GALACTOSYLCERAMIDE, GLUCOSYLCERAMIDE, LACTOSYLCERAMIDE, AND SPECIFIC CATCHERS THEREFOR FOR USE IN THE PROPHYLAXIS OR THERAPY OF PREDIABETES, DIABETES AND/OR ASSOCIATED COMPLICATIONS

Abstract: The use of glycolipids, in particular galactosylceramide, glucosylceramide and lactosylceramide, and specific catchers therefor (antibodies or lectins), in particular monoclonal antibodies, for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual and for use in the production of pharmaceutical preparations for treatment of said conditions is disclosed.

Description: GALACTOSYLCERAMIDE, GLUCOSYLCERAMIDE, LACTOSYLCERAMIDE, AND SPECIFIC CATCHERS THEREFOR FOR USE IN THE PROPHYLAXIS OR THERAPY OF PREDIABETES, DIABETES AND/OR ASSOCIATED COMPLICATIONS Technical field of the invention The present invention relates to the use of glycol- ipids and specific catchers therefore in treatment of diabetes . Background of the invention Galactosylceramide is a glycolipid consisting of ce- ramide to which galactose is attached. It is made by the enzyme ceramide galactosyltransferase which binds its two parts together. Galactosylceramide is a precursor of sul- fatide and is present in the neural system and in islets of Langerhans in small amounts. A sulfatransferase enzyme is able to attach sulfate to the galactose group and thereby to convert galactosylceramide to sulfatide . It is a possibility that galactosylceramide given in vivo is converted to sulfatide. However, galactosylceramide may also act by itself and, indeed effects of galactosylceramide have been described in vitro (Buschard, K. , Dia- mant, M., Bovin, L. F., Fredman, P., Bendtzen, K. , Sul- phatide and its precursor, galactosylceramide, influence the production of cytokines in human mononuclear cells. APMIS 104: 938-944, 1996). It has been shown that galactosylceramide can modulate and mainly enhance the production of different cytokines from both monocytes and T- cells after stimulation with LPS and PHA, respectively. Most importantly TNF and IL-6 production is increased compared to incubation with LPS and PHA without galactosylceramide. Glucosylceramide and lactosylceramide are related glycolipids which likely have the same effects as galactosylceramide . Summary of the invention The present invention relates to the use of a glycolipid or a specific catcher thereof for the production of a pharmaceutical preparation for treatment of predia- betes, diabetes and/or associated complications in an individual . Furthermore, the invention relates to glycolipids, in particular galactosylceramide, glucosylceramide and lactosylceramide, and specific catchers therefore (antibodies or lectins) for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual. The invention also relates to a specific catcher for the glycolipids according to the invention, said catcher being a monoclonal antibody against galactosylceramide, glucosylceramide or lactosylceramide. The invention also relates to a method for preventing the development of prediabetes, diabetes and/or asso- ciated complications in an individual, wherein a glycolipid, in particular galactosylceramide, glucosylceramide or lactosylceramide, is administered to said individual, preferably at its perinatal stage. The characterising features or the invention will be evident from the following description and the appended claims . Detailed description of the invention As stated above, the invention relates glycolipids, in particular galactosylceramide, glucosylceramide and lactosylceramide, and specific catchers therefore (antibodies or lectins) for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual, as well as to the use of a glycolipid or a specific catcher thereof for the production of a pharmaceutical preparation for treatment of prediabetes, diabetes and/or associated complications in an individual. The expression "treatment" used herein relates to both the prophylaxis of said conditions in an individual being in risk of developing any of the conditions and to the therapeutic treatment of an individual who have already developed any of the conditions. The prophylactic treatment is performed by inducing tolerance to the antigenic glycolipids. When the glycol- ipids, the specific catchers therefore and/or the pharmaceutical preparation according to the invention is used for this purpose they are preferably administered perina- tally to said individual. The glycolipid used according to the invention is preferably galactosylceramide, glucosylceramide or lactosylceramide. The specific catcher used according to the invention is preferably an antibody or a lectin, and more preferably a monoclonal antibody against galactosylceramide, glucosylceramide or lactosylceramide. The glycolipid, the specific catcher or the pharmaceutical preparation according to the invention may be administered in any suitable way known to the man skilled in the art. Preferably, they are administered nasally, orally, subcutaneously, intramuscularly, or