Peptide-based immunization therapy for treatment of atherosclerosis and development of peptide-based assay for determination of immune responses against oxidized low density lipoprotein

Abstract

The present invention relates to fragments of apolipoprotein B, in particular defined peptides thereof, for immunization or therapeutic treatment of mammals, including humans, against ischemic cardiovascular diseases, as well as diagnosing the presence or absence of antibodies related to increased or decreased risk of developing ischemic cardiovascular diseases, using one or more of said peptides in an ELISA (Enzyme Linked Immuno Sorbent Assay).

1 . Fragments of apo-lipoprotein B intended for immunization or therapeutic treatment of mammals, including humans, against ischemic cardiovascular diseases and having immunogenic or therapeutic properties against ischemic cardiovascular diseases, and/or diagnosing the presence or absence of antibodies related to increased or decreased risk of developing ischemic cardiovascular diseases.  
     
     
         2 . Fragments according to  claim 1 , wherein the fragments are haptens of aldehydes.  
     
     
         3 . Fragments according to  claim 2 , wherein the fragments are modified using malone dealdehyde or hydroxynonenal.  
     
     
         4 . Peptides being fragments according to claims  1 - 3 , native or as aldehyde-derivatives comprising the group 
 FLDTVYGNCSTHFTVKTRKG    PQCSTHIQWLKRVHANPLL    VISIPRLQAEARSEILAHWS    KLVKEALKESQLPTVMDFRK    LKFVTQAEGAKQTEATMTFK    DGSLRHKFLDSNIKFSHVEK    KGTYGLSCQRDPNTGRLNGE    RLNGESNLRFNSSYLQGTNQ    SLTSTSDLQSGIIKNTASLK    TASLKYENYELTLKSDTNGK    DMTFSKQNALLRSEYQADYE    MKVKIIRTIDQMQNSELQWP    IALDDAKINFNEKLSQLQTY    KTTKQSFDLSVKAQYKKNKH    EEEMLENVSLVCPKDATRFK    GSTSHHLVSRKSISAALEHK    IENIDFNKSGSSTASWIQNV    IREVTQRLNGEIQALELPQK    EVDVLTKYSQPEDSLIPFFE    HTFLIYITELLKKLQSTTVM    LLDIANYLMEQIQDDCTGDE    CTGDEDYTYKIKRVIGNMGQ    GNMGQTMEQLTPELKSSILK    SSILKCVQSTKPSLMIQKAA    IQKAAIQALRKMEPKDKDQE    RLNGESNLRFNSSYLQGTNQ    SLNSHGLELNADILGTDKIN    WIQNVDTKYQIRIQIQEKLQ    TYISDWWTLAAKNLTDFAEQ    EATLQRIYSLWEHSTKNHLQ    ALLVPPETEEAKQVLFLDTV    IEIGLEGKGFEPTLEALFGK    SGASMKLTTNGRFREHNAKF    NLIGDFEVAEKINAFRAKVH    GHSVLTAKGMALFGEGKAEF    FKSSVITLNTNAELFNQSDI    FPDLGQEVALNANTKNQKIR    ATRFKHLRKYTYNYQAQSSS    or an active site of one or more of these peptides.    
     
     
         5 . A peptide according to  claim 3 , selected from the group 
 HTFLIYITELLKKLQSTTVM    ALLVPPETEEAKQVLFLDTV    FLDTVYGNCSTHFTVKTRK    PQCSTHILQWLKRVHANPLL    LLDIANYLMEQIQDDCTGDE    CTGDEDYTYKIKRVIGNMGQ    GNMGQTMEQLTPELKSSILK    SSILKCVQSTKPSLMIQKAA    IQKAAIQALRKMEPKDKDQE    IEIGLEGKGFEPTLEALFGK    VISIPRLQAEARSEILAHWS    KGTYGLSCQRDPNTGRLNGE    RLNGESNLRFNSSYLQGTNQ    SLTSTSDLQSGIIKNTASLK    TASLKYENYELTLKSDTNGK    DMTFSKQNALLRSEYQAPYE    GSTSHHLVSRKSISAALEHK    IALDDAKINFNEKLSQLQTY    IENIDFNKSGSSTASWIQNV    NLIGDFEVAEKINAFRAKVH    IREVTQRLNGEIQALELPQK    GHSVLTAKGMALFGEGKAEF    KTTQSFDLSVKAQYKKNKH    FPDLGQEVALNANTKNQKIR    ATRFKHLRKYTYNYQAQSSS    or an active site of one or more of these peptides.    
     
