthesis or biological activity of the affected molecule(s) compared to suitable controls.

These changes are typically measured relative to controls that lack a F3C or that use known agonists or antagonists of one or more relevant molecules. Assays can be based on measuring decreases or increases in, e.g., one or more of protein levels, RNA or mRNA levels, a ligand binding activity, transcription of a target gene(s) and the like. Suitable assay protocols include any suitable polymerase chain reaction assay to measure an RNA or mRNA, any suitable blotting protocol for nucleic acid or for protein such as a Northern or Western blot method or any transcription assay, including DNA footprinting or a gene expression or gene function assay. Typically the F3Cs will effect detectable changes in the synthesis or one or more biological activities in a concentration range of about 0.5.times.l0.sup.-9 M to about 3.times. 10.sup.-5 M. Exemplary compositions that comprise a F3C for use in, e.g., in vivo animal assays, in vitro cell or tissue culture assays or in cell free assays, will comprise one or more suitable solvents or vehicles including DMSO, ethanol, water and a suitable tissue culture medium.

One or more of these transcription factors, receptors or complexes can be a component in methods when, e.g., they are used with a F3C in cell-free assays or in tissue culture assays. Formation of these complexes in cells or analysis of the effects of F3Cs on one or more of their biological activities is facilitated by inserting into the cells a DNA construct! s) that expresses one or more of these proteins, e.g., mammalian or yeast cells containing a stable -¾ DNA construct or a construct used for transient transfection assays. Methods to perform assays or to induce biological responses in vitro or in vivo using the F3Cs as agonists, antagonists or as reference standards are essentially as described, see, e.g., U.S. Pat. Nos. 5,080,139, 5,696,133, 5,932,431, 5,932,555, 5,935,968, 5,945,279, 5,945,404, 5,945,410, 150 IE 0 e o j j 2 ,945,412, 5,945,448, 5,952,319, 5,952,371, 5,955,632, 5,958,710, 5,958,892 and 5,962,443; International Publication Numbers WO 96/19458, WO 99/41257 and WO 99/45930. The complexes or assay systems, that comprise a F3C and one or more of these molecules are embodiments of the invention, as are the use of these compositions when employed in the practice of any ofthe assay methods or in any of the clinical treatment methods disclosed herein or in the cited references.

Invention embodiments include a method comprising contacting a F3C(s) with a cell(s), whereby the F3C(s) forms a complex with a steroid hormone receptor or results directly or indirectly in the modulation of a biological activity of the steroid hormone receptor or a gene that it regulates. The steroid hormone receptor may be an orphan nuclear hormone receptor or a characterized receptor such as the glucocorticoid receptor, estrogen receptor or the androgen receptor that displays a moderate or high binding affinity for the F3C(s). In some embodiments, the nuclear hormone receptor is a known receptor. Biological effects from interaction of a F3C and a receptor can lead to interference with the replication or development of a pathogen or the cell(s) itself, e.g., detectably inhibited proliferation of cancer cells. For example, expression of HIV transcripts in HIV-infected cells may be altered. The receptor-F3C complex may directly interfere with LTR-dependent transcription of HIV genes, leading to reduced viral replication. Alternatively, such effects can include the decreased synthesis or biological activity of a protein or gene product that is associated with the establishment, maintenance or progression of a disease condition described herein or in the cited references.

An aspect of F3C biological activity is their capacity to modulate the opacity of cells or tissues described herein to express one or more enzymes that mediate phase 11 detoxification and reduction of damaging or reactive species such as xenobiotics, including electrophiles and chemical carcinogens, and superoxide radicals or hydrogen peroxide. Modulation of these genes is mediated by one or more transcription factors or complexes of factors that include bZip transcription factors such as Nrf2 (NF-E2 related factor 2, Unigene symbol Nfe2L2) and Maf proteins such as MafG, MafK or MafF. These factors bind to cisIE 06o 3 12 151 elements such as EpRE (electrophile response element) or ARE (antioxident response element). EpRE and/or ARE elements present in the promoters of phase II detoxification enzymes including NAD(P)H:quinine oxidoreductase-1 (NQO1) and glutathione-Stransferase (GST) as well as cellular defensive enzymes such as thioredoxins, heme oxygenase 1 (HO 1, or HMOX1), the catalytic and regulatory subunit .gamma.glutamylcycteine syhthetase (.gamma.GCS or GCLM) and and xCT (SLC7A11), a subunit of the cystine/glutamate exchange transporter. EpRE and ARE mediate upregulation of these genes following exposure ofthe cells to many xenobiotics. In situations where enhanced expression of these genes or transcription factors is desirable, the F3C upregulate the activity or levels of one or more of these factors and/or enzymes.

Thus, in some embodiments the F3C are used to modulate the level or activity of one or more ofNrf2, a thioredoxin, NQO1, GST, HO 1, the catalytic subunit of .gamma.GCS, the regulatory subunit of .gamma.GCS and xCT in cells or tissues that are exposed to a F3C. In some embodiments, the cells or tissues are treated with a F3C when an unwanted acute or chronic condition such as toxin exposure or elevated oxidative stress is present in the cells or tissues. Such conditions can occur, e.g., as described elsewhere herein, including in acute or chronic pathological inflammation conditions, acute or chronic infections and trauma conditions. The effect of the F3Cs is restoration of normal expression or establishment of desired levels of expression of one or more of these transcription factors or enzymes, e.g., decreased expression in situations where chronic over-expression occurs. Thus, the F3C can be used to modulate these or other genes described herein, e.g., to decrease expression or mRNA levels or protein levels of one or more of these or other genes in clinical conditions where excess or unwanted expression or levels ofthe gene is associated with establishment, maintenance, severity or progression of the clinical condition resulting in clincal improvement in the disease or an unwanted symptom.

Other genes in cells or tissues in vitro or in vivo that the F3Cs can modulate include 1, 2, 3, 4, 5, 6 or more of the following genes (which are listed by their Unigene symbols): LOC55967, 37149, LOC64148, LOC57823, AGPAT1, PHACS, OAS1, OAS3, OASL, Ιί 0og312 152 IFRG28, C(27)-3BETA-HSD, LOC55831, HMGCR, HMGCS1, PDPK1, HPD, AIRP, PFKFB3, DHCR7, OGGI, ADAM9, ADAMTS1, ADAMTS10, AKAP1, AKAP10, AKAP3, AKAP4, AKAP8, TOB3, ABO, AIM1, LOC55902, ACAT2, ASMTL, ACP5, OA48-18, ACO1, ACO1, ACR, ABLIM, ACF7, LOC81569, ARPC4, ACTC, ACTG2, ACTL6, ALCAM, PC4, ATF1, ATF3, ATF4, AICDA, ABR, ACVR1, ACVR2, ACADS, ACOX1, ACOX2, AOAH, ACYP1, AD-003, LOC55829, AP1B1, AP1G2, AP1S1, AP2M1, AP3B2, AP3M1, ADD3, ADORA2A, ADORA2B, ADA, ADAR, ADAT1, AMPD1, ADCY3, ADCY7, AK1, ADSL, ARF4, ARF4L, ARF6, ARL4, ADPRTL2, ART3, LOC51578, ADRB2, ADRB3, ADM, ADMR, AGER, AFG3L1, AFG3L2, AGC1, AGRP, ALDH1 Al, ALDH1B1, ALDH2, AKR1C1, AKR1C2, AKR1C3, AKR7A2, AKR7A3, ALDOC, ALG5, ALPI, ALPL, ABH, AF1Q, AIF1, KIAA1017, ATRX, A2M, A2M, AMACR, ASPSCR1, ABP1, ACCN1, ACCN2, AOC2, ACY1, ALAD, ALAS1, LOC64167, AMPH, AGL, APPBP1, ALS2CR2, AGTRL1, ANK2, ASB1, ASB2, ANXA1, ANXA11, ANXA9, MOX2, KIN, AMHR2, KIAA0106, APELIN, APG5L, API5L1, APXL. APOARGC, APOC4, LOC51275, APAF1, APAF1, APEXL2, AQP1, AQP2, LOC51326, HSU52521, ARG2, AVPR1B, ARMET, ASS, ALX3, ALX4, ACTR3, ARTN, ARNTL, AIP, ARSA, ARSB, ARSD, ASH2L, AMSH, AKNA, ACLY, ATP5F1, ATP5I, ATP5J, ATP5A1, ATP5C1, ATP50, ATP2A1, ATP2B1, ATP2B2, ATP2C1, ATP7B, ATP6D, ABC A3, ABCA4, ABCA5, ABCA7, ABCC5, ABCD1, ABCE1, ABCF1, ABCF2, ABCG4, AUH, BAL, B29, B7-H3, BPI, BIRC1, BIRC3, BIRC4, BIRC7, BIRC8, BAF53A, BAIAP2, BARX2, BHLHB2, BLZF1, BATF, BTEB1, BCL11A, BCL2, BCL3, BCL6, BCL7A, BCL9, BTG1, BBC3, BMF, BMF, BNIP3, BAG2, BIK, BCL2L1, BCL2L11, BOKL, SBBI42, BFSP1, BBP, ICAT, GAL3ST-4, BACE, BACE2, BACE2, BTRC, BF, BID, BIC, BLVRA, BTD, BPHL, BM88, BN51T, BPOZ, BRI3, BAI2, BSMAP, BCAT2, BCKDHB, BCRP2, BCAR3, BIG1, BIG2, BAZ1A, BAZ2A, BAZ2B, BP75, BRD1, BRD2, BRD3, BRUNOL5, BRUNOL6, BTK, BTBD2, BTG2, BUB3, BUP, BRF2, BBOX1, BTN3A1, BYSL, LOC81558, C2F, ZFP26, C6.1A, C9orfl0, CABP1, CHP, CACNB1, CALM2, CAPN1, CAPN2, CAPN5, LOC57118, H_GS165L15.1, CHST2, CA3, CA11, CA12, CPM, CLC, CROT, CBL, CSNK2A2, CSN2, CFLAR, CASP10, CASP2, CASP6, CASP8, CARD 14, CECR5, CECR6, CAT56, CTSC, CTSE, CTSL, CTSS, CTSW, CTSZ, 153 IE 0 6 0 3 12 LOC’56996, CITED1, CEBPE, CBF2, CNOT2, CD14, CD1A, CD1D, CD2AP, CD33, CD36L1, CD3D, ASE-1, CD3G, CD4, CD44, CD5L, CD69, CD72, CD79A, CD8A, CD8B1, CD83, CD84, CDC14A, CKS2, CEP2, CDC45L, CLK2, TOK-1, CDV-1, CDC25A, CDC25C, CGR19, GP110, CREG, CRABP1, CENTA1, CENTB2, CENTG1, CETN3, CENPA, CENPB, CENPF, CEP1, CER1, CCM1, LOC51148, CLN2, CLN3, FIGF, LOC50999, CGI-152, CGI-152, CHES1, CCR1, CCR8, CCRL2, CX3CR1, CMKLR1, CHN1, CLCN2, CLCN3, CLCA1, CLCA4, CLIC1, CHK, CHKL, LOC56994, CHRNA3, CHRNA4, C4S-2, C4ST, CSPG4, CSPG6, CHAC, CGB8, CSH1, CHAF1B, CBX4, CHD1L, CHD3, CHD3, Cl lorfl4, C21orfll, C22orf5, CTRL, CIG30, CIZ1, LUC7A, CS, CLTCL1, CLDN12, CLDN15, CLDN17, CPSF3, CPSF5, CPSF6, CLU, MERTK, LOC55907, VI, IRLB, JP01, CIA, CLP, F3, F9, F8, F8A, LOC51137, COPA, CKN1, COQ7, CRSP6, CRSP7, CIAS1, CLPS, COL2A1, COL4A3BP, COL5A3, COL7A1, COL19A1, COL15A1, COL16A1, COL18A1, MIC1, CSF1R, CSF3, C1QA, C1R, C2, C5R1, C8A, C8G, CPLX2, GJA10, CGTHBA, CHUK, CNTN5, CNTNAP1, COPS4, CPNE3, CBFB, CBFA2T3, CORO2A, COX 10, CPXCR1, CRY1, CRYGA, CRYZL1, CSK, CTDP1, CLECSF6, CLECSF9, LOC51266, CUGBP2, CUL1, CUL2, CUL3, C.UL4B, CUL5, HUMMHCW1A, CNGB1, CCNB3, DMTF1, CCNE2, CCNG2, CNNM2, CNNM4, CDK10, CDK3, CDK7, CDKN1A, CDKN1B, CDKN2B, CDKN2C, CDKN2D, CDKN3, CTH, CBS, CST8, CSTA, CST3, CSRP1, CSRP3, CHORDC1, CCBLl, CRIP2, CHIC2, CYSLT1, CARS, CTNS, CMAH, CYB5, HCS, COX5A, COX7C, LOC57404, CYP51, CYP1A1, CYP1B1, CYP2A6, CYP2D6, CYP2E, CYP2J2, CYP4F3, CYP11B1, CYP21A2, CYP24, CYP27B1, CRLF2, CREME9, CNK, N-PAC, KIAA0068, PIR121, #22, LOC81501, DRIL2, DDXBP1, DDX10, DDX16, DDX19, DDX8, DELGEF, DAF, DEFB2, DSS1, DLL1, DLL3, DSIPI, KIAA1365, DRPLA, DNASE1, DTYMK, #23, D4ST-1, #24, DDEF1, DRG2, DGKZ, DGAT, DIAPH1, DEF6, DGS-D, DGS-H, DHODH, DPYSL2, DPYD, HUM2DD, DDAH1, DDAH2, HSA249128, DTR, DLG4, DISCI, SAS10, DLX2, '< UBD, DKFZP434A236, DKFZP434B1O3, DKFZP434B168, DKFZP434C212, DKFZP4341216, DKFZP434NO14, DKFZP434N043, DKFZP434N161, DKFZP564C1940, DKFZP564M1416, DKFZP564O123, DKFZP566C134, DKFZP566F0546, DKFZP566F2124, DKFZP566K023, DC8, CTRP5, DKFZP586M1523, DKFZP727C091, 154 ,£ 0 6 0 3 12 DNMT1, DNMT2, DNMT3B, DFFB, DNAJA1, DNAJB11, DNAJB4, DNAJB5, DNAJB6. DNA.TB6, DNAJB9, DNAJC3, DOK2, DPMI, DO, DRD1, DRD5, DUX4, DORFIN, ADAR3, DSCR4, DONSON, DUOX1, DUSP1, DUSP11, DUSP12, DUSP2, DUSP3, DUSP4, DUSP5, DUSP7, DYRK3, DYRK4, DNCI1, DKC1, DMWD, EP300, E2F5, SMURF1, ELF1, EAF1, EAP30, EBF, EDR1, EGR1, EGR2, EGR4, EVI2A, EVI2B, ENC1, ENTPD1, ENTPD2, EMR1, EMR3, EGLN1, EHD1, EHD2, ETFB, HSA277841, ELK4, EMK1, ELKS, ELL, EVC, ELL2, ELOVL4, EMD, KIAA0709, ENG, PODLX2, EROl-L(BETA), ENDOFIN, EDG4, EPAS1, EZFIT, EDN1, ET(B)R-LP-2, EN1, HEF1, ERH, EZH1, ENO3, ENO1B, EFNA2, EFNB1, LOC84648, EPS15R, EPM2A, EPIM, EMP2, EPLIN, DD96, EPHX1, EBI3, ERGL, EPB41L2, EPB72, LOC51145, EPOR, ECRG4, EBAG9, EBBP, ETV1, ETV5, EEF1B2, EIF1AY, EIF2S3, EIF3S10, EIF3S7, EIF4G3, EIF4B, EIF4EL3, EIF5, EIF5A, EIF5A2, ETF1, EXTL1, EXTL2, LAK-4P, KIAA0165, EMILIN-2, XLKD1, ECM2, EYA2, FGD1, FHR5, FANCE, FANCF, FNTA, FNTB, FADD, FABP5, FADS3, FAAH, FACL2, FACL3, FACL5, FOSB, FBXL11, FBXL3A, FBXL4, FBXL9, FBXW2, FBXW3, FBXO11, FBXO2, FBXO22, FBXO24, FBXO3, FBXO7, FBXO8, FBX30, FCAR, FCGBP, FCGR2B, FCGR3A, FE65L2, FES, FER, FTHL17, FTL, FALZ, HSRNAFEV, #25, FOP, FHOS, FBN1, FGL1, FGF11, FGF9, FGFR1, FGFRL1, FLRT1, FBLN1, FLNB, FLNC, M83, FRAP1, LOC51303, LOC51661, FKSG42, FKSG58, FKSG64, FKSG87, FMO1, FMO3, FMO5, LOC51167, FLJ00005, LOC51066, FLT3LG, FLT4, FOLR2, FSTL1, FSTL3, FOXC2, FOXFI, FOXG1A, FOXO1A, FOX03A, FPRL1, FS, FOSL2, FOSL1, FHL1, FHL2, FHIT, FRDA, FPGT, FUCA1, FUT5, FUT8, FUSIP2, FXYD1, FYN, FYB, FZR1, ZSIG37, #26, RDC1, GPR19, GPR31, GPR34, GPR35, GPR41, GPR50, GPR61, GPR64, GPR65, GPR68, GPR7, GPR84, GPR91, GPRC5C, GPRC5D, GSPT2, GTSE1, LOC51161, G5C, G6C, GABARAPL1, GABARAPL2, GAJ, GLA, GALR2, LOC85329, HSY11339, GGCX, GGH, GDAP1, GJA3, GJA8, GJB2, FGR, GRPR, GATA2, GATA6, GCN1L1, GCN2, GMDS, LOC51291, < #27, HSA250839, #28, RES4-25, GOA, GTF2A1, GTF2B, GTF2F1, GTF2F2, GTF3C1, GPHN, GGPS1, LOC51087, LOC64396, LOC51738, #29, GK003, GL002, GMFB, GBAS, GLTSCR1, GLP1R, GMEB2, #30, GPI, GLUT11, G6PC, G6PD, GAD1, GLUD1, GLUD2, GRIP1, GRIA2, GRIN1, GRIN2A, GRIN2C, GRM4, GRM6, GCLM, GLS, GLRX, GCDH, IE 0 60 312. .

