| PATENT NUMBER | This data is not available for free |
| PATENT GRANT DATE | September 28, 1999 |
| PATENT TITLE |
Method for treating multiple sclerosis |
| PATENT ABSTRACT | A method for treating multiple sclerosis, through the administration of anti-tumour necrosis factor antibody, of soluble tumour necrosis factor receptor or of a compound capable of blocking tumour necrosis factor production, its effects and/or tumour necrosis factor receptor signal transduction, is disclosed. The method can be used to aid in therapy for humans and other mammals |
| PATENT INVENTORS | This data is not available for free |
| PATENT ASSIGNEE | This data is not available for free |
| PATENT FILE DATE | March 13, 1996 |
| PATENT CT FILE DATE | July 30, 1993 |
| PATENT CT NUMBER | This data is not available for free |
| PATENT CT PUB NUMBER | This data is not available for free |
| PATENT CT PUB DATE | February 9, 1995 |
| PATENT REFERENCES CITED |
Ruddle, N. H., et al., "An Antibody to Lymphotoxin and Tumor Necrosis Factor Prevents Transfer of Experimental Allergic Encaphalomyelitis", J. Exp. Med., 172:1193-1200 (Oct. 1990). Selmaj, K., et al., "Anti-Tumor Necrosis Factor Therapy Abrogates Autoimmune Demyelination", Ann Neurol., 30(5):694-700 (1991). Brennan, F. M. and Feldmann, M., "Cytokines in autoimmunity", Current Opinion in Immunology, 4(6):754-759 (Dec. 1992). Gray, P.W., et al., "Cloning of Human Tumor Necrosis Factor (TNF) Receptor cDNA and Expression of Recombinant Soluble TNF-Binding Protein", Proc. Natl. Acad. Sci. USA, 87:7380-7384 (1990). Brennan, F.M., et al., "Enhanced Expression of Tumor Necrosis Factor Receptor mRNA and Protein in Mononuclear Cells Isolated from Rheumatoid Arthritis Synovial Joints", Eur. J. Immunol., 22:1907-1912 (1992). Deleuran, B.W., et al., "Localization of Tumor Necrosis Factor Receptors in the Synovial Tissue and Cartilage-Pannus Junction in Patients with Rheumatoid Arthritis", Arthritis and Rheumatism, 35(10):1170-1178 (1992). Williams, R.O., et al., "Anti-Tumor Necrosis Factor Ameliorates Joint Disease in Murine Collagen-Induced Arthritis", Proc. Natl. Acad. Sci. USA, 89:9784-9788 (1992). Cope, A.P., et al., "Increased Levels of Soluble Tumor Necrosis Factor Receptors in the Sera and Synovial Fluid of Patients with Rheumatic Diseases", Arthritis and Rheumatism, 35(10):1160-1169 (1992). Brennan, F.M., et al., "TNF.alpha..sup.-- A Pivotal Role in Rheumatoid Arthritis?", British J. of Rheumatology, 31:293-298 (1992). Sharief, New England Journal of Medicine 325:467-472 (1991). Teuscher, Clin. Immunol and Immunopathol. 54:442-453 (1990). |
| PATENT PARENT CASE TEXT | This data is not available for free |
| PATENT CLAIMS |
We claim: 1. A method of treating multiple sclerosis in a mammal, comprising administering to said mammal a therapeutically effective amount of a monoclonal anti-tumor necrosis factor alpha antibody which ameliorates the effects of multiple sclerosis, wherein said antibody is administered after onset of the multiple sclerosis. 2. A method of claim 1 wherein the mammal is a human being. 3. A method of claim 2 wherein the anti-tumor necrosis factor alpha antibody is administered in a pharmaceutically-acceptable vehicle. 4. A method of claim 2 wherein a therapeutically effective amount of an anti-tumor necrosis factor alpha antibody is administered directly to the central nervous system of the human being. 5. A method of claim 4 wherein the anti-tumor necrosis factor alpha antibody is administered intrathecally. 6. A method of claim 2 wherein the anti-tumor necrosis factor alpha antibody is an antigen binding antibody fragment. 7. A method of treating multiple sclerosis in a mammal, comprising administering to said mammal a therapeutically effective amount of a soluble tumor necrosis factor alpha receptor which ameliorates the effects of multiple sclerosis, wherein said receptor is administered after onset of the multiple sclerosis. 8. A method of claim 7 wherein the mammal is a human being. 9. A method of claim 8 wherein the soluble tumor necrosis factor alpha receptor is administered in a pharmaceutically-acceptable vehicle. 10. A method of claim 8 wherein a therapeutically effective amount of a soluble tumor necrosis factor alpha receptor is administered directly to the central nervous system. 11. A method of claim 8 wherein the soluble tumor necrosis factor alpha receptor is administered intrathecally. 