     
         6 . Peptide according to claims  4 - 5 , in native form.  
     
     
         7 . Peptide according to claims  4 - 5 , in oxidized form.  
     
     
         8 . Peptide according to  claim 7 , wherein the peptide has been oxidized using copper.  
     
     
         9 . Peptide according to claims  4 - 5 , wherein the peptide is present in a combination with phospholipid liposomes.  
     
     
         10 . Peptide according to claims  4 - 5 , in the form of a malone dealdehyde (MDA) derivative thereof.  
     
     
         11 . Peptide according to claims  4 - 5 , in the form of a hydroxynoneal-derivative thereof.  
     
     
         12 . The use of one or more of the fragments/peptides of  claims 1  to  11 , in native or MDA or hydroxynoneal derivative form, in the preparation of pharmaceutical compositions intended for immunotherapy or therapy for the treatment of ischemic cardiovascular diseases, optionally in combination with an adjuvant.  
     
     
         13 . Use according to  claim 12 , wherein an immunization dose is 1 to 100 mg of the fragments/peptides.  
     
     
         14 . Pharmaceutical preparation comprising a therapeutically effective amount of one or more of the fragments/peptides of claims  1 - 11 , optionally in combination with one or more pharmaceutically innocuous fillers, and/or adjuvants.  
     
     
         15 . Pharmaceutical preparation according to  claim 14 , wherein the fragments/peptides are present as linked to cationized bovine serum albumine, and using aluminum hydroxide as an adjuvant.  
     
     
         16 . Pharmaceutical composition according to  claim 15 , wherein the composition is present as an injectable composition.  
     
     
         17 . Vaccine for immunization of mammals, including humans, against ischemic cardiovascular diseases comprising one or more fragments/peptides of claims  1 - 11 , optionally in combination with an adjuvant  
     
     
         18 . Vaccine for immunization of mammals, including humans, against ischemic cardiovascular diseases comprising a therapeutically effective amount of purified or recombinantly produced antibodies against one or more of said native and/or MDA modified sequences of claims  1 - 11 .  
     
     
         19 . Vaccine for immunization according to  claim 17 , wherein the fragments/peptides present for immunization are present as linked to cationized bovine serum albumine, and using aluminum hydroxide as an adjuvant.  
     
     
         20 . Method of prophylactic or therapeutic treatment of a mammal, including a human being, suffering from atheroschlerosis or facing the risk of developing ischemic cardiovascular diseases, whereby a therapeutically effective amount of one or more of the fragments and/or peptides of claims  1 - 11 , either in native form or in the form of a malone dealdehyde or a hydroxynoneal derivative, is administered to said mammal suffering from atheroschlerosis, or facing the risk of developing ischemic cardiovascular diseases, in particular myocardial infarction.  
     
     
         21 . Method of prophylactic or therapeutic treatment of a mammal, including a human being, suffering from atheroschlerosis or facing the risk of developing ischemic cardiovascular diseases, whereby a therapeutically effective amount of one or more of purified, or recombinantly produced antibodies against native and MDA-modified sequences to produce a passive immunization is administered.  
     
     
         22 . Method according to  claim 20 , wherein the conditions are one or more of unstable atherosclerotic plaques in which oxidized LDL is likely to contribute to inflammation, cell toxicity and risk for plaque rupture, as well as coronary heart disease in older individuals.  
     
     
         23 . Method of diagnosing the presence or absence of antibodies related to increased or decreased risk of developing ischemic cardiovascular diseases, using one or more of the fragments and/or peptides of claims  1 - 11  in an assay.  
     
     
         24 . Method according to  claim 23 , wherein the assay is an immunoassay.  
     
     
         25 . Method according to  claim 24 , wherein the immunoassay is an ELSA, RIA, Western blotting, Southern blotting