intrave- nously. The glycolipids or the pharmaceutical preparation may lead to increased levels of suppressor or regulator cells or antibodies against lymphocytes recognising the antigenic glycolipids in said individual. Alternatively, they may lead to the removal of antibodies and/or lymphocytes recognising the antigenic glycolipids from the blood stream of the individual. It may be suitable to administer the glycolipids according to the invention together with bacterial adju- vants. The pharmaceutical composition according to the invention may therefore also comprises a least one bacte- rial adjuvant, such as cholera, staphyloccoc or galactosylceramide (alpha-form) of bacterial origin. The pharmaceutical preparation according to the invention may also comprise substances used to facilitate the production of the pharmaceutical preparation or the administration of the preparations. Such substances are well known to people skilled in the art and may for example be pharmaceutically acceptable adjuvants, carriers and preservatives. When antibodies are used according to the invention they will lead to an increase of anti-antibodies in said individual . The invention also relates to a method for preventing the development of prediabetes, diabetes and/or asso- ciated complications in an individual, wherein a glycolipid, in particular galactosylceramide, glucosylceramide or lactosylceramide, is administered to said individual, preferably at its perinatal stage. The method may be performed by removing lymphocytes from the individual, contact the lymphocytes with a glycolipid, in particular galactosylceramide, glucosylceramide or lactosylceramide, in vitro to make them recognise this antigen, irradiating them to inhibit their cytotox- icity, and (a) returning them to the individual to raise suppressor or regulator cells or antibodies against lymphocytes reactive with this antigen, or (b) administering them parenterally to another mammal in order to raise antibodies against lymphocytes reactive with this antigen in said mammal and then isolating serum containing the antibodies from said mammal and administering it to the individual. The method may also be performed by contacting the blood stream of the individual with an immobilised glycolipid, in particular galactosylceramide, glucosylcera- mide or lactosylceramide, to remove antibodies and/or lymphocytes recognising the antigenic glycolipids from the individual. Finally, it is also possible to perform the method by parenterally administer an antibody against glycolipids, in particular galactosylceramide, glucosylceramide or lactosylceramide, (a) to said individual in a suffi- cient amount to raise anti-antibodies in said individual, or (b) to another mammal in order to raise anti- antibodies in said mammal and then isolating serum containing the anti-antibodies from said mammal and administering it to the individual . The invention will now be further explained in the following example. This example is only intended to illustrate the invention and should in no way be considered to limit the scope of the invention. Example - Treatment of NOD mice with the intrathymic injections of galactosylceramide in order to modulate the later diabetes incidence Ma terials and methods The NOD mouse model is an animal model of type 1 diabetes. The mice develop spontaneously diabetes within 200 days of life with an incidence of 50% or more. In the present study the animals were examined daily and once a week checked for glucosuria. They were diagnosed as diabetic if their blood glucose values was higher than 200 mg glucose per 100 ml. When diagnosed as diabetics the animals were sacrificed. Otherwise the remaining non- diabetic animals were sacrificed at the end 5 of the study after 200 days of age. Four groups of animals each containing between 30 and 40 animals were investigated. The animals were injected intrathymically when 3 weeks old. There was injected 50 μl galactosylceramide in 100 μl vehicle (PBS) . The control mice were treated with the 100 μl PBS alone. In another experiment liposome preparation of galactosyl- ceramide was made using phosphatidylcholin and galactosylceramide. The control group was treated with phosphatidylcholin and PBS alone. .Results In the first study with treatment of galactosylceramide in pure form 17 of 37 mice (=45.9%) developed diabetes, whereas 21 of 37 (=56.8%) of the control (PBS- treated) mice developed the disease. In the second study with galactosylceramide in liposome form 14 of 33 (=42.4%) of the galactosylceramide liposome treated mice developed diabetes whereas 18 of 31 (=58.1%) of the control (PBS-treated) mice develop the disease. Taking the two studies together 31 of 70 (=44.3%) of the galactosylceramide treated mice developed diabetes whereas 39 of 68 (=57.4%) of PBS mice developed the disease. In the first study the diabetes development in the galactosylceramide treated group occurred later than among the control mice.

