Albumin-based colloid composition having at least one protected thiol region, methods of making, and methods of use

Abstract

A composition comprising an albumin-based colloid composition having at least one protected thiol region, method of making the same, and method for use, including treating hypovolemic conditions such as capillary leak syndrome and shock, are disclosed. The composition also is modified with an indicator reagent such as chromophores.

1 - 17 . (canceled) 
     
     
         18 . A method for making a composition comprising a polyethylene glycol-albumin colloid composition having Cys-34 preserved as a thiol (PEG-Alb Cys-34 ), wherein PEG-Alb Cys-34  has a hydrodynamic radius that is sufficiently large to preclude the composition from leaking through a patient's capillaries wherein the PEG-Alb- Cys-34  is human albumin or bovine serum albumin modified with an indicator reagent;
 comprising the steps of:   modifying albumin or PEG-albumin with at least one thiol selective reagent without perturbing the disulfide structure of the albumin protein;   modifying the thio-modified protein with indicator reagent; and   purifying the albumin or PEG-albumin to remove unmodified protein, excess reagent and reaction byproducts.   
     
     
         19 . A method according to  claim 18  wherein the step of modifying albumin or PEG-albumin with a thio reagent is carried out by treating with an excess of dithiothreitol. 
     
     
         20 . A method according to  claim 18  including the step of basing the method on a multiple chromophore technique. 
     
     
         21 . A method according to  claim 18  including the step of basing the method on a double chromophore technique. 
     
     
         22 . A method according to  claim 18  including the steps of:
 tagging the albumin and PEG-albumin with spectroscopically distinct chromophores; and   repeatedly assessing the concentrations of the chromophores over time.   
     
     
         23 . A method according to  claim 22  including the steps of predicting the development of multiorgan dysfunction before it happens or early in the development of the dysfunction. 
     
     
         24 . A method of predicting the development of multiorgan dysfunction before it happens or early in the development of the dysfunction comprising the step of:
 providing a first protein with a known molecular weight;   providing a second protein with a significantly different molecular weight;   tagging the first and second proteins with chromophores having distant emission and excitation wavelengths; and   repeatedly assessing the concentrations of the chromophores over time.   
     
     
         25 . A method according to  claim 18  wherein the indicator reagent is a dye. 
     
     
         26 . A method according to  claim 25  wherein the dye is a combination of dyes having widely and significantly distant emission and exitation wavelengths. 
     
     
         27 . A method according to  claim 21  wherein the dye is a red maleimide dye or indocyanine. 
     
     
         28 . A method according to  claim 18  wherein the indicator reagent is a fluorescein. 
     
     
         29 . A method according to  claim 28  wherein indocyanine green is used in combination with the fluorescein. 
     
     
         30 . A method according to  claim 28  wherein the fluorescein is a 5-iodoacetamidofluorescein. 
     
     
         31 . A method of accurate identification of patients at risk of developing multiorgan dysfunction syndrome comprising the steps of:
 using multiple proteins or molecules with different molecular weights and tagged with different fluorophores each with distinct and distant emission and excitation wavelengths;   administering the multiple proteins or molecules to a patient at risk of developing multiorgan dysfunction; and   following the concentration of the different fluorophores serially at multiple times under the same pathophysiological processes.   
     
     
         32 . A method according to  claim 31  wherein those pathophysiological processes are hemoconcentration and capillary leak. 
     
     
         33 . A method for the prevention of mammalian tissue injury from at least on hpovolemic condition comprising the administration of a therapeutic amount to a mammal of a composition comprising a polyethylene glycol-albumin colloid composition having Cys-34 preserved as a thiol (PEG-Alb Cys-34 ), wherein PEG-Alb Cys-34  has a hydrodynamic radius that is sufficiently large to preclude the composition from leaking through a patient's capillaries wherein the PEG-Alb- Cys-34  is human albumin or bovine serum albumin modified with an indicator reagent;
 the composition being incapable of leaking through the mammal's capillaries and being present in an amount sufficient to protect said tissue from injury.   
     
     
         34 . A method for the prevention of mammalian tissue injury from at least one hypovolemic condition comprising the administration of a therapeutic amount to a mammal of a composition comprising a polyethylene glycol-albumin colloid composition having Cys-34 preserved as a thiol (PEG-Alb Cys-34 ) wherein PEG-Alb Cys-34  has a hydrodynamic radius that is sufficiently large to preclude the composition from leaking through a patient's capillaries, wherein the PEG-Alb- Cys-34  is human albumin or bovine serum albumin modified with an indicator reagent:
 the composition being incapable of leaking through the mammal's capillaries and being present in an amount sufficient to protect said tissue from injury.   
     
     
         35 . The method of  claim 34  where the injury is due to sepsis, shock, burn, trauma, surgery, predisposition to capillary leak, hyperviscosity states, hypoalbuminemia, leukopheresis, nutritional albumin deficiency, nephritic syndrome, liver failure, and/or anoxia. 
     
     
         36 . The method of  claim 34  including the steps of:
 using the composition as a marker to measure and quantify vascular leak which is a surrogate for multiple organ failure; and   predicting patients in danger of developing organ failure.