EP0994717A1 - Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate - Google Patents

Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate

Info

Publication number
EP0994717A1
EP0994717A1 EP98908452A EP98908452A EP0994717A1 EP 0994717 A1 EP0994717 A1 EP 0994717A1 EP 98908452 A EP98908452 A EP 98908452A EP 98908452 A EP98908452 A EP 98908452A EP 0994717 A1 EP0994717 A1 EP 0994717A1
Authority
EP
European Patent Office
Prior art keywords
methotrexate
disease
autoimmune
antigen
combination
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP98908452A
Other languages
German (de)
English (en)
Other versions
EP0994717A4 (fr
Inventor
Howard L. Weiner
Ahmad Al-Sabbagh
Patricia A. Nelson
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Autoimmune Inc
Original Assignee
Autoimmune Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Autoimmune Inc filed Critical Autoimmune Inc
Publication of EP0994717A1 publication Critical patent/EP0994717A1/fr
Publication of EP0994717A4 publication Critical patent/EP0994717A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0008—Antigens related to auto-immune diseases; Preparations to induce self-tolerance
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B41—PRINTING; LINING MACHINES; TYPEWRITERS; STAMPS
    • B41J—TYPEWRITERS; SELECTIVE PRINTING MECHANISMS, i.e. MECHANISMS PRINTING OTHERWISE THAN FROM A FORME; CORRECTION OF TYPOGRAPHICAL ERRORS
    • B41J2/00—Typewriters or selective printing mechanisms characterised by the printing or marking process for which they are designed
    • B41J2/005—Typewriters or selective printing mechanisms characterised by the printing or marking process for which they are designed characterised by bringing liquid or particles selectively into contact with a printing material
    • B41J2/01—Ink jet
    • B41J2/21—Ink jet for multi-colour printing
    • B41J2/2107—Ink jet for multi-colour printing characterised by the ink properties
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • A61K2039/541—Mucosal route
    • A61K2039/542—Mucosal route oral/gastrointestinal
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06K—GRAPHICAL DATA READING; PRESENTATION OF DATA; RECORD CARRIERS; HANDLING RECORD CARRIERS
    • G06K2215/00—Arrangements for producing a permanent visual presentation of the output data
    • G06K2215/0082—Architecture adapted for a particular function
    • G06K2215/0094—Colour printing
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06K—GRAPHICAL DATA READING; PRESENTATION OF DATA; RECORD CARRIERS; HANDLING RECORD CARRIERS
    • G06K2215/00—Arrangements for producing a permanent visual presentation of the output data
    • G06K2215/0082—Architecture adapted for a particular function
    • G06K2215/0097—Printing on special media, e.g. labels, envelopes