12. A method of claim 8 wherein the soluble tumor necrosis factor alpha receptor is a binding fragment thereof. 13. A method of claim 8 wherein the soluble tumor necrosis factor alpha receptor is a soluble human p55-tumor necrosis factor alpha receptor. 14. A method of claim 13 wherein the soluble human p55-tumor necrosis factor alpha receptor is a binding fragment thereof. 15. A method of treating multiple sclerosis in a human being, comprising administering directly to the central nervous system of said human being a therapeutically effective amount of an anti-tumor necrosis factor alpha antibody, wherein said antibody is administered after onset of the multiple sclerosis. |
| PATENT DESCRIPTION |
BACKGROUND Multiple sclerosis (MS) is an autoimmune demyelinating disease of the central nervous system which usually presents in the form of recurrent attacks of focal or multifocal neurologic dysfunction. Attacks occur, remit, and recur, seemingly randomly over many years. Remission is often incomplete and as one attack follows another, a stepwise downward progression ensues with increasing permanent deficit. Clinical disease is associated with blood-brain barrier dysfunction; infiltration of the central nervous system by mononuclear cells, mainly macrophages and T lymphocytes, and serum products; and demyelination (Harris J. O., et al., Ann. Neurol. 29:548 (1991); Kermonde A. G., et al., Brain 113:1477 (1990)). Presently the nature of autoantigens responsible for multiple sclerosis is not known, nor is the action which triggers the autoimmune response. One popular theory involves the similarity of a viral protein to a self antigen, which results in autoreactive T cells or B cells recognizing a self antigen. Whereas B-lymphocytes produce antibodies, thymus-derived or "T-cells" are associated with cell-mediated immune functions. T-cells recognize antigens presented on the surface of cells and carry out their functions in association with "antigen-presenting" cells. Currently no practical and efficacious treatments for multiple sclerosis exist. Thus, the development of a method for treating multiple sclerosis would be of immense benefit. SUMMARY OF THE INVENTION The present invention relates to a method of treating multiple sclerosis in a mammal. The invention is based on the discovery that tumour necrosis factor (TNF) has a role in the pathogenesis of multiple sclerosis and experimental allergic encephalomyelitis (EAE). The method comprises administering to a mammal a therapeutically effective amount of an anti-tumour necrosis factor (anti-TNF) antibody which ameliorates the effects of multiple sclerosis. A therapeutically effective amount can be administered in the form of a single dose, or a series of doses separated by intervals of days, weeks or months. Another method comprises administering to a mammal a therapeutically effective amount of a soluble TNF receptor which ameliorates the effects of multiple sclerosis. A therapeutically effective amount can be administered in the form of a single dose, or a series of doses separated by intervals of days, weeks or months. Another method comprises administering to a mammal a therapeutically effective amount of a compound which is capable of blocking TNF production, its effects and/or tumour necrosis factor receptor signal transduction (anti-TNF compound). The anti-TNF antibody, soluble TNF receptor or anti-TNF compound can be administered together with a pharmaceutically-acceptable vehicle. In a preferred embodiment administration of said antibody, soluble receptor or anti-TNF compound is by injection directly into the central nervous system of a human being. Injection directly into the central nervous system can be by injection directly into the lumbar cerebrospinal fluid (intrathecally). In another embodiment administration of said antibody, soluble receptor or anti-TNF compound is intravenously. The present invention further relates to a pharmaceutical composition comprised of a pharmaceutically-acceptable carrier and a multiple sclerosis-therapeutically effective amount of anti-TNF antibody which ameliorates the effects of multiple sclerosis, soluble TNF receptor which ameliorates the effects of multiple sclerosis or anti-TNF compound. The benefit of the method of therapy of the subject invention is that it provides an efficacious treatment for multiple sclerosis. BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a