GPX2, GPX3, GSTA2, GSTA4, GSTM1, GSTM2, GSTM5, GSTP1, GSTT1, GSTT2, GSS, GAPDS, GNPAT, GATM, GCAT, GNMT, GYS1, GYG2, GYPA, GYPE, GP3ST, GP1BB, GP9, GARS, GLO1, GPC1, GLG1, GOLGA1, GOLGA2, GOLGA4, GOLGB1, GOLPH2, GOLPH4, GOSR1, GOLTC1, KIAA0855, GLP, GNRH1, GNRHR, GNLY, GZMB, GAB2, GRO1, GRO2, GRO3, PLA2G13, GADD45B, GRB10, GFER, LOC84649, HUMGT198A, GTF21RD1, GCH1, GRAF, GEM, RAGB, GTT1, GNAL, GNAI1, GNAZ, GNB5, GNAQ, GNG10, KIM0277, GUCY1B2, GUCA1A, GUCA2B, #31, #32, H1F2, H1F4, HLX1, H2BFQ, H2BFR, H3FD, H3FK, H3FM, H3F3B, H326, H4FH, H4FM, HRY, HP, LOC51773, #33, HCR, #34, #35, HSP105B, HSPE1, HSPB7, HSPA1A, HSPA1B, HSPA6, HSPA1L, HSPCB, APG-1, HIP, KIAA0054, HELLS, KIM0928, HEMI, HHEX, HMOXl, HMOX2, HEBP, HBE1, HPX, HCK, HS3ST3B1, HS6ST, HSPG2, HPSE, XIP, LOC63928, LOC51339, HPS, HHLA2, HNRPAO, HNRPA2B1, HNRPF, HNRPM, HNRPU, HDLBP, HGRG8, HMG14, HMG2, HMGIY, HIRIP3, HIRIP5, HRH1, HRH2, HBOA, #36, HDAC7A, HRB, ΗΤΑΤΙΡ, BAT8, TCF-3, HNK-1ST, HESX1, HOXA1, HOXA11, HOXA6, HOXA7, HOXB5, HOXC10, #37, HERPUD1, RRS1, HOOK2, HOOK3, HCF-2, HP1-BP74, LOC51202, HRASLS, HSKM-B, HSPBP1, HSPCO18, HSPCO22, HSPCO25, HSPCO30, LOC51669, HSPCO39, LOC51125, LOC51122, HSPCO49, HSPCO55, HSPCO63, HSPCO71, HSPCO73, HSPC125, HSPC142, HSPC144, HSPC154, HSPC156, HT014, HTGN29, LOC58514, #38, HYPK, HMMR, HYAL3, HSD11B2, CG018, FLJ00060, LOC54557, LOC51235, HSPC228, HSPC192, #39, LOC56270, AF140225, BK1048E9.5, 13CDNA73, DJ1057B20.2, DJI 181N3.1, DJ465N24.2.1, DJ796117.1, FELL, DKFZP434F0272, DKFZp434G0522, DKFZP434K046, DKFZP434N1429, DKFZP434N185, DKFZp434Pl 115, DKFZP566G1424, DKFZP761L0424, DKFZp762B226, KIAA1630, DKFZp7620076, FLJ00001, FLJ10074, FLJ10081, FLJ10201, FLJ10357, FLJ10407, FLJ10420, FLJ10512, FLJ10656, FLJ10697, FLJ10707, FLJ10719, FLJ10826, FLJ10830, FLJ11113, FLJ11183, FLJ11560, FLJ12221, FLJ12572, FLJ12895, FLJ20033, FLJ20097, FLJ20203, FLJ20281, FLJ20297, FLJ20333, FLJ20689, FLJ20793, FLJ20886, FLJ20986, FLJ21877, FLJ22035, FLJ22263, FLJ22341, FLJ22376, FLJ22593, FLJ22955, FLJ23059, FLJ23119, FLJ23132, FLJ23316, FLJ23563, MGC10485, MGC14961, MGC20255, MGC20496, MGC4856, MGC5618, PRO1546, LOC63923, IE 060 3 12 156 H1F1A, HIG2, IDN3, IDS, LNIR, IK, IKKE, 1LVBL, IER3, HIVEP2, IGHM, ISLR, IGSF2, IPW, INHBA, INHBC, IBTK, ID1, ING1, ING3, IKBKB, IKBKAP, ITPR1, ITPR2, ITPKA, ITPKB, INPP4B, INPP5B, INSIG1, LOC55971, IGF1R, IGF2AS, IGFBP6, IGFBP4, INSM1,1TM1, ITGA1, ITGA2B, ITGA3, ITGA5, ITGA6, ITGB3, ITGB4, ITGB7, ITGB7, ICAM1, ICAM2, ICAM5, ICSBP1, IFNGR1, IFNGR2, IRF1, IRF2, IRF4, ISG20, IFNA17, IFNA6, IF127, IF130, IF141, IF141, IFRD1, IL1RN, IL1RL2, ILLA, IL1B, FIL1 (EPSILON), FIL1, IL10, IL10RA, ILI IRA, IL13RA1, IL15RA, IL17E, IL18BP, IL18R1, IL18RAP, IL2RA, IL2RB, #40, IL23A, IL6, IL7R, IL8, IL8RA, IL1HY2, #41, SYNCOILIN, ITSN1, IHABP4, INVS, IRX5, IDI1, ICMT, IVD, JDP1, JAG1, JAG2, SSI-1, JAK1, JM4, JUNB, JUND, KLK4, KLK6, KLK8, KLKB1, KAI1, KPNB2, KPNA3, KATNA1, KATNB1, #42, AB026190, KEL, KRT3, KRT6A, KAP4.10, KAP4.2, KRTHB1, KRTHB6, KIAA 1274, KIAA0007, KIAA0008, KIAA0009, KIAA0014, KIAA0020, KIAA0036, KIAA0074, KIAA0100, KIAA0101, KIAA0133, KIAA0135, KIAA0140, KIAA0143, KIAA0146, KIAA0166, KIAA0172, KIAA0185, KIAA0189, KIAA0191, KIAA0194, KIAA0196, KIAA0202, KIAA0211, KIAA0225, KIAA0229, KIAA0232, KIAA0240, KIAA0244, stabl, KIAA0247, KIAA0251, KIAA0256, KIAA0265, KIAA0268, KIAA0290, KIAA0295, KIAA0306, KIAA0321, KIAA0323, KIAA0328, KIAA0332, KIAA0335, KIAA0342, KIAA0346, KIAA0356, KIAA0365, KIAA0368, KIAA0370, KIAA0404, KIAA0408, KIAA0410, KIAA0417, KIAA0418, KIAA0419, KIAA0429, KIAA0430, KIAA0433, KIAA0467, KIAA0469, KIAA0470, KIAA0535, KIAA0552, KIAA0561, LOCI 14659, KIAA0586, KIAA0590, KIAA0595, KIAA0605, KIAA0615, KIAA0618, KIAA0635, KIAA0637, KIAA0645, KIAA0649, KIAA0663, KIAA0671, KIAA0680, KIAA0685, KIAA0710, KIAA0711, KIAA0713, KIAA0716, KIAA0726, KIAA0728, KIAA0737, KIAA0752, KIAA0769, KIAA0773, KIAA0775, KIAA0792, MAST205, KIAA0843, KIAA0860, KIAA0871, KIAA0872, KIAA0874, KIAA0893, KIAA0907, KIAA0913, KIAA0914, KIAA0922, KIAA0924, KIAA0939, KIAA0945, KIAA0957, KIAA0963, KIAA0964, KIAA0971, KIAA0982, KIAA0990, KIAA1001, KIAA1008, KIAA1009, KIAA1018, KIAA1023, KIAA1041, KIAA1042, KIAA1043, KIAA1049, RAP140, KIAA 1116, KIAA1128, KIAA 1143, KIAA1150, KIAA1171, KIAA1199, KIAA1203, KIAA1204, KIAA1224, KIAA1228, KIAA1234, ΙΕ υ 6 0 3 12 157 KIAA1235, KIAA1237, KIAA1240, KIAA1243, KIAA1244, KIAA1246, KIAA1253, KIAA1255, KIAA1266, KIAA1271, KIAA1278, KIAA1288, KIAA1292, KIAA1295, KIAA1298, KIAA1301, KIAA1303, KIAA1305, KIAA1306, KIAA1311, KIAA1320, KIAA1332, KIAA1337, KIAA1339, KIAA1340, KIAA1345, KIAA1350, KIAA1363, KIAA1376, KIAA1387, KIAA1388, KIAA1402, KIAA1409, KIAA1414, KIAA1424, KIAA1430, KIAA1441, KIAA1451, KIAA1457, KIAA1460, KIAA1464, KIAA1467, KIAA1483, KIAA1484, KIAA1485, KIAA1486, KIAA1509, KIAA1512, KIAA1513, KIAA1522, KIAA1524, KIAA1527, KIAA1528, KIAA1535, KIAA1538, KIAA1542, ΚΊΑΑ1547, KIAA1549, ΚΊΑΑ1553, KIAA1554, KIAA1557, KIAA1559, KIAA1563, KIAA1571, KIAA1587, KIAA1598, KIAA1599, KIAA1617, KIAA1638, KIAA164L KIAA1656, KIAA1666, KIAA1668, KIAA1673, KIAA1677, FLJ10898, KIAA1694, KIAA1698, KIAA1701, KIAA1705, KIAA1708, KIAA1710, KIAA1725, KIAA1726, KIAA1727, KIAA1728, KIAA1737, KIAA1741, KIAA1742, KIAA1758, KIAA1762, KIAA1771, KLAA1785, KIAA1789, KIAA1795, KIAA1798, KIAA1802, KIAA1811, KIAA1813, KIAA1814, KIAA1819, KIAA1821, KIAA1832, KIAA1842, KIAA1858, KIAA1862, KIAA1870, KIAA1887, KIAA1896, KIAA1899, K1AA1908, KIAA1917, KIAA1919, K.IAA1937, KIAA1938, KIR3DL1, KIR3DL2, KIR3DS1, KIR2DL4, KIR2DS5, KLRF1, KIF13A, KIF13B, KIF3C, KIF3A, KIF5B, KIFC3, KIF1B, KNSL5, CENPH, KITLG, KR18, SZF1, SERHL, KRML, LOC51045, KLF1, KLF4, ZK1, KUB3, KCNIP2, KYNU, KMO, HADHSC, LDHL, LMNA, LMNB1, LMNB2, LAMA2, LAMA5, LTBP1, LTBP3, LTBP4, LBP-9, LCHN, LOC51157, LGALS3, LGALS3BP, LGALS9, KIAA0821, HSOBRGRP, LRRFIP2, LZTFL1, LETM1, LRPPRC, LRRC3, RNO2, LZTR1, LIF, LILRA3, ILT10, LILRB2, LILRB4, IRC1, LENG1, LENG3, LAIR2, LIG3, LIG4, KIAA0175, FLJ23293, KIAA0203, SLY, SV2, E2-230K, DKFZP564M112, ARV1, LASPI, LDB2, LIMK2, LMO2, LMO4, LMO6, LHX2, LIM, LIN-7-C, LIN-7B, LIPA, LPIN2, LHFP, LHFPL2, LSR7, NUDE1, LIV-1, LOC88523, LDLR, #43, LDLB, LDLC, LRP3, QP-C, #44, LSM1, LUC7L, LFNG, LABH1, Tl A-2, LW-1, LOC51088, LNK, LY117, LY64, LC-P2, LBC, LRMP, LTB, LPAAT-delta, LYSAL1, LALP1, LAPTM5, LOXL3, LOC51300, HML2, MACMARCKS, MARCO, MST1, MADH4, MADH7, MAD2L1, MADP-1, MEF2A, MAGEF1, FLJ20798, MAGOH, LOC51678, HLA-DRB5, IE 0 6 0 3 12 158 MVP, K1AA0936, MPI, MPDU1, MAN2A2, MGAT1, MGAT3, MKP-7, MPN, MRG, MAML1, MMP1, MMP10, MMP11, MMP14, MMP15, MMP19, MMP2, MMP3, MMP7, MMP8, MNT, MAD, MAX, MBLR, MEFV, MRE1 IA, MEIS2, MEIS1, MC1R, MAGEA6, MAGED1, MAGEE1, MDA5, MS4A1, MS4A4A, MS4A6A, MS4A7, LOC51336, LOC51337, MSLN, CPX-1, MTF1, MSRA, MBD4, MTHFD2, MGST1, MAP4, MAPT, MAPRE2, MAPRE3, MIP-T3, MID2, LOC55972, LOC56993, MRP63, MRP64, MRPL1, MRPL12, MRPL13, MRPL15, MRPL16, MRPL17, MRPL32, MRPS24, MRPS25, MRPS33, MRPS36, TOPI MT, MTRF1, MAPK1, MAPK7, MAPK8, MAP2K4, MAP3K1, MAP3K13, MAP3K14, MAP3K2, MAP3K5, MAP3K6, MAP3K8, MAP3K9, MAP4K1, MAP4K2, MKP-L, MLN51, MRF-1, MAIL, MAOA, MAOB, MMD, MRG 15, MDM4, MPHOSPH9, #45, FLJ00029, #46, #47, #48, #49, #50, #51, #52, #53, #54, #55, #56, #57, #58, #59, #60, #61, #62, #63, MSTP031, MSTP043, MUC6, MCOLN1, MALT1, MADCAM1, MUL, MEN1, MINPP1, BMI1, MLH1, MAG, MPZL1, MNDA, MLL, MLLT1, MLLT6, MLLT7, MYOC, MB, MYO1E, MYO9B, MIR, MYO10, MYL5, MYL6, MLCB, MYBPC3, MYBPH, MTMR1, MYOZ, MACS, MX1, NAGA, HSA242910, NAT2, NDUFA4, NDUFA5, NDUFA7, NDUFA9, NDUFB1, NDUFB2, NDUFB3, NDUFB4, NDUFB6, NDUFS6, NDUFV3, NOX1, JFC1, #64, NPPA, CD244, BY55, NAP4, NDRG4, WWP2, LOC51667, NAF1, ΝΕΟΙ, NESCA, NESH, NAPG, LOC51162, LOC84687, NEDD4, NXPH2, NBL1, NAG, NEUD4, NF1, NEUROD1, NEUROD2, NEUROD4, NEUROD6, NGB, NRGN, NLGN2, NMU, NPAS1, NPAS2, NRP2, NTRK3, NSMAF, NCF2, NNT, NPC2, NBS1, NEK2, NEK3, NIN, NINJI, NIT2, KIR-023GB, NME7, NMNAT, NDRG3, NMT2, NOD2, NIMP, #65, NOT56L, NOTCH3, HSNOV1, DJ434014.5, NSPi, NSP3, SGSH, NTHL1, NTT5, GS2NA, SP140, NFE2, NFE2L2, NFAT5, NFATC3, NFKB1, NFKB2, NFKBIA, NFKBIE, NFKBIL1, P84, MDM1, NRBF2, NCOA1, NCOA4, AIB3, NR1D1, NR1D2, NR1H3, NR2C1, NR2F6, NR3C1, NR4A1, NR4A2, NR5A2, NRF1, NXF2, NXF3, NFYA, NFYB, NFX1, SC65, NOL3, NOLA2, NOLA3, NPM1, NSBP1, NUMBL, NRM, NYD-SP12, LOC51133, OA1, OR11A1, OR12D3, OR2C1, OR7E24P, OGT, OSM, OIP5, OPRL1, OPRM1, OPN1LW, OPN1SW, ORC2L, OAZ2, OAZIN, ORNT2, ORM1, OSRF, OSF-2, OSTF1, OCIA, OXR1, RODH, OLR1, OSBP, OSBPL2, FLJ10217, BITE, PAK6, PCAF, PIGPC1, p53DINPl, p53R2, IE 060 3 12 159 P53AIP1, PAI-RBP1, PACE, PAX1, PAX4, PILR(BETA), PMX1, PRX2, P1TX1, KIAA0992, PANX1, PAPA-1, PRCC, LOC55893, PARD6A, HUMPPA, PON2, PTMS, PORI, PVALB, PAXIP1L, PCQAP, PC3-96, PSIP1, PSIP2, PAF65A, PAF65B, PCTK3, PDLIM1, PDZK1, PDZ-GEF1, LOC51735, ΤΟΡΚ, PTX3, PMPCB, PYY2, PDI2, PAM, PPIL2, PPIF, PRF1, PCM1, PLIN, PRAX-1, PEX1 ΙΑ, PEX16, PXMP3, PEX1, PPARA, PPARG, PPARGC1, PET112L, PHF1, PMAIP1, MDS019, PCYT2, PPAP2A, PIK4CA, PICALM, PIGB, PIGC, PIGF, PIGH, PIP5K1B, PIP5K2B, PDE1B, PDE1C, PDE2A, PDE4B, PDE4D, PDE5A, PDE6A, PDE6C, PDE8A, PDE8B, PGAM1, PGAM2, PIK3CG, PIK3C2A, PIK3C2B, PIK3R1, P101-PI3K, PEPP3, PLA2G6, PLCE, PLCG1, PLD1, PLD2, PLSCR1, PLSCR3, LHPP, C8FW, #66, PSPH, PIM1, PIM2, PINX1, LOC96626, PIR, PTTG3, PL6, PGCP, PLAT, PLAU, PLGL, PLS1, PAFAH2, PDGFA, PDGFB, PDGFRA, PLEK, PSD, PLEKHA1, PSCD2, PSCDBP, PHLDA1, PHLDA3, PLEC1, PLAGL1, PLAGL2, PLXNB2, PLXNB3, PLXNC1, #67, #68, #69, #70, PVRL2, PAPOLA, PABPC1, PKD2L1, PQBP1, POLI, POLK, POLA, POLD3, POLL, POLM, POLQ, POLS, POLR2D, POLR2E, POLR2K, RPC39, POLR3K, #71, PMSCL2, POP4, POP2, POP3, MG61, PMS2, PMS2L3, CRIPT, HERG-3, KCNK15, KCNK3, KCNK5, KCNK9, KCNN2, KCNN3, KCNJI, KCNJI1, KCNJ15, KCNJ16, KCNJ5, KCNJ6, KCNK4, KCNMB3, KCNMB1, KCNS1, KCNQ1, KCNQ2, KCNQ3, KCNA10, KCNA3, KCNA5, KCND1, KCNH3, POU2F1, POU6F1, #72, PRDM13, PRDM2, PRDM8, SET07, PBEF, PRAME, PFDN2, OKL38, PRP17, PSEN1, PRIM1, PRO0233, PRO0365, PRO0641, PRO0644, PRO1073, #73, PRO 1900, PR02000, #74, #75, PCOLCE2, PLOD, PGRMC1, PDCD4, PDCD5, PDCD6, PDCD61P, PDCD7, PIP, B4-2, PML, PPBP, PCSK7, PTGDR, PTGER2, PTGER4, PTGES, PTGIR, PTGIS, PARTI, PSK, #76, LOC85414, PRM1, PRM2, PRSS11, PRSS16, SPUVE, PSMD4, PSMD9, PSMA3, PSMA5, PSMA6, PSMB10, AWP1, PCCX2, PDI, PIAS3, PKIG, PACSIN2, PRKCABP, PRKCG, PRKCQ, PRKCZ, PRKCL2, PKD2, NJMU-R1, NYD-SP15, PRKAA1, PRKACA, PRKAR2B, PRKWNK1, P3, LOC51207, PPP1CB, PPP1R12A, PPP1R14C, PPP1R15A, PPP1R2, PPP1R3B, PPM1G, PPP2R3, PPP2R5B, PR48, PPP3CA, PPP3CC, PPP4R1, PME-1, PPEF1, HSU79274, LOC84518, PROS1, PTK9, PTPN11, PTPN12, PTPN3, PPFIA1, PTPRG, PTPRO, PTPLA, LOC51184, PRKRIR, PCMT1, POMT1, AD013, PRG1, PRG2, LOC51685, U5-100K, PTD011, IE 0 6 0 J 12 160 PPFIBP2, #77, P2RX2, P2RX7, PURA, HM74, M96, #78, GPR, GPCR150, #79, SHI20, PHTF1, KIAA0436, M6A, MRS3/4, N6AMT1, HRIHFB2122, LOC51051, C3F, LOC51086, P2Y10, RABEX5, RNASE3L, FJX1, SIG11, LOC64172, HS1-2, P2RY6, PC, PDHB, PDK3, QDPR, RABL2B, RABI 1 B, RAB33A, RAB38, RAB31 Ll, RAB5B, RAB5C, GAPCENA, RAB6C, RAB7, RAB7L1, #80, RAB5EP, RAD23B, RAD51C, PIR51, RAD54L, RAD54B, RAD9, RSP3, RDX, RAE1, RALGPS1A, REPS2, LOC83859, RGL, RANBP1, RANBP6, EPAC, RAP1GDS1, RPIP8, RAMP, LOC54453, #81, ARHF, ARHH, RIN1, GAP1IP4BP, RASAL1, RASAL2, RASGRF1, RREB1, G3BP, RIS1, RAC3, RRP22, LOC51655, RBAK, REST, RLF, RAMP3, RIPK1, RIPK2, RIPK3, RBPSUHL, RCV1, RECQL, RECQL4, RGPR, RGS14, RGS16, RGS8, RFX4, RFX5, RFXAP, RSC1A1, RNTRE, RENBP, RFC3, RFC4, RFC5, RIP60, RPA3, RSN, REP, RFP2, RFPL2, RCN2, RTN2, RTN4, LOC51170, RP1, RP2, RPGR, RBBP1, RBBP4, RAI1, RAI15, RAI2, RARA, RARB, RARG, LOC51036, RXRG, RDH5, RDHL, RBP1, REV1L, REV3L, RECK, RGC32, RHAG, RTKN, RNASEHI, RNASEHI, RPP14, RPP38, RNASE1, RNASE4, RPN1, RPIA, RPL14, RPL19, RPL21, RPL23, RPL23A, RPL24, RPL26, RPL27, RPL4, RPL44, RPL8, RPS12, RPS14, RPS15, RPS21, RPS23, RPS25, RPS27A, RPS6KB1, RPS6KA1, RPS6KA3, RPS6KA4, RIT, RING1, RNF17, RNF2, RNF20, RNF23, RNF24, RNF25, RNF30, RYBP, LOC51285, RNMT, RPC, RBM10, RBM7, RBM8A, RBM9, RBMS1, RBMS2, LOC84549, RNAHP, RNUT1, RNU17D, RNPS1, RMP, RU2, RUNX1, RUNX2, RUVBL1, RUVBL2, RYR2, RYK, S100AI0, S100A12, S100A13, S100A3, S100A5, LOC57402, SAC2, AHCY, SAMHD1, SAMSN1, SARCOSIN, SBBI31, SAFB, SCHIP1, SOST, SEC10L1, SEC13L1, SEC22A, SEC22L1, SEC24A, FLJ10578, SEC.61G, SFRP5, SCAMP3, SEL1L, SEPN1, SEMA3B, SEMA3C, SEMA4B, SEMA4D, SEMA4F, SEMA4G, SEMA4C, SEMG2, SENP2, 37137, SQSTM1, SERPINB4, SERPINB8, SERPINDL SERPINE1, SERPINF2, SERPING1, SERPINH1, SR-A1, SDS, SHMT1, SPTLC2, SPINK5, SPINT1, SPINLW1, SRR, NDR, STK10, STK11, STK13, STK16, STK17A, STK17B, STK18, STK19, STK2, STK.6, MST4, MGC4809, SDCCAG10, SDCCAG28, SDCCAG3, SDCCAG31, PK428, SES2, SETDB1, SET, SIAH2, SCML2, SRY, HSSEXGENE, SRPK1, SH2D2A, SPAP1, SH3BGR, SH3BGRL2, SH3BP2, SH3GL2, SH3GLB2, SHB, HRIHFB2072, SHOX2, HEP27, SIT, SIGLEC10, S1GLEC11, 161 SIGLEC8, S1GLEC9, NEU2, SN, STHM, SIAT1, SIAT4B, SIAT4C, SIAT6, SIAT8D, SIAT8B, SIAT8A, SIAT8C, SIAT9, SRP19, SRPR, SSR1, SSR2, STAT2, STAT5A, STAT5B, STAT6, SIPA1, SLAM, SILV, #82, #83, LOC57168, KEO4, #84, #85, #86, MGC14386, #87, Rpol-2, #88, #89, LOC92797, #90, #91, #92, #93, #94, #95, #96, #97, #98, #99, LOC91151, #100, #101, #102, #103, #104, #105, #106, #107, #108, #109, #110, #110, #112, LOC90522, #113, #114, #115, #116, #117, #118, #119, #120, #121, #122, #123, #124, #125, HS1119D91, LOC57167, #126, #126A, UPF3A, UPF3B, #127, #128, SAP30, SIX2, SIM1, SSBP2, SIRT2, SIRT5, SSA1, SSB, XP5, #129, SMA3, HspB9, SCYA1, SCYA3, SCYA4, SCYA5, SCYA16, SCYA18, SCYA20, SCYA22, SCYA24, SCYA8, SCYB10, SCYB14, SCYB6, SMP1, SNRPA1, SNRPE, SNRPF, SNRPG, SOLH, SHARP, SMCX, SMOH, SNAI1, SNAPAP, SRCAP, SCNN1A, SCNN1G, SLC1A6, SLC12A4, SLC12A6, SLC12A5, SLC13A3, SLC16A5, SLC18A2, SLC19A1, SLC2A1, SLC2A3, SLC2A6, SLC20A1, SLC21A12, SLC21A9, SLC22A8, SLC22A1, SLC22A1LS, SLC22A5, SLC24A3, SLC25A20, SLC25A5, SLC25A6, SLC25A10, SLC26A2, SLC4A1AP, SLC28A3, SLC29A1, SLC29A2, SLC3A2, SLC3A1, SLC30A1, SLC30A4, SLC4A1AP, SLC4A7, SLC4A10, SLC5A3, SLC6A8, SLC6A1, SLC6A13, SLC6A9, SLC6A7, SLC6A6, SLC7A10, SLC7A11, SLC9A1, SLC9A5, SSTR5, SON, S0S1, SORL1, SNX12, SNX15, SNX16, SNX2, HSSOX6, SP2, SP3, SPOCK, SATB1, SSH3BP1, SPTAN1, SPTBN1, SPAG1, SPAG4, SPAG9, #130, STRBP, SPATA2, SAT, SKP2, SMPDI, SPHK2, SF3A1, SF3A2, SF3B1, SF3B2, SFRS10, SFRS3, SFRS5, SFRS6, SFRS7, SPON2, SPR1, SPRY1, SPRY2, SART-2, SLA, SOX13, SOX2, SOX22, SOX4, SSI-3, STATO, CIS4, STMN3, SLK, SCGF, SLU7, ZAK, SREBF1, SREBF2, STG, STOML1, STCH, STAG1, SDF1, STIM1, SNT-1, SDHC, SDHD, SUOX, SULT2B1, STE, SUSP1, SOD2, SVIL, ST5, ST7, SUFU, SKD1, SKD3, SURF5, SURF6, SMARCAL1, SMARCB1, SMARCD1, SMARCF1, SYNE-1B, SV2B, SYNGR4, SYTL2, SDC4, SS18, STX10, STX11, STX16, STX5A, STX7, STXBP1, STXBP2, SNTB1, TACTILE, TRA@, TRD@, TRG@, TAF9L, SIL, TBK1, TARBP1, TOM1L2, TAS2R13, TAS2R14, TAS2R7. TAF3C, TAF2A, TAF2C2, TAF2N, TAX1BP1, TBX21, TBX6, TALI, TIAM1, TCP10, TEKT3, TEX13A, TEX13B, FLJ20499, TSGA10, TSGA14, TSKS, TETRAN, TSPAN-1, TIAF1, TGIF, TGIF2, TH1L, ΤΡΜΤ, AOE372, TXNRD1, TR2, THBD, THBS3, TK2, 162 TRIP10, TRIP11, TRIP12, TRIP13, TRIP3, TRIP4. TRIP6, THRA, TRAP240, TRHR, TIA1, TIAL1, TIGA1, TJP2, TSTA3, TLH29, TRAF1, TRAF3, GG2-1, TLR1, TLR2, T0P2A, T0P3A, T0P3B, TRF4-2, TPARL, AD022, TANK, KIAA0057, ICBP90, TCF17, TCF19, TCF3, TCF6L1, TFAP4, TFE3, TFCP2, TFEC, NRF, TCFL1, TCFL4, TCFL5, TTF1, TTF2, CR0C4, ALY, TIF1, TReP-132, T0B1, TLE4, TF, TFRC, HSU53209, TGFB1I1, TAB1, TGFA, TGFB1, TGFBR3, TGM3, TGM5, TRPC6, TERE1, GC20, IF2, TIM17, TIMM8A, TIMM8B, KIAA0016, TOMM70A, TRAM, #134, TLOC1, TM4SF1, TM4SF5, TM7SF2, TACI, TMG4, TMEM1, TMEM2, TMEM5, TMEM7, TMPIT, TMEFF1, TAPI, LOC58486, #135, RAP1, THH, TREM1, TNRC11, TNRC12, TNRC4, TPI1, TRIM14, TRIM22, TRIM26, TRIM33, TRIM34, TRIM5, TRIM6, TPP2, TRIO, TFG, IPT, SECP43, TGT, TRO, TROAP, TMOD2, TMOD3, TPM1, TPM2, TPM4, LOC51149, WARS2, TSFM, TSPYL, TTK, TUFM, TULP3, TSC2, TUBA3, TUBB, TDRKH, PCTAIRE2BP, TUFT1, HCC8, TEM8, TNFSF13, TNFSF15, TNFSF7, TNFSF9, TNF, FIP2, TNFRSF10D, TNFRSF21, TNFRSF4, TNFRSF6, TNFRSF6B, TNFAIP1, TNFAIP2, TNFAIP3, TNFAIP6, TPD52L1, TP53BPI, DLM1, TSSC1, TSSC3, TUCAN, TSG, 14, PSK-1, TYRO3, TYRP1, YWHAE, YWHAH, YWHAG, TIE, U2AF65, HPRP8BP, LSM5, LOC51691, UQCRB, USP10, USP12, USP14, USP16, USP18, USP9X. UBE4A, UBE2A, UBE2C, UBE2E1, UBE2E3, UBE2H, UBE2L3, UBE2N, UBL1, UBL3, NEDD4L, B4GALT5, B3GNT5, B3GNT6, UGCGL2, UGDH, GALNT2, GNE, UAP1, ULBP3, BM036, MDS028, MDS030, MDS032, HARP11, HT007, HT008, VDUP1, UCC1, UBTF, UREB1, USF1, UNG2, UMPH1, UP, UCN, UPK2, HSHUR7SEQ, V1RL1, ABL1, ABL2, VPS26, VPS4, AKT2, VCP, VARS2, VANGL2, VR1, OTRPC4, VNN3, VCY2, VCAM1, VEGFC, VIP, VIPR1, CRK, CRKL, VAX2, ERBB3, VAMP1, VAMP4. ETS1, ETS2, FOS, SCAM-1, VMD2, VIT1, MYB, MYBL1, MYC, KIAA1329, BRAF, RALA, RALB, REL, RELA, RELB, SKI, YES1, WASF1, WDR10, WDR3, WDR4, WDF2, WDR11, WHIP, KIAA0105, WAS, WHSC2, WW45, WWOX, MDS009, XAGE-1, XBP1, XPR1, XPA, XPNPEP2, HSXQ28ORF, XRCC2, YAF2, ZF5128, ZFP, ZNF-U69274, ZFD25, ZNF131, ZNF146, ZNF151, ZNF16, ZNF165, ZNF185, ZNF187, ZNF19, ZNF193, ZNF195, ZNF200, ZNF205, ZNF213, ZNF22, ZNF221, ZNF230, ZNF236, ZNF237, ZNF238, ZNF257, ZNF258, ZNF26, ZNF264, ZNF268, ZNF278, ZNF297, ZNF302, ZNF313, 163 ZNF33B, ZNF36, ZNF44, ZNF45, ZNF79, ZNF83, ZNF85, ZNF9, ZNF91, ZFP161. ZNFN1A1, PEGASUS, ZFX, ZNRD1, ZHX1, ZYG and ZYX.