Definitions

  • This invention pertains to the field of treatment of autoimmune diseases by tolerization. This invention also relates to the field of treatment of autoimmune diseases by administration of methotrexate.
  • Autoimmune diseases are characterized by an abnormal immune response directed against normal autologous (self) tissues.
  • autoimmune diseases are currently being treated with drugs that suppress immune responses systemically in a non-specific manner, i.e., drugs incapable of selectively suppressing the abnormal immune response.
  • drugs that suppress immune responses systemically in a non-specific manner
  • drugs incapable of selectively suppressing the abnormal immune response are
  • One such drug is methotrexate, a biological response modifier that selectively inhibits fast growing cells.
  • methotrexate has significant toxic and other side effects and eventually induces "global" immunosuppression in the subject being treated.
  • prolonged treatment with a normal course of methotrexate downregulates the normal protective immune response against pathogens, thereby increasing the risk of infection.
  • patients subjected to prolonged global immunosuppression have an increased risk of developing severe medical complications from the treatment, such as malignancies, liver, and kidney failure.
  • the present invention relates to a method for treating autoimmune disease.
  • the method involves mucosally administering an autoantigen.
  • the method also involves orally, enterally, or parenterally administering methotrexate.
  • the amounts of autoantigen and methotrexate are effective in combination to suppress autoimmune response associated with the disease.
  • Figure 2 is a graph that shows the effects on EAE clinical scores of administering either an optimal dose of methotrexate or a 1 mg dose of MBP.
  • Figure 3 is a graph that shows the effects on EAE clinical scores of combining an optimal dose of methotrexate with MBP as compared to administering an optimal dose of methotrexate alone.
  • Figure 5 is a graph showing the effects on EAE of administering a combination of a sub-optimal dose of 20 ⁇ g methotrexate and a 1 mg dose of MBP as compared with administering either alone.
  • Figure 6 is a graph showing the swollen joint count measured in rheumatoid arthritis patients who, beginning at week 8 of a clinical study, were administered either a combination of oral type ⁇ collagen and oral methotrexate (open diamonds) or only oral type U collagen (closed boxes).
  • the present invention allows a lower amount of methotrexate to be administered for treatment of autoimmune disease than would otherwise be necessary (a suboptimal dose). This is highly advantageous because of methotrexate' s high toxicity.
  • the suppressive effect of administering a combination of methotrexate and MBP has been found to be more effective than the effect of administering either alone.
  • Methotrexate has been found not to prevent treatment by mucosal tolerization, even though methotrexate is known to inhibit cell division, i.e., it has not detrimentally affected the cells mediating treatment of autoimmune disease by tolerance. Furthermore, administration of tolerizing agent has been found not to interfere with methotrexate' s inhibitory effects.
  • methotrexate and tolerizing agent can be frequently administered at lower doses per administration. While methotrexate is normally administered once a week in a high dose to treat autoimmune disease, the combination of the invention has been found to be effective when administered three times a week at lower doses. As described above, administering an overall lower dose of methotrexate according to the invention acts to reduce toxicity. Administering that lower dose in subdivided form at intervals during the week further acts to reduce toxicity.
  • methotrexate is normally administered by itself at an optimal dose of 10 mg per week to treat an autoimmune disease
  • a suboptimal dose of 2 mg, three times a week can be administered with an effective amount of tolerizing agent.
  • Autoimmune disease is defined herein as a spontaneous or induced malfunction of the immune system of mammals, including humans, in which the immune system fails to distinguish between foreign immunogenic substances within the mammal and/or autologous substances and, as a result, treats autologous tissues and substances as if they were foreign and mounts an immune response against them.
  • the term includes human autoimmune diseases and animal models therefor.
  • Autoantigen is any substance or a portion thereof normally found within a mammal that, in an autoimmune disease, becomes the primary (or a primary) target of attack by the immune system.
  • the term also includes antigenic substances that induce conditions having the characteristics of an autoimmune disease when administered to mammals.
  • the term includes peptic subclasses consisting essentially of immunodominant epitopes or immunodominant epitope regions of autoantigens. Immunodominant epitopes or regions in induced autoimmune conditions are fragments of an autoantigen that can be used instead of the entire autoantigen to induce the disease.
  • Treatment is intended to include both prophylactic treatment to prevent or delay the onset of an autoimmune disease (or to prevent the manifestation of clinical or subclinical, e.g. , histological, symptoms thereof), as well as therapeutic suppression or alleviation of symptoms after the manifestation of autoimmune disease, by abating autoimmune attack and preventing or slowing down autoimmune tissue destruction.
  • “Abatement”, “suppression” or “reduction” of autoimmune attack or reaction encompasses partial reduction or amelioration of one or more symptoms of the attack or reaction.
  • a “substantially” increased suppressive effect (or abatement or reduction) of autoimmune reaction means a significant decrease in one or more markers or histological or clinical indicators of autoimmune reaction or disease.
  • Parenteral administration includes subcutaneous, intradermal, intramuscular, intravenous, intraperitoneal or intrathecal administration.
