JPH07506372A - Compositions and methods for modulating IL-6 production in vivo - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Compositions and methods for modulating IL-6 production in vivo TECHNICAL FIELD The present invention provides methods and compositions for use in modulating cytokines in vivo, and more. In particular, infusion with a composition consisting of DHEA congener. Concerning the reduction of abnormally elevated levels of tarleukin-6.

background Interleukin 6 (rlL-6J) is produced by a variety of cells and has a wide range of Acts on tissues and has proliferation-inducing, proliferation-inhibiting, and differentiation-inducing effects depending on the properties of camphor cells. It is a pleiotropic cytokine that exhibits IL-6 is involved in immune responses, acute phase reactions and and hematopoiesis, and may play a central role in host defense mechanisms. It is believed that

Because of its pleiotropic effects, IL-6 has previously been used to treat β2-interferon (IFN) -β2), B cell stimulating factor 2 (BSF-2), 26KDa protein, hybrid plasmacytoma growth factor (HPGF or IL-HPI), hepatocyte stimulator (H3P) and type 2 mononuclear granulocyte inducer (MCI-2). It was However, molecular cloning studies have revealed that all of these factors have been identified. A common identity of the lipeptidic chains was demonstrated.

IL-5 induces BSF cell differentiation, acute phase protein induction in hepatocytes, and myeloma. / Plasmacytoma / Hybridoma cell growth promotion, IL-2 and IL-2 receptor expression induction, proliferation and differentiation of T cells, inhibition of proliferation of certain myeloid leukemia cell lineages and induction of their differentiation into macrophages, hematopoietic stem cells induced by IL-3 Promotion of pluripotent colony cell formation and megakaryocyte maturation as a thrombogenic blade induction, mesangial cell proliferation induction, neural differentiation of PCI2 cells and keratinocyte increase It is thought to be necessary for reproductive induction. Dysregulation or overproduction of IL-6 , polyclonal BSF cell abnormalities and autoantibody production (e.g., cardiac myxoma, Castleman's Disease ) disease, AIDS, alcoholic cirrhosis, systemic erythema lupus (SLE)), proliferative disease disease (proliferative glomerulonephritis of the interglomerular interstitium, psoriatic meningitis, bacterial meningitis, tuberculous meningitis, herpes simplex meningitis), and malignant Tumors (plasmacytoma and myeloma, lymphoma and leukemia, and renal cell carcinoma) It is thought to be associated with a variety of pathological conditions, including: non-surgical Serum of an individual after injury (e.g. during acute rejection observed in kidney transplants) and abnormally high levels of IL-6 in the urine have also been observed. Regarding IL-6 For reviews, see Hirano, T. et al. Autology Today (1+++aunology Today) Volume 11 (1 990), Wolvekamp, M.C.G.I. M, C.

J,) and R,L,Marquet (R,L,Marquet), Immuno Immunology Letters Volume 24 1-1 Page 0 (1990), and Pan Snick 1. Jay (Van 5nick, J,) Annual Review of Immunology (^nnu, Rev , Imwunol, Vol. 8, pp. 253-278 (1990). that Therefore, it is desirable to reduce and regulate abnormal levels of IL-6.

Summary of the invention Applicants' invention addresses elevated levels of IL-6 associated with pathological conditions in individuals. Lowering (“downregulation”) elevated levels of IL-6, including IL-6 The present invention is directed to methods and compositions useful for. Applicants surprisingly discovered that DHE Dehydroepiandrosterone (rDHEAJ) containing A and DHEA-3 symptoms may be induced by trauma or related to aging or autoimmune diseases. The associated elevated IL-6 levels are restored to those seen in normal adult individuals. It was discovered that it is an effective drug for

Accordingly, one aspect of the invention consists of a pharmacological dose of DHEA concinina. The procedure for correcting abnormally elevated IL-6 levels consists of administering to an individual a suitable composition. This is a method of treating individuals by lowering their levels.

Another embodiment of the invention provides a pharmacological dose of DH in a pharmaceutically acceptable excipient. Reduce the abnormally elevated IL-6 level consisting of EA conner to improve the individual's health. A composition for treating.

Yet another embodiment of the invention provides a pharmacological dose of DHEA concinina. to reduce abnormally elevated IL-6 levels by mixing with above-accepted excipients. A method of making a composition for treating an individual by reducing

A further embodiment of the invention is a suitable drug comprising a pharmacological dose of a DHEA compound. abnormally elevated I associated with a B-cell disease, comprising administering the composition to an individual. A method of treating an individual by lowering L-6 levels.

Another embodiment of the invention provides a suitable composition comprising a pharmacological dose of a DHEA compound. reducing abnormally elevated IL-6 levels, It is a method of treating an individual by restoring cellular responsiveness to growth factors.

Related technology Therapeutic use of DHEA and certain analogs has been shown to treat diabetes, xeroderma, high intraocular pressure, and hypertrophy. It has been reported in relation to cancer and retrovirus infections. The explanation of these reports is , U.S. Pat. No. 4,395.408 (Tore U.S. Pat. No. 4,518.595 (May 21, 1985) Coleman et al.), U.S. Pat. No. 4.542.129 (Granted to Orentrejch on September 17, 1985) U.S. Pat. No. 4,617,299 (Knepper (K. No. 4.628.052 (December 1986) U.S. Pat. No. 4.666.898 (No. 19 Coleman et al. on May 19, 1987), European Patent Application Publication No. 0 133 9 No. 95 A2 (dated February 8, 1984: nine inventor Schwartz ) et al.), British Patent Application Publication Tube GB 2 204 237 A (April 1988) 1 Inventor Prendergast), USA Patent No. 4.898.694 (granted to Prendergast on September 11, 1990) ), and U.S. Pat. No. 4,898,694 (Schwartz on February 6, 1990) (granted to).

Brief description of the drawing Figure 1 shows mature adults, aged adults, and Mucosal (deep layer, peri-aortic) (of the circumference, epithelium, Beyer's patches) and non-mucosal (of the viscera, groin, upper arm, armpit) draining lymphoid organ IL-6 levels found in the supracellular groove of activated 1 Luva cells isolated from It is a bar graph.

Bar graph showing IL-6 levels found in plasma of adult mice and untreated old mice. It is f.

FIG. 3 shows that mature adult mice, untreated old mice, and long-term DHEA-8 supplemented mice. FIG. 3 is a bar graph showing serum IL-5 levels found in aged mice that have undergone replenishment. Figure 3 A shows the results for C3H mice, and FIG. 3B shows the results for Balb/C mice.

