Amyloid conjugate and uses and methods thereof

Abstract

A composition includes aluminum hydroxide gel and a conjugate of at least one CysA13(33-40) peptide linked to the keyhole limpet hemocyanin (KLH). Maleimidobutyric acid Nhydroxysuccinimide ester (SM) serves as cross-linking agent. The composition can produce an effective and specific immune response against Aβ40. The antibodies produced are specific for Aβ40 without significantly binding to Aβ42. The composition can increase the response against Aβ40 compared with the response produced by other conjugates that include CysAβ(33-40) peptide and KLH, and are bound or conjugated by other crosslinking agents.

What is claimed is: 
     
         1 . A composition comprising aluminum hydroxide gel and a conjugate of at least one CysA13(33-40) peptide (SEQ ID NO: 1) linked to the keyhole limpet hemocyanin (KLH), wherein the crosslinking agent connecting each CysA13(33-40) peptide to the keyhole limpet hemocyanin (KLH) of the conjugate is the maleimidobutyric acid Nhydroxysuccinimide ester (SM). 
     
     
         2 . A pharmaceutical composition comprising i) a conjugate comprising at least one CysA13(33-40) peptide (SEQ ID NO: 1) linked to keyhole limpet hemocyanin (KLH) via a crosslinking agent, and ii) an aluminum hydroxide gel, wherein the crosslinking agent connecting each CysA13(33-40) peptide to the keyhole limpet hemocyanin (KLH) of the conjugate is maleimidobutyric acid N-hydroxysuccinimide ester (SM), and wherein a pH of the composition ranges from 5.8 to 7.0. 
     
     
         3 . The pharmaceutical composition according to  claim 2 , wherein the pH of the composition ranges from 6.2 to 7.0. 
     
     
         4 . The pharmaceutical composition according to  claim 2 , wherein the pH of the composition ranges from 5.8 to 6.2. 
     
     
         5 . The pharmaceutical composition according to  claim 2 , wherein the concentration of the at least one CysA13(33-40) peptide (SEQ ID NO: 1) is at least 100 μg. 
     
     
         6 . The pharmaceutical composition according to  claim 2 , wherein the concentration of the at least one CysA13(33-40) peptide (SEQ ID NO: 1) is at least 150 μg. 
     
     
         7 . The pharmaceutical composition according to  claim 2 , wherein the concentration of the at least one CysA13(33-40) peptide (SEQ ID NO: 1) is from 150 μg to 400 μg. 
     
     
         8 . The pharmaceutical composition according to  claim 2 , wherein the concentration of the at least one CysA13(33-40) peptide (SEQ ID NO: 1) is from 160 μg to 240 μg. 
     
     
         9 . The pharmaceutical composition according to  claim 2 , wherein the conjugate comprises a ratio of at least 45 CysA13(33-40) peptides (SEQ ID NO: 1) linked to each keyhole limpet hemocyanin (KLH). 
     
     
         10 . A glass ampule comprising the pharmaceutical composition of  claim 2 . 
     
     
         11 . A method of manufacturing a pharmaceutical composition, the method comprising:
 a. adding maleimidobutyric acid N-hydroxysuccinimide ester to a composition comprising keyhole limpet hemocyanin (KLH) in a buffer at a pH between 7.0 to 9;   b. removing an excess of maleimidobutyric acid N-hydroxysuccinimide ester from the solution of step a);   c. adding CysA13(33-40) peptides (SEQ ID NO: 1) in DMSO at a pH of between 6.6 to 7.0 to produce the conjugates comprising at least one CysA13(33-40) peptide (SEQ ID NO: 1) linked to keyhole limpet hemocyanin (KLH) via maleimidobutyric acid N-hydroxysuccinimide ester;   d. removing unconjugated peptide from step c) to produce a solution;   e. adjusting a pH to between 5.8 to 6.2; and   f. adding aluminum hydroxide gel once the pH has been adjusted.   
     
     
         12 . The method of  claim 11 , wherein the excess of step b) is removed by using a 0.02 M Na-Phosphate buffer at a pH of about 6.6 to 7.0. 
     
     
         13 . The method of  claim 11 , wherein the excess unconjugated peptide of step d) is removed by using a 0.01 M PBS-buffer at a pH of 6.6 to 7.0. 
     
     
         14 . The method of  claim 11 , wherein the solution of step d) is filtered. 
     
     
         15 . The method of  claim 11 , wherein the pH of step e) is adjusted to a pH of about 6.0. 
     
     
         16 . The method of  claim 11 , wherein the solution of step d) is filtered using a 0.2 μm filter. 
     
     
         17 . A method of delaying onset and/or treating an amyloid disease, the method comprising:
 identifying a subject with the amyloid disease; and   providing a pharmaceutical composition of  claim 2  to the subject,   thereby delaying onset and/or treating the amyloid disease.   
     
     
         18 . The method of  claim 17 , wherein the amyloid disease is selected from the list consisting of Alzheimer's disease, Parkinson's disease, cerebral amyloid angiopathy, vascular dementia of an amyloid origin, inclusion-body myositis, and dementia with Lewy bodies. 
     
     
         19 . The method of  claim 18 , wherein the amyloid disease is Alzheimer's disease.