Homologs of these genes or proteins in other species, e.g., primates, are also modulated by the F3C. Modulation of these genes can be observed, e.g., as increased expression or mRNA levels or protein levels of the biomolecules in clinical conditions where insufficient or suboptimal expression or levels of the gene is associated with establishment, maintenance, severity or progression of the clinical condition to produce a desired clinical improvement.

Other therapeutic and biological applications and activities. The F3Cs are useful for preventing, slowing the progression of or treating certain chronic conditions in a subject such as a mammal or a human. Chronic conditions include diseases and conditions that arise or develop over a relatively long time period, e.g., over about 3 months to 10 years or more. Such conditions include chronic renal failure, which may result from polycystic kidney disease, from, e.g., an autoimmune condition such as acute or chronic glomerulonephritis, or from diabetes, interstitial nephritis, hypertension and other conditions discussed elsewhere herein. Chronic conditions include chronic pulmonary conditions such as chronic bronchitis, lung fibrosis, right ventricular hypertrophy, pulmonary hypertension, emphysema, asthma and chronic obstructive pulmonary disease, which may be treated with a F3C. These conditions or their symptoms may be mild, moderate or severe. The subject may be suffering from the disease or condition or may be subject to developing the condition, e.g., the subject may display early signs or a predisposition to develop a chronic condition. Such treatment will generally facilitate prevention of the disease, delay the onset or severity of the disease or condition, ameliorate one or more symptoms, e.g., reduce shortness of breath, coughing or dyspnea, or slow progression of the disease or condition. In these and other chronic conditions described herein, the F3Cs will generally be administered to a subject such as a human for a relatively long time period, e.g., for at least about 3 months to about 10 years or \ more. Dosages, routes of administration and dosing protocols for the F3Cs are essentially as described herein. Dosing of the compound can be daily or intermittent using a dosing protocol using dosages as described herein, e.g., about 0.1 to about 20 mg/kg of a F3C administered to a subject once or twice per day daily or intermittently. The use of the F3Cs IE 0 6 0 j j 2 164 can be combined with other treatments, e.g., .beta.-agonists such as metaproterenol or albuterol, or corticosteroids, e.g., prednisone, for asthma or chronic obstructive pulmonary disease.

The F3Cs can modulate the biological activity of cytokines or interleukins that are associated with various immune deficiency or dysregulation conditions, which may be transient or chronic. They can thus be used to ameliorate, treat or prevent naturally occurring age-related decline in immune function in a subject or immune deficiency or dysregulation resulting from trauma, stress, bums, surgery, autoimmunity or infections as described herein. Such immune deficiency dysregulation may be associated with, e.g., an age-related increase in production of one or more of IL4, IL-5 and IL-6 or an age-related decrease in production of one or more of IL-2, IL-3, .gamma.-IFN, GM-CSF or antibodies. In these embodiments, the F3C is administered to the subject to detectably decrease production or levels of one or more of IL4, IL-5 and IL-6 or to detectably increase production or levels of one or more of IL-2, IL-3, IL-5, IL-12, GM-CSF and .gamma.-IFN. These cytokine changes facilitate normalization of the subject's immune responses. Such nonnalization can be observed by various means. These means include monitoring appropriate cytokine RNA or protein level(s) in the subject or by measuring biological responses such as restoration or detectable improvement of contact hypersensitivity in a subject with depressed or suboptimal contact hypersensitivity response. The F3Cs can thus be used to enhance or restore a deficient or suboptimal immune response such as contact hypersensitivity response in a subject with a chronic or transient state of immune deficiency or dysregulation. In these embodiments, the F3C is administered using the dosages, routes of administration and dosing protocols for the F3Cs essentially as described herein. Treatment with the F3Cs is optionally combined with other appropriate treatments or therapies essentially as described herein, e.g., a antibacterial or antiviral agent(s) is coadministered with a F3C to treat, prevent or ameliorate an infection in an infected subject or a subject suffering from, e.g., a bum. Methods to measure changes in cytokine levels or contact hypersensitivity are known and can optionally be applied in these embodiments, see, e.g., U.S. Pat. Nos. 5,919,465, 5,837,269, 5,827,841, 5,478,566.

IE Ο 6 ο j 12 165 The capacity of the F3Cs to modulate immune functions permits their use for treating, preventing, slowing the progression of or alleviating the a symptom(s) of subjects with psychological disorders, metabolic disorders, chronic stress, sleep disorders, conditions associated with sexual senescence, aging, or premature aging. Metabolic disorders include parathyroidism, pseudoparathyroidism, hypoparathyroidism, hypercalcemia, hypocalcemia and detectable symptoms thereof such as fatigue, constipation, kidney stones and kidney malfunction. Chronic stress and related disorders include fibromyalgia, chronic fatigue syndrome, hypothalamic-pituitary axis dysregulation. Other related pathological conditions that can be treated with the F3Cs include hormone deficiency associated with aging or with a pathological condition, hypogonadism, vaginal atrophy, diminished libido, urinary incontinence, skin collagen loss, loss or impairment of skin, organ or joint connective tissue and menopause or its symptoms such as hot flashes, unwanted mood changes, fatigue and insomnia. In these embodiments, treatment of subjects witli a F3C is optionally combined with other suitable agents such as triiodothyronine, tetraiodothyronine, an insulin-like growth factor, insulin-like growth factor binding protein-3, an estrogen or a progestin.