Figure 4 shows mature adult mice, untreated old mice, and long-term DHEA-3 supplemented mice. FIG. 2 is a bar graph showing serum amyloid protein level 1 in aged mice subjected to hydration.

FIG. 4A shows the results for C3H mice; FIG. 4B shows the results for Ba1b/C mice.

Figure 5 shows that mature adult mice, untreated old mice, and long-term DHEA-8 supplemented mice. FIG. 2 is a bar graph showing serum 1gG isotitility levels observed in old mice subjected to hydration. .

Figure 6 shows bacterial infection before bacterial infection, after bacterial infection, and when pretreated with DHEA. FIG. 2 is a bar graph showing blood fiIL-6 levels in adult mice at later stages. Ru.

Figure 7 shows all Listeria-infected mice, but control , showing fluctuations in serum IL-5 levels of burnt and DHEA-treated burnt mice. It is a graph.

Figure 8 shows MRL/Ipr mice at 10 weeks (untreated) and 14 weeks ( untreated), and serum at 14 weeks (received chronic DHEA-3 supplementation) Figure 2 is a bar graph showing IL-6 levels.

Figure 9 shows the activation of T cells with and without pretreatment with IL-6. The effects of PDGF-BH on IL-2, IL-4 and γ-IFN production are shown. This is a graph.

Figure 10 shows young mice (untreated), old mice (untreated) and old mice. IL-2, IL-4 and T cells from mice (treated with DHEA-8) It is a graph showing the effect of PDGF-BH on γ-IFN production/activation.

Detailed description of the invention As used herein, the term "individual" refers to a propellant and preferably a DHEA-S Includes members of species that exhibit lufatase activity, including livestock, competition animals and It is not limited to primates including humans.

The term "effective amount" refers to the amount by which the DHEA concinin is administered to determine the level of IL-6 in the individual to whom it is administered. to restore normal levels and/or associated with elevated IL-6 levels. The amount of concinin that is sufficient to alleviate one or more pathological symptoms.

As used herein, the term "immunocompromised individual" refers to An individual whose answer is significantly smaller than the average of normal individuals of the same species. “immunodeficiency Methods for determining total the ability of an individual to exhibit contact hypersensitivity, the humoral response to antigenic challenge, or tests the ability of an individual to develop resistance to infection by microorganisms. .

"Treatment" is the administration of a composition to an individual to reduce abnormally elevated IL-6 levels. It refers to things that reduce the effects of cancer, and includes prevention and/or treatment.

An "antigen" is a secretory, humoral and/or cellular antigen that stimulates the host's immune system to produce secretory, humoral and/or cellular antigens. A molecule that has one or more epitopes that elicit a different response.

This term is also used interchangeably with the term "immunogen."

An "immunological response" to a composition or vaccine consisting of an antigen is defined as of a cellular and/or antibody-mediated immune response to a composition or vaccine that Refers to development in the host. Typically, such responses are related to the composition or composition in question. are antibodies, B cells, and helper cells produced by test animals against antigens contained in vaccines. -consist of T cells, suppressor T cells, and/or cytotoxic T cells.

As used herein, the term "prohormone" refers to a water-soluble precursor of DHEA, e.g. For example, DHEA derivatives (e.g. DHE A-5 (and other pond precursors known in the art).

As used herein, "pharmacological dose" means a desired physiological dose when administered to an individual. effects (e.g., reducing abnormally elevated IL-6 levels, particularly in treated individuals) ).

As used herein, ``abnormally elevated levels of IL-6J may be associated with a disadvantage in trauma.'' negative responses (e.g. immunosuppression), adverse responses to infections, and in autoimmune conditions. IL-6 levels associated with the presence of autoantibodies or circulating I associated with e.g. aging Refers to the structural level of L-6.

The term ``decreased levels of abnormally elevated IL-6'' refers to normal adult individuals of the same species. low levels associated with elevated IL-6 levels and/or to levels seen in 1L to a level equal to that associated with the reduction of at least one pathological symptom. -6 level is lowered.

As used herein, "rDHEA compound" refers to the general formula (1): [wherein R is water Elementary atom or halogen atom (including chlorine, bromine or fluorine): R1 is a hydrogen atom or 3020M group or PO20M group (where M is hydrogen or sodium or potassium atom), Or phosphatide group, (In the formula, R1 and R8 are the same or different, and i having 1 to 14 carbon atoms) chain or branched alkyl group) or glucuronide group The broken line in the formula may be a double bond if desired, and the hydrogen atom at the 5th position is α or β configuration] or both configurations. A composition consisting of a mixture of figurines.

The practice of this invention involves the use of molecular biology, microbiology, biochemistry, chemistry, and the like, unless otherwise indicated. Conventional methods of immunology and medicinal chemistry may be used and are known to those skilled in the art. There is. Such methods are well explained in the literature. For example, Animal Center Le Culture (＾NIMAL CELL CtlLTURE) Lee Fre/Uni (R, I.

Freshney), 1986), Immobilized Cells & E. IMMOBILIZED CELLS AND ENZYMES (IRL Press, 1986): Methods I METHODS IN ENZYMOLOGY Kademic Press Inc. , Immunological Meso, Z in Cell and Molecular Biology G (IMMUNOLOGICAL METHODS INCELL AND ll0LECULARBTOLOGY) (Academic Break in London (＾ academicPress, London), Scopes , (1987), Remington's Pharmaceutical Sciences ( Remington's Pharmaceutical 5 Sciences ) 17th edition (1985, Easton, Penn.) Gu Haflishing Company - (liack Publishing C Company): and Handbook of I’) Suhe’) Men 9 ・Imu／　o　　　、；　(HANDBOOli　OF　EXPERIMENT AL　IMMIINOLOGx) Volumes ■ to IV, (D, M, Weir) and N. /--Blackwell (C.C. Blackwell 1), ed., 1986. . All patents, patent applications, and publications cited above and below herein are cited as sources. Incorporated herein by express reference.

In one embodiment, the invention provides a pharmacological dose of a set of DHEA compounds. Abnormally elevated levels of IL-6 in an individual by treating the individual with a compound provide a method to reduce The exact amount required depends on the subject and control Depending on the species, age, and general symptoms, the severity of the symptoms to be treated, the method of administration, etc. It will change. Therefore, it is not possible to specify a precise effective amount. However However, appropriate effective amounts can be determined by those skilled in the art using only routine experimentation. can. As a method for measuring IL-6 levels, for example, immunoassay is known in the art. Some methods are described below. In addition, pathological symptoms In particular, a method to examine the reduction of decrease in the degree of immunodeficiency due to It is known in the art to examine the reduction in physiological evidence caused by autoimmune diseases. There is.