histogram and a graph illustrating the kinetics of weight changes and clinical signs during acute phase chronic relapsing experimental allergic encephalomyelitis (CREAE) induced in Biozzi AB/H mice. FIG. 2 is a histogram illustrating blood-brain barrier permeability to cells and protein during acute phase CREAE. FIG. 3 is a graph illustrating the effect on EAE of a single injection of TNF-specific monoclonal antibody. FIGS. 4A and 4B are a pair of graphs illustrating the inhibition of the development of clinical disease following the injection of TNF-specific monoclonal antibody. FIG. 5 is a pair of graphs showing that anti-TNF, unlike anti-CD4, is not immunosuppressive and does not diminish the proliferative response to the contact sensitizer oxazolone. FIG. 6 is a graph illustrating the dose-dependent inhibition of the progression of clinical EAE following injection of TNF-specific monoclonal antibody directly into the central nervous system. FIG. 7 is a graph illustrating the inhibition of the development of clinical disease following injection of TNF-specific monoclonal antibody directly into the central nervous system. FIGS. 8A-8C are a set of three histograms illustrating the individual clinical grades of 5 different animals (in each group) following injection of TNF-specific monoclonal antibody directly into the central nervous system and intraperitoneally. FIG. 9 is a graph illustrating the greater inhibition of EAE by intracerebral injection of a TNF-specific monoclonal antibody or of a soluble human p55-TNF receptor, than by intraperitoneal injection. FIG. 10 is a graph illustrating the inhibition of EAE by systemic injection of a soluble human p55-TNF receptor. DETAILED DESCRIPTION OF THE INVENTION The present invention concerns the treatment of multiple sclerosis through the administration of anti-TNF antibody, of soluble tumour necrosis factor receptor (TNF-R) or of a compound capable of blocking tumour necrosis factor production, its effects and/or tumour necrosis factor receptor signal transduction (anti-TNF compound). Multiple sclerosis is an autoimmune disease of the central nervous system. The disease is associated with blood-brain barrier dysfunction, infiltration of the central nervous system by mononuclear cells (mainly macrophages and T lymphocytes, and serum products), and demyelination (Harris, J. O., et al., Ann. Neurol. 29:548 (1991); Kermonde, A. G., et al., Brain 113:1477 (1990)). Although CD4.sup.+ T lymphocytes are involved in the induction of the disease (Mokhtarian, F., et al., Nature 309:356-358 (1984); Waldor, M. K., et al., Science 227:415 (1985)), the effector mechanisms mediating pathogenesis of MS are unknown. Tumour necrosis factor (TNF) has been implicated as an important effector molecule in the pathogenesis of various human diseases and animal models such as gram negative sepsis and rheumatoid arthritis (Tracey, K. J., et al., Nature 330:662 (1987); Brennan, F. M., et al., Lancet 2:244 (1989); Williams, R. O., et al., Proc. Natl. Acad. Sci. 89:9784 (1992)). TNF.alpha. is a protein secreted primarily by monocytes and macrophages in response to endotoxin or other stimuli as a soluble homotrimer of 17 kD protein subunits (Smith, R. A., et al., J. Biol. Chem. 262:6951-6954 (1987)). A membrane-bound 26 kD precursor form of TNF has also been described (Kriegler, M., et al., Cell 53:45-53 (1988)). The expression of the gene encoding TNF.alpha. is not limited to cells of the monocyte/macrophage family: TNF is also produced by CD4+ and CD8+ peripheral blood T lymphocytes, and by various cultured T and B cell lines (Cuturi, M. C., et al., J. Exp. Med. 165:1581 (1987); Sung, S. -S. J., et al., J. Exp. Med. 168:1539 (1988); Turner, M., et al., Eur. J. Immunol. 