Methods and Apparatus for Creating Particle Derivatives of HDL with Reduced Lipid Content

Abstract

The present invention is directed to systems, apparatus and methods for creating derivatives of at least one form of HDL without substantially affecting LDL. These derivatives of HDL are particles with reduced lipid content, particularly reduced cholesterol content. These particles have the capacity to bind cholesterol and are administered to a patient to enhance cellular cholesterol efflux and reduce cholesterol levels in cells, tissues, organs, and blood vessels. The present method is useful for treating atherogenic vascular disease and may be combined with other therapies such as statins, inhibitors of cholesterol absorption, niacin, anti-inflammatories, exercise and dietary restriction.

1 - 29 . (canceled) 
     
     
         30 . A method for making a particle derivative of at least one form of high density lipoprotein wherein the particle derivative comprises a protein shell and a lipid bilayer comprising the steps of:
 a. connecting a patient to a device for withdrawing blood;   b. withdrawing blood containing blood cells from the patient;   c. separating the blood cells from the blood to yield a blood fraction containing high density lipoprotein and low density lipoprotein;   d. separating the low density lipoprotein from the blood fraction;   e. mixing the blood fraction with a lipid removing agent which removes lipids associated with the lipid bilayer of the high density lipoprotein to yield a mixture of lipid, the lipid removing agent, and the particle derivative;   f. separating the particle derivative from the lipid and the lipid removing agent; and,   g. delivering the particle derivative to the patient.   
     
     
         31 . The method of  claim 30 , wherein steps c through f occur remote from the subject. 
     
     
         32 . The method of  claim 30 , wherein the separation of the low density lipoprotein from the blood fraction is performed using an apheresis device. 
     
     
         33 . The method of  claim 30 , wherein the lipid removing agent is at least one of an ether, di-isopropyl ether, sevoflurane, a combination of an alcohol and an ether, or a combination of a sevoflurane and an alcohol. 
     
     
         34 . The method of  claim 33 , wherein the lipid removing agent is a mixture of sevoflurane and n-butanol. 
     
     
         35 . The method of  claim 34 , wherein the mixture comprises 95 parts sevoflurane and 5 parts n-butanol. 
     
     
         36 . The method of  claim 30 , wherein the step of separating the particle derivative from the lipid and the lipid removing agent is achieved by at least one of an absorbent, separator, centrifuge, or charcoal column. 
     
     
         37 . The method of  claim 36 , wherein the at least one of an absorbent, separator, centrifuge, or charcoal column does not modify the protein shell of the particle derivative. 
     
     
         38 . The method of  claim 30 , wherein the particle derivative is first recombined with the blood cells before delivering the particle derivative to the patient. 
     
     
         39 . The method of  claim 30 , wherein the mixing is performed using at least one of a static mixer, vortexer or centrifuge. 
     
     
         40 . A method for making a particle derivative of at least one form of high density lipoprotein wherein the particle derivative comprises a protein shell and a lipid bilayer comprising the steps of:
 a. connecting a patient to a device for withdrawing blood;   b. withdrawing blood containing blood cells from the patient;   c. separating the blood cells from the blood to yield a blood fraction containing high density lipoprotein and low density lipoprotein;   d. mixing the blood fraction with a lipid removing agent which removes lipids associated with a lipid bilayer of the high density lipoprotein without substantially modifying the low density lipoprotein to yield a mixture of lipid, the lipid removing agent, the particle derivative, and the low density lipoprotein;   e. separating the particle derivative and the low density lipoprotein from the lipid and the lipid removing agent; and,   f. delivering the particle derivative and the low density lipoprotein to the patient.   
     
     
         41 . The method of  claim 40 , wherein steps c through e occur remote from the subject. 
     
     
         42 . The method of  claim 40 , wherein the lipid removing agent is at least one of an ether, di-isopropyl ether, sevoflurane, a combination of an ether and an alcohol, or a combination of sevoflurane and an alcohol. 
     
     
         43 . The method of  claim 42 , wherein the lipid removing agent is a mixture of sevoflurane and n-butanol. 
     
     
         44 . The method of  claim 43 , wherein the mixture comprises 95 parts sevoflurane and 5 parts n-butanol. 
     
     
         45 . The method of  claim 40 , wherein the step of separating the particle derivative and low density lipoprotein from the lipid and lipid removing agent is achieved by at least one of an absorbent, separator, centrifuge, or charcoal column. 
     
     
         46 . The method of  claim 45 , wherein the at least one of an absorbent, separator, centrifuge, or charcoal column does not modify the protein shell of the particle derivative. 
     
     
         47 . The method of  claim 40 , wherein the particle derivative and the low density lipoprotein are first recombined with the blood cells before delivering the particle derivative and the low density lipoprotein to the patient. 
     
     
         48 . The method of  claim 40 , wherein the mixing is performed using at least one of a static mixer, vortexer or centrifuge. 
     
     
         49 - 72 . (canceled)