Individuals to be treated are those with elevated levels associated with pathological conditions. for example, polyclonal B-cell abnormalities or autoantibody production (e.g. cardiac mucus). tumor, rheumatoid arthritis, Giersleman's disease, AIDS, alcoholic cirrhosis, whole body Individuals with proliferative disease (proliferative glomerulonephropathy of the mesangium) individuals with acute infectious neurological diseases (viral meningitis, bacterial meningitis) , tuberculous meningitis, herpes simplex meningitis), and individuals with malignant tumors (plasma meningitis). in individuals with cystomas and myelomas, lymphomas and leukemias, and nephrocytosis). be. Other individuals to treat include those with trauma and/or stress and immunity. Individuals with elevated IL-6 levels as a result of epizootic failure, e.g. those with burn injuries , surgical patients, and animals with transport fever. and other individuals to be treated. These include aging individuals with constitutive circulating IL-6 levels. So Additionally, individuals with abnormally high levels of rL-6 associated with allografts may also be treated. (In such an individual, the specimen should be modeled by means known to those skilled in the art.) There are symptoms of rejection of the graft, which can be monitored). Abnormally high IL- Those with 6 levels, those that do not respond to growth factors (below) should be treated Included in the individual. Growth factors are known in the art, particularly PDGF, TG F-β and intulin (in response to these growth factors) Associated diseases are impaired wound healing, osteoporosis, and diabetes, respectively). .

If desired, DHEA can be converted into precursors that are later metabolized to DHEA or its metabolites. It may also be administered to an individual as a body substance. For example, the sulfonated form of DHEA DHEA-3 can be administered. However, tissue-related DHEA administered to individuals who can metabolize prohormones to DHEA by fatase. give The presence or absence of DHEA-sulfatase in specific tissues Probably causes localization of DHEA in specific anatomical sites .

The properties of DHEA concininase and the effects required for regulating IL-6 levels to be obtained. Depending on the amount of time available, ease and/or effectiveness may be considered for method selection. , administration is carried out using the most appropriate method to obtain the desired effect. Therefore, especially oral and intravenous (1,V,), subcutaneous, topical, intranasal and rectal routes known to those skilled in the art. It can be seen that administration is carried out by the specified route.

Generally, a person receiving a composition consisting of DHEA concinina Select the appropriate form of steroid based on its oxidative effect and the resulting metabolites. Choose. For example, when the indication for administration is prophylaxis or long-term treatment. to avoid the side effects associated with long-term administration of high levels of DHEA. is preferably a prohormone DHEA connor (eg DHEA-3).

In this case, the level of DHEA-3 is relatively low (about 5 to about 100 mg per day). range, preferably about 10 to about 80 mg per day, even more preferably 1 It may range from about 15 to about 60 mg per day.

Alternatively, if the indication for treatment is acute trauma or acute infection, a single Treatment with high doses of DHEA or higher is preferred. Giant circle Drug administration ranges from about 1 to about 20 mg per kg body weight, more usually per kg body weight. ranging from about 2 to about 10 mg per kg, and preferably about 3 mg per kg of body weight. It can be administered in a range of 8 to 8 mg.

In some conditions, two or more methods of administration and/or two or more More DHEA compounds can be used simultaneously or sequentially to achieve the desired concentration of active ingredient. Systemic administration is preferred.

Administration of DHEA concinina, which is an IL-6 antagonist in vivo Other drugs and/or elevated IL- Simultaneously or sequentially with other drugs for the reduction of symptoms of diseases related to level 6 It is also understood that anything that can be done is within the scope of the invention.

Other embodiments of the invention provide methods for reducing abnormally elevated levels of IL-6 in an individual. This is a composition of various types. Some compositions include a pharmaceutically acceptable dose of DHEA. (excluding DHEA or DHEA-5). Other compositions include DHE A concina- (including DHEA or DHE A-S) and elevated IL -6 levels at least effective in reducing or preventing pathological responses associated with levels. Consists of one or more other drugs. Both concinina and other drugs Present in a pharmaceutically acceptable amount. Antagonists of rL-6 belong to these drugs Ru.

As an antagonist of IL-6 activity, it can be used to treat diseases associated with increased IL-6 activity. Drugs that do not cause IL-6 to respond to normal receptors that contribute to physical responses, and Examples include drugs that inhibit IL-6 synthesis. Therefore, the antagonism of IL-5 activity Examples of antagonists include those that bind to normal IL-6 receptors but are associated with pathological responses. IL-6 analogs lacking or having reduced IL-6 activity, IL-6 Pathology that binds to receptors other than -6 receptors but is associated with elevated l-6 levels Other drugs that reduce the physiological response (e.g., steroids, immunomodulators, growth factors, among others) ): Anti-IL-6 antibodies, IL-6 receptor fragments or analogs, etc. It will be done. DHEA concinina belongs to the antagonists of IL-6 activity. Therefore , some compositions consist of at least two different DHEA conciniars. Small A composition consisting of two different DHEA concinnatants is an IL-6 antagonist. It may consist of strikes. In addition, at least one kind of DHEA concinina-1 and and/or allow the reduction of abnormally elevated IL-6 levels and/or from synergists to reduce pathological symptoms associated with elevated IL-6 levels. Also within the scope of this invention are compositions that include: Examples of the latter include, in particular, growth factors (e.g. P DGF and TGF-β).

Typically, the composition also comprises pharmaceutically acceptable excipients. pharmaceutically acceptable The excipients used will depend, in part, on the mode of administration, but these are known in the art. There is. For example, Remington's Pharmaceutical Sciences (Remington) Inton's Pharmaceutical 5 Sciences), No. 17th edition (Easton, Pennsylvania) llack Publishing Cospany (1985). Examples of suitable pharmaceutically acceptable carriers include, among others, common pharmaceutical carriers. iline and non-toxic saline solutions, glucose solutions, and oils at or near physiological concentrations. Liquid carriers such as fats, as well as human-use substances such as talc or sucrose. solid carriers, and even livestock feed.