17:1807-1814 (1987)). The term antibody is intended to encompass both polyclonal and monoclonal antibodies. The term antibody is also intended to encompass mixtures of more than one antibody reactive with TNF (e.g., a cocktail of different types of monoclonal antibodies reactive with TNF). The term antibody is further intended to encompass whole antibodies, biologically functional fragments thereof, and chimeric antibodies comprising portions from more than one species, bifunctional antibodies, etc. Biologically functional antibody fragments which can be used are those fragments sufficient for binding of the antibody fragment to TNF. The chimeric antibodies can comprise portions derived from two different species (e.g., human constant region and murine variable or binding region). The portions derived from two different species can be joined together chemically by conventional techniques or can be prepared as single contiguous proteins using genetic engineering techniques. DNA encoding the proteins of both the light chain and heavy chain portions of the chimeric antibody can be expressed as contiguous proteins. Monoclonal antibodies reactive with TNF can be produced using somatic cell hybridization techniques (Kohler and Milstein, Nature 256:495 (1975)) or other techniques. In a typical hybridization procedure, a crude or purified protein or peptide comprising at least a portion of TNF can be used as the immunogen. An animal is vaccinated with the immunogen to obtain anti-TNF antibody-producing spleen cells. The species of animal immunized will vary depending on the species of monoclonal antibody desired. The antibody producing cell is fused with an immortalizing cell (e.g., myeloma cell) to create a hybridoma capable of secreting anti-TNF antibodies. The unused residual antibody-producing cells and immortalizing cells are eliminated. Hybridomas producing desired antibodies are selected using conventional techniques and the selected hybridomas are cloned and cultured. Polyclonal antibodies can be prepared by immunizing an animal with a crude or purified protein or peptide comprising at least a portion of TNF. The animal is maintained under conditions whereby antibodies reactive with TNF are produced. Blood is collected from the animal upon reaching a desired titre of antibodies. The serum containing the polyclonal antibodies (antisera) is separated from the other blood components. The polyclonal antibody-containing serum can optionally be further separated into fractions of particular types of antibodies (e.g., IgG, IgM). Murine hybridomas which produce TNF specific monoclonal antibodies are formed by the fusion of mouse myeloma cells and spleen cells from mice immunized against a TNF positive T cells, purified TNF, or other biological preparations comprising TNF. To immunize the mice, a variety of different protocols may be followed. For example, mice may receive primary and boosting immunizations of TNF. The fusions are accomplished by standard procedures well known to those skilled in the field of immunology. Kohler and Milstein, Nature, 256:495 (1975) and Kennet, Monoclonal Antibodies (Kennet, et al., Eds. pp. 365, Plenum Press, N.Y., 1980). The co-transfected resulting clones are then screened for production of antibody reactive with TNF or biological preparations comprising TNF. Those which secrete antibodies of the appropriate reactivity and specificity are cloned to yield a homogeneous cell line secreting anti-TNF antibody. Human hybridomas which produce monoclonal anti-TNF antibodies are formed from the fusion of B cells from an individual producing anti-TNF antibodies and a human B lymphoblastoid cell line. Alternatively, the fusion partner for the myeloma cell may be a peripheral blood anti-TNF producing lymphocyte. The fusion and screening techniques are essentially the same as those used in the production and selection of murine anti-TNF generating hybridomas. Also mouse and human hybridomas which produce human anti-TNF antibody may be formed from the fusion of a human antibody producing cell and a murine plasmacytoma cell or a cell which itself is a hybrid having the appropriate properties such as the ability to fuse with human lymphocytes at high frequency; support the synthesis and secretion of high levels of antibody; support the secretion of antibody for prolonged periods of time in culture. Another way of forming the anti-TNF producing cell line is by transformation of antibody producing cells. For example, an anti-TNF producing B lymphocyte may be infected and transformed with a virus such as Epstein-Barr virus in the case of B lymphocytes to yield an immortal anti-TNF producing cell. See e.g., Kozbor and Roder, Immunology Today, 4(3):72 (1983). Alternatively the B lymphocyte may be transformed by a transforming gene or transforming gene product. The TNF specific monoclonal antibodies are produced in large quantities by injecting anti-TNF antibody producing hybridomas into the peritoneal cavity of mice or other appropriate animal hosts