As noted above, in some embodiments a treatment with a F3C. is combined with a corticosteroid or glucocorticoid. Corticosteroids are used in a number of clinical situations to, e.g., decrease the intensity or frequency of flares or episodes of inflammation or autoimmune reactions in conditions such as acute or chronic rheumatoid arthritis, acute or chronic osteoarthritis, ulcerative colitis, acute or chronic asthma, bronchial asthma, psoriasis, systemic lupus erythematosus, hepatitis, pulmonary fibrosis, type I diabetes, type II diabetes or bachexia. However, many corticosteroids have significant side effects or toxicities that can limit their use or efficacy. The F3Cs are useful to counteract such side effects or toxicities without negating all of the desired therapeutic capacity of the corticosteroid. This allows the continued use, or a modified dosage of the corticosteroid, e.g., an increased dosage, without an intensification of the side effects or toxicities or a decreased corticosteroid dosage. The side-effects or toxicities that can be treated, prevented, ameliorated or reduced include one or more of bone loss, reduced bone growth, enhanced IE 0 ββ 312 166 bone resorption, osteoporosis, immunosuppression, increased susceptibility to infection, mood or personality changes, depression, headache, vertigo, high blood pressure or hypertension, muscle weakness, fatigue, nausea, malaise, peptic ulcers, pancreatitis, thin or fragile skin, growth suppression in children or preadult subjects, thromboembolism, cataracts, and edema. Dosages, routes of administration and dosing protocols for the F3C would be essentially as described herein. An exemplary dose of F3C of about 0.5 to about 20 mg/kg/day is administered during the period during which a corticosteroid is administered and optionally over a period of about 1 week to about 6 months or more after dosing with the corticosteroid has ended. The corticosteroids are administered essentially using known dosages, routes of administration and dosing protocols, see, e.g., Physicians Desk Reference 54.sup.th edition, 2000, pages 323-2781, ISBN 1-56363-330-2, Medical Economics Co., Inc., Montvale, N.J. However, the dosage of the corticosteroid may optionally be adjusted, e.g., increased about 10% to about 300% above the normal dosage, without a corresponding increase in all of the side effects or toxicities associated with the corticosteroid. Such increases would be made incrementally over a sufficient time period and as appropriate for the subject's clinical condition, e.g., daily corticosteroid dose increases of about 10% to about 20% to a maximum of about 300% over about 2 weeks to about 1 year.

Such corticosteroids include hydrocortisone (cortisol), corticosterone, aldosterone, ACTH, triamcinolone and derivatives such as triamcinolone diacetate, triamcinolone hexacetonide, and triamcinolone acetonide, betamethasone and derivatives such as betamethasone dipropionate, betamethasone benzoate, betamethasone sodium phosphate, betamethasone acetate, and betamethasone valerate, flunisolide, prednisone, fluocinolone and derivatives such as fluocinolone acetonide, diflorasone and derivatives such as diflorasone diacetate, halcinonide, dexamethasone and derivatives such as dexamethasone dipropionate and dexamethasone valerate, desoximetasone (desoxymethasone), diflucortolone and derivatives such as diflucortolone valerate), fluclorolone acetonide, fluocinonide, fluocortolone, fluprednidene, flurandrenolide, clobetasol, clobetasone and derivatives such as clobetasone butyrate, alclometasone, flumethasone, and fluocortolone.

IE οΰ3ΐ8 167 In some applications, the F3C(s) may directly and/or indirectly interfere with replication, development or cell to cell transmission of a pathogen such as a virus or a parasite (malaria). Improvement in a subject's clinical condition may arise from a direct effect on an infectious agent or on a malignant cell. Interference with cellular replication can arise from inhibition of one or more enzymes that a parasite or an infected cell uses for nonnal replication or metabolism, e.g., glucose-6-phosphate dehydrogenase, which affects cellular generation of NADPH (see, e.g., Raineri et al., Biochemistry 1970 9: 2233-2243). This effect may contribute to cytostatic effects that some F3Cs can have. Modulation of cellular enzymes expression or activity may also interfere with replication or development of a pathogen, e.g., HIV or malaria parasites or with replication or development of neoplastic cells, e.g., inhibition of angiogenesis. Clinical improvement will also generally result from an enhanced immune response such as an improved Thl response.

Administration of a F3C can lead to a decrease in adenosine levels in a subject's tissue(s), e.g., lung or central nervous system tissue. This effect can be used to treat, prevent, ameliorate one or more symptoms of or slow the progression of a disease(s) or clinical condition(s) where a relatively high level of adenosine is a factor in or can contribute to the disease or condition, e.g., in asthma.

Adenosine is associated with the symptoms of bronchial asthma, where it can induce bronchoconstriction or contraction of airway smooth muscle in asthmatic subjects, see, e.g., J. Thome and K. Broadley, American Journal of Respiratory & Critical Care Medicine 149(2 pt. 1):392-399 1994, S. Ali et al., Agents & Actions 37:165-167 1992, Bjorck et al., American Review of Respiratory Disease 145:1087-1091 1992. This effect is not observed in non-asthmatic subjects. In the central nervous system, adenosine can inhibit the release of ., neurotransmitters such as acetylcholine, noradrenaline, dopamine, serotonin, glutamate, and GABA. It can also depress neurotransmission, reduce neuronal firing to induce spinal analgesia and it possesses anxiolytic properties, see, e.g., A. Pelleg and R. Porter, Pharmacotherapy 10:157 1990. In the heart, adenosine suppresses pacemaker activity, slows IE 0 ο ύ 3 12 168 AV conduction, possesses antiarrhythmic and arrhythmogenic effects, modulates autonomic control and triggers the synthesis and release of prostaglandins. In addition, adenosine has vasodilatory effects and can modulate vascular tone.

The unwanted effects of excess adenosine can be ameliorated or reduced by administering sufficient amounts of a F3C to a subject who is subject to developing or who has an unwanted level of adenosine in one or more tissues or organs. In typical embodiments, one will administer about 10 mg/kg/day to about 100 mg/kg/day of a F3C to a subject over a period of about 1 week to about 4 months to effect detectable changes in adenosine levels or amelioration in one or more symptoms associated with high adenosine in one or more of the subject's tissues. Such changes may be determined by comparing the subject's adenosine levels before treatment with the F3C is started. Alternatively, for subjects with symptoms that are consistent with high adenosine levels, the decrease can be inferred by comparing the normal level of adenosine in the target tissue(s) for subjects of the same species and similar age or sex with the level that is observed after treatment. Methods to measure adenosine levels in mammalian tissue are known and can optionally be used in these embodiments, e.g., U.S. Pat. No. 6,087,351.

In some clinical conditions, the F3Cs can inhibit activated T lymphocytes in vivo, and they can inhibit the expression or biological activity of one or more of TNF-.alpha., IFN21 .gamma., IL-6, IL-8 or insulin like growth factor-1 receptor (IGF-IR) or IL-6 receptor. The compounds are thus useful to treat, prevent or ameliorate conditions where this is a component of pathology. Such conditions include inflammation conditions such as psoriasis, psoriatic arthritis, osteoarthritis, and rheumatoid arthritis. The compound can thus ameliorate the inflammation, e.g., by inhibiting expression of one or more of TNF-.alpha., IFN26 .gamma., IL-6, IL-8 or IGF-1 R. Also, the compounds can inhibit unwanted T cell activity.

They can thus ameliorate one or more psoriasis symptoms such as skin scaling, skin thickening, keratinocyte hyperproliferation, deficient filaggrin expression (B. Baker et al., Br. J. Dermatol. 1984, 111 :702), deficient strateum comeum lipid deposition or they can improve a clinical assessment such as the Psoriasis Activity and Severity Index. The F3Cs IE 0 β ξ) j j £ 169 can be delivered to a subject with psoriasis using topical or systemic formulations as described herein. Topical formulations include gels, lotions and creams, e.g., as described herein. Daily or intermittent administration of the compound can be used essentially as described herein. The use of the F3Cs is optionally combined with one more current psoriasis treatments, e.g., topical emollients or moisturizers, tars, anthralins, systemic or topical corticosteroids, vitamin D analogs such as calcitriol, methotrexate, etretinate, acitretin, cyclosporin, FK 506, sulfasalazine, ultraviolet B radiation optionally combined with one or more of a topical corticosteroid, tar, anthralin, emollient or moisturizer or ultraviolet A plus psoralen. Such additional treatments essentially would use known dosages and routes of administration, which are applied, e.g., within a month before, during or within a month after a treatment course with a F3C.

Other desirable modulation effects of the F3Cs on cells or tissues include (1) inhibition of one or more of bone resorption or calcium release or gp80, gpl30, tumor necrosis factor (TNF), osteoclast differentiation factor (RANKL/ODF), RANKUODF receptor, IL-6 or IL-6 receptor expression or biological activity in, e.g., bone loss or osteoporosis conditions or in osteoclasts, or in cancers such as prostate cancer, metastatic breast cancer or metastatic lung cancer (e.g., with bone metastases), (2) inhibition of osteoclastogenesis or osteoclast development front progenitor cells, (3) enhancement of NF.kappa.B inhibition that is mediated by nuclear hormone receptors, e.g., enhanced inhibition of estrogen receptor.alpha. or estrogen receptor-.beta. mediated inhibition of NF.kappa.B in inflammation, rheumatoid arthritis or osteoporosis, (4) enhancement of osteoblastogenesis, osteoblast, bone callus or bone development, e.g., from progenitor cells in bone fractures, depressed bone healing situations (e.g., in a bum patient or in a patient being treated with a glucocorticoid), bone growth or osteoporosis or other bone loss conditions, by, e.g,, modulation or enhancement of osteoblast replication or development or modulation or enhancement of the \ synthesis or biological activity of a transcription factor such as Cbfal, RUNX2 or AML-3 (5) normalization of hypothalamic-pituitary-adrenal axis function in conditions where there is dysregulation such as in chronic inflammatory diseases, chronic asthma or rheumatoid arthritis (increased cortisol to ACTH ratio), (6) modulation of ligand-gated ion channels in ΙΕ ο 6 0 3 12 170 neurons in, e.g., depression, sleep or memory disorders, (8) modulation of G-protein coupled receptors in neurons in, e.g., depression, sleep or memory disorders, (9) modulation, e.g., induction or inhibition, of the synthesis or biological activity of metabolic enzymes such as a cytochrome or a hydroxylase (e.g., 1 l.beta. hydroxylase, a CYP enzyme such as CYP1A1, CYP2B1, CYP2blO, CYP4A, CYP7A, CYP7A1, CYP7B, CYP7B1, CYP11A1, CYP11B1, CYP17, P450 3A4, P450cl7, P450scc, P450c21 or an isozyme, homolog or mutant of any of these) in cells or tissues such as liver cells, neurons, neuron precursor cells, brain, breast, testes or colon, (10) enhancement of collagen synthesis or levels in, e.g., skin in aging or skin damage from, e.g., trauma, thermal injury or solar radiation, (11) inhibition of nitric oxide production in cells or tissue, e.g., in nervous system tissue or in microglial cells in dementias such as Alzheimer's disease, (12) enhancing glucose-stimulated insulin synthesis in hyperglycemia or diabetes conditions, (13) modulation of gamma-aminobutyric acid (GABA), dopamine or N-methyl-D-aspartate (NMDA) receptor activity or levels in, e.g., brain tissue or neurons, (e.g., decreased GABA-mediated chloride currents or potentiation of neuronal response to NMDA in the hippocampus) in, e.g., conditions such as a dementia (Alzheimer's Disease), depression, anxiety, schizophrenia or memory loss due to, e.g., aging or another condition described herein, (14) modulating (e.g., enhancing) the expression or activity of a transcription factor(s), or a homolog(s) or isofonn(s), such as SET, nerve growth factor inducible protein B, StF-IT, SF-1 in cells or tissues such as nerve cells, neuronal precursor cells or liver cells, (15 ) inhibition of eosinophil infiltration or reduction IgE levels in allergic responses or in lung or other tissue, (16) modulation, e.g., a decrease, in serum or blood of leptin levels in, e.g., obese subjects such as humans with a body mass index of about 27, 28, 29, 30, 31, 32, 33, 34 or greater, (17) increased corticotropin releasing hormone synthesis or activity in, e.g., elderly subjects such as humans at least about 60 years of age or at least about 70 years of age, (18) enhancement of memory or reduction of memory loss or disorientation in aging or dementias such as Alzheimer's Disease, (20) enhancement of the synthesis or activity of one or more enzymes responsible for thermogenesis, e.g., liver gIyceroI-3-phosphate dehydrogenase or malic enzyme, in subjects such as obese or diabetic humans, (21) modulation, e.g., reduction, of the synthesis or biological activity of the CXCR4 receptor or the CXCL1 2 chemokine in hyperproliferation IB θ 6 Ο 3 12 171 conditions such as breast cancers or precancers, (22) modulation of the synthesis or biological activity of one or more of holocytochrome c, cytochrome c, second mitochondria3 derived activator of caspase, Apaf-1, Bax, procaspase-9, caspase-9, procaspase-3, caspase-3, caspase-6 and caspase-7, e.g., enhanced translocation of these molecules from mitochondria to cytosol or activation of these molecules in the cytosol in cancer precancer cells, cancer cells or cells that mediate autoimmunity, (23) modulation of the synthesis or biological activity of one or more of tumor necrosis factor-, alpha., interleukin-1.beta, converting enzyme, IL-6, IL-8, caspase4 and caspase-5, e.g., decreased activation of these molecules in injured cells or cells subject to injury from, e.g., ischemia or infarction (e.g., vascular, cardiac or cerebral), reperfusion of hypoxic cells or tissue or an inflammation condition such as rheumatoid arthritis, ulcerative colitis, viral hepatitis, alcoholic hepatitis, or another inflammation condition disclosed herein, (24) decrease of the synthesis, biological activity or activation of one or more of phospholipase A2, caspase-1, caspase-3 and procaspase-3 in neurodegeneration disorders or dementias such as Alzheimer's disease, Huntington’s disease, or another neurological condition disclosed herein, (25) regulation or normalization of dysregulated protein kinase B, phosphatidylinositol 3-kinase and Forkhead transcription factors, e.g., a class O Forkhead transcription factor, in immune dysregulation or oxidative stress conditions such as immune suppresion, allergy or autoimmune conditions. The F3Cs can thus be used where one or more of these conditions or their symptoms is present.

Methods to measure the synthesis or biological activity of these molecules has been described, see, e.g., U.S. Pat. Nos. 6,200,969, 6,187,767, 6,174,901, 6,110,691, 6,083,735, 6,024,940, 5,919,465 and U.S. Pat. No. 5,891,924.

Specific embodiments. Aspects of the invention and related subject matter include the following specific embodiments. l.A method to prevent, treat, ameliorate or slow the progression of cystic fibrosis, blood cell deficency disorder, neutropenia or thrombocytpoenia in a subject, or to treat a symptom of the neutropenia or thrombocytpoenia, comprising administering to a subject, or delivering to the subject's tissues, an effective amount of a Formula 1 compound and/or a Formula 2 £ 060 3 12 172 compound and/or a Formula 3 compound, collectively within the text referenced as F3C or a metabolic precursor or a metabolite of any of Formula 1, and/or Formula 2 compounds alone or in combination with each other.

The method of embodiment 2, wherein one or more symptoms or syndromes are ameliorated, or wherein the progression of the disease is reduced.

The method of embodiment 2, wherein the one or more symptoms or syndromes are 1, 2, or more Haemophilus influenzae, Pseudomonas or Burkholderia respiratory tract or lung infection or propensity to develop a detectable infection or colonization, coughing, wheezing, cyanosis, bronchiolitis, bronchospasm, pneumothorax, hemoptysis, pancreatic exocrine insufficiency, bronchiectatic lung disease, atelectasis-consolidation, pulmonary edema, increased lung vascular hydrostatic pressure, increased lung vascular permeability, sinusitis, respiratory insufficiency, bronchial wall or interlobular septa thickening, reduction of forced expiratory volume in 1 second, dyspnea, impaired male fertility, elevated sweat chloride, mucous plugging, tree-in-bud sign, mosaic perfusion pattern, glucose intolerance or abnormal elevation of one or more of IL-4, IL-8, RANTES, neutrophil elastase, eosinophils, macrophages, neutrophils, eosinophil cationic protein or cysteinyl leukotrienes.

The method of embodiment 2,3 wherein the method further comprises another suitable treatment, which is optionally selected from oral or aerosol corticosteroid treatment, ibuprofen treatment, DNAse treatment, IL-10 treatment, diet control, vaccination against pathogens, or chest physical therapy.

. A method to prevent, treat or ameliorate neutropenia in a human comprising administering to the human an effective amount of a formula 1 compound of any of embodiments 1-4 wherein the neutropenia is postinfectious neutropenia, autoimmune neutropenia, chronic idiopathic neutropenia or a neutropenia resulting from or potentially resulting result from a cancer chemotherapy, chemotherapy for an autoimmune disease, an antiviral therapy, radiation exposure, tissue or solid organ allograft or xenograft rejection or ΙΕ °δ0312 173 immune suppression therapy in tissue or solid organ transplantation or aging or immunesenescence. 6. A method to prevent, treat or ameliorate neutropenia in a human comprising administering to the human an effective amount of a fonnula 2 compound of any of embodiments 1-4 wherein the neutropenia is postinfectious neutropenia, autoimmune neutropenia, chronic idiopathic neutropenia or a neutropenia resulting from or potentially resulting result from a cancer chemotherapy, chemotherapy for an autoimmune disease, an antiviral therapy, radiation exposure, tissue or solid organ allograft or xenograft rejection or immune suppression therapy in tissue or solid organ transplantation or aging or immunesenescence 7. A method to prevent, treat or ameliorate neutropenia in a human comprising administering to the human an effective amount of a formula 1 compound combined with a formula 2 compound ,of any of embodiments 1-4 wherein the neutropenia is postinfectious neutropenia, autoimmune neutropenia, chronic idiopathic neutropenia or a neutropenia resulting from or potentially resulting result from a cancer chemotherapy, chemotherapy for an autoimmune disease, an antiviral therapy, radiation exposure, tissue or solid organ allograft or xenograft rejection or immune suppression therapy in tissue or solid organ transplantation or aging or immunesenescence. The word combined shall mean at any time duration apart that is within 14 days. 8 . A composition comprising a compound of formula 1 as defined in any preceding embodiment and one, two, three, four, five or more excipients. 9.. A composition comprising a compound of formula 2 as defined in any preceding embodiment and one, two, three, four, five or more excipients.