The composition comprising the DHEA compound of the present invention can be administered enterically, parenterally or locally. For topical administration, or for implantation, or for nasal spray or spray, or It may be formulated for transdermal application.

Examples of suitable oral dosage formulations include, for example, hard or soft gelatin capsules, sugar drugs, pills, tablets including dragees, elixirs, suspensions, syrups or sprays and and their sustained release forms.

Suitable topical formulations include creams, gels, jellies, serums, pastes and Examples include ointments. The compound is formulated for transdermal administration, e.g., in the form of a transdermal batch. Good too.

Suitable injection solutions include subcutaneous and intramuscular injection solutions.

Another embodiment of the invention is for use in treating individuals with abnormally elevated IL-5 levels. a set of one or more DHEA containers in the manufacture of a medicament for use in It is the use of synthetic products. Furthermore, the present invention provides a method for treating abnormally elevated IL-6 levels and/or or in the manufacture of pharmaceuticals used for the treatment of pathological symptoms related thereto. more than one DHEA antagonist and at least one IL-6 antagonist Provided is the use of a composition comprising:

Examples of the invention are described below, but are only for illustrating the invention, This is not intended to limit the scope of the invention. In this disclosure, a number of items that fall within the scope of the claims are Many specific examples will be obvious to those skilled in the art.

! ! Yutaka Example I IL-6 lymphokine production ability in responsive lymphocytes of old mice induced by DHEA restoration of normal regulation of This study demonstrated that the lymphoid organs obtained from various mucosal or non-mucosal lymphoid organs of old mice The ability of activated cells to produce IL-5 is probably due to aging-related endogenous DHEA production. was shown to be associated with a decrease in

C57BL/6 strain mice were purchased from the National Cancer Institute ( The bait was purchased from National Cancer Institute (National Cancer Institute). Vergity of Utah Vivarium The animals were raised in a tah (Vivarium). In the Kono experiment, mature adult mice The old mice were 22 weeks old and the old mice were 117 weeks old. Furthermore, anti-CD3. after stimulation 100 weeks for 17 weeks before lymphatic vessel isolation for analysis of IL-5 production. Groups of aged C57BL/6 mice were supplemented with 1 oug/ml DHEA in their drinking water. I gave it to you. Single cell suspensions were isolated from specific lymphatic vessels obtained from groups of 3-4 mice. 1% Neutride-7 (Neut), washed twice with sterile balanced salt solution, ridoma) -NS (Boehringer Mannheim Mannheim), antibiotics, L-glutamine 200mM and 2-methane 1X10' in RPM 11640 supplemented with 5X10'M lucaptoethanol. Re-turbidate the cells/m+/well in a 24-well partitioned plate (maper well). Coas, Cambridge, Sachusetts (manufactured by Ter). Anti-CD3. (1.5μg/ml) Antibody polyclonal Used as a T cell activation agent and then incubated at 37°C for 24 hours in a humidified incubator. Incubated for a period of time. The culture supernatant was removed, clarified by centrifugation, and IL- Stored at 4°C until 6 assays.

Van Sunisok et al. (Journal of Experiments) Tal Medicine 0. Exp, Med, ) Vol. 165, p. 2641 (198 IL-6 bioactivity was evaluated by the method 7)). T L-6 dependent indicator The cell line B9 was incubated in predetermined concentrations of recombinant IL-6. I transplanted it every 3 days.

The test supernatant was washed 1X106 times in RPM11640 medium supplemented with 10% FC3. 9 cells were serially diluted into culture wells. 72 hours of incubation During the last 2 hours, 5 μg of MTT was added to each culture. Then each Solubilize the contents of the culture well and place it in a Vmax microplate. spectrophotometer (Menlo Park, California) Absorbance was measured using a device (manufactured by Recular Devices). Standard human recombinant IL-6 The amount of IL-6 in the culture supernatant was determined based on the response of B9 cells to the dose-response curve of p Expressed in g/m+.

Mature adult mice, old mice, and old mice receiving long-term DHEA-S supplementation. Donor mucosal (deep neck, peri-aortic, epithelial, Beyer's patch) and non-mucosal ( Activated isolated from the draining lymphoid ducts of the pancreas, groin, upper arm, armpit) Representative experimental results showing IL-6 levels found in the cell supernatant of lymphoid cells are shown in the figure. Shown in 1. In vivo anti-CD3. Most of the phosphorus from old mice after activation A clear increase in T cell IL-6 production was observed in the papilla-like organs. related to aging Most dramatic changes in IL-6 production associated with mucosal draining lymphoid Occurred in the organs and pancreas. DHEA-5 supplementation in old animals can improve this aging relationship. Phenotypic changes are facilitated and seen in mature adult lymphoid cell donors. The level was completely the same as l-6 production.

DHEA-3 replacement therapy induces I (Daynes) and Araneo ( ^raneo), in press), as well as IL-6 production, which increased significantly with aging. This complete anatomical analysis is intended to correct changes at the raw level. This is strongly suggested by the results of Moreover, supplemental use of DHEA -6 production capacity appears to be restored to its normal localization (thereby, this hormone tissue-specific end organ metabolism occurs). Applicants believe that this localization of cellular functions is the most effective form of irritation in both mucosal and non-mucosal tissues. It is assumed that a state response is provided to the host.

Example 2 Short-term treatment of old mice with either DHEA or DHEA-3 normally restores plasma IL-6 levels As an important stimulant of the acute phase response, IL-6 contributes to the host's natural defense mechanisms. It plays an important role in However, constitutive IL- The presence of 6 indicates that this lymphokine is abnormally regulated. . This results in either an increase in the rate of biosynthesis or a decrease in the rate of utilization and catabolism. caused by more. The presence of constitutive IL-6 in the blood is not normal. HIV infection Patients (Honda et al., Journal of Immunology (J, I) +wuno1. ) Volume 145: p. 4059 (1990)), Burn patients (Both Pos et al., Clinical and Experimental Immunology ( CI in, Exp, Immunol, Volume 82: 579 (1990 ), tumor patients (Hirano et al., Immunology Today), tumor patients (Hirano et al. m+muno1. Today) Volume 11: 443 (1990)), and Autoimmune disease patients (Hirame et al., Immunology Today Vol. 11: p. 443) The presence of IL-6 in plasma was reported in 1990). The presence of L-6 may be responsible for some of the clinical evidence associated with these diseases. Ru. Applicants' studies show that "normal" aging is associated with constitutive IL-6 in serum/plasma. associated with the presence of DHEA and that abnormally elevated levels may be due to DHEA This indicates that the process will return to its original state.