  • Administration "in conjunction with” encompasses simultaneous and sequential administration, as well as administration in combined form or separately.
  • Immunization with Mycobacterium tuberculosis or with CFA in oil into the dorsal root tail of susceptible mammals induces a disease used as a model for human rheumatoid arthritis.
  • immunization with Type U collagen with an adjuvant will also induce a disease (collagen-induced arthritis or "CIA") that serves as a model for human rheumatoid arthritis.
  • CIA collagen-induced arthritis
  • TGF- ⁇ transforming growth factor-beta
  • Th2 -enhancing cytokines such as interleukin 4 (IL-4); and/or interleukin 10 (IL-10) at the locus of the autoimmune attack.
  • IL-4 interleukin 4
  • IL-10 interleukin 10
  • the immunoregulatory cytokines released by the elicited regulatory cells are antigen-nonspecific, even though these regulatory T-cells release (or induce the release of) immunoregulatory cytokines only when triggered by an antigenic determinant identical to one on the mucosally administered antigen.
  • Recruitment of the immunoregulatory T-cells to a locus within a mammal where cells contributing to the autoimmune destruction of an organ or tissue are concentrated allows for the release of immunoregulatory substances in the vicinity of the autoimmune attack and suppresses all types of immune system cells responsible for such attack.
  • suppressor (CD8+ or CD4+) T-cells are induced, released into the blood or lymphatic circulation, and then recruited to the area of autoimmune attack, where they cause the release of TGF-]S and/or other immunoregulatory substances that downregulate the activated helper T-cells as well as the B-cells directed against the mammal's own tissues. Chen, Y. et al. , Science. 1994 supra. Suppression induced in this manner is antigen-nonspecific.
  • Bystander antigens and autoantigens can be purified from natural sources (the tissue or organ where they normally occur) and can also be obtained using recombinant DNA technology, in bacterial, yeast, insect (e.g. baculovirus) and mammalian cells using techniques well- known to those of ordinary skill in the art.
  • Amino acid sequences for many potential and actual bystander antigens are known: See, e.g., Hunt, C. et al PNAS (USA). 82:6455-6459, 1985 (heat shock protein hsp70); Burkhardt, H. , et al. , Eur. J. Immunol.
  • IRBP IRBP
  • bovine and mouse PLP The amino acid sequences for bovine and mouse PLP; bovine, human, chimpanzee, rat, mouse, pig, rabbit, guinea pig MBP; human and bovine collagen alpha- 1(11) and bovine collagen alpha- 1(1); chicken collagen U, and human insulin are well-known and published and these antigens can be synthesized by recombinant techniques, as is well-known in the art. Fragments of these antigens can be chemically synthesized or also synthesized by recombinant techniques.
  • tissue-specific antigens are commercially available: e.g., insulin, glucagon, myelin basic protein, myelin, collagen I, collagen ⁇ , proteolipid protein, etc.
  • T-cells can be similarly tested for secretion of IL-4 or IL-10 (antibodies for IL-4 and IL-10 are commercially available, e.g., from Pharmingen, San Diego, CA).
  • Tissue-specific antigens that are not effective bystanders are those so segregated from the inflammatory locus (of autoimmune attack) so that the immunoregulatory cytokines released are too far removed from the locus of inflammation to exert a suppressive effect.
  • the efficacy of mucosally induced bystander suppression can be assessed, e.g.
  • the tolerance induced by the bystander antigens of this invention is dose-dependent over a broad range of oral (or enteral) or inhalable dosages. However, there are minimum and maximum effective dosages. In other words, active suppression of the clinical and histological symptoms of an autoimmune disease occurs within a specific dosage range, which, however, varies from disease to disease, mammal to mammal, and bystander antigen to bystander antigen. For example, when the disease is PLP-induced EAE in mice, the suppressive dosage range when MBP is used as the bystander is from about 0.1 to about 1 mg/mouse/feeding (with feedings occurring about every other day e.g.
  • a most preferred dosage is 0.25 mg/mouse/feeding.
  • the MBP suppressive dosage range is from about 0.5 to about 2 mg/rat/feeding and the most preferred dosage is 1 mg/rat/feeding.
  • An effective dosage range for humans with MS, when MBP is used as the oral tolerizer, is between about 1 and about 100, preferably between about 1 and about 20 mg MBP per day (administered every day or on alternate days for a period of time ranging from several months to several years) with the optimum being between about 7 and 10 mg per day.
  • EAE limb paralysis which can be scored as follows: 0-no disease; 1- decreased activity, limp tail; 2-mild paralysis, unsteady gait; 3- moderate paraparesis, limbs splayed apart; 4-tetraplegia; and 5- death.
  • RA joint swelling, joint tenderness, morning stiffness, grip strength, joint imaging techniques.
  • AUR visual acuity; number of T-cells in the eye and "cloudiness" in the eye.
  • Type I Diabetes pancreatic beta cell function (assessed, e.g. , by OGTT glucose tolerance test).
  • NOD Model insulitis and delay of diabetes onset.
  • the total arthritis score is the sum of the scores for all paws.
  • Maximum arthritis score is the highest score for an animal over the course of the disease. According to this grading method the highest arthritis score possible is 16 (4 paws X 4 score-per-paw). Stabilization of symptoms, under conditions wherein control patients or animals experience a worsening of symptoms, is one indicator of efficacy of a suppressive treatment.
  • Another measure of improvement is the ability to reduce or discontinue other medications, e.g., steroids or other anti-inflammatory medications, and biologic response modifiers.
  • the optimum dosage of a bystander antigen or autoantigen is one generating the maximum beneficial effect assessed as described above.
