Method for In-Vivo Binding of Chromatin Fragments

Abstract

A process for substantially reducing levels of circulating chromatin fragments (CCFs) from a medium using binding agents such as antibodies or antibodies complexed with haemocompatible natural polymer substrates like as alginates, chitosan and pullulan to form complexed antibodysubstrate nano-particulates (CNP) to bind and/or inactivate CCFs is disclosed. The amount of antibody bound to the polymer varies from 30% to 100% of activated sites in the polymer. Elevated levels of CCFs can be substantially reduced following administration of tissue damaging agents that generate apoptotic chromatin fragments by the concomitant administration of CNPs or concomitant administration of H4 antibody alone. A method of treatment is disclosed wherein therapeutic dose of CNPs, or H4 antibody alone, are administered systematically, or orally, in a delivery system to curb pathological conditions that are associated with increased burden of circulating chromatin fragments.

What is claimed is: 
     
         1 . A method of making a nanoparticle comprising a polymer having a selective affinity to liver cells of a mammal, and an antibody that binds or inactivates circulating chromatin fragments (CCFs), the method comprising:
 forming an activated polymer,   forming an activated antibody,   forming complexed antibody substrate nanoparticulates (CNPs) by reacting the activated polymer and the activated antibody.   
     
     
         2 . The method of  claim 1 , wherein forming the activated polymer comprises introducing reactive functional groups comprising thiols, carboxyl or amino groups to the polymer, wherein forming the activated antibody comprises activating the amino groups in the antibody by introducing hetero-bifunctional cross-linking agents. 
     
     
         3 . A nanoparticle comprising a polymer having an affinity for Asialo Glycoprotein Receptor (ASGPR) to selectively transport the nanoparticle in vivo to liver in a mammal. 
     
     
         4 . The nanoparticle of  claim 3 , further comprising an antibody capable of binding and/or inactivating a chromatin fragment thereto. 
     
     
         5 . The nanoparticle of  claim 4 , further comprising the chromatin fragment. 
     
     
         6 . The nanoparticle of  claim 4 , wherein the antibody comprises anti-histone antibodies selected from the group of those against histones H1, H2A, H2B, H3 and H4; and those producing antibodies against other chromatin proteins comprising High Mobility Group Proteins. 
     
     
         7 . The nanoparticle of  claim 4 , wherein the antibody comprises anti-histone antibodies comprising those against histones H1, H2A, H2B, H3 and H4, that are activated using maleimide reactive groups present in hetero-bifunctional crosslinking agents at pH of 6.5-7.5. 
     
     
         8 . The nanoparticle of  claim 4 , wherein an amount of the antibody bound to the polymer varies from 30% to 100% of activated sites in the polymer. 
     
     
         9 . The nanoparticle of  claim 3 , wherein the polymer is a polymer substrate to form a complex with an antibody. 
     
     
         10 . The nanoparticle of  claim 3 , wherein the nanoparticle comprises monodisperse nanoplexes having a size of less than 100 nm for efficient endocytosis. 
     
     
         11 . The nanoparticle of  claim 3 , wherein the polymer comprises biodegradable low molecular weight polymers. 
     
     
         12 . The nanoparticle of  claim 3 , wherein the polymer comprises natural polymers having a molecular weight of 30000 to 100000 Daltons. 
     
     
         13 . The nanoparticle of  claim 3 , wherein the polymer is haemocompatible. 
     
     
         14 . The nanoparticle of  claim 3 , wherein the polymer comprises an alginate, chitosan, and pullulan. 
     
     
         15 . The nanoparticle of  claim 3 , wherein the polymer is chitosan activated using galactose, maleimide functional groups. 
     
     
         16 . The nanoparticle of  claim 3 , wherein the polymer is a cationic polymer. 
     
     
         17 . A method for substantially reducing levels of circulating chromatin fragments (CCFs) from a medium, comprising introducing into the medium a nanoparticle comprising (a) a polymer having a selective affinity to liver of a mammal, and (b) an antibody that inactivates CCFs, wherein the antibody is covalently bound to the polymer. 
     
     
         18 . A method of treatment comprising:
 systematically or orally administering therapeutic dose of a nanoparticle is in a delivery system to aid in their removal from circulation thereby to curb diverse pathological conditions that are associated with increased burden of circulating chromatin fragments in conditions comprising cancer, systemic autoimmune disorders, diabetes, cerebral stroke, myocardial infarction, inflammation, sepsis, critical illness, trauma, pulmonary embolism, inflammatory bowel disease, organ transplantation and pre-eclampsia,   wherein the nanoparticle comprises: a) a polymer having an affinity to selectively transport the nanoparticle in vivo to liver in a mammal, and (b) an antibody that has an affinity to inactivate CCFs.