and, after appropriate time, harvesting the resulting ascitic fluid which contains a high titre of antibody and isolating the monoclonal anti-TNF antibody therefrom. Allogeneic or xenogeneic hybridomas should be injected into immunosuppressed, irradiated or athymic nude mice. Alternatively, the antibodies may be produced by culturing anti-TNF producing cells in vitro and isolating secreted monoclonal anti-TNF antibodies from the cell culture medium. Chimeric anti-TNF antibodies are produced by cloning DNA segments encoding the heavy and light chain variable regions of a non-human antibody specific for TNF and joining these DNA segments to DNA segments encoding human heavy and light chain constant regions to produce chimeric immunoglobulin encoding genes. The fused gene constructs coding for the light and heavy chains are assembled in or inserted into expression vectors. The genes are co-transfected into a lymphoid recipient cell (e.g., a myeloma cell) where the immunoglobulin protein can be synthesized, assembled and secreted. The transfected recipient cells are cultured and the expressed immunoglobulins are collected. A more detailed description of anti-TNF antibodies and their use in treatment of disease is contained in the following references, the teachings of which are incorporated by reference: U.S. application Ser. No. 07/943,852, filed Sep. 11, 1992; Rubin, et al., EPO Patent Publication 0218868, Apr. 22, 1987; Yone, et al., EPO Patent Publication 0288088, Oct. 26, 1988; Liang, C. -M., et al., Biochem. Biophys. Res. Comm. 137:847 (1986); Meager, A., et al., Hybridoma 6:305 (1987); Fendly, et al., Hybridoma 6:359 (1987); Bringman, T. S., et al., Hybridoma 6:489 (1987); Hirai, M., et al., J. Immunol. Meth. 96:57 (1987); Moller, A., et al., Cytokine 2:162 (1990); Mathison, J. C., et al., J. Clin. Invest. 81:1925 (1988); Beutler, B., et al., Science 229:869 (1985); Tracey, K. J., et al., Nature 330:662 (1987); Shimamoto, Y., et al., Immunol. Lett. 17:311 (1988); Silva, A. T., et al., J. Infect. Dis. 162:421 (1990); Opal, S. M., et al., J. Infect. Dis. 161:1148 (1990); Hinshaw, L. B., et al., Circ. Shock 30:279 (1990). Particular preferred antibodies are TNF-specific antibodies with a high binding affinity, i.e., with an association constant K of at least 10.sup.8 liters per mole. The association constant K can be determined by equilibrium dialysis as described in Kuby, J., Immunology, W.H. Freeman & Co., New York, 1992, pp. 122-124. A more detailed description of antibody affinity and he association constant K is contained in the following references, the teachings of which are incorporated by reference: Kuby, J., Immunology, W.H. Freeman & Co., New York, 1992, pp. 122-124; Hood, L. E., et al., Immunology, Second Edition, The Benjamin/Cummings Publishing Co., Menlo Park, Calif., 1984, pp. 58-60; Abbas, A. K., et al., Cellular and Molecular Immunology, W.B. Saunders Co., Philadelphia, 1991, pp. 53-54. The term soluble receptor is intended to encompass cloned soluble whole receptors, biologically functional fragments thereof, and cloned soluble chimeric receptors. The term soluble receptor is further intended to include all cloned soluble molecules which neutralize tumour necrosis factor (i.e, bind to TNF) or which inhibit TNF biological activity. Biologically functional receptor fragments which can be used are those fragments sufficient for binding of the tumour necrosis factor or those fragments capable of inhibiting TNF biological activity. Cloned soluble chimeric receptors include those molecules capable of binding TNF which are made by fusion of a portion of a TNF receptor to at least one immunoglobulin heavy or light chain. The portion of the TNF receptor present in the cloned chimeric receptor consists of at least a portion of the extracellular region of the TNF receptor. Other types of fusions which result in molecules that are capable of binding TNF are also included. The chimeric receptors can comprise portions derived from two different species (e.g., extracellular domain of human TNF receptor and C.sub.H 2 through C.sub.H 3 domains of murine IgG1 heavy chain). The portions derived from two different species can be joined together chemically by conventional techniques or can be prepared as single contiguous proteins using genetic engineering techniques as described in Peppel, K., et al., J. Exp. Med. 174:1483-1489 (1991). DNA encoding both portions of the chimeric receptor can be expressed as contiguous chimeric receptor proteins. The chimeric receptors can comprise two portions derived from the same species (e.g., extracellular domain of human TNF receptor and constant domains of human IgG heavy chain). The two portions can be joined together chemically by conventional techniques or can be prepared as single contiguous proteins using genetic engineering techniques as described in Lesslauer, W., et al., Eur. J. Immunol. 