According to this institute, the active metabolite T3HEA or the precursor form DHEA-3 Acute treatment by giving 100 μg of either to old mice by subcutaneous injection over a short period of time. The results showed that blood levels of IL-6 decreased.

Naive Balb/C mice at 24 and 120 weeks of age were randomly selected. -Together The same number of old Ba1b/C mice were given 0.1 ml of propylene glycol. Inject 100 μg DHEA or 100 μg DHEA-8 subcutaneously in a gel carrier. Ta. After 24 hours, samples were obtained from each treated and untreated mouse. Serum IL-6 concentration was assessed.

Rat/anti-murine IL-6 antibody was added to appropriate cells adapted to grow in serum-free conditions. PharMingen ) (San Diego, California). child These reagents were prepared according to the manufacturer's instructions and according to Schumacher's instructions. (Journal Male Immunology Vol. 141: p. 1576 (1988) ) was used for the quantification of murine IL-6 by ELISA. Simple To simply explain, 2ug/m in 0.05m Tris-HCI (pH 9,6) Aspirate 100 μl of the appropriate capture antibody of I into a 96-well microtest plate. The cells were dressed, washed, and treated with PBS/1% BSA. Test supernatant and appropriate control Dispense 2-fold dilution of tokine (100 μm/well) into the wells and incubate thoroughly. After washing, each well was biotinylated at 100 μm/well. Detection antibody (1 μg/ml) was dispensed. Anti-avidin-HRP and ABTS groups ELISA was performed using the quality. V max 96 unit micro test play Spectrometer (Molecular Devices, Menlo Park, California) The absorbance at 405 nm was measured using a commercially available commercially available product. The detection limit for this cytokine is 1 It was 5 to 30 pg/mI.

The results are shown in Figure 2. 60 pg/m1 or less of IL-6 in mature adult Ba1b/ found in the plasma of CV mice, but more than 600 μg/ml of IL-6 was found in untreated mice. Measured in old Ba1b/c mice. D HE A and DHEA-3 treatment The analytical value of IL-6 in the serum of treated old Ba1b/c mice was similar to that of mature adult mice. The amount was similar to that of IL-6. Due to decreased TL-6 serum levels in old mice As can be seen, both DHEA and DHEA-3 have been shown to be differentially active in old mice. This means having the ability to modulate the kinetics of normal IL-6 production and/or utilization. This is shown by the findings of

Example 3 Mice supplemented with DHEA-3 for a long period of time have lower serum IL-6 and serum amino acids. Shows normal levels of Roid P (SAP) and serum immunoglobulins.

IL-6 is an effective stimulator of acute phase protein production by hepatocytes. Moreover, there is increased serum immunoglobulin and autoantibody production associated with pathological conditions in species. There is also increasing evidence that IL-6 plays a role. Consequences of normal aging As a result, acute phase proteins increase in plasma. This is a natural defense mechanism It has been reported that it is associated with impaired function. In addition, elderly y) Levels of all types of serum immunoglobulins in donor serum is on the rise. Each of these clinical findings suggests that age-related IL-6 synthesis may changes in developmental regulation (which Applicants found to be present in elderly donors) more likely to be caused. This study tested mature adult mice, untreated old mice, and Quantitative measurement of IL-6 in the serum of old mice that received long-term DHEA-8 supplementation It shall be determined. For serum samples, serum amyloid P (subacute or (indicative of persistent or repeated acute phase responses) and immunoglobulin content; Quantitative analysis of immunoglobulin subclasses was also performed.

C3H/HeN MTV- and Balb/C strain mice were used in these experiments. Ta. At the time of blood collection, female adult C3H mice were 22 weeks old and female B a1b/c mice were 24 weeks old. First set of female old C3H and B The a1b/c mice were 93 weeks old, while the second set of Ba1b/c mice He was 12,020 weeks old. - all C3H and Ba1b/C reared together; Mice were given 100 μg/ml DHEA-3 in their drinking water for 9 weeks (at 93 weeks of age). subjects) or for 52 weeks (12,020 week old a1b/c mice). of each experimental value The n value is shown. Schuwacher (Journal O by a modification of the method of Sus Immunology Vol. 141: p. using reagents purchased from Pharmingen (San Diego, CA). Serum IL-6 was quantified by performing a capture ELISA. previously published Reports (Gahrfng, Dayne S, et al.) Roshidinges Os National Academy Os Sciences (Pr. oc, Natl, Acad, Sci, ) Volume 81: 1198-1202 (1984)) serum amyloid P was quantified using radial immunodiffusion. . The amount of globulin subclasses in serum was determined using standard isotype reagents (Arabic Southern Biological in Birmingham, MA (manufactured by Tern Biological), using the protocol indicated by the manufacturer. Capture with horseradish peroxidase-conjugated antibody with slight modification of call Measured by EL I SA.

The results of this study demonstrate that dysregulated IL-6 production in elderly mice may occur during the acute phase. The idea that it is responsible for increased levels of reactants and increased immunoglobulin levels. is supported. Normal old mice have a very clear concentration of constitutive protein in the plasma. -6 levels and serum amyloid P content (Figures 3 and 4). moreover , aging animals show a significant increase in the abundance of all immunoglobulin isotypes. (Figure 5). Three of these clinical trials after 9 weeks of DHEA-3 supplementation of drinking water All findings are similar in phenotype to untreated adult mice. It turns out that it is possible to go backwards. These results indicate that DHEA-3/DHEA Suggested that it is necessary for the regulation of physiologically normal endogenous IL-6 biosynthesis It is. Furthermore, these data indicate that long-term DHEA-3 administration common in elderly animals, including natural defense mechanisms and the presence of autoantibodies. There are some clinical features that are good (meaning that they can be treated).

Example 4 DHE A treatment of normal and burn-injured mice Induced by infection with Listeria monocytogenes increases resistance to elevated serum IL-6 levels Bacterial or viral infection results in stimulation of IL-6 production in the plasma. got burns infection results in elevated plasma IL-6 levels that persist for a long time. Continue. In fact, sustained IL-6 levels in burn patients are associated with mortality. (Lephant et al., Journal Male Clinical End) Clinology and Metabolism (J, Cl1n, Endocrino Metab, Vol. 64: 842 (1986)). According to the research below By administering DHEA, burn-injured mice were cured after infection with Listeria mononotogae. It was confirmed that it is possible to regulate the plasma IL-6 level of the human body.