. The method of embodiments 1 -9 wherein the level or activity of IgE in the subject is at least transiently detectably reduced, e.g., shortly after allergen exposure (such as within ΙΕ ο 6 Ο 3 12 174 about 1 hour to about 1 week) or at one or more later times. 11. A method to increase the efficacy of an immune response to dendritic cells in a subject, comprising (1) contacting for a sufficient time an effective amount of a Formula 1 and/or Formula 2 compounds alone or in combination with any of each other compound of any of embodiments 1-10 and an effective amount of a non-self antigen with the subject’s dendritic cells in vitro, (2) optionally expanding the dendritic cells in vitro in the presence or absence of the formula 1 compound and/or the antigen, (3) infusing the dendritic cells into the subject, (4) optionally administering an effective amount of the formula 1 compound to the subject and/or optionally administering an effective amount of the non-self antigen to the subject. 12. The method of embodiment 11 wherein the non-self antigen comprises an antigen derived or obtained from an infectious agent or from a malignant cell or a pre-malignant cell, wherein the malignant or pre-malignant cell is from the subject or is from another individual of the same species as the subject. 14. A method to increase the efficacy of allergy vaccinations in a subject having an allergy, comprising (1) at an effective time before or during vaccination of the subject with an allergen, administering to the subject an effective amount of a F3C compound of embodiment 1, (2) optionally administering to the subject an effective amount of the Formula 1 compound and/or a Formula 2 compound, daily or intermittently for 2, 3, 4, 5, 6, 7,14 or more days after the vacination of step (1), (3) after passage of sufficient time, repeating step (1) and (4) after passage of sufficient time, optionally repeating steps (1), (3 ) and/or (2).

. The method of embodiment 14 wherein the subject has a chronic allergy or atopic disease, optionally selected from allergic rhinitis, psoriasis, eczema, gastrointestinal allergies, atopic dermatitis conditions, allergic asthma, food allergies and hay fever and (1) wherein the level of IgE in the subject is at least transiently detectably reduced during or ΙΕ θ δ 0 3 f 2 175 after exposure to the allergen, or (2) wherein the total number of anti-allergic vaccinations that are needed to reduce allergy reactions or symptoms to allergen exposure is reduced or there is an increase in the quality or length of an effective response to allergen vaccination or there is an increase the proportion of subjects in which allergy vaccination is effective. 16. A method to prevent, treat or to reduce the severity of vascular or micro vascular occlusions in human sickle cell or thalassemia diseases, comprising administering to the patient an effective amount of a F3C composition of any of embodiments 1-14. 17. The method of embodiment 16 further comprising daily or intermittent administration oftheF3C composition. 18. The method of embodiment 16 or 17 further comprising monitoring the subject's blood cell status to determine if further administration of the F3C compound is desirable, and, if further treatment is desirable, administering to the patient an effective amount of the F3C compound. 19. The method of embodiment 16,17 or 18 wherein the frequency of observed vascular occlusion events in the subject or the severity of the subject's symptoms of vascular occlusions is at least transiently detectably reduced.

. A method to identify a compound that modulates the expression in a cell of the level of or an activity of 1, 2, 3,4, 5, 6, 7, 8, 9, 10 or more genes or gene products or gene transcripts in the cell, comprising contacting an effective amount of the compound with the cell under suitable conditions and for a sufficient time to detectably modulate the activity or level of the genes, or gene products in the cell, wherein the compound is a F3C composition. 21. The method of embodiment 20 wherein the genes or gene products are optionally selected from the group consisting of the genes or gene products disclosed herein. ΙΕ 312 176 22. The method of embodiment 20 or 21 wherein the genes or gene products are selected from the group consisting of USF1, c-Fos, EGR1, Cull, RIPK2,1.kappa.Ba, I.kappa.BKb, NF-.kappa.Bl p50, FCAR, c-Fos/C/EBP.beta., RANTES, ICAM1, TSG (TNFAIP6), IL-2 receptor .alpha., GRO2, GRO3, HOI, Jun B, c-Fos/JunB complex, JunB/ATF3 complex, cJun, c-Fos/c-Jun complex, ATF-3, MMP1, TSG-6 (TNFAIP3), EGR1, TGF.beta., ATF-3/cJun complex, c-Fos, MMP3, IL-8, STAT5A, STAT5B, CDKN1 A, IFN.gamma. receptor 2 (IFN.gamma.R2), T-bet, C reactive protein, immunoglobulin E, an AP-1 family protein, SOD2, GATA-3, Jak2, Tyk2, stall, stat3, stat4, stat5, stat6, MIP-la, MIP-2, IP-10, MCP-1, TNF-.alpha., TNF-.beta., IFN-.alpha., IFN-.beta., TGF-.beta.l, NF-.kappa.B, IL-1.alpha., IL-l.beta., IL-4, IL-6, IL-10, IL-12 receptor .beta.l, IL-12p35 IL-12p40, IL-23 and IL-23 receptor. 23. The method of embodiment 20,21 or 22 wherein the level or activity of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13,14,15,16,17, 18,19, 20, about 30, about 50, about 100, about 150, about 200, about 250, about 300, about 500, about 700 or about 1000 of the genes or gene products is detectably modulated at least transiently. 24.. The method of any of embodiments20,21,22, or 23 wherein the level or an activity of the gene product is a detectable increase in the level of the mRNA, the protein or one or more biological activities associated with the gene product.

. The method of embodiment 24 wherein the increase is a transient increase of about 1 hour to about 12 hours. 26. The method of embodiment 24 wherein the level or an activity of the gene product is a detectable decrease in the level of the mRNA, the protein or one or more biological activities associated with the gene product. 27. The method of embodiment 26 wherein the increase is a transient decrease of about 1 If 060 J12 177 hour to about 12 hours. 28. The method of any of embodiments 20-24 wherein the modulation is an increase or a decrease of at least about 2-fold to about 1000-fold in the level or an activity of the mRNA, the protein or at least one biological activity associated with the gene product. 29. A method to enhance the healing of a trauma or an acute injury in a subject who has experienced or who is expected to experience a trauma or an acute injury comprising administering an effective amount of a compound to the subject, wherein the compound is a F3C compound of any of embodiments 1-28.

. The method of embodiment 29 wherein the subject will experience or has experienced an immune suppressive event within about 2-3 weeks or about 34 weeks of the occurrence of the trauma or acute injury, wherein the immune suppressive event is exposure of the subject to an immune suppressive amount of ionizing radiation. 31. The method of embodiment 30 wherein the ionizing radiation exposure is about 0.2 Gy to about 30 Gy of the ionizing radiation. 32. The method of embodiment 30 wherein the radiation exposure is about 0.5 Gy to about 15 Gy of the ionizing radiation. 3.The method of embodiment 30 wherein the subject has experienced an immune suppressive event within 3 weeks of the occurrence of the trauma or acute injury, wherein the immune suppressive event is selected from an immune suppressive amount of an immunosuppressive chemotherapy. 34. The method of embodiment 33 wherein the immunosuppressive chemotherapy is an immunosuppressive cancer chemotherapy, an immunosuppressive amtimicrobial therapy or an immunosuppressive glucocorticoid therapy.

IE oea 12 178 . The method of embodiment 33 wherein the immunosuppressive cancer chemotherapy is treatment of the subject with an immunosuppressive amount of cyclophosphamide, 5fluorouracil or a platinum compound optionally selected from cisplatin and carboplatin. 36. The method of embodiment 33 wherein the immunosuppressive glucocorticoid chemotherapy is treatment of the subject with an immunosuppressive amount of dexamethasone, prednisone, hydrocortisone or cortisol. 37. The method of any of the embodiments wherein the subject is a human or a primate, 38. A method to modulate the expression of one or more transcription factors or enzymes in a subject who has a dysregulated oxidative stress condition comprising administering an effective amount of a compound to the subject, wherein the compound is F3C compound or a derivative or analogs of any of the components of an F3C composition that can convert to these compounds or analogs by hydrolysis or by metabolism. 39. Tlie method of embodiment 38 wherein the compound modulates the level or activity of a transcription factor or enzyme selected from one or more of Nrf2, a Maf protein, a thioredoxin, NQO1, GST, HO 1, SOD2, the catalytic subunit of .gamma.GCS, the regulatory subunit of .gamma.GCS and xCT. 40. The method of embodiment 38 or 39 wherein the dysregulated oxidative stress condition is associated with exposure of the subject to a toxin that can cause cell or tissue damage or wherein elevated oxidative stress is present in one or more of the subject's cell types or tissues. 41. The method of embodiment 38,39 or 40 wherein the subject has an acute or chronic inflammation condition, an acute or chronic infection or a trauma.

IE 0 6 0 3 1 2 179 42. The method of embodiment 38,39,40 or 41 wherein the levels or activity of one, two or more of Nrf2, a Maf protein, a thioredoxin, NQO1, GST, HO 1, the catalytic subunit of .gamma.GCS, the regulatory subunit of .gamma.GCS and xCT is increased. 43. The method of embodiment 38,39,40,41 or 42 wherein the compound is a F3C composition. 44. A product produced by the process of (1) contacting a F3C(s) and an excipient or (2) contacting a composition comprising a F3C(s) and one or more excipients with one or more additional excipients. 45. Use of a compound, composition, formulation or product of any of embodiments 1-44 or of any species or group of F3Cs disclosed herein for the preparation of a medicament.

The medicament can be for the prophylaxis or treatment of a disease or condition disclosed herein in a subject having or susceptible to developing the disease or condition. 46. The use of a compound, composition, formulation or product of any of embodiments 1 45 or of any species or group of F3Cs disclosed herein to prepare a medicament for use to prevent or to treat, or to ameliorate one or more symptoms associated, with one, two or more acute or chronic diseases or conditions disclosed herein, e.g., an infection, an immunesuppression condition, an allergy, a cardiovascular condition, a metabolic disorder, a pulmonary condition, a skin condition, aging, a trauma such as a bum or a bone fracture, immune suppression, a neurological or centeral or peripheral nervous system condition or disorder, an unwanted or pathological inflammatory condition, toxicity or unwanted sideeffects of a chemotherapy or of radiation exposure such as a glucocorticoid treatment or a cancer chemotherapy, an autoimmune disease or condition, a malignancy or cancer, a premalignant condition or to modulate a mammal's immune response, such as enhancing a Thl response or decreasing a Th2 response, in a subject, e.g., a human or a mammal, having the acute or chronic disease or condition or subject to developing the acute or chronic disease or condition. fe V6 0 3 12 180 47. Use according to embodiment 46 wherein the medicament is for intermittently administering the F3C(s) to a subject or delivering to the subject's tissues the F3C(s), e.g., using an intermittent dosing protocol disclosed herein. 48. Use of a compound, composition, formulation or product of any of embodiments 1-45 or of any species or group of F3Cs disclosed herein to prepare a medicament for use to enhance a specific or an innate humoral or cellular response to vaccination or to the presence of 1, 2, 3, 4, 5, 6 or more endogenous antigens or epitopes associated with establishing or maintaining a disease or pathogenic agent such as a tumor antigen or an antigen associated with a pathogen. 49. Use according to embodiment 49 wherein the subject's innate immunity is enhanced. 50. Use according to embodiments wherein the subject's innate or adaptive immunity is enhanced or wherein an unwanted immune response is decreased, or wherein number or activity of one, two or more of the subject's Thi cells, tumor-infiltrating lymphocytes (TIL cells), NK cells, peripheral blood lymphocytes, phagocytes, monocytes, macrophage, neutrophils, eosinophils, dendritic cells, fibrocytes, astrocytes, gilal cells or stromal cells, e.g,, bone marrow, lymph node or spleen stroma, are increased or activated at least transiently (e.g., for at least 10 minutes to 10 days or more), which is optionally as measured by, e.g., enhanced ,sup.3H-thymidine uptake compared to untreated controls or by an increase in the number of the cell type in circulation or demonstrable movement of the cell type from one tissue or compartment (e.g., skin or blood) to another tissue or compartment (e.g., blood, lymph node, spleen, Peyer's patches, GALT or thymus), or wherein the transcription rate, protein level or biological activity of one or more genes in the subject's NK cells, TIL cells, phagocytes, monocytes, macrophages, neutrophils, eosinophils, dendritic cells, fibrocytes, astrocytes, gilal cells or stromal cells are detectably modulated, e g., increased (e.g., as measured by increased enzyme or biological activity of a ‘'evj 12 181 biomolecule such as a nuclear hormone receptor such as an orphan nuclear hormone receptor, a transcription factor, a chemokine or a cytokine, which is optionally compared to suitable control cells or tissues). 51. A method to (a) modulate (detectably increase or decrease) the expression of at least one immune cell antigen by an immune cell in a subject, whereid the immune cell antigen is selected from CD3, CDllc, CD14, CD16, CD19, CD25, CD38, CD56, CD62L, CD69, CD45RA, CD45RO, CD123, HLA-DR, IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, TNF.alpha., IGF.sub. 1 and .gamma.IFN, or (b) activate CD8.sup.+ T cells or CD8.sup.- T cells in a subject, wherein the activation comprises at least transiently enhanced expression of CD25 or CD69 by the T cells, or (c) increase the proportion of CD8.sup.+ or CD8.sup.lymphokine activated killer cells in a subject's CD16+ cells (e.g., CD8.sup.+, CD16.sup.+, CD38.sup.+ or cells CD8.sup.-, CD16.sup.+, CD38.sup.+), or (d) increase the proportion of (i) CD8.sup.-, CD16.sup.+ natural killer cells, (ii) CD8.sup.+, CD16.sup.+ natural killer cells or (iii) CD8.sup.-, CD16.sup.+ cells that mediate antibody-dependent cell-mediated cytotoxicity, or (iv) CD8.sup.+, CD16.sup.+ cells that mediate antibody-dependent cellmediated cytotoxicity, or (e) increase the proportion of dendritic cell precursors in a subject's circulating white blood cells (e.g., Lin.sup.-, HLA-DR.sup.+, CD123.sup.+ or Lin.sup.- HLA-DR.sup.+, CD1 lc.sup.+ cells) or (f) increase the proportion of CD45RA.sup.+ T cells or CD45.sup.+, R0.sup.+ T cells in a subject's circulating white blood cells, or (g) change (increase or decrease) the proportion or relative numbers of CD62L.sup.+ T cells in a subject's circulating white blood cells, or (h) increase the proportion of CD8.sup.+ or CD4.sup.+ T cells that express CD62L in a subject's circulating CD8.sup.+ or CD4.sup.+ T cells, or (i) decrease the proportion of CD8.sup.+ or CD4.sup.+ T cells that express CD62L in a subject's circulating CD8.sup.+ or CD4.sup.+ T cells, or (j ) increase the proportion of HLA-DR.sup.+, CD8.sup.+, CD38.sup.+ cells in a subject's \ circulating white blood cells, or (k) decrease the level of IL4 or IL-10 that is expressed by or present in a subject’s white blood cells or in a subject's plasma (or that is expressed after the subject's white cells are stimulated in vitro), (1) at least transiently increase the number of dendritic cell precursors or dendritic cells that are present in a subject's white blood cells or 182 in a subject's plasma, or (m) enhance the capacity of an immune cell, e.g., macrophages, CD4.sup.+ T cells, CD8.sup.+ T cells to express IL-2, IL-12 or .gamma.IFN or to activate such cells, the method comprising administering to the subject an effective amount of a F3C, which is optionally present in a composition or a formulation comprising 1, 2, 3, 4, 5, 6 or more phanuaceutically acceptable excipients. The F3Cs is any compound as described herein, e.g., a compound of embodiments 1-40 or in any chemical structure or any compound group. 52. A method to detect or to characterize a biological response (e.g., increased or decreased cytokine or cell surface antigen expression or activity, increased numbers of circulating neutrophils, increased phagocytic activity by phagocytic cells or modulation of a disease or symptom described herein) associated with the administration of a F3C to a subject comprising (1) obtaining a first biological sample from the subject and analyzing the sample to obtain a baseline value for the response, (2) administering the F3C to the subject to obtain a treated subject (3) within about 15 minutes to about 28 days after administering the F3C, obtaining a second biological sample from the treated subject and analyzing the sample to obtain a treated value for the response, and (4) optionally comparing the control information with the experimental information to detect the presence, absence, relative magnitude or absolute magnitude of the biological response. 53, The method of embodiment 137 wherein the biological response is modulation of CD8.sup.+ T cells, CD4.sup.+ T cells, CD8.sup.+ lymphokine activated killer cells, CD8.sup.-, CD16.sup.+ natural killer cells, circulating dendritic cell precursors, circulating dendritic cells, tissue dendritic cell precursors, tissue dendritic cells, CD8.sup.+ lymphokine activated killer cells, CD8.sup.- lymphokine activated killer cells, CD8.sup.-, CD16.sup.+ natural killer cells, CD8.sup.+, CD16.sup.+ natural killer cells, CD8.sup.-, CD16.sup.+ cells -< that mediate antibody-dependent cell-mediated cytotoxicity, CD8.sup.+, CD16.sup.+ cells that mediate antibody-dependent cell-mediated cytotoxicity, CD45RA.sup.+ T cells, CD45RA.sup.+, CD45RO.sup.+ T cells, CD45RO.sup.+ T cells, CD8.sup.+, CD62L T cells, C.D4.sup.+, CD62L.sup.+ T cells or HLA-DR.sup.+, CD8.sup.+, CD38.sup.+ T cells, l£ υ δ ν J 12 183 monocytes, macrophages, glial cells or astrocytes, or wherein the biological response is at least transient modulation of an immune cell antigen or an immune accessory cell antigen (e.g., an adhesion molecule at the surface of endothelial cells or a cytokine receptor at the surface of T cells or B cells or a CD molecule, an interleukin or a cytokine, optionally selected from CD16, CD25, CD38, CD62L, CD69, CD45RA, CD45RO, IL-1, IL-2, IL4, IL6, IL-8, IL-10, TNF.alpha., IGF.sub.l and .gamma.IFN). 54. A kit comprising a formulation that comprises a unit dosage or a multiple dosage comprising a F3C, e.g., any compound described or within any structure disclosed herein, and one or more excipients wherein the formulation is dispensed in a suitable closed container, wherein the kit optionally further comprises a label that provides information about one or more of (1) the F3C's chemical structure, (2) any recommended dosing regimen, (3) any adverse effects of administering the F3C to a subject that are required to be disclosed and (4) the amount of the F3C that is present in each unit dose or in the entire container. 55. A method to treat a symptom or condition associated with one or more delayed adverse or unwanted effects -of radiation exposure or therapy in a subject in need thereof comprising administering to the subject, or delivering to the subject's tissues, an effective amount of a compound of formula 1, wherein the F3C is administered or delivered to the subject's tissues beginning at least 2 weeks after the subject has been exposed to a dose of radiation that will cause or could potentially cause the one or more delayed adverse or unwanted effects of the radiation exposure, or wherein the F3C is administered or delivered to the subject's tissues beginning at least 2 weeks after the subject has been exposed to at least one subdose of a planned course of radiation exposures that will cause or could potentially cause the one or more delayed adverse effects or unwanted effects of the radiation exposure. 56. The method of embodiment 55 wherein the subject has received, or is anticipated to receive, a total radiation dose of at least about 0.25 Gy to about 300 Gy, wherein the subject received the radiation dose in a single dose or in two or more divided doses, e.g., a total ϋ b U 3 12 184 radiation dose of at least about Gy 1 to about 15Gy, or at least about Gy 0.5 to about 12 Gy. 57. The method of embodiment 55 or 56 wherein the symptom or condition associated with one or more delayed adverse effect of radiation is one or more of encephalopathy, myelopathy, nausea, vomiting, diarrhea, acute inflammation, e.g., of the lung, chronic inflammation, e.g., of the lung, edema, pain, headache, depression, fever, malaise, weakness, hair loss, skin atrophy, skin ulceration, skin lesion, keratosis, telangiectasia, infection, hypoplasia, atrophy, fibrosis, pneumonitis, bone marrow hypoplasia, hemorrhage, leukopenia or thrombocytopenia. 58. The method of embodiment 55, 56 or 57 wherein the symptom or condition associated with one or more delayed adverse or unwanted effect of the radiation exposure or therapy is caused by or associated with radiation damage to one or more of bone marrow cells, bowel epithelium, bone marrow, testicles, ovaries, brain nerves or tissue, peripheral nerves, spinal cord nerves or tissue or skin epithelium.