21:2883-2886 (1991), and Ashkenazi, A., et al., Proc. Natl. Acad. Sci. USA 88:10535-10539 (1991). DNA encoding both portions of the chimeric receptor can be expressed as contiguous chimeric receptor proteins. Other chimeric TNF receptor compositions are possible and can be employed in the subject invention. (See e.g., Scallon, B., et al., U.S. application Ser. No. 08/010,406, filed Jan. 29, 1993). A more detailed description of chimeric TNF receptors and their ability to bind TNF is contained in the following references, the teachings of which are incorporated by reference: Scallon, B., et al., U.S. application Ser. No. 08/010,406, filed Jan. 29, 1993; Peppel, K., et al., J. Exp. Med. 174:1483 (1991); Lesslauer, W., et al., Eur. J. Immunol. 21:2883 (1991); Ashkenazi, A., et al., Proc. Natl. Acad. Sci. USA 88:10535 (1991). Compounds and ligands capable of blocking TNF production, its effects and/or tumour necrosis factor receptor signal transduction but which are not receptors can also be employed in the subject invention. Such compounds include, but are not limited to, peptides, anti-TNF drugs and anti-TNF signal transduction compounds. Anti-TNF antibodies, soluble TNF receptors or anti-TNF compounds are useful if, upon administration to the host in an effective amount, they ameliorate the clinical symptoms or causes of multiple sclerosis. The symptoms or causes are ameliorated if they are significantly reduced or eliminated. It is desirable to administer the anti-TNF antibodies, soluble TNF receptors, and anti-TNF compounds employed in the subject invention directly to the central nervous system. However, the existence of the blood-brain barrier limits the free passage of many types of molecules from the blood to cells of the central nervous system (e.g., potentially useful and therapeutic agents such as anti-TNF antibodies and soluble TNF receptors). During the active phase of inflammatory diseases such as MS and EAE, blood brain leakage is known to occur and will permit entry of anti-TNF antibody, soluble TNF receptors or anti-TNF compounds to the central nervous system. Nevertheless, there are several techniques that either physically break through the blood-brain barrier or circumvent it to deliver therapeutic agents. Examples of these techniques include intrathecal injections, surgical implants, and osmotic techniques. In addition, the permeability of the blood-brain barrier to anti-TNF antibodies, soluble TNF receptors, and anti-TNF compounds can be increased by administering a bradykinin agonist of blood-brain permeability (e.g., N-acetyl ›Phe.sup.8 (CH.sub.2 -NH)Arg.sup.9 ! bradykinin). A more detailed description of these techniques that either physically break through the blood-brain barrier or circumvent it are contained in Malfroy-Camine, U.S. Pat. No. 5,112,596, May 12, 1992, the teachings of which are incorporated by reference. A preferred embodiment for the administration of the antibodies, soluble receptors and anti-TNF compounds is by intrathecal injection, i.e., directly into the cerebrospinal fluid by puncturing the membranes surrounding the central nervous system. Puncturing of the membranes surrounding the central nervous system is usually by lumbar puncture. Sustained dosages of agents directly into the cerebrospinal fluid can be attained by the use of infusion pumps that are implanted surgically. Another embodiment for the administration of anti-TNF antibodies, soluble TNF receptors, and anti-TNF compounds is by injection directly into the lumbar cerebrospinal fluid (intrathecally) or by injection intravenously. Other methods and modes of administration can also be employed. The pharmaceutically-acceptable form in which the anti-TNF antibody, soluble TNF receptor, or anti-TNF compound is administered will depend, at least in part, on the route by which it is administered. For example, in the case of administration by injection, the