  • An effective dosage causes at least a statistically or clinically significant attenuation of at least one marker, symptom or histological evidence characteristic of the disease being treated as described above. (Clinically significant-attenuation is one observed by a clinician of ordinary skill in the field of a particular autoimmune disease.)
  • the maximum effective dosage of a bystander antigen can be ascertained by testing progressively higher dosages in animals and then extrapolating to humans. For example, based on the dosages given above, for rodents, the maximum effective dose of MBP for humans has been estimated between 50 and 100 mg/feeding. Similarly, the maximum effective amount of Collagen Type U for humans has been estimated at about 1 mg/day.
  • the present invention can also be advantageously used to prevent the onset of an autoimmune disease in susceptible individuals at risk for an autoimmune disease.
  • methods for the identification of patients who are at risk for developing Type 1 diabetes are extant and reliable and have been recently endorsed by the American Diabetes Association (ADA).
  • ADA American Diabetes Association
  • Various assay systems have been developed which (especially in combination) have a high predictive value assessing susceptibility to Type 1 diabetes (Diabetes Care 13: 762-775, 1990). Details of one preferred screening test are available to those of ordinary skill in the art (Bonifacio, E. et al. , The Lancet 335: 147-149, 1990).
  • Bystander antigens and autoantigens can also be administered by inhalation.
  • the bystander amounts that need to be inhaled are generally smaller than those for oral administration. Effective amounts for inhalation therapy can be assessed using the same methodologies provided above.
  • Type-I, Type-II and Type-in collagen have activity as mucosal tolerizers. Other collagens are likely to be similarly active.
  • T-cell has been identified in mice orally tolerized against EAE as a CD4+ suppressor T-cell, and a CD8+ suppressor T-cell has been identified in rats.
  • immunodominant epitopes of autoantigens e.g. MBP
  • Additional such epitopes can be identified by feeding a bystander antigen to a mammal and isolating from the mammal T-cells that recognize a fragment of the antigen (and thus identifying suppressive fragments), or by identifying T-cells from a bystander fed mammal that can adoptively transfer protection to naive (not-fed) animals.
  • Autoantigen administration is carried out as disclosed in PCT
  • cytokine and non-cytokine synergists can be conjoined in the treatment to enhance the effectiveness of oral tolerization with methotrexate.
  • Oral and parenteral use of other cytokine synergists has been described in PCT/US95/04120, filed 04/07/95.
  • Administration of Th2 enhancing cytokines is described in PCT application no. PCT/US95/04512, filed 04/07/95.
  • IL-4 and IL-10 can be administered in the manner described in PCT/US95/04512.
  • immunoregulatory lipoproteins such as peptides covalently linked to tripalmitoyl-S-glycarylcysteinyl-seryl-serine (P 3 C55) which can be obtained as disclosed in Deres, K. et al. (Nature. 342:561-564, 1989) or "Braun's" lipoprotein from R coli which can be obtained as disclosed in Braun, V. , Biochim. Biophys.
  • LPS cholera toxin /5-chain
  • the effective dosage range for noncytokine synergists for mammals is from about 15 ⁇ g to about 15 mg per kg weight and preferably 300 ⁇ g - 12 mg per kg weight.
  • the effective dosage range for oral Type I interferon for mammals is from 1,000 - 150,000 units with no maximum effective dosage having been discerned.
  • the optimum regimen for administering both the tolerizing agent and methotrexate is determined in light of the information disclosed herein and well known information concerning administration of bystander antigens, autoantigens, and methotrexate. Routine variation of dosages, combinations, and duration of treatment is performed under circumstances wherein the severity of autoimmune reaction can be measured. Useful dosage and administration parameters are those that result in reduction in autoimmune reaction, including a decrease in number of autoreactive T-cells, or in the occurrence or severity of at least one clinical or histological symptom of the disease.
  • the tolerizing agent is preferably administered within 24 hours of administration of methotrexate. More preferably, it is administered at the same time as methotrexate. Most preferably, both are administered in a combined oral formulation.
  • Methotrexate for use in the invention is commonly commercially available in both oral and parenteral dosage forms. Alternately, methotrexate is conventionally formed in an oral or parenteral dosage form, using carriers and other agents as further described below.
  • Each oral (or enteral) formulation according to the present invention may comprise inert constituents including pharmaceutically acceptable carriers, diluents, fillers, solubilizing or emulsifying agents, and salts, as is well-known in the art.
  • tablets may be formulated in accordance with conventional procedures employing solid carriers well-known in the art.
  • Capsules employed in the present invention may be made from any pharmaceutically acceptable material, such as gelatin, or cellulose derivatives.
  • Sustained release oral delivery systems and/or enteric coatings for orally administered dosage forms are also contemplated, such as those described in U.S. Patent No. 4,704,295, issued November 3, 1987; U.S. Patent No. 4,556,552, issued December 3, 1985; U.S. Patent No. 4,309,404, issued January 5, 1982; and U.S. Patent No. 4,309,406, issued January 5, 1982.
  • solid carriers examples include starch, sugar, bentonite, silica, and other commonly used carriers.
  • carriers and diluents which may be used in the formulations of the present invention include saline, syrup, dextrose, and water.
  • formulations for tolerizing agents that are administered by inhalation are provided in PCT/US90/07455, filed 12/17/90.