Claims: CLAIMS 1. Use of a glycolipid or a specific catcher thereof for the production of a pharmaceutical preparation for treatment of prediabetes, diabetes and/or associated complications in an individual. 2. Use according to claim 1, wherein said glycolipid is galactosylceramide, glucosylceramide and lactosylceramide . 3. Use according to claim 1 or 2, wherein said specific catcher is an antibody or a lectin. 4. Use according to any one of the claims 1-3, wherein said pharmaceutical preparation is formulated for nasal, oral, subcutaneous, intramuscular, or intravenous administration. 5. Use according to any one of the claims 1-4, wherein said pharmaceutical preparation is intended for the prophylaxis of prediabetes, diabetes and/or associated complications in an individual being in risk of developing said disease, by inducing tolerance to the antigenic glycolipids. 6. Use according to claim 5, wherein said pharmaceutical preparation is intended for perinatal administration thereof to said individual. 7. Use according to any one of the claims 1-6, wherein said pharmaceutical preparation upon administra- tion to an individual will lead to increased levels of suppressor or regulator cells or antibodies against lymphocytes recognising the antigenic glycolipids in said individual. 8. Use according to any one of the claims 1-6, wherein said pharmaceutical preparation upon administration to an individual will lead to the removal of antibodies and/or lymphocytes recognising the antigenic glycolipids from the blood stream of the individual. 9. Use according to any one of the claims 1-8, wherein said pharmaceutical preparation further comprises a least one bacterial adjuvant. 10. Use according to claim 9, wherein said bacterial adjuvant is cholera, staphyloccoc or galactosylceramide (alpha-form) of bacterial origin. 11. Glycolipids, in particular galactosylceramide, glucosylceramide and lactosylceramide, and specific catchers therefore (antibodies or lectins) for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual. 12. Galactosylceramide, glucosylceramide or lactosylceramide for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual given nasally, orally, subcutaneously, intramuscularly, intravenously or otherwise. 13. Glycolipids according to claim 11 or 12, for use in the prophylaxis of prediabetes, diabetes and/or associated complications in an individual being in risk of developing said disease, by inducing tolerance to the antigenic glycolipids, for example by perinatal administration thereof to said individual. 14. Glycolipids according to claim 11 or 12, for use in the prophylaxis or therapy or prediabetes, diabetes and/or associated complications in an individual by raising suppressor or regulator cells or antibodies against lymphocytes recognising the antigenic glycolipids in said individual. 15. Glycolipids according to claim 11 or 12, for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual by removing antibodies and/or lymphocytes recognising the antigenic glycolipids from the blood stream of the individual . 16. Glycolipids according to any one of the claims 11-15 given to an individual together with bacterial ad- juvants, for example cholera, staphyloccoc or galactosylceramide (alpha-form) of bacterial origin. 17. A specific catcher for glycolipids according to any of claims 11-16, said catcher being a monoclonal an- tibody against galactosylceramide, glucosylceramide or lactosylceramide . 18. Antibody according to any of claims 11 and 17, for use in the prophylaxis or therapy of prediabetes, diabetes and/or associated complications in an individual by raising anti-antibodies in said individual. 19. A method for preventing the development of prediabetes, diabetes and/or associated complications in an individual, wherein a glycolipid, in particular galactosylceramide, glucosylceramide or lactosylceramide, is ad- ministered to said individual, preferably at its perinatal stage. 20. A method for preventing or treating prediabetes, diabetes and/or associated complications in an individual, wherein lymphocytes are removed from the individual, contacted with a glycolipid, in particular galactosylceramide, glucosylceramide or lactosylceramide, in vitro to make them recognise this antigen, irradiating the lymphocytes to inhibit their cytotoxicity, and (a) returning them to the individual to raise suppressor or regulator cells or antibodies against lymphocytes reactive with this antigen, or (b) administering them parenterally to another mammal in order to raise antibodies against lymphocytes reactive with this antigen in said mammal and then isolating serum containing the antibodies from said mammal and administering it to the individual. 21. A method of preventing or treating prediabetes, diabetes and/or associated complications in an individual, wherein the blood stream of the individual is contacted with an immobilised glycolipid, in particular ga- lactosylceramide, glucosylceramide or lactosylceramide, to remove antibodies and/or lymphocytes recognising the antigenic glycolipids from the individual. 22. A method of preventing or treating prediabetes, diabetes and/or associated complications in an individual, wherein an antibody against glycolipids, in particular galactosylceramide, glucosylceramide or lactosylcera- mide, is parenterally administered (a) to said individual in a sufficient amount to raise anti-antibodies in said individual, or (b) to another mammal in order to raise anti-antibodies in said mammal and then isolating serum containing the anti-antibodies from said mammal and ad- ministering it to the individual. 23. A method according to any on of the claims 19- 21, wherein the glycolipid is selected from the group consisting of galactosylceramide, glucosylceramide, or lactosylceramide .

Inventors: BUSCHARD KARSTEN, FREDMAN PAM, BUSCHARD, KARSTEN, FREDMAN, PAM
Applicants: A & SCIENCE INVEST AB, BUSCHARD KARSTEN, FREDMAN PAM, A+ SCIENCE INVEST AB, BUSCHARD, KARSTEN, FREDMAN, PAM
Application year: 1998
Main classification: A61K31
IPC codes: A61K31, A61K39, A61P3