anti-TNF antibody, soluble TNF receptor, or anti-TNF compound can be formulated with conventional pharmaceutically-acceptable vehicles into pharmaceutical compositions in the usual way for that route of administration. Such vehicles are inherently nontoxic and nontherapeutic. A therapeutically effective amount of anti-TNF antibody, soluble TNF receptor, or anti-TNF compound is that amount necessary to significantly reduce or eliminate symptoms associated with multiple sclerosis. An efficacious amount of anti-TNF antibody for mice is in the range of 150 .mu.g -1 mg/injection. Therefore, a reasonable and preferred therapeutically effective amount of anti-TNF antibody for humans is in the range of 0.1-50 mg/kg/dose. Similarly, a preferred therapeutically effective amount of soluble TNF receptor for mice is in the range of 15-150 .mu.g/injection. Therefore, a reasonable and preferred therapeutically effective amount of soluble TNF receptor for humans is in the range of 0.1-10 mg/kg/dose. The therapeutically effective amount will be determined on an individual basis and will be based, at least in part, on consideration of the individual's size, the severity of symptoms to be treated, the result sought, etc. Thus, the therapeutically effective amount can be determined by one of ordinary skill in the art employing such factors and using no more than routine experimentation. The therapeutically effective amount can be administered in the form of a single dose, or a series of doses separated by intervals of days, weeks or months. Once the therapeutically effective amount has been administered, a maintenance amount of anti-TNF antibody, of soluble TNF receptor, or of anti-TNF compound can be administered. A maintenance amount is the amount of anti-TNF antibody, soluble TNF receptor, or anti-TNF compound necessary to maintain the reduction or elimination of symptoms achieved by the therapeutically effective dose. The maintenance amount can be administered in the form of a single dose, or a series of doses separated by intervals of days, weeks or months. Like the therapeutically effective amount, the maintenance amount will be determined on an individual basis. Other anti-inflammatory or anti-immune drugs, such as methotrexate or cyclosporin A, or antibodies, such as anti-CD4 antibodies, can be administered in conjunction with the anti-TNF antibody, the soluble TNF receptor, or the anti-TNF compound. (See e.g., Feldmann, M., et al., U.S. application Ser. No. 07/958,248, filed Oct. 8, 1992). The method of the present invention can be used to treat multiple sclerosis or any related disease in any mammal. In a preferred embodiment, the method is used to treat multiple sclerosis in human beings. Described herein is work which illustrates the effect of anti-TNF antibody and soluble TNF-R IgG fusion protein in EAE, the results of which indicate that blocking TNF biological activity or other TNF effects, or blocking TNF receptor signal transduction is useful in treating MS. The experiments described herein utilizes EAE as an experimental model of the human demyelinating disease multiple sclerosis. Chronic relapsing EAE is an autoimmune demyelinating disease of the central nervous system used as an experimental model of MS. This mouse model of induced EAE has similarities to human MS in its clinical signs. In both EAE and MS, clinical disease is associated with blood-brain barrier (BBB) dysfunction, infiltration of central nervous system by mononuclear cells (mainly macrophages and T lymphocytes, and serum products), and demyelination (Baker, D., et al., J. Neuroimmunol. 28:261 (1990); Butter, C., et al., J. Neurol. Sci. 104:9 (1991); Harris J. O., et al., Ann. Neurol. 29:548 (1991); Kermonde A. G., et al., Brain 113:1477 (1990)). Thus, the mouse model serves as a good approximation to human disease. To facilitate the determination of whether TNF is important in the pathogenesis of neuroimmunological diseases such as EAE and MS, a TNF-specific monoclonal antibody (TN3.19.12) was administered as described in Examples 5, 6 and 7 during actively-induced EAE (Example 1), shortly before (1-2 days) pre-clinical weight loss, when BBB dysfunction and infiltration of the central nervous system became apparent (Example 2) (Butter, C., et al., J. Neurol. Sci. 104:9 (1991)), and during active clinical disease when neurological signs were manifested (Example 1), and a soluble human TNF receptor (human p55 TNF-R) was administered as described in Examples 8 and 9 during actively-induced EAE, shortly following the onset of clinical signs. As described in Examples 5, 6, 7, 8 and 9, TNF immunotherapy was found to inhibit the progression of chronic relapsing EAE, and thus has implications for the therapeutic strategies in the human disease multiple sclerosis. As described in Example 2, the disease episodes of chronic relapsing EAE are associated with BBB dysfunction (FIG. 2) and marked cellular infiltration of the central nervous system (Baker, D., et al., J. Neuroimmunol. 