  • the pharmaceutical formulations for administration by inhalation of the present invention may include, as optional ingredients, pharmaceutically acceptable carriers, diluents, solubilizing and emulsifying agents, and salts of the type that are well-known in the art.
  • pharmaceutically acceptable carriers such as pharmaceutically acceptable saline solutions, such as physiologically buffered saline solutions, and water.
  • Dry aerosol in the form of finely divided solid particles of tolerizing antigens that are not dissolved or suspended in a liquid are also useful in the practice of the present invention.
  • the tolerizing antigens may be in the form of dusting powders and comprise finely divided particles having an average particle size of between about 1 and 5 microns, preferably between 2 and 3 microns.
  • Finely divided particles may be prepared by pulverization and screen filtration using techniques well known in the art.
  • the particles may be administered by inhaling a predetermined quantity of the finely divided material, which can be in the form of a powder.
  • EAE Induction of EAE.
  • Lewis rats were immunized in the foot pads with 50 ⁇ l of emulsion each.
  • 0.1 ml of emulsion contained 25 ⁇ g of guinea pig MBP and 200 ⁇ g of Mycobacterium tuberculosis in complete Freund's adjuvant (CFA).
  • CFA complete Freund's adjuvant
  • methotrexate was administered by itself, and in combination with 1 mg of GP-MBP (guinea pig MBP).
  • the 80 ⁇ g of methotrexate administered at a frequency of three times a week corresponds to the optimal dosage of methotrexate administered to humans, when adjusted for weight.
  • the recommended dose for a 60 Kg person is 10 mg of methotrexate per week.
  • EXAMPLE 2 Suppression of EAE in Rats with a Combination of a Sub-Optimal Dose of Methotrexate and Tolerizing Agent
  • Example 1 The procedures described in Example 1 were followed, for the protocol shown below. Sub-optimal dosages of 40 ⁇ g and 20 ⁇ g were administered in combination with 1 mg of MBP over a period of 14 days. An optimal dosage of 80 ⁇ g was also administered.
  • % TOLERANCE MMCS (CONTROL) - MMCS (TEST) X 100
  • FIGS 1-5 show clinical data from this experiment for 20 days after immunization with MBP in Complete Freund's Adjuvant.
  • Figure 1 shows the results of administration of methotrexate alone. Suppression of EAE with the optimal dose (80 ⁇ g) and suboptimal doses (40 ⁇ g and 20 ⁇ g) of methotrexate can be seen to follow a dose dependent response.
  • the graph shows a rebound effect associated with methotrexate administration, wherein, following the highest degree of suppression, at day 18, the clinical symptoms of the disease reappear.
  • Figure 2 shows the effects on clinical scores of administering an optimal dose of methotrexate compared with an optimal dose of MBP. A rebound effect is seen at day 20 for rats administered methotrexate, but not for rats administered MBP.
  • Figure 4 shows the results of administering MBP alone and a sub- optimal dose of 40 ⁇ g of methotrexate alone as compared with administering a combination of the two.
  • the combination achieves greater suppression than that achieved with either alone, and avoids the rebound effect seen with methotrexate alone.
  • the degree of suppression is essentially the same as that observed with the an optimal dose of methotrexate alone.
  • Figure 5 shows the results of administering MBP alone and a suboptimal dose of 20 ⁇ g of methotrexate alone as compared with administering a combination of the two.
  • the combination achieves greater suppression than that achieved with either alone.
  • the degree of suppression is similar to that observed with an optimal dose of methotrexate alone.
  • a lower dose of methotrexate can be administered than is optimal, but, as a result of the combination with MBP, a suppressive effect is achieved that is similar to that of an optimal dose of methotrexate alone.
  • the combination eliminates the rebound effect associated with methotrexate administration alone.
  • DBA/J1 mice were orally dosed once a day for 5 consecutive days with type JJ collagen (300 ⁇ g in 0.25 ml), methotrexate (10 ⁇ g in 0.25 ml), a combination of the two, or with a control solution of 0.1 N acetic acid (0.25 ml).
  • the type JJ collagen was chicken sterna type JJ collagen (1.2 mg/ml) (Sigma Chemical Co., St. Louis, MO).
  • the methotrexate was obtained as 2.5 mg tablets (Roxane Lab. Inc.). The tablets were mixed with 0.1 normal acetic acid to form a 2.5 mg/ml solution.
  • the amount of methotrexate administered was determined based on an optimal dosage to a 60 Kg human of lOmg/week, using the converting factor of 12 for mice (Chemotherapy National Cancer Institute), and based on an average weight of 26 g per mouse.
  • each animal was injected intradermally into its shaved back with 0.1 ml of an emulsion containing 100 ⁇ g of type U collagen and 100 ⁇ g of Mycobacterium butyricum.
  • All mice were boosted with an intraperitoneal injection of 100 ⁇ g of type JJ collagen in 0.1 N acetic acid.
  • Animals were monitored for onset of arthritis for 60 days. Beginning on day + 10, animals are scored for signs of arthritis on a scale of 0 to 4. The arthritis score for each animal was the sum of the score for each of the four paws.
  • Serum free culture supematants are collected from cells from test animals. See, for example, Kehri, et al. J. Exp.Med.163: 1037-1050, 1986; and Wahl, et al.
  • modulator cells are first cultured for 8 hours with the antigen (50 ⁇ l/ml) in proliferation medium. Thereafter cells are washed three times and resuspended in serum-free medium for the remainder of the 72 hour culture, supematants collected, then frozen until assayed. Determination of TGF- ⁇ content and isoform type in supematants is performed using a mink lung epithelial cell line (American Type Culture Collection, Bethesda, MD #CCL-64) according to Danielpour et al. (Danielpour, D., et al. J. Cell. Phvsiol.
  • This assay can be adapted to test any antigen which is a candidate for use as an inducer of bystander suppression. Those antigens, antigen fragments and/or amounts of antigen which produce the highest concentration of TGF- ⁇ as measured by this assay are suitable for use in the treatment method of the present invention.