Variations and modifications of these embodiments, the claims and the remaining portions of this disclosure will be apparent to the skilled artisan after a reading thereof. Such variations and modifications are within the scope and spirit of this invention. All citations herein are incorporated herein by reference in their entirety. All citations herein are incorporated herein by reference with specificity.

EXAMPLES The following examples further illustrate the invention and they are not intended to limit it in any way. Variations of these examples that are included in the invention may include, \ e.g., any of the F3Cs described herein or parts or all of any of the methods, formulations, treatment protocols and/or assays described herein.

Example 1 185 Chemicals Riboflavin, Nitroblue tetrazolium (NBT), reduced glutathione (GSH), S-S' dithiobis (24 nitrobenzoic acid) (DTNB), and 1- chloro-2, 4-dinitrobenzene (CDNB) were obtained from Sisco Research Laboratories Pvt. Ltd, Mumbai, India. Thiobarbituric acid was purchased from Hi-media laboratories, Mumbai, India, 1,1,3,3-tetramethoxy propane was supplied from Sigma-Aldrich USA. All other chemicals and reagents used in this study were of analytical grade. Oltipraz, a Formula 2 compound, was supplied by the contractor Canopus Corp.

Inbred 4-6 weeks old female Swiss albino mice (20-30g) were obtained from Small animal breeding station, Kerala Agricultural University, Mannuthy, Thrissur. Animals were kept in well-ventilated cages under standard conditions of temperature, pressure and humidity. The animals were provided with normal mouse chow and water ad libitum. All animal experiments conducted during the present study got prior permission and followed the guidelines of Institutional Animal Ethics Committee (IAEC).

Irradiation Animals were treated with a single dose of radiation of 700 rads (7Gy). The source of radiation was a 6°Co Theratron-Phoenix teletherapy unit (Atomic Energy Ltd, Canada).

Animals were restrained in specially designed, well-ventilated cages without anesthesia and exposed to whole body radiation at a rate of 1.33 Gy/min in a field size of 25 x 25 cm2 and at a distance of 80 \ cm from the source.

Determination of effect of Oltipraz on haematological parameters of irradiated animals 186 Twenty-four mice were randomly divided into three groups of 8 animals each. Group I was treated as irradiated control served with vehicle. Group II was treated with Oltipraz (250mgs/Kg b wt) ten days prior to irradiation, Group III was administered Oltipraz (250mgs/Kg b. wt) ten days prior to irradiation and continued for another fifteen days after irradiation. All the three groups were irradiated with a single dose of 750 rads.

Body weights of all the animals were determined one day prior to irradiation and every third day thereafter. Blood was collected from tail vein into heparinized tubes and the following parameters were analyzed one day before radiation and every third day thereafter. The parameters analyzed were total WBC count (haemocytometer method), differential count (Leishman's staining method) and haemoglobin by Drabkin's method.) Determination of effect of Oltipraz on bone marrow viability and antioxidant parameters of irradiated animals.

Thirty six animals were divided into four groups of nine animals each. For groupl to 4 treatment protocol were similar as described above. Group 4 was treated as normal animals without any treatment. On days 5,10 and 15 after irradiation (750) rads three animals from each group was sacrificed. Blood was collected to heparinized tubes, and plasma was removed and following parameters were assayed in the blood. Activity of the enzyme SOD was measured by NBT reduction method of McCord and Fridovich. CAT activity was estimated by the method of Aebi by measuring the rate of decomposition of hydrogen peroxide at 240nm. Level of GSH was assayed by the method of Moron et al based on the reaction with DTBN. Assay of GPX followed the method of Hafeman based on the degradation of H2O2 in the presence of GSH. The method of Habig was followed to assay the activity of GST based on the rate of increase in the conjugate formation between GSH and CDNB.

The femurs of three above animals were dissected out and bone marrow cells were flushed into phosphate buffered saline (ph7.4) containing 2% fetal calf serum. The cells /£ veojf2 187 were washed and bone marrow viability was determined by the method of Sredni. The results were expressed as number of live bone marrow cells xl0(6)/femur.

The liver of the sacrificed animals were excised quickly washed in ice-cold saline and kept at -70°C till the day of analysis. On the day of analysis 25% homogenate was pre-pared in tris-HCl buffer (0.1 M, pH7.4). The homogenate was centrifuged at 12000 rpm for 30 minutes and supernatant was used to determine the tissue lipid peroxide levels(LPO) using the TBA method of Okhawa et al.

Data Analysis.

Data was expressed as mean ± standard deviation (SD). Significance levels for comparison of differences were deter-mined using Student's t test. The mean of Oltipraz treated group was compared with that of radiation alone treated group. The radiation alone treated group was then further compared with untreated group. The differences between means were considered to be statistically significant if p <_ 0.001.

Radiation treatment at the dose level used here did not produce a statistically significant reduction in the body-weight of the exposed animals. Initial body weight of animals were 26.27 ± 3.76, 23.92 ± 4.61 and 24.98 ± 3.76 respectively for group I, II and III. On day 6 body weight was reduced to 21.82 ± 3.34, 21.02 ± 4.14 and 21.10 ± 2.51 respectively (p > 0.05). Radiation significantly lowered the total leukocyte count in irradiated animals.

Administration of Oltipraz was found to increase the count. In the initial days after irradiation both group II (Oltipraz pre treated group) and group ΙΠ (Oltipraz continuously administered group) showed almost similar number of WBC. But at later days after irradiation, group III showed a significantly elevated WBC as compared with group I (radiation alone treated group) and group II. This indicated that continuous Oltipraz administration stimulated the hemopoetic system in a concentration dependent manner.

This observation is further supported by the increased bone marrow viability found in-group III (Fig. 1 B). Bone marrow viability in normal animals was (Group IV) was 16.21 ± 0.45 x 188 ΙΟ6 cells/femur. Bone marrow viability was significantly decreased after irradiation. After the 15th day of post- irradiation group II possessed a value of 6.1 x 10fe cells/ femur where as group II and III showed 5.82 x 106 and 14.32 x 106 cells/ femur respectively. The hemoglobin levels were significantly reduced after irradiation. On day 6 radiation alone treated group had a hemoglobin level of 10.37 ± 3.19 where as Oltipraz continuously administered group had a value of 12.42 ± .2.76. The differential count did not show any significant variation.

The activity of both SOD and CAT, two of the major enzymes involved in the antioxidant defense mechanism were found to be decreased after irradiation The continuous administration of Oltipraz enhanced SOD activity, which showed the maximum value on the 15lh day after irradiation and CAT on tenth day after irradiation.

Activity of GPX was also found to be decreased after whole body irradiation. Continuous administration of Oltipraz elevated the activity of GPX. On the 15th day after irradiation group I had an activity of 1254.00 ± 116.23 U/L of haemolysate whereas group III showed an activity of 1927.87 ± 136.06 demonstrating that Oltipraz administration stimulated GPX activity (p<_ 0.001). The levels of the major cellular antioxidant GSH increased after Oltipraz administration. The levels of GSH were brought down after irradiation. On the 15th day the levels of GSH increased almost three times in Oltipraz continuously administered group as compared wit radiation alone treated group indicating that Oltipraz administration elevated the GSH levels (p < 0.001), It could be presumed that an increased level of antioxidant enzymes and GSH is a direct consequence of Oltipraz administration and could be seen in un-irradiated animals as well. Oltipraz administration also elevated the activity of GST, an enzyme involved in the glutathione mediated detoxification system. On the 15th day group III showed an activity of 2.38 ± 0.25 (nano-moles of CDNB-GSH conjugate formed) (p <_ 0.01) where as group I had an activity of 1.51 ± 0.37 only. Radiation increased the levels of lipid peroxidation in all the radiation treated animals. On day 15, it was 3.47± 0.31 (nano-moles ofMDA formed/mg of protein for group I, whereas in-group III it was significantly reduced to a level of 2.12 ± 0.21 (p <_ 0.001).

II 189 Example 2 CASE STUDY OF PATIENT T.W.

BACKGROUND-The patient was treated with a Formula 1 compound and his blood cell parameters were monitored.

T.W. an immune comprised patient with multiple recurrent infectious bouts requiring hospitalization while in a nearly moribund condition began treatment with the experimental drug IPA. Mr. T.W. had been hospitalized with avert life-threatening infections. Mr. T.W.'s total WBC counts were in the range of 2200-4900 for the three month period preceding hospitalization. A T4/T8 cell ratio of 50/270 (0.22) was observed two months earlier. Patient was hospitalized from depression, exhaustion, bronchial infection, severe diarrhoea, severe weight loss and complete loss of appetite and spiked fever. Weight was 98 pounds.

The patient, although initially was very co-operative relative to receiving experimental IPA therapy, for no rational reasons at various times stopped IPA.

IPA THERAPY - IMMUNOLOGY STUDIES --------------------------------22 Patient initially received IPA (see Charts 1 and 2). T and B lymphocyte competence as measured by the ability of these cells to proliferate in response to specific stimulating mitogens (PHA for T cells and PWM for B cells)which was monitored on a weekly basis until death. As shown in Charts 1 and 2 the patient's T and B lymphocyte competence on was well below normal. Indeed, the competence of both these immune cell populations was less than the 1st percentile of normal donors; that is, greater than 99% of all normal donors historically tested (> 1460 normal donors in our laboratory) had responses greater than the patient demonstrated. 190 A major rapid recovery of both T and B lymphocyte competence was experienced within less than 7 days after initiation of IPA therapy. The levels of competence within 7 days of drug therapy initiation were above the lower limits'of the normal donors reactivity. This represented a remarkable recovery since patients receiving. Most immune stimulating agents (e.g., BCG, Levamisole) in our laboratory's experience exhibit a slower and less dramatic immune recovery capacity. The patient conscientiously continued to take constant oral doses of IPA.

The patient by his words he felt cured and abruptly stopped taking the drug. No one except the patient and his friend were aware of this unilateral decision until several weeks later. He was in great spirits, Fever and diarrhoea had completely subsided and the patient exhibited a 15 pound weight gain during this time.

As shown on Charts 1 and 2, at the time that the patient proclaimed self-cure and thus stopped all IPA medication, his T and B lymphocyte competence again dropped to less than 1% or normal donors' competence. This was most dramatically observed with T lymphocyte function but was also observed in somewhat lagged fashion with B lymphocyte function. Total WBC counts were in the range of 1650 to 2550 during the period. His peripheral blood showed 50 T4 cells and 220 T8 cells (0.23 ratio).

The patient stopped IPA therapy on a few days. During this time both T and B lymphocyte competence rapidly bottomed out, the patient was hospitalized in for a 7 -10 day period for severe bronchial infections and fever, diarrhoea, anxiety and weight loss.

Mr. T.W. resumed IPA therapy at recommended dose levels. Minor T lymphocyte competence returned during the following week. ; B lymphocyte competence continued to decrease. Within two weeks T lymphocyte competence was again nearly within the normal range. Although B lymphocyte competence was still somewhat depressed, this cell's competence was also improving. Within a month T lymphocyte competence had risen to IE 191 well within the range of normal donor reactivity, while B lymphocyte competence was somewhat below lower normal limits. The patient's personal status demonstrated a dramatic improvement; he was periodically dining out with a large appetite, diarrhoea had lessened, fever had subsided, he was riding his bicycle daily and going dancing at night.He again stopped taking medication for a week without anyone's knowledge until later. T lymphocyte competence again demonstrated a major sudden decrease to less than 1% of normal donors' activity. The patient again exhibited spiked fever, acute diarrhoea and exhaustion. He again resumed IPA therapy the following week, for one week, with one9 half the recommended dosage taken. His T lymphocyte competence spiked dramatically to above the lower limit of the normal range. B lymphocyte actively demonstrated a slight rise. During the next week the dosage of IPA was greatly (> 3X) increased. T lymphocyte competence demonstrated depression suggesting immunotoxicity by IPA; since a 33% decreased dosage the following week resulted in T lymphocyte competence increasing once again.

The patient stopped therapy during the next month , both T and B lymphocyte function fell to nil, as a direct consequence.. The patient died approximately two weeks later suffering from acute bronchial infection accompanied with high fever, diarrhoea, exhaustion and weight loss.

Conclusions 1. Direct correlations were observed during the period of administration of IPA with quality of life and depressed or elevated T and B lymphocyte function. 2. Enhanced T and B lymphocyte competence correlated well with continued IPA medication; optimum levels of competence appeared to be experienced at optimum IPA dose levels administered. 3. Since competent T and B lymphocyte function is required for combating microbial infections by the host, major infectious bouts correlated with times when competence was most severely compromised.

It !92 4. The patient experienced 5 months of life with periods of excellent quality after initiation of IPA therapy at a time when he was terminally moribund.

. It would appear that the patient would have enjoyed appreciable therapeutic benefits from IPA, if he had conscientiously and continuously received IPA medication. If optimum dose levels had been more clearly established this would have presumably optimized quality and extension of life to a greater extend than was actually observed in this patient.

Study Director: Signed James L. McCoy, Ph.D. Immunologist, ImmuQuest Laboratories, Inc. ιε Oo 193 The present invention employs one or more compound according to Formula I: -.5 Fonnula I wherein: R, = H, Ej = CH3, R3 = CH3 and R4 = H, or R1 = H or CH3S and R4 - CH2 and Rg = CH^ cl, OH or a monophosphate group Rg = CH3, CH2OH or Cl R7 - H or Br Formula 1 Compound it and X-, and X2 are independently selected from H, methyl, ethyl, hydroxyl, the halogens and carboxyl v or R4 or R, O « or R4 = CNH-Rg and Cl or Rg = and R2 = OH and R3 = OH, monophosphate, diphosphate or triphosphate group or R2 and R3 are linked to form a 3’, 5’-cyclic monophosphate derivative, or a physiologically acceptable salt of any such compound. Formula I” is used herein to refer to all of such compounds and salts, as well as (when referring to administration against one or more parasite) polymer of IF A — identified herein as Poly N6- Isopentenyl Adenosine”—, -preferably comprising 2-3 monomers.-----30 Formula 1 Compound /£ 195 Listed below are chemical groups Rj-R4 for — especially preferred compounds la-Xu according to this invention.

Xa: R, = H, Rg = OH, Rj = OH and s ____CH3 η CH3 β 2 ' Ν -(Δ -isopentenyl) adenosine lb: R, H, Rg = OH, R3 = monophosphate, and R« “ CH2083 H <»3 N6- (Δ2- isopentenyl) adenosine-5 ‘ -monophosphate Xc: R, = H, Rg and R3 are linked to form a 3*, 5*-cyclic monophosphate derivative, and R* B CH3 H ' CK3 N6- ( A2-isopentenyl) adenosine-3 ·, 5 ‘ cyclic monophosphate Id: Rt = H, Rg = OH, R3 = OH, and_ R4 = CH2C6H6 N6-benzyladenosine le: R, = H, Rg = OH R3 = monophosphate, and R4 = CHgCgHg H6-benzyladenosine-5 ’ -monophosphate.

If: R1 = H, Rg and R3 are linked to form a 3’, 5‘-cyclic monophosphate derivative and R4 « 0¾¾¾ N6-benzyladenosine-3 ‘, 5 ’ -cyclic monophosphate Ig: R1 = H, Rg = OH, Ra = OH, R4 = CHgFonnula 1 Compound l£ u 196 Furfuryladenosine Ih: R-j = .H, .Bj.;*·' OH, R3 = monophosphateand R4 = CHj Ii: phosphate Ij: Ik: N6-furfuryladenosine-5 ’ -monophosphate R, s H, Rj and R3 are linked to fora a 3', 5 * -cyclic monophosphate derivative, and K, lr-furfuryladenosine-3', 5 * -cyclic- monoR, — H, Rj 8 OH, Rj 8 OH and O M _ZZ\ R* 8 CNH-( Q > Cl N- ( purin- 6-ylcarbamoyl) -o-chloroaniline ribonucleoside R1 e h, Rj = OH, R3 e monophosphate and O Jt4 = OTH -(6> Cl N- (purin-6-ylcarbamoyl) -o-chloroaniline ribonucleoside-5 ’ -monophosphate II: R, 8 H, Rj = OH, Rj β OH and «4 N6-adamantyladenosine Im: Rj β H, 1^2 = OH, R3 = monophosphate and «4 = Formula 1 Compound IE 197 N°-adamantyladenosine-5 * -monophosphate In: R, = H, Rg = OHy R3 - OH and Io: Ip: Ig: o M .

R4 « CNH(CH2 N- ( purin-6-ylcarbamoyl) -n-oc t ylamine ribonucleoside R, = H, Rj = OH, R3 = monophosphate and O It R< = CNH(CH2)7CH3 N- ( purin-6-ylcarbamoyl, -n-octy lamine ribonucleoside-5*-monophosphate Et, «= H, Rjj and R3 are linked to form a 3 *-5’-cyclic monophosphate derivative, and ? R< « N- (purin-6-ylcarbamoyl) -n-octy lamine ribonucleoside-3', 5 * -cyclic monophosphate R, “ CH3S, Rj b OH, Rj · OH and R, Ir: CH* < --H CH3 N6 - (A2-isopentenyl)-2-methyl thioadenosine R, = H, = OH, R3 = OH and R4 = CH2 CH3 CHgOH N6- (4-hydroxy-3-methyl- trans-2-butenyl) -¾ adenosine Is: R, = H, Rj = OH, R3 = OH,, and R4 = 0¾ Cl H CH3 N6 -(3-chioro-trans-2-butenyl) adenosine Formula 1 Compound IE V ό ϋ 3 1 ϊ 198 It: .5 ....