28:261 (1990); Butter, C., et al., J. Neurol. Sci. 104:9 (1991)). As described in Examples 1, 2 and 3, these parameters which are modulated by anti-TNF antibody (Table 2), correlated with progressive weight loss which occurs shortly before the detection of clinical neurological deficit (FIG. 1). The ability of TNF to augment clinical disease (Kuroda, Y., et al., J. Neuroimmunol. 34:159-164 (1991)) and induce the production of other proinflammatory cytokines (Beutler, B., et al., Science 229:869 (1985); Brennan, F. M., et al., Lancet 2:244 (1989)), and the inhibition of EAE following TNF-neutralization as described herein, implicates an important proinflammatory role for TNF in the pathogenesis of EAE. As described in Examples 5, 6 and 7, clinical EAE developed following the cessation of antibody therapy, indicating that TNF immunotherapy is not exerting an effect through generalized immunosuppression. As described in Example 4, in contrast to the immunosuppressive action of CD4-specific monoclonal antibody on EAE and T cell proliferation, inhibition of TNF activity exhibited minimal effects on T proliferative responses (FIG. 5), indicating that TNF-directed immunotherapy targets effector cell function rather than the induction of disease and consistent with the inability of in vitro treatment of encephalitogenic cells to inhibit adoptive transfer of disease (Selmaj, K., et al., Ann. Neurol. 30:694 (1991)). Therefore the relative timing of antibody administration is important for an inhibitory effect to be observed. For example, in contrast to the inhibitory effect of multiple doses observed when treatment was administered during the anticipated development of disease (Table 1), similar treatment (3.times.250 .mu.g TN3.19.12 injected intraperitoneally) terminated prior to development of anticipated clinical disease failed to prevent the development of clinical EAE (8 of 8 affected with a mean group score of 3.3.+-.0.5). As described in Examples 5, 6 and 7, although systemic administration of neutralizing TNF antibodies inhibited EAE (FIGS. 4A-4B, 7 and 8A-8C), significantly increased benefit was observed when TNF was administered directly into the central nervous system (FIGS. 7 and 8A-8C), indicating that the majority of TNF activity is generated within the central nervous system. Antibodies have a limited potential to cross the intact blood brain barrier (Hafler, D. A., et al., Ann. Neurol. 21:89 (1987)). However, if TNF-specific monoclonal antibody is administered systemically to multiple sclerosis patients, increased targeting of antibody into the central nervous system would occur when blood-brain barrier dysfunction is present, which frequently occurs in clinically silent MS (Harris, J. O., et al., Ann. Neurol. 29:548 (1991); Kermonde, A. G., et al., Brain 113:1477 (1990)), as well as during clinical episodes. The Examples described herein demonstrate the important role of TNF in the demyelinating disease EAE, an experimental model of MS, thereby indicating that TNF is a suitable target for immune intervention and indicating a method for treating multiple sclerosis. Further, the work described herein indicate the advantages of administering TNF antibodies, soluble TNF receptors or anti-TNF compounds directly into the central nervous system. In addition, unlike previous studies using anti-TNF in EAE where the effect on EAE was limited only if the antibody was given before the onset of clinical manifestations, i.e., prophylactically (Selmaj, K., et al., Ann. Neurol. 30:694 (1991); Ruddle, N., J. Exp. Med. 172:1193 (1990)), the work described herein demonstrate therapy after the onset of clinical manifestations, a situation which is relevant to treating multiple sclerosis in human beings |
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