  • a transwell culture system described below, can be used to indicate the level of TGF- which is being produced. This culture system measures the production of TGF-/3 as a function of suppression of cell proliferation.
  • IL-4 and/or IL-10 in culture supematants of antigen- stimulated cells can also serve as an indicator that the antigen is suitable for use as an inducer of bystander suppression.
  • IL-4, IL-10 (and TGF-0) can be assayed by ELISA using commercially available antibodies to each polypeptide as described in Chen, Y. et al. , Science. 1994, supra.
  • Transwell Cultures A dual chamber transwell culture system (Costar, Cambridge, MA), which is 24.5 mm in diameter and consists of two compartments separated by a semi-permeable polycarbonate membrane, with a pore size of 0.4 ⁇ m, is used. The two chambers are 1 mm apart, allowing cells to be co-incubated in close proximity without direct cell-to-cell contact.
  • 5 x 10 4 antigen specific line cells raised and maintained for example, as previously described (Ben-Nun, A. et al., Eur. J. Immunol. 11: 195.
  • thymocytes are cultured with 10 6 irradiated (2,500 rad) thymocytes, in 600 ⁇ l of proliferation media in the lower well.
  • Spleen cells from orally tolerized animals (e.g. rats) or controls (fed BSA) are added to the upper well (5 x 10 s cells in 200 ⁇ l).
  • Spleens are removed 7-14 days after the last feeding, and a single cell suspension is prepared by pressing the spleens through a stainless steel mesh.
  • the antigen 50 ⁇ g/ml is added in a volume of 20 ⁇ l. Because modulator cells are separated from responder cells by a semi-permeable membrane, they do not require irradiation.
  • modulator cells are added in the lower well together with responder cells, and in these instances modulator cells are irradiated (1,250 rad) immediately before being placed in culture.
  • Proliferation media consisted of RPMI 1640 (Gibco Laboratories, Grand Island, NY) supplemented with 2 x 10 5 M 2- mercaptoethanol, 1 % sodium pyruvate, 1 % penicillin and streptomycin, 1 % glutamine, 1 % HEPES buffer, 1 % nonessential amino acids, and 1 % autologous serum. Each transwell is performed in quadruplicate. The transwells are incubated at 37° C in a humidified 6% CO 2 and 94% air atmosphere for 72 hours.
  • each lower well is pulsed with 4 ⁇ Ci of [ 3 H]thymidine and at 72 hours split and reseeded to three wells in a round-bottomed 96-well plate (Costar) for harvesting onto fiberglass filters and counting using standard liquid scintillation techniques.
  • Percent suppression 100 x (1 - ⁇ cpm responders cultured with modulators/ ⁇ cpm of re- sponders).
  • Patients suffering from rheumatoid arthritis were administered either oral type U collagen alone, or a combination of oral type JJ collagen and oral methotrexate, as part of a clinical study designed to compare administration of oral type JJ collagen with administration of placebo following abrupt withdrawal from methotrexate treatment.
  • Functional Class IV includes patients who are limited in their ability to perform usual self-care, vocational and mobilizional activities. All patients exhibited at least 6 tender joints, and at least 3 swollen joints. Stable steroids, NSAIDS (non-steroidal anti-inflammatory agents, and certain analgesics, were administered to those who wished to use them).
  • Type JJ collagen Methods for preparation of such type JJ collagen are described in WO 97/25435.
  • the type JJ collagen used in this study had been subjected to conventional pepsin treatment in acetic acid in order to remove telopeptides.
  • Compositions containing 20 ⁇ g of type JJ collagen, methods of administration of such a dosage, and beneficial results of such administration, are described in WO 97/02837.
  • Figures 6-8 show the results of the study.
  • Figure 6 shows the swollen joint count measured during the period of the study.
  • the group of patients with exacerbated symptoms at week 8 exhibited increased swollen joint count at that time.
  • this group of patients exhibited a decrease in the number of swollen joints, and after eight weeks of combination therapy (which corresponds to the 16 week mark in Figure 6) exhibited a greater decrease in swollen joints (from their baseline number) than patients receiving only type JJ collagen.
  • the results shown in Figure 7 with respect to tender joint count followed a similar pattern.
  • the percent of patients achieving cumulative Paulus 20 response shown in Figure 8 was higher for patients receiving combination therapy than for those receiving only oral type JJ collagen.
  • the Paulus response for the patients receiving the colloral-methotrexate combination was calculated from the time that methotrexate was combined with type JJ collagen at week 8.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Diabetes (AREA)
  • Epidemiology (AREA)
  • Rheumatology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Transplantation (AREA)
  • Pain & Pain Management (AREA)
  • Obesity (AREA)
  • Ophthalmology & Optometry (AREA)
  • Biomedical Technology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Hematology (AREA)
  • Endocrinology (AREA)
  • Emergency Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne une combinaison d'un antigène de voisinage susceptible d'être administré par voie muqueuse et d'un méthotrexate susceptible d'être administré par voie parentérale, entérale ou orale. Cette combinaison est utilisée pour mettre en oeuvre une formulation pharmaceutique et pour traiter ou prévenir des maladies auto-immunes. Les quantités d'antigène de voisinage et de méthotrexate sont efficaces en combinaison pour supprimer une réponse auto-immune associée à la maladie auto-immune.
EP98908452A 1997-01-24 1998-01-26 Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate Withdrawn EP0994717A4 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US3672297P 1997-01-24 1997-01-24
US36722P 1997-01-24
PCT/US1998/001648 WO1998032451A1 (fr) 1997-01-24 1998-01-26 Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate

Publications (2)

Publication Number Publication Date
EP0994717A1 true EP0994717A1 (fr) 2000-04-26
EP0994717A4 EP0994717A4 (fr) 2000-07-26

Family

ID=21890248

Family Applications (1)

Application Number Title Priority Date Filing Date
EP98908452A Withdrawn EP0994717A4 (fr) 1997-01-24 1998-01-26 Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate

Country Status (10)

Country Link
EP (1) EP0994717A4 (fr)
JP (1) JP2001511134A (fr)
KR (1) KR20000070460A (fr)
AU (1) AU6648898A (fr)
BR (1) BR9807112A (fr)
CA (1) CA2278152A1 (fr)
HU (1) HUP0001960A2 (fr)
IL (1) IL131025A0 (fr)
NO (1) NO993600L (fr)
WO (1) WO1998032451A1 (fr)

Families Citing this family (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20040103930A (ko) 2002-02-11 2004-12-09 앤태어스 파머, 인코퍼레이티드 피내 주사기
BRPI0614025A2 (pt) 2005-01-24 2012-12-25 Antares Pharma Inc injetores de jato
GB0523576D0 (en) * 2005-11-18 2005-12-28 Theradeas Ltd Drug composition and its use in therapy
US8251947B2 (en) 2006-05-03 2012-08-28 Antares Pharma, Inc. Two-stage reconstituting injector
US9144648B2 (en) 2006-05-03 2015-09-29 Antares Pharma, Inc. Injector with adjustable dosing
EP2268342B1 (fr) 2008-03-10 2015-09-16 Antares Pharma, Inc. Dispositif de sécurité pour injecteur
JP5611208B2 (ja) 2008-08-05 2014-10-22 アンタレス・ファーマ・インコーポレーテッド 多数回服用量の注射装置
AU2010226442A1 (en) 2009-03-20 2011-10-13 Antares Pharma, Inc. Hazardous agent injection system
EP2582373A4 (fr) 2010-06-16 2013-10-30 Bruce Chandler May Utilisation de la lévocétirizine et du montélukast dans le traitement de la grippe, du rhume et d'une inflammation
US9220660B2 (en) 2011-07-15 2015-12-29 Antares Pharma, Inc. Liquid-transfer adapter beveled spike
US8496619B2 (en) 2011-07-15 2013-07-30 Antares Pharma, Inc. Injection device with cammed ram assembly
JP6165786B2 (ja) 2012-03-06 2017-07-19 アンタレス・ファーマ・インコーポレーテッド 離脱力特徴を備える充填シリンジ
CN104487114A (zh) 2012-04-06 2015-04-01 安塔雷斯药品公司 针头辅助喷射注射给予睾酮组合物
WO2013169800A1 (fr) 2012-05-07 2013-11-14 Antares Pharma, Inc. Dispositif d'injection ayant un ensemble piston à came
EP2953667B1 (fr) 2013-02-11 2019-10-23 Antares Pharma, Inc. Dispositif d'injection à jet assistée par aiguille ayant une force de déclenchement réduite
WO2014164786A1 (fr) 2013-03-11 2014-10-09 Madsen Patrick Injecteur de dose avec système à pignon
WO2014165136A1 (fr) 2013-03-12 2014-10-09 Antares Pharma, Inc. Seringues pré-remplies à volume constant et leurs trousses
CA2901413A1 (fr) 2013-03-13 2014-10-09 Bruce Chandler May Utilisation de levocetirizine et de montelukast dans le traitement de la vascularite
RU2672871C2 (ru) 2013-03-13 2018-11-20 Инфламматори Респонс Ресёрч, Инк. Применение левоцетиризина и монтелукаста при лечении травматических повреждений
RU2015134423A (ru) * 2013-03-13 2017-04-26 Инфламматори Респонс Ресёрч, Инк. Применение левоцитиризина и монтелукаста при лечении аутоиммуных расстройств
EP3193875B1 (fr) 2014-09-15 2022-02-16 Inflammatory Response Research, Inc. Lévocétirizine et montélukast dans le traitement de troubles médiés par une inflammation