Iu: R, « H, Rg - OH, R3 = OH and = “ .CH3 >=< H Cl H6 - (3-chloro-cls-2-bubenvl) adenosine R1 = H, Rg = CH3, R3 CH3 and R* = H The present invention also employs, one or more metabolite of the family of compounds of Formula I. For v example, preferred metabolites include: Ι^-ίΔ2-isopentenyl) adenine; 6-N-(3-raethyl-3-hydroxybutylamino) purine; Adenine; Hypoxanthine; Uric Acid; and Methylated xanthines» . \ ' x Formula 1 Compound N^-Benzyl-Adenosine-S’-monophosphate 199 '·* .1 ·. I Molecular Weight 437.215 Molecular Formula: CiyH^NgOyP Formula 1 ιε * .t 200 (N6-Benzyl)Adenyl-p-(N6-Benzyl)Adenyl-p4N6-Benzyl) Adenosine •Η Molecular Weight 1373.39 Formula 1 “ονj 201 The present invention employs one or more compound according to Formula 2 : OltiprazFig 1 ADO-Fig3 Formula 2 Compoui li u 202 1,2-DHHIOLE 3-TfflONE Kg4 Lipoamide (1,2-dithiolane) - Fig 5 13-dithiole 2-thione-Fig6 [l^JWduolo[4,3-c]-l^-dithiole-3,6dithione Fig 6a Λ λ Malotilate-Fig7 Formula 2 Compou 1^2-Difhiolane Class 1 203 1,2-Dithiole Class 2 . 1,3-Dithiole Class 3 1,3-Dithiolane Class 4 Wherern 1^/),NR&2,CR2 and Z can have lhe designations optionally and independently for al the classes. R in this case includes, H, alkyl(Cl-C5), alkoxy (C1-C5) alkoxycarbonyl (C1-C5) R2 can form spiro rings about the ring carbon atom.

For the thiolane classes tiie ring carbon atoms can be doubly substituted.

Formula 2 Compo 204 £ ύ 6 o j j 2 R1-R4 are the main ring substituents for all classes and in order to cover a wide variety of substituents should include optionally and independently H, alkyl, aryl, heterocyclic, halogen, alkoxycarbonyl (C1-C5) or carboxyl.

Rl, R2 or R3, R4 can form can form a spiro ring around the carbon atom to which they are attached or they can form fused or bridged rings to adjacent carbon atoms ^he following definitions cover the majority of compounds.

Alkvl defined as C1-C10 linear or branched chain, saturated or unsaturated which can optionally singly or multiply substituted by halogen, alkyl (C1-C5), hydroxyl, alkoxy (C1-C5X alkoxycarbonyl, (C1-C5), caiboxyk amido, alkyl amido (CI-C5), amino, inono and dialkyl amino (C1-C5),alkyl carbamoyl (C1-C5), thiol, alkylthio (C1-C5) or benzenoid aryl.

Aryl defined as any optionally singly or multiply substituted benzenoid group (C6-C14).

Substituents defined below.

Heterocvlic radical defined as any 4,5 or 6 membered, optionally substituted heterocyclic ring, saturated or unsaturated, containing 1-3 ring atoms of which are N,0 or S, the remaining ring atoms being carbon.

Formula 2 Compc 205 Substituents on the aryl or heterocyclic radical indude: ...... -. . halogen, alkyl (C1-C5), hydroxyl, alkoxy (C1-C5), alkoxycarbonyl, (C1-C5), carboxyl, amido, alkyl amido (C1-C5), amino, mono and dialkyl amino (C1-C5),alkyl carbamoyl (Cl-CS), thiol, alkyl thio (C1-C5) or benzenoid aryl, cyano, nitro, halo alkyls, alklsulfonyl (C1-C5), sulfonate.

. . · Two of such substituents can he part of a fused ring, which can be either saturated, or unsaturated, heterocyclic or carbo cyclic.

Kg. 8 s-s Fig. 10 Kg. 11 in which Formula 2 Compou 206 X is chosen from =s, =0.

—N—OH, . ' =N—Rs Rs beingaCrC6 alkyl oranaiyl group, —N—NH—CO—14¾ and K ==N—NH—CS-NH2, and ssC Z and Z’ being electron-attracting groups such as ester or cyano groups.

A is chosen from a >C=N—OH group. A group of formula >C==N—OR3. (where R3 is Chosen ftom hydroxyl, amino, chloro and Ci,-C<, alkoxy groups, an aryL(Ci,-C6 lkyl) group, a (C i-Q alkyl)Cafbonl group and an aiyl(Ci-C6 alkyl)caibonyl group). a may also be chosen from a >0=0 group, a X>N—R4, group. R4 being a Ci-Ce alkyl group or Λ an aryl group, and a CHOH group, λ Ri and R2 are chosen, independently of one another, from hydrogen, a halogen, a nitro group, a nitroso group, a thiocyano group, a Ci-Q alkyl group, a CyC6 alkenyl group, an aryl group, aryl(Ci-C6 alkyl) group, an aryl (C2-C6 alkenyl) group, a carboxyl group, a (Ci-Ce alkyl)carbonyl ad Formula 2 Compou IE 312 207 group, aa arylcarbonyl group, a (CrCe alkoxy)carbonyl group, a (Q-C6 alkoxy)carbonyl (Ci.-Cs alkyl) group, a Cj-Q alkoxy group, a trifluoromethyl group, aa amino group, a di(Cj-C6 alkyl) , amino(C}-Cc alkyl) group, an acylamino group of formula -NHCOCnflhn+i with n from 0 to 6, a group -NH—CSCJi 2oH with n from 0 to 6, a terpenyl group, a cyano group, a C2-C5 alkynyl ... · . ' group, a C2-C6 alkynyl group substituted with a CrCe, alkyl or an aryl group, a hydroxy(C3-Cc alkyl) group, a (Ci-C« acyl) oxy (Ci-Ce alkyl) group, a (Cj-Ce alkyl) thio group and an aiylthio group. tl· or alternatively Ri and R2 together form a mono- or polycyclic C2-C20 alkylene group optionally comprising one or more hetero atoms, with the exception of the 2^dimethyltrimethylene group, or a C3-C12 cycloalkylene group.

R is chosen from a Cr-Ce, alkyl group, and their pharmaceutically acceptable salts, In the foregoing definition, aryl group or aryl fraction of an arylalkyl group denotes an aromatic carbon-based group such as a phenyl or naphthyl group or an aromatic heterocyclic group such as a thienyl of furyl group it being possible for these groups to bear one or more substituents chosen from a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a trifluoromethyl group, a nitro group and a hydroxyl group.

Formula 2 Compoi IE 208 Oximes of l,2-dithiole-3-thione derivatives such as shown in figure 12,13 &14 Fig 12 ecu Fig 13 tf* Fig 14 Additionally Aldehydes or Ketones of previoulsy identified compounds are incorporated such as shown in Fig. 15 k Formula 2 Compour οϋ 3 12 209 one or more of the following compounds according to embodiment, wherein A (figure 10,1 l) is a group C=N—OR’3 where RT is an optionally substituted Cj-Cgalkyl group, in particular substituted with one or more groups chosen from hydroxyl, aminn chloro bromo, flure, iodo and Ci-C4 alkoxy groups, or an aiyl (Ci-Cgalfcyl) group, that is to say compounds of fonnula T* ? Fig. 16 *> OR Fig. 18 in whichTRs has the meaning given above *5 one or more of the following compounds as described in embodiment (Fig 10 & 11) in which A is a group C=N-0-C0-Rw3, R”3 being chosen from a hydrogen atom, an optionally substituted Cj-Cg alkyl group, an aryl group and an aiyl (Cj-Cg alkyl) group, that is to say compounds of formula Formula 2 Compo IE 0 6 0 3 12 210 in which R”3 being chosen fiom a hydrogen atom, an optionally substituted Qj-Ce alkyl group, an aryl group. v Another group of compounds is formed toy fhe compounds of embodiment (Fig, 10 & 11) in which A is a CH-OH group, that is to say the compounds of formula ao, Fig 20 I 1» Another group of compounds is formed by the compounds of embodiment (Fig. 10 & 11) in which A is a group ΟΝ-R, R, toeing a Ci-Q alkyl or an aryl group, that is to say compounds of formula A, FIG 21 Formula 2 Compoi ΙΕ Οθο3ί2 211 Another group of compounds include compounds of embodiment (Fig 10 & 11) in wliieh A is a 0=0 group and X is an oxygen atom, that is to say compounds of formula: Kfs2215 in which Ri is chosen, from hydrogen, a halogen, a nitro group, a nitroso group, a thiccyano group, a Cj-Q alkyl group, a C^-Cs alkenyl group, an aryl group, an aryl (Cj-Q allyl), group, an aryl (Cz-Ce alkenyl) group, a carboxyl group, a (Cj-Ce alkyl) carbonyl group, an arylcaibonyl group a (Ci-Q alkoxy)carbonyl group, a (Cj-Q alkoxy) carbonyl (Ci-Q alkyl) group, a Cj-Ce alkoxy group, a trifluoromethyl group. An amino group, a di (Ci-Ce alkyl) amino (Ci-C& alkyl) group. an acylamino group of formula -NHC0C πΗμ with n from 0 to 6 . a group -NH-CSCnH 2n+i with n from 0 to 6, a texpenyl group, a cyano group, a Ci-Q alkynyl group, a C2-^6 alkynyl 25 λ group substituted with a Cj-Gg alkyl or an aryl group, a hydroxy (C1-C6 alkyl) group, a (Ci-Cc acyl)-oxy(Ci-C6 alkyl) group, a (Cj-Ce 6 alkyl)thio group and an aiylthio group.

' Formula 2 Comp* 212 R2 is chosen from a nitro group, a nitroso group, a thiocyano group, a Cj-Q allyl group, a C2-Cg alkenyl group, an aryl group, an aryl (Ci-Q allyl) group, an aryl (Cj-Ce alkenyl) group, a carboxyl g&up. a (Cj-Q alkyl)carbonyl group, an arylcarbonyl group, a (Ci-Ce alkoxy)catbonyl group, a/Ci-Ce alkoxy)carbonyl (Ci-Q alkyl) group, a trifluoromcthyl group, a di(Ci-C<; allyl)amino(Ci-C6 alkyl) group, an acylamino group of formula -NIlCOQJl^+i with « from 0 to 6, a group -NH—CSCaHz+i with n from 0 to 6. a terpenyi group, a cyano group: a CrCs alkynl group, a C2-C6 alkynyl group substituted with a Cr-C$ allyl or an aryl group, a hydroxy (Cj-Cc allyl) group, a Cr-Q acyl)-oxy(Ci-C6 allyl) group, a (Ci-Ce alkyl)thio group and an aiylthio group, or alternatively Ri and R2 together form a mono- ot polycyclic C2-C20 alkylene group optionally comprising one or more hetero atoms one or more of tbe following compounds of formula shown in Figure 23 t on ε ,~T COOCHj Fig. 23 Formula 2 Comps IE 213 1« Ri and R2 are chosen, independently of one another, from hydrogen, a halogen, a nitro group a nitroso group, a thiocyano group, a Cj-Q allyl group, a Qz-Q alkenyl group, an aiyl group, aryl(Ci-C6 alkyl) group, an aiyl (Q-Q alkenyl) group, a carboxyl group, a (Cj-Q alkyl)caibonyl group. A aiyleaibonyl group, a (Cj-Q alkoxy)carbonyl group, a (Q-Ce alkoxy)carbonyl (Ci,-Ce alkyl) group, a Ci-C$ alkoxy group, a trifluoromethyl group, an amino group, a di(Ci-Cs alkyl) amino(Ci-Cs alkyl) group, an acylamino group of formula-NHCOCnH^n with n from 0 to 6, a group -NH—CSCaH 2n+i with nfrom 0 to 6, a terpenyl group, a cyano group, a C2-Ce alkynyl group, a CrQ alkynyl group substituted with a Ci-Ce, alkyl or an aryl group, a hydroxy^Cj^g alkyl) group, a (Cj-Cg acyl) oxy (Cj-Cg alkyl) group, a (Ci-Cg alkyl) thio group and an arylthio group. or alternatively Ri and Ri together form a mono- or polycyclic Q-C» alkylene group optionally comprising one or more hetero atoms.

R is chosen fiom a Ci-Cg, alkyl group, and their pharmaceutically acceptable salts, lh the foregoing definition, aryl group or aiyl fraction of an aiylalkyl group denotes qn aromatic carbon-based group such as a phenyl or naphthyl group or an aromatic heterocyclic group such as a thienyl of finyl group, it being possible for these groups to bear one or more substituents chosen from ahalogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a trifluoromethyl grihip, a nitro group and a hydroxyl group.

Formula 2 Compoi ιε °b(l3l2 214 one dr more of tiie following isobenzothiazolone derivative having the structure: Fig 24 o B N—»’ I s In tiiis structure at least one of R1and R2 is preferably nitro, arylazo, substituted aiylazo, 10 benzylideneamino or substituted benzylideneairiino. When only one of R* and R2 is so substituted, one of R1and R2 may be hydrogen. The R3 substituent is selected from alkyl groups in less than about 7 carbon atoms, amino, hydroxyl, alkoxyl, and aryl groups (and functionalized forms thereon,) Preferred species of tiie isobenzothiazole derivative of the present invention comprise R1 as nitro or aiylazo and R2 as hydrogen, for example. Examples include compounds where R2 is hydrogen . and R1 is phenylazo; substituted arylazo such as 4-hydroxyphenylazo; 4— nitio-2-methylphenylazo; 2-hydroxy-l-napthylazo; 2- hydroxy-5-metbylphenylazo; 2-hydroxy-4-methyl-5-nitrophenylazo; 4-hydroxy-l-napthylazo; 4-hydroxy-3-methyl- 1 -napthylazo; 4-hydroxy-5-aza-l -napthylazo; 2-amino-l-napthylazo; l-hydroxy-2-napthylazo; 3-N,Ndimethylaminopropylcaiboxyamido-l-hydroxy-4-naph-thylazo; \ l-hydroxy-4-methoxy-2-naphthylazo, 2_ hydroxy-3-carboxy-l-naphthylazo; l-liydroxy-3, 6disulfonato-2-naphthylazo; 2, 3-dihydroxy-l-naphthylazo; or 2-hydroxy-3, 5-dimethyl-l-phenylazo. In one particular embodiment R1 is the substituted ben zylideneamino, 2,4-dinitrobenzylideneamino and R2 is hydrogen. Additionally R1 as hydrogen and R2 as 2-hydroxy-l-naphthylazo or 4-hydroxy-lphetiylazo.

Formula 2 Compel νό03,2 215 In one aspect, R3 substituents witii sufficient polarity to confer aqueous solubility upon the compound. For example, R3 may be where n is from 2 to 6 and R4 and R5 are simple alkyls or hydrogens. Other possible water solubilizing side chains include 3-carboxypropyl, sulfonatoetbyl and polyethyl ethers of the type -CH2(CH20CH2),CH3 where n is less than 10. Preferred compounds include R3 side chains containing aminoalkyl, carboxyalkyl, omega amino polyethyl ethers and N-haloacetyl derivatives. In a broader sense, for various utilities R3 may be alkyl, aryl, heteroaryl, alkoxy, hydroxy or amino groups. When including substitutions for solubility or reactivity purposes, R may be aminoalkyl, carboxyalkyl, hydroxyalkyl or haloalkyl. The aryl or heteroar R3 moieties may be substituted, for example as aminoaiyl, caihoxyiyl orhydroxyaiyl. one or more of the following Isobenzothiazolone derivative having the structure: Fig 25 wherein at least one of R1 and R2 is nitro, aiylazo, substituted aiylazo, benzylideneamino or 25 substituted benzyfideneamino and one of Rl and R2 may be hydrogen and R3 is a aminoall^yl, aminoaryl and aminoheteroaryl, carboxyalkyl, carboxyaryl or carboxyheteroaryl covalently linked to a polymer comprising amino or hydroxy groups. The spacer arm R3 can comprise oligmers or polyethylene-glycol and its derivatives, hi one aspect, R3 may be 17-chloracetamido-3,6,9,12, 1 5-pentaoxyhep- tadecyl where hexaethylene glycol has been cWoroacetamidatedL When lhe polymer groups, Y and R Formula 2 Compor 216 v ti o 3 ί ζ , comprises ccftoxyl groups, Π» covalent linkage is preferably through an ester bond. When the polymer comprise amino groups, the analog covalent linkage is through an amide bond. The a„,i„dxOTng polymer, when coupled to R’, may be a polymer suoh as chitosan, pdyalkytamine, 5 arninodcxtranpolyethyleneimine, polylysine or amityrene.

The R3 substituents of the present invention may also comprise an alkyj linked to an aminebearing polymer by amine displacement of a halogen from an alpha-haloalkyl or JO alpha-lialoalkylcarbox amido R3 precursor. In the case of aminoalkyl or aminoaiyl groups the R3 substituent may also be covalently linked to a polymer such as polyepichlorohydrin, chloromethylpolystyrene, polyvinylalcohol or polyvinylpyridine. The R3 substituent ofthe present invention may generally be an aminoalkyl, hydroxyalkyl, aminoaryl or hydroxyaiy] group linked to a polymer comprising carboxyl groups through amide or ester linkages. &hen polymers are involved in the R3 structure, the polymer may be one such as polyacrylic acid, polymethacrylic acid, polyitaconic add, oxidized polyethylene oxide, poly(methyhnethacrylate/me(haciylic acid), carboxyinethyl cellulose, caiboxymethyl agarose or caiboxymethyl dextran. When such a carboxyl polymer is involved, the R3 may be aminoalkyl (such as $ aminohexyl, for example), hydroxyalkyl, aminoaiyl or hydroxyaiyl linked to the · ' · \ polymer through amide or ester linkages. In such cases, an R3 precursor iunction may bearan amine or hydroxyl group to be covalently linked to a polymer by reaction with an acid anhydridebearing polymer or by coupling wilh a caiboxylatebearing polymer through carbodimideinduced bond formation Formula 2 Compota The R3 substituent or precursor thereto in the compound of tiie present invention may also be a haloalkyl or carboxylialoalkyl moiety such as chloracetamido. Such a substituent may readily JESS’ coupled to an aminebearing polymer by amine displacement of the halogen. 0312 217 Aryl, as used herein, is intended to include organic residues derived from aromatic hydrocarbon or aromatic heterocyclic ring systems. Accordingly aryl groups include tiie uribbsfjtuted ring residues such as phenyl and naphthyl and substituted forms thereof Heterocyclic or heteroaryl residues may be those comprising one or more heteroatoms (e.g., nitrogen, oxygen, sulfur) in the ring system sueh as pyridyl, oxazolyl, quinoly), thiazolyl and substituted forms thereof.