Male and female C3H/HeN MTV- From National Cancer Institute University Male Utah Vivarium (University Male Utah Vivarium) with purchased bait The animals were raised in Utah Vivarium. 4 in the 6-8 week age range Groups of ~6 animals were used in all experiments, age and sex matched.

The virulent strain of L. monocytogenes is Keith Bishop (Keith B15h). OP) was a gift from Dr. (University of Utah). The bacteria were treated with tributicase-soy broth. (trypticase-soy broth) (Cokesbee, Maryland) BBL-Microbiorono Systems in Cockeysville (manufactured by BBL Microbiology Systems), 10' at -70 °C in a saline until use as colony-forming units (CFU). saved.

Guide for Care and Use of Male Laboratory Animals (G uide for the Care and Use of Laborat ory Ania + als) (D HEW)”/<bu lagoon■ Noyon NIH78-23, revised in 1985) regarding animal trials Hot water burns were inflicted using standard methods according to guidelines. Chloral After general anesthesia with hydrate, body hair was removed using clippers and a depilatory agent. 2 days Afterwards, the animals were deeply anesthetized with methquinoflurane, which covered 20% of the total surface area (TBSA). caused burns. Apply hot water maintained at a constant temperature of 70℃ to the skin of the back for 6 seconds using a mold. Burns were caused to the exposed areas. spanning the entire skin thickness using this protocol Burn injuries are disclosed by histological examination. Size of the burn relative to the entire surface is the measurement of dry leprosy and Meeh's formula.

A=KW! /3 [where A=area expressed in square centimeters, K=8.95. W = table in Durham ] Calculated using Liquid consciousness recovery agent in regular sterile saline to burn mouse belly Intracavitary injection. 2ml for the first 24 hours, then 1ml on the first and second day. In addition, the mice were given food and water ad libitum during the experiment. There is no odor for several hours after the burn. in a controlled environment under a warming lamp. All animals were kept at home to help maintain body temperature.

The experiment illustrated by the data in FIG. Infection of normal adult mice with S. monocytogenes results in significant IL- 6 responses. In addition, due to DHEA To investigate whether the increase in IL-6 levels that accompanies such infection can be prevented. It belonged to Crystallized IL of a group of adult C3H/HeN mice before experimental manipulations. -6 level was quantitatively evaluated. Mice were then further treated with untreated and DHEA divided into treatment groups. Subcutaneous injection of 100 μg DHEA in propylene glycol was given to the DHEA-treated group. Treated and untreated mice were then exposed to 2X infected with 106 live L. mono/togenes, and 24 hours after infection, one individual Blood samples were obtained from mice. The mean serum IL-6 levels in the plasma of mice in the untreated group It was found to be approximately 3000 pg/ml as a result of the infection. But long The group received 100 μg of DHEA by subcutaneous injection 24 hours before infection. of mice whose IL-6 levels are at levels observed in pre-infected mice. It was close.

Mice of the C3H/HeN strain naturally carry a small amount of Listeria monocytogenes. It is resistant to the deadly effects of infection by However, infection and burn injuries Together, this strain of animals significantly increases the susceptibility to this pathogen. (data not shown). After infection with this organism, there is a sudden increase in plasma IL-6. Because of the sexual response, Applicants have shown that normal C3H/HeN mice are susceptible to burn injury. It was hypothesized that normal blood regulation of IL-6 would be impaired. normal and burn-injured mice were previously described (Merril et al., American Jar Naru of Surgery (^+a, J, Surg,) Volume 156: 62 3 (1987)). Half of all scalded animals have post-scald 1 Within hours, one injection of 100 μg DHEA in propylene glycol was given.

After 3 days, all mice were inoculated with 2 x 10 live L. monotogenes. Infected.

Listeria infection in control, burnt, and DHEA-treated burnt mice. Kinetic evaluation of serum IL-6 levels induced by control and DHE It was shown that there was a similar response in the A-treated burn group (Figure 7). Targetally, Serum IL detected in burnt mice infected with Listeria monocytogenes -6 levels were significantly elevated and maintained significantly.

Example 5 Abnormally elevated serum IL-6 levels in a murine model of autoimmune disease IL-6, which can be effectively regulated by HEA-5, is involved in hematopoiesis, acute phase response, and It is a multifunctional cytokine that plays a central role in immunoglobulin production. It has been proven that (Van 5nick), annual Review of Immunology (^nn, Rev.

1 dragon uno1. ) Volume 8 = 253 pages (1990)). This lymphokine have been implicated in contributing to the pathology of certain autoimmune diseases (Flounder (Hfra) no) et al., Immunology Today (I+1Iluno1.Today) No. 11:443 (1990)).

MRL/Ipr is an inbred strain of mice, all of which were born young (approximately 14 The disease is characterized by hyperplasia, arteritis, myocardial infarction, arthritis, and premature death. autoimmune diseases (Thiofilapoulos) ) and Dixon, Advanced Immunology (Adv, I Immunol, Volume 37: Page 269 (1985)). The site of injury to the glomerulus is , contains monocytes with both endothelial and mesangial proliferation (thiophila Poulos and Dixon, Advanced Immunology Volume 37: p. 269 (1) 985)). The role of IL-6 in glomerular development is currently being investigated; Autocrine growth factors for glomerular mesangial cells It is believed that this is evidenced by the ability of IL-6 to serve as a child (Holey). Horii) et al., Journal of Immunology (J.

Immunol, Volume 143: Page 3949 (1989)). This thing is , IL-6 transgenic mice develop mesangial proliferative glomerulonephritis. have a tendency to rub (Hirano et al., Immunoronau toudie). munol, Today) Volume 11 = Page 443 (1990)) This is supported by the findings in Bo. The MRL/Ipr strain is associated with clinical disease. It is believed to provide a useful animal model for the 6 levels. This stock's ma long-term DHEA supplementation before clinical changes occur in these animals. Determine whether it is possible to suppress the increase in serum IL-6 levels seen in Provide a model for 10 week old MRL/I pr has no disease. It seems that it is. However, by 14 weeks of age, significant lymph node damage is evident. Histological examination of the kidney revealed a clear glomerular kidney with mesangial proliferation. A flame is shown.