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992004049A1 (fr) * 1990-09-06 1992-03-19 Rijksuniversiteit Te Utrecht Inhibiteurs de la reponse de lymphocytes et de maladies immunitaires
WO1993016724A1 (fr) * 1992-02-28 1993-09-02 Autoimmune, Inc. Suppression de maladies auto-immunes par des antigenes en attente
EP0666080A1 (fr) * 1987-06-24 1995-08-09 Brigham & Women's Hospital Traitement de maladies auto-immunitaires par administration orale d'auto-antigènes
WO1996040235A1 (fr) * 1995-06-07 1996-12-19 Nexstar Pharmaceuticals, Inc. Procede d'utilisation de superantigenes pour cibler des sous-populations de lymphocytes t
WO1996041644A1 (fr) * 1995-06-13 1996-12-27 Nippon Meat Packers, Inc. Medicament a administration par voie orale, contre la polyarthrite rhumatoide et aliment actif

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5571500A (en) * 1987-06-24 1996-11-05 Autoimmune, Inc. Treatment of autoimmune diseases through administration by inhalation of autoantigens
US5571499A (en) * 1987-06-24 1996-11-05 Autoimmune, Inc. Treatment of autoimmune diseases by aerosol administration of autoantigens

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0666080A1 (fr) * 1987-06-24 1995-08-09 Brigham & Women's Hospital Traitement de maladies auto-immunitaires par administration orale d'auto-antigènes
WO1992004049A1 (fr) * 1990-09-06 1992-03-19 Rijksuniversiteit Te Utrecht Inhibiteurs de la reponse de lymphocytes et de maladies immunitaires
WO1993016724A1 (fr) * 1992-02-28 1993-09-02 Autoimmune, Inc. Suppression de maladies auto-immunes par des antigenes en attente
WO1996040235A1 (fr) * 1995-06-07 1996-12-19 Nexstar Pharmaceuticals, Inc. Procede d'utilisation de superantigenes pour cibler des sous-populations de lymphocytes t
WO1996041644A1 (fr) * 1995-06-13 1996-12-27 Nippon Meat Packers, Inc. Medicament a administration par voie orale, contre la polyarthrite rhumatoide et aliment actif

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
AL-SABBAGH, A.M., ET AL.: "Combination of oral antigens with oral methotrexate enhances suppression of autoimmune disease" JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, vol. 99, no. 1(PART 2), Abs. 99, January 1997 (1997-01), page s24 XP000909440 *
See also references of WO9832451A1 *

Also Published As

Publication number Publication date
NO993600L (no) 1999-09-23
IL131025A0 (en) 2001-01-28
NO993600D0 (no) 1999-07-23
AU6648898A (en) 1998-08-18
HUP0001960A2 (hu) 2000-10-28
CA2278152A1 (fr) 1998-07-30
BR9807112A (pt) 2001-09-18
EP0994717A4 (fr) 2000-07-26
JP2001511134A (ja) 2001-08-07
KR20000070460A (ko) 2000-11-25
WO1998032451A1 (fr) 1998-07-30

Similar Documents

Publication Publication Date Title
US5935577A (en) Treatment of autoimmune disease using tolerization in combination with methotrexate
AU720695B2 (en) Bystander suppression of autoimmune diseases
US6645504B1 (en) Bystander suppression of type I diabetes by oral administration of glucagon
WO1998032451A1 (fr) Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate
WO1998032451A9 (fr) Traitement des maladies auto-immunes a l'aide d'une tolerance induite en combinaison avec du methotrexate
Miller et al. Epitopes of myelin basic protein that trigger TGF-beta release after oral tolerization are distinct from encephalitogenic epitopes and mediate epitope-driven bystander suppression.
EP1033998B1 (fr) Suppression des modifications liees aux beta-amyloides dans la maladie d'alzheimer
AU9023791A (en) Treatment of autoimmune diseases by oral administration of autoantigens
AU695883B2 (en) Treatment of autoimmune disease using oral tolerization and/or Th2-enhancing cytokines
AU686797B2 (en) Treatment of autoimmune disease using oral tolerization and/or type I interferon
US20010007758A1 (en) Treatment of multiple sclerosis using COP-1 and Th2-enhancing cytokines
US20040115217A1 (en) Bystander suppression of autoimmune diseases
WO1994004121A1 (fr) Suppression par un antigene tiers d'une maladie neurologique associee a un retrovirus
WO2008045488A2 (fr) Traitement de l'inflammation, de la démyélinisation et de la perte neuronale/axonale
JP2001502306A (ja) インシュリン依存性糖尿病の処置方法および組成物
JP2003512435A (ja) 予防及び治療方法
JP2003512435A5 (fr)

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19990818

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

A4 Supplementary search report drawn up and despatched

Effective date: 20000614

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

RIC1 Information provided on ipc code assigned before grant

Free format text: 7A 61K 31/505 A, 7A 61K 39/00 B, 7A 61P 37/00 B, 7A 61K 39/00 J, 7A 61K 31:505 J

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20000801