Alkyl,” as used herein, is intended to include aliphatic and cyclic organic residues having a carbon at a point of attachment. According, alkyl groups indude unsubstituted hydrocarbon residues of tiie formula C s Η Μ and substituted and cyclic forms thereof. Such hydrocarbons are usually of tiie lower allyl class which have six carbons or less. It is understood that larger alkyl groups may be used. Alkyl indudes substituted residues which are intended to include tbe hydrocarbon residues bearing one or more, same or different, functional groups as described below.

The alkyl and aryl group previously described may be substituted with functional groups. Such functional groups include essentially all chemical groups which can be introduced synthetically ,, ' Λ \ 3 and result in stable compounds. Examples of these functional groups are hydroxyl, halogen (fluoro, chloro, bromo), amino (including alkylamino and dialkylamino), cyano, nitro, carboxy (including earbalkoxy), carbamoyl (including N and N,N alkyl), sulfb, alkoxy, alkyl, aryl, and arylazo.

Formula 2 Compo«« IE U () y j | 2 218 one or more of the following compounds Fig26 . wherein Rj and R^re independently (=0) or -OR, where R is H or (Q-C*) alkyl; and R3 is 11 or 10 (C1-C4) alkyl. Preferably, R3 is BL Preferably Rj and Ra are (=0) or OH. one or more of the following compounds OR, Fig27 . v •ϊ wherein X is H or both Xs represent a direct bond between the two sulfur atoms; Ri is (=o) or -OH; and R2is H, Na, K or (Q-C4)alkyl In particular die compound maybe 3-keto lipoie add, 3-hydroxy lipoic acid, 3-keto dihydrolipoic acid or 3-liydroxy dihydrolipoic acid.

Formula 2 Colour IE 219 one or more of tiie following 1,2-dithioiethione derivative of a formula shown in Fig. 28 / Y\ Fig. 28 R wherein R denotes hydrogen, halogen, lower alkoxy group, lower alkyl group, amino group, lower.alkylsubstituted amino group or lower alkoxycarbonyl group.

In tiie above-described formula Fig 28, the term lower means methyl, ethyl, propyl and butyl, as well as its structural isomers such as isopropyl, isobutyl and tertiaiybutyl.

Among tiie compounds of the formula shown in Figure 28, preferred compounds include 5-(4-phenyl-l,3-butadienyl)-l,2-dithiol-3-thione, -4(4-chlorophenyl)-13-butadienyl-l^-dithtol-3-thione, -{4-(4-methoxyiAenyl)*-13-butadxenyl)-1^2'dithiol-3-tiuone, t -{4-{p-toluyl)-13’butadienyl}-l^-dithiol-3-thione, / -{4l,3-butadienyl}-l,2-dithi-o]-3-thione,and 5-{4-(m-methylphenyl)-l,3butadienyl}-l,2-fiSthiol-3-thione * The following compounds are also included: Formula 2 Compoun 220 Fig 31a Kg 32 Kg. 33 Fonnula 2 Compow ιε υ 6ί ο j 12 221 one or more of the following; 1,2-dithiole of tiie formula (Fig 34).

BetFig 34 wherein Het represents pyrimidin-2-yl, pyrimidin-4-yl, or pyrimidin-5-yl, or a said pyrimidin-2-yd, pyrimidin4-yl or pyrimidin-5-yl substituted by halogen, alkyl of 1 throu^i 4 carbon atoms, alkoxy of 1 through 4 carbon atoms, mecapto, alkylthio of 1 throu^i 4 carbon atoms, or dialkylamino having 1 through 4 carbon atoms in each alkyl, and R represents halogen, alkyl of 1 through 4 carbon atoms, alkyl of 1 through 4 carbon atoms substituted by . alkoxycarbonyl having 1 through 4 carbon atoms in the alkoxy, carboxy, alkoxycaibonyl having 1 through 4 carbon atoms in the alkoxy, carbamoyl, N-alkylcaibamoyl having 1 through 4 carbon atoms in the alkyl, or R,-CH(OHl· in which R, represents hydrogen or alkyl of 1 through 3 carbon atoms.

Formula 2 Compom Claims What is claimed is: 1. A method to prevent, treat, ameliorate or slow the progression of blood cell deficiency disorder, neutropenia or thrombocytopenia or anemia in a subject, or to treat a symptom of the neutropenia or thrombocytopenia or anemia, comprising administering to a subject, or delivering to the subject's tissues, an effective amount of a Formula 1 cytokinin compound

Claims (21)

1. And/or an effective amount of a Formula

2. Compound , alone or in combination.

3. , 4, 5, 6, 7, 8, 9,10 or more gene products or gene transcripts in the cell, comprising contacting an effective amount of the compounds of this patent with the cell under suitable conditions and for a sufficient time to detectably modulate the activity or level of the genes, or gene products in the cell, wherein the compound is a compound of any of Formula 1 or Formula 2 and the gene products or gene transcripts are selected from USF1, c-Fos, EGR1, Cull, RIPK2, G-CSF, CCAAT/enhancer binding protein alpha, I.kappa.Ba, I.kappa.BKb, NF-.kappa.Bl p50, FCAR, c-Fos/C/EBP.beta., RANTES, ICAM1, TSG (TNFAIP6), IL-2 receptor .alpha., GRO2, GRO3, HOI, Jun B, cFos/JunB complex, JunB/ATF3 complex, c-Jun, c-Fos/c-Jun complex, bcl-2, ATF-3, MMP1, TSG-6 (TNFAIP3), AP-1, EGR1, TGF.beta., ATF-3/c-Jun complex, c-Fos, MMP3, IL-8, STAT5A, STAT5B, CDKN1A, IFN.gamma. receptor 2 (IFN.gamma,R2), T-bet, C reactive protein, immunoglobulin E, an AP-lfamily protein, GATA-3, Jak2, Tyk2, statl, stat3, stat4, stat5, stat6, MIP-1.alpha., MIP-2, IP10, MCP-1, TNF-.alpha., TNF-.beta., LT-.beta., IFN-.alpha., IFN-.beta., TGF-.beta.l, NF-.kappa.B, IL-l.alpha., IL-l.beta., IL-4, IL-6, IL-10, IL-12 receptor .beta.l, ILo0 3 12 12p35, IL-12p40, IL-23, IL-23 receptor, Nrf2, a Maf protein, a thioredoxin, NQO1, GST, HO 1, SOD2, the catalytic subunit of .gamtna.GCS, the regulatory subunit of .gamma.GCS and xCT. 3 2. The method of claim 1 where the Fonnula 1 compound used is N6-Isopentyl Adenosine

4. - [3,5-bis(l,l,3,3-tetramethylbutyl)-4-hydroxyphenyl]-l,2-dithole-3-thion- e; 4-(3,5-bis(lmethylcyclohexyl)-4-hydroxyphenyl]-l,2-dithiole-3-thione; 4-(3,5-bis(l,ldimethylbenzyl)-4-hydroxyphenyl]-l,2-dithiole-3-thione; 4-(3t-butyl-4-hydroxy-Sisopropylphenyl)-1,2-dithiole-3-thione; 4-(3t-butyl-4-hydroxy-5-methylphenyl)-1,2dithiole-3-thione; 4-(3(1, l-dimethylpropyl)-4-hydroxy.-5-isopropyiphenyl]-l,2-dithiole-3thi-one; 4-(3(1,1 -dimethylbenzyl)-4-hydroxy-5-isopropylphenyl]-1,2-dithole-3-thione; 4 and the Fonnula 2 compound used is Oltipraz..

5. - benzylthio-4-(3,5-di-t-butyl-4-hydroxyphenyl)-l ,2-dithole-3-thion- e; 5-benzylthio-4[3,5-bis(l,l-dimethylpropyl)-4-hydroxy-phenyl]-1,2-dithi- ole-3-thione; 5-hexylthio-4-(3,5di-t-butyl-4-hydroxyphenyl)-1,2-dithole-3- -thione; 5-hexylthio-4-[3,5-bis(l,ldimethylbutyl)-4-hydroxy-phenyi]-l,2-d- ithole-3-thione; 5-octadecylthio-4-(3,5-di-t-butyl4-hydroxyphenyl)-l,2-di- thiole-3-thione; 5-octadecylthio-4-(3,5-bis(l,l-dimethylbenzyl)-4hydroxyp- henyl]-l,2-dithiole-3-thione; 5-allylthio-4-(3,5-di-t-butyl-4-hydroxypheny- ΟΙ ,2-dithiole-3-thione; 5-cyclohexylthio-4-(3,5-di-t-butyl-4-hydroxyphenyl)-1,2-dithiole-3°603j 2 thione; and 4-(3,5-di-sec-butyl-4-hydroxyphenyl)-l,2-di- thiole -3-thione. 35. The method of claim 1-34 wherein the compounds of Formula 2 chelates with, or forms a complex with, one or more divalent or trivalent radioactive metal ions, whereby the divalent or trivalent radioactive ions in the subject's cells or tissues are redistributed or sequestered such that the ions are limited in their capacity to participate in unwanted tissue destruction. 36. The method of claim 35 wherein the divalent or trivalent metal ions are selected from Fe, Cu, Ni, Ca, Mg, Mn, Cd, Pb, Al, Hg, Co, I, Se, Cs, U, Pa, Th, Ra, Ce, and Zn ions. 37. The method of claims 1-35 wherein the Formula 2 compound is an oxime or a derivative of said compound. 38 . The method of claims 1-35 wherein said compound is formulated into a composition that further comprises a pharmaceutically acceptable carrier. 39. The method of claims 1-38, in which Formula 2 compounds comprise Oltipraz, and/or 3H-l,2-dithiole-3-thione, and/or sulforaphane, narigin. 40. Use of one or more of the compounds of claims 1-39 for the manufacture of a medicament for to prevent, treat, ameliorate or slow the progression of blood cell deficiency disorder, neutropenia or thrombocytopenia or anemia in a subject, or to treat a symptom of the neutropenia or thrombocytopenia or anemia. 42 . The method of claim 1 wherein the subject has, or is subject or susceptible to developing, Thrombocytopenia 43. The method of claim 1 wherein the subject has, or is subject or susceptible to developing anemia 222 /£ () /J Λ 223 -LYMPHOCYTE FUNCTION of an AIDS PATIENT RECEIVING EXPERIMENTAL THERAPY - IPA

6. 3. The method of claim 2 wherein the Formula I compound is selected from the analogues

7. And derivatives as outlined in the text of this patent application.

8. The method of claim 1, wherein one or more symptoms or syndromes of radiation organism or tissue damage are ameliorated, or wherein the progression of the blood cell deficiency disorder is reduced. r .-*-***. Λ (jo IE 0 60 3 12

9. The method of claim 1 wherein the formula 1 listed cytokinin compounds is a prodrug of the cytokinins that can convert to the biologically active compound by metabolism or hydrolysis. 9 4. The method of claim 2 wherein the Fonnula 2 compound is selected from the analogues

10. The method of claim 1 wherein the subject is a human who has a blood cell deficiency disorder. 10 and derivatives as outlined in the text of this patent application.

11. The method of claim 10 wherein the treatment reduces (1) the severity of pain during vascular or microvascular occlusions, (2) the severity of vascular or microvascular occlusions or (3) the frequency of vascular or microvascular occlusions.

12. The method of claim 11 wherein the formula 1 compound is administered by an intermittent administration protocol. 12 5. The method of claim 1 wherein the compound is selected from any combination of a

13. A method to modulate the expression in a cell ofthe level of or an activity of 1,2, 13 Formula 1 compounds and/or a Fonnula 2 compounds together or alone or in any

14. The method of claim 13 wherein there is a detectable increase in the level of the mRNA, the protein or one or more biological activities associated with the gene product. 14 combination,.or an analog or derivitive of any of the foregoing compounds that is suitably

15. The method of claim 12 wherein the formula 1 compound is N6 isopentyl adenosine and the Formula 2 compound is Oltipraz. 15 substituted to fall within the scope of the claim.

16. A method to enhance the healing of a trauma or an acute injury in a subject who has experienced or who is expected to experience a trauma or an acute injury comprising administering an effective amount of a Formula 1 compound and/or a Formula 2 compound, compound to the subject, wherein the Formula 1 compound is N6 isopentyl adenosine and the Formula 2 compound is Oltipraz.

17. A method of claim 16 wherein wherein the Formula 1 compound is N6 isopentyl adenosine and the Formula 2 compound is Oltipraz. 17 6. The method of claim 1 wherein the subject has, or is subject or susceptible to developing, 18. The method of claim 18 wherein the ionizing radiation exposure is about 0.2 Gy to about \ 15 Gy of ionizing radiation

18. The method of claim 16 wherein the subject will experience or has experienced an immune suppressive event within about 2-3 weeks or about 34 weeks from the occurrence of the trauma or acute radiation injury, wherein the immune suppressive event is exposure of the subject to an immune suppressive amount of ionizing radiation. 18 neutropenia.

19. The method according to claims 1-16 wherein the treatment comprises administering said formulations of Compounds of Formula 1 or Compounds of Formula 2 contained in a 0 6 0 3 12 enterically coated capsules, or intravenous solution or trans-dermal patch .

20. . The method according to claims 1-19 wherein the treatment comprises administering said formulation of Formula 1 alone or in combination with Formula 2 compounds in unit doses of 0.01 mg. to 1000 mg/per Kg of bodyweight. 21. The method according to claim 1-19 wherein the treatment comprises administering said form ulation of Formula 1 compounds alone or in combination with Formula 2 compounds intravenously in a daily unit dose of 0.01 mg to 80 mg per kilogram of patient body weight. 22. A method of treatment of a patient, either animal or human, for establishing improved immune response for patients in which immune-deficiency resulting from any disturbance to the immune system by exposure to radiation, trauma or environmental factors the treatment comprising administering an effective dosage of a pharmaceutical formulation comprising compounds of Formula 1 or a physiologically acceptable salt thereof alone and/or in combination with compounds of Formula 2 or a physiologically acceptable salt thereof.. 23. A method of claim 22 wherein the compound to be administered is a Formula 2 compound or a physiologically acceptable salt thereof 24. The method according to claim 22 wherein the treatment further comprises administering an effective amount of an adenosine deaminase enzyme inhibitor to the patient. This administration of a deaminase inhibitor is to prevent the hydrolysis of compounds of Formula 1, an example of an adenosine deaminase inhibitor suitable for use in this patented application is Pentostatin.. 25. The method of claim 1-16, and claim 22, claim 23, wherein the compound of Formula 2 include the following Oltipraz, 5-(4-methoxyphenyl)-3H-l,2-dithiole-3-thione, ADT, ADO, l,2-dithiole-3-thione, 1,2-dithiolane, l,3-dithiole-2-thione, and malotilate. 0 6 0 3 j 2 26. The method wherein the Formula 2 compound, to be administered in claims 1-24, chelates with, or fonns a complex with, one or more divalent or trivalent radioactive metal ions, whereby the divalent or trivalent radioactive ions in the subject's cells or tissues are redistributed or sequestered such that the ions are limited in their capacity to cause serious vital organ or tissue damage. 27. The method of claim 26 wherein the divalent or trivalent radioactive metal ions are selected from Fe, Cu, Ni, Ca, Mg, Mn, Cd, Ph, Al, Hg, Co ,1 ,Se, Cs, U, Pa, Th ,Ra, Ce, and Zn ions. 28. The method of claims 1 -26 wherein said compound of Formula 2 enhances one or more phase II detoxification enzymes. 29. The method of claim 28 wherein said phase II detoxification is selected from the group enzymes consist of glutathione S transferase, .gamma.-glutamylcysteine synthetase, glutathione reductase, glutathione peroxidase, epoxide hydrase, AFB.sub.l aldehyde reductase, glucuronyl reductase, glucose-6-phosphate dehydrogenase, UDPglucuronyl transferase, and AND(P)H:quinone oxidoreductase. 30. A method to treat, prevent or to reduce the level of radioactive iron or to reduce free transition radioactive metal ion levels in the body or in certain body compartments, in a subject in need thereof, the method comprising administering to the subject or delivering to the subject's tissues a therapeutically effective amount of a Fonnula 2 compound. 31. The method of claims 1 - 30 wherein the compound of Formula 1 and/or Fonnula 2 are -< micronized or the compounds is present in a composition that comprises a pharmaceutically acceptable carrier, the carrier optionally selected from phosphatidylcholine, diphosphatidylcholine, vitamin E, a cyclodextrin, magnolol, a microbial preservative, water or a liquid excipient suitable for ophthalmic phannaceutical formulations., or formulated in a θβθ3, 2 tissue decomposition matrix to allow slow delivery from a polymeric delivery such as with the use of silicone polymers. 32 . The method of claims 1-31 wherein said compound of Formula 2 enhances the cellular production of phase II detoxification enzymes. 33. The method of claim 32 wherein said phase II detoxification enzyme is selected from the group consisting of glutathione S transferase, .gamma.-glutamylcysteine synthetase, glutathione reductase, glutathione peroxidase, epoxide hydrase, AFB.sub.l aldehyde reductase, glucuronyl reductase; glucose-6-phosphate dehydrogenase, UDP-glucuronyl transferase and AND(P)H:quinone oxidoreductase. 34. The method of claim 1 -33 which comprises administering or delivering to the subject a therapeutically effective amount of a compound of Formula 2 some of which include : 4(3,5-diisopropyl-4-hydroxyphenyl)-l,2-dit- hiole-3-thione; 4-(3,5-di-t-butyl-4hydroxyphenyl)-l,2-dithiole-3-thione; 4-(3,5-bis(l,l-dimethylpropyl)-4-hydroxyphenyl]l,2-dithi-ole-3-thione; 4-[3,5bis(l,l-dimethylbutyl)-4-hydroxyphenyI]-l,2-dithiole-3-thione; 20 7 . The method of claim 6 wherein the subject is a human and wherein the neutropenia is 21 postinfectious neutropenia, autoimmune neutropenia, chronic idiopathic neutropenia or a 22 neutropenia resulting from or potentially resulting from a cancer chemotherapy, 23 chemotherapy for an autoimmune disease, an antiviral therapy, direct radiation 24 exposure,secondary radiation exposure through environmental contamination, tissue or solid 25 organ allograft or xenograft rejection or immune suppression therapy in tissue or solid organ 26 transplantation or aging or immunesenescence.

21. ^2 28/2 3/3 14/3 28/3 18/4 25/4 1/5 8/5 15/5 22/5 30/5 6/8 12/8 19/8 28/8 10/7 TEST DATES (DAY/MONTH) CHART 2