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 . A fluid comprising a modified protein distribution,
 wherein the protein distribution in the fluid prior to modification has a first state, the first state having more alpha high density lipoproteins than pre-beta high density lipoproteins, and   wherein the fluid comprising the modified protein distribution is obtained by a method comprising the steps of:
 exposing the fluid to a lipid removing agent wherein the exposure modifies the protein distribution from the first state into a second state, the second state having more pre-beta high density lipoproteins than alpha high density lipoproteins; and, 
 removing the lipid removing agent from the fluid, 
 wherein the lipid removing agent comprises a C4 to C8 alcohol, a C4 to C8 ether, a fluoroether, trifluoroethane or a combination thereof. 
   
     
     
         2 . The fluid of  claim 1 , wherein the fluid is a biological fluid. 
     
     
         3 . The fluid of  claim 1 , wherein the fluid is plasma. 
     
     
         4 . The fluid of  claim 1 , wherein the lipid removing agent further comprises ethanol. 
     
     
         5 . The fluid of  claim 1 , wherein the lipid removing agent comprises isoflurane, sevoflurane, diisopropyl ether, diethyl ether, propylether or n-butanol. 
     
     
         6 . The fluid of  claim 1 , wherein the lipid removing agent further comprises ethanol. 
     
     
         7 . The fluid of  claim 1 , wherein the lipid removing agent comprises a combination of a C4 to C8 alcohol and a C4 to C8 ether, a combination of a fluoroether and a C4 to C8 alcohol, or a combination of C4 to C8 ether and a fluoroether. 
     
     
         8 . The fluid of  claim 1 , wherein the lipid removing agent comprises a combination of n-butanol with at least one of hexanol, diethyl ether, propylether, diisopropyl ether, isoflurane, or sevoflurane. 
     
     
         9 . The fluid of  claim 1 , wherein the lipid removing agent comprises a combination of diisopropyl ether with at least one of n-butanol, hexanol, ethanol, isoflurane, sevoflurane or trifluoroethane. 
     
     
         10 . The fluid of  claim 1 , wherein the lipid removing agent comprises a combination of sevoflurane with at least one of n-butanol, hexanol, ethanol, isoflurane, diisopropyl ether, or trifluoroethane. 
     
     
         11 . The fluid of  claim 1 , wherein the fluid comprises low density lipoproteins prior to exposing the fluid to the lipid removing agent. 
     
     
         12 . The fluid of  claim 11 , wherein the low density lipoproteins are not removed from the fluid prior to exposing the fluid to the lipid removing agent. 
     
     
         13 . The fluid of  claim 12 , wherein the low density lipoproteins are substantially unaffected by the lipid removing agent. 
     
     
         14 . The fluid of  claim 11 , wherein apolipoprotein B (ApoB) proteins are associated with low density lipoproteins in the fluid. 
     
     
         15 . The fluid of  claim 14 , wherein the ApoB proteins are substantially unaffected by the lipid removing agent. 
     
     
         16 . The fluid of  claim 11 , wherein the fluid is exposed to the lipid removing agent using an extracorporeal process. 
     
     
         17 . The fluid of  claim 11 , wherein exposing the fluid to the lipid removing agent leaves low density lipoproteins substantially unmodified, as compared to the low density lipoproteins in the fluid prior to exposing the fluid to the lipid removing agent. 
     
     
         18 . The fluid of  claim 17 , wherein the extracorporeal process transforms at least some of the alpha high density lipoproteins into pre-beta high density lipoproteins. 
     
     
         19 . The fluid of  claim 18 , wherein the pre-beta high density lipoproteins transformed from the alpha high density lipoproteins have a lower cholesterol content than the alpha high density lipoproteins. 
     
     
         20 . A method for modifying the protein distribution in a patient, comprising administering to the patient the fluid of  claim 1 . 
     
     
         21 . The method of  claim 20 , wherein the fluid with the modified protein distribution is autologous to the patient. 
     
     
         22 . The method of  claim 20 , wherein the fluid with the modified protein distribution is not autologous to the patient. 
     
     
         23 . A kit for performing a method of modifying protein distribution in a fluid,
 wherein the protein distribution in the fluid prior to modification has a first state, the first state having more alpha high density lipoproteins than pre-beta high density lipoproteins,   wherein the method comprises the steps of:
 exposing the fluid to a lipid removing agent wherein the exposure modifies the protein distribution from the first state into a second state, the second state having more pre-beta high density lipoproteins than alpha high density lipoproteins; and, 
 removing the lipid removing agent from the fluid, 
   and wherein the kit comprises:
 a. a container for the fluid prior to modification; 
 b. a lipid removing agent source container; 
 c. a mixer comprising at least one of a static mixer, vortexer, or centrifuge; 
 d. a separator comprising at least one of an absorbent, separator, centrifuge, or charcoal column; 
 e. an output container for storing the fluid having the modified protein distribution; and 
 f. a plurality of tubing and a plurality of valves for controlling the flow of high density lipoprotein from the container (a) and lipid removing agent from the container (b) to the mixer (c), for controlling the flow of the mixture of the fluid and the lipid removing agent to the separator (d), and for controlling the flow of the fluid having the modified protein distribution to the output container.