Ten MRL/lpr mice, 11 weeks old, were divided into two groups. These maw DHEA-3 supplementation (100 μl/mL in drinking water) was started in half of the patients, while also Half of the mice received no treatment. When these mice reached 14 weeks of age Blood was collected from all treated and untreated mice at points. Additionally, phenotypically normal Blood was collected from 5- to 10-week-old retired L/Ipr mice of the group to determine IL-6 levels. was measured.

Serum IL-6 levels were measured using a standard capture ELISA (Sch. umacher) et al., Journal of Immunology (J, Immunol , Volume 141, page 1576 (1988)).

Before signs of disease appear, young MRL/ipr mice (10 weeks old) have minimal serum T showed L-6 levels (Figure 8). 14 week old mice without DHEA-3 treatment exhibits characteristic signs of an autoimmune syndrome and at the same time has very high levels of serum 1L -6 was expressed. 14-week-old MR who received long-term DHEA supplementation from 11 weeks onwards. Serum IL-6 level of L/l pr showed no signs at the time of serum collection. It was within the normal range for the 10 week old group.

This finding suggests that DHEA-3 supplementation may be associated with certain types of autoimmune conditions. effective therapeutic forms used to prophylactically modulate elevated blood ff11L-6 levels. This shows that it can be useful as a mode of action.

Example 6 IL-6 affects cellular responsiveness to PDGF; DHEA- 8 administration restores PDGF responsiveness.

Platelet-derived growth factor (PDGF) is produced and released after stimulation of various cells. A small group of effective cytokines (Ilart et al., J.D. Journal of Investigative Dermatology (J, Invest , Dermatol, Volume 94: 53s (1990)). PDGF too It is also present in platelet granules. Main PDGF released during inflammation The main source is platelets. The most abundant PDGF isoform (i) in human platelets. sofor++) are PDGF-BB and PDGF-AB (Bowen Boven-Pope et al., Journal of Biological Chemistry Tory (J. Biol, Chem.) Vol. 264, p. 2502 (1989 Year)). PDGF can be produced in specific tissue microenvironments (e.g. tumors). Ru. All three isoforms of PDGF play a role in chemotaxis and proliferation of various forms of target cells. have been studied for their ability to induce reproductive responses (Hart et al., Journal of Investigative Dermatology Volume 94: 53s (1990)) .

These molecules also contribute to angiogenesis (Hart et al., Journal of Investigation Tibetan Dermatology Volume 94, 53s (1990)) and Wound Healing ( Hart et al., Journal of Investigative Dermatology No. 94 Vol. 53s (1990)).

Growth factor-dependent wound healing and vascular morphology in aging or severely injured individuals formation and other repair processes are moderately to fairly well coordinated. There is. These clinical symptoms may be due to maintained blood levels of this lymphokine. Because of the correlation with the apparent age-related dysregulation of IL-6, Applicants proposed that IL-6 We decided to investigate whether there is a relationship between the presence of -6 and growth factor responsiveness. Ta.

C57BL/6 donor mice were sacrificed and their lymph node cells were divided into IL-6 pretreated groups and and an untreated group. The treatment consisted of long/ml human recombinant IL-6 and short The procedure consisted of an incubation period of 1 hour at 37°C. before this Following treatment, cells were washed in balanced salt solution and serum-free RPM11640 (1 % Nutrisome SR (Boehringer-Mannheim) supplement), lX1 Reconstituted into 07 pieces. lX107 cells were incubated with a specified concentration of human recombinant (hr ) Anti-C with or without PDGF-BB (Boehringer Mannheim) It was distributed into 24-well macrowells along with 1.5 μg of D3. PDGF-B The range of the amount of B was 2.0 ng/m+ or 2.0 ng/m+. at 37℃ After 24 hours of incubation, cell-free supernatants were obtained and cytokines l-2, I L-4 and γIFN were quantified. IL-2 value (unit/ml) was determined by bioa IL-4 and γI FN (pg/m) in the culture supernatant were measured by assay, Previously reported (Daynes et al., Journal of Experimental ・Medicine (J.

EX+), Med, Volume 174: 1323 (1991)) It was measured by capture ELI SA as follows.

The data in Figure 9 show the effect of PDGF-BH on murine T cells upon activation (de Ing et al., Journal of Experimental Medicine, Vol. 174, 13. 23 (1991)). IL-4 of activated T cells and γIFN production capacity decreases, while IL-2 production by activated lymphocytes decreases. It is clear that there will be an increase. These effects are based on the effects published by this laboratory. , indicating that PDGF has a significant effect on T cell function. It is. IL-6 pretreatment in this in vivo model results in increased response to stimulation. The effect of PDGF-BB on T cell production of lymphokines was completely suppressed. Ru. These results demonstrate that target cells in the microenvironment where inflammatory cytokines are present , especially cells that exert receptor function for both cytokines and growth factors. If IL-6 is present, it suggests that scales have lost their responsiveness to growth factors. .

Because the presence of IL-6 suppresses T cell responsiveness to PDGF, Thus, Applicants have demonstrated that the physiological symptoms are associated with IL-6 dysregulation. We decided to investigate whether or not T lymphocytes derived from humans can respond to growth factors. heavier In summary, Applicants demonstrate that short-term DHEA administration does not result in any growth factor response failure. I decided to find out if it was possible to recover.

Lymphocytes were cultured in mature adult C3H/HeN donor mice (13 weeks old) and 1 of 2 groups. Prepared from 00 week old aged C3HeN. Group 1 received 10 doses 24 hours before the start of the experiment. 0 μg of DHEA-5 was injected subcutaneously. have dysregulation in IL-6 production Lymphocytes from mice of the old C3H/HeN strain, which is known to be - Anti-CD3 with or without BB. Cultures were performed as described above.

After 24 hours of culture, specific cytokine levels were measured in the supernatant. Figure 10 shows normal T cells from adult animals are sensitive to the effects of PDGF; - shows that normal T cells derived from - are not sensitive to the effects of this growth factor. .

The results of this experiment also demonstrated that a single injection of 1 ooμg DHEA-3 24 hours before sacrifice Normal lymphokine and Figure 3 shows a dramatic recovery of PDGF and PDGF responsiveness.

Hitoku Toyoyu DHEA-8 supplementation in all mice reduces serum autoantibody activity during aging. The effect of long-term DHEA supplementation on autoantibody activity in three groups of C This was investigated by comparing the autoantibody activity of 3H/HeN mice. The three groups are ( 1) Untreated normal animals (12 to 14 weeks old, n=8): (2) Untreated old animals ( (22 months of age or older, n=8); and (3) DHEA for 8-9 weeks before serum collection. -3 (100 μg/ml in drinking water) old animals (22 months of age and older).

n; 8).

Autoantibody activity in serum was determined from brain, lung, liver, and Tested by immunohistological method using multi-organ frozen section specimens including kidney and thymus. I put it out. Frozen sections were cut on a cryostat and stored at -20°C until use. .

Frozen sections were washed 3 times with cold saline and washed at a ratio of 1:100 to 1:6000. Various serum dilutions were placed in the sample and kept at room temperature for 2 hours before being washed with cold saline. Washed. Peroxidase-labeled goat/anti-mouse immunoglobulin at desired dilution. The blood was reacted with Robulin reagent for 1 hour at room temperature, and then developed with appropriate color. Autoantibody activity in the serum was measured. Next, Kato and Hiro Kawa ( Hirokawa) (Aging (＾ging): Immunology and... Ivaunology and Infect ious Disease) Volume 1 = Page 117 (1988)) The presence of autoantibodies was examined microscopically using the same method.

The results in Table 1 show that normal Confirmed the existence of a significant amount of autoantibodies that reacted with multi-organ tissue specimens from mice. It was done. In contrast, patients received DHEA-3 supplementation for 8-10 weeks before serum collection. Older animals (22 months of age) have much lower antibody titers (lower than those seen in mature adult control donors). (close to the same value).

Table 1 of mature adult mice, untreated old mice, and old mice receiving DHEA-8 supplementation. Normal organ-reactive autoantibodies in serum age age age Average autoantibody titer 1:250 1:5500 1:400 Commercial utility The compositions and methods of the present invention can be used to reduce abnormally elevated IL-6 levels in individuals. Can be used for treatment. Additionally, one or more DHEA concinina - A composition of the present invention comprising: treatment of individuals with abnormally elevated IL-6 levels; It is useful in the production of medicines used in Additionally, one or more DHEA components Compositions comprising a TL-6 antagonist and at least one TL-6 antagonist also include , for the manufacture of medicaments for use in the treatment of individuals with abnormally elevated IL-6 levels. Useful.

Ku Mucosal lymphoid tissue Non-mucosal lymphoid tissue n=6 n=6 n=6 Mature adult, elderly, elderly DHEAS long-term supplementation FIG, 3A n=19 n=12 n=10 Mature adult, elderly, elderly DHEAS long-term supplementation FIG. 4A DHEAS long-term supplementation FIG. 4B treatment person treatment old FIG. 6 Days after burn FIG.7 DHEAS long-term supplementation Age of MRL/lpr mice FIG.8 PDGF-88 Ko Iyi (nglml) PDGF-883 degrees (nc7m PDGF-BBJff (ng/m1) Ffj’r　8’Ej’r　8’E? 'z 8 'E' Ω ~ to - − Continuation of front page (81) Designated countries EP (AT, BE, CH, DE.

DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE) , 0A (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN.

TD, TG), AT, AU, BB, BG, BR, CA.

CH, CZ, DE, DK, ES, FI, GB, HU, JP, KP, KR, LK, LU, MG, MN, MW, NL, No, NZ, PL, PT, RO, RU, SD, S.E.

SK, UA, VN

Claims (32)

[Claims] 1. Administering to an individual a suitable composition comprising a pharmacological dose of a DHEA congener. A method for treating an individual by reducing an abnormally elevated IL-6 level, characterized by . 2. Additionally, determining whether an individual has abnormally elevated IL-6 levels The method according to claim 1, characterized in that: 3. 2. The method of claim 1, wherein the elevated level of IL-6 is associated with trauma. 4. The person according to claim 3, wherein the trauma is caused by a burn and is associated with an immunodeficiency. Law. 5. 2. The method of claim 1, wherein the elevated levels of IL-6 are constitutive. 6. 6. The method according to claim 5, wherein the individual is old. 7. 3. The method of claim 2, wherein the elevated levels are associated with an autoimmune disease. 8. consisting of a pharmacological dose of a DHEA congener in a pharmaceutically acceptable excipient. A composition for treating an individual by reducing abnormally elevated IL-6 levels. 9. 9. The composition of claim 8, wherein the elevated levels of IL-6 are associated with trauma. 10. 9. The method of claim 8, wherein the trauma is caused by a burn and is associated with an immunodeficiency. Composition. 11. 9. The method of claim 8, wherein the elevated level of IL-6 is constitutive. 12. 9. The method of claim 8, wherein the individual is old. 13. 9. The method of claim 8, wherein the elevated levels are associated with an autoimmune disease. 14. Mixing a pharmacological dose of DHEA congener with a pharmaceutically acceptable excipient To treat an individual by reducing abnormally elevated IL-6 levels consisting of A method for producing a composition. 15. 14. The method of claim 13, wherein the composition is used in the method of claim 2. 16. 14. The method of claim 13, wherein the composition is used in the method of claim 3. 17. 14. The method of claim 13, wherein the composition is used in the method of claim 4. 18. 14. The method of claim 13, wherein the composition is used in the method of claim 5. 19. 14. The method of claim 13, wherein the composition is used in the method of claim 6. 20. Administering to an individual a suitable composition comprising a pharmacological dose of a DHEA congener. abnormally elevated IL-6 levels associated with B-cell disorders, characterized by Methods of treatment for individuals that reduce. 21. 21. The method of claim 20, wherein the B cell disorder is malignant. 22. 22. The method of claim 21, wherein said disorder is a myeloma. 23. 22. The method of claim 21, wherein the B cell disorder is B cell lymphoma. 24. Administering to an individual a suitable composition comprising a pharmacological dose of a DHEA congener. It is characterized by reducing abnormally elevated IL-6 levels and acting on growth factors. A method of treating an individual that restores cellular responsiveness. 25. 25. The method of claim 24, wherein the growth factor is PDGF. 26. The method according to claim 25, wherein inhibition of wound healing is removed by restoration of cellular responsiveness. Law. 27. 25. The method of claim 24, wherein the growth factor is TGF-β. 28. 27. The method of claim 26, wherein osteoporosis is reduced by restoring cellular responsiveness. 29. Claim 6: Treatment with a DHEA congener reduces autoantibody activity. the method of. 30. Claims that immunoglobulin levels decrease due to treatment with DHEA congeners The method described in Section 6. 31. Claims that treatment with DHEA reduces metabolites associated with acute phase responses The method described in 6. 32. 33. The method of claim 32, wherein the metabolite is serum amyloid P.