Formulations useful for immunotherapy for cancers containing bacterial component as an active ingredient

Abstract

An oil-in-water emulsion that comprise a bacterial component as an active ingredient as well as essentially an oil, surfactant, and a stabilizer, and that can be used as an agent for immunotherapy, and the preparation therefor are provided.

1 - 10 . (canceled) 
     
     
         11 . A process for preparation of an oil-in-water emulsion wherein the emulsion is negative for agglutination reaction with lectin, and a bacterial component that is selected from a BCG-CWS and  Nocardia rubra -CWS is encapsulated in oil, which comprises the following steps:
 (a) stirring a mixture of a bacterial component that is selected from a BCG-CWS and  Nocardia rubra -CWS, an oil, and a dispersion-aiding solvent to disperse the bacterial component in the mixture;   (b) evaporating off the dispersion-aiding solvent to form an oil wherein the bacterial component is homogeneously dispersed, or an oil droplet wherein the bacterial component is encapsulated in the oil; and then,   (c) adding an aqueous solution containing a surfactant thereof, and emulsifying the mixture.   
     
     
         12 . (canceled) 
     
     
         13 . The process for preparation of an oil-in-water emulsion of  claim 11 , wherein the bacterial component is a BCG-CWS, and the oil is squalane. 
     
     
         14 . The process for preparation of an oil-in-water emulsion of  claim 32  or  33 , wherein the dispersion-aiding solvent is ethanol or toluene. 
     
     
         15 . The process for preparation of an oil-in-water emulsion according to any one of  claims 11 ,  13 ,  32  and  33 , wherein the oil droplet is dispersed in a manner that the diameter of the particle is about 100 μm or less. 
     
     
         16 . A process for preparation of an oil-in-water emulsion, which comprises the following steps:
 (a) stirring a mixture of a bacterial component, an oil, and a dispersion-aiding solvent to disperse the bacterial component in the mixture;   (b) evaporating off the dispersion-aiding solvent; and then;   (c) performing the following two-step emulsification process which comprises adding an aqueous solution containing a surfactant:
 i) adding an aqueous solution containing a surfactant at a low concentration to the evaporated material, and stirring gently the mixture, thereby performing rough emulsification; and 
 ii) optionally, adjusting the concentration of the surfactant in the roughly emulsified solution, and stirring vigorously the mixture, thereby performing adequate emulsification in order to provide a desired particle diameter distribution. 
   
     
     
         17 . The process for preparation of an oil-in-water emulsion of  claim 16 , wherein an amount of the surfactant in the aqueous solution containing the surfactant at a low concentration used in rough emulsification of the two-step emulsification process is 10% or less of the oil. 
     
     
         18 . The process for preparation of an oil-in-water emulsion of  claim 16  or  17 , wherein the surfactant is Polysorbate 80 (Tween80). 
     
     
         19 . The process for preparation of an oil-in-water emulsion of  claim 16 , wherein the bacterial component is a BCG-CWS or a CWS of  Nocardia rubra.    
     
     
         20 . The process for preparation of an oil-in-water emulsion of  claim 16 , wherein the bacterial component is a BCG-CWS, and the oil is squalane. 
     
     
         21 - 26 . (canceled) 
     
     
         27 . A process for preparation of an oil or an oil droplet wherein a bacterial component that is selected from a Bacillus Calmette-Guerin cell wall skeleton and  Nocardia rubra  cell wall skeleton having a particle diameter of about 100 μm or less is homogeneously dispersed, which compromises the following steps:
 (a) stirring a mixture of the bacterial component, an oil, and a dispersion aiding solvent to disperse the bacterial component in the mixture;   (b) evaporating off the dispersion aiding solvent to form an oil wherein the bacterial component is homogeneously dispersed, or a droplet wherein the bacterial component is encapsulated in the oil.   
     
     
         28 . The process according to  claim 27 , wherein the particle diameter is about 25 μm or less. 
     
     
         29 . The process according to  claim 27 , wherein the oil is squalane. 
     
     
         30 . The process according to  claim 27 , wherein the dispersion aiding solvent is selected from the group consisting of an aromatic hydrocarbon, an aliphatic hydrocarbon, a halogenated hydrocarbon, a lower alcohol, an acetate an ether and a ketone. 
     
     
         31 . The process according to  claim 30 , wherein the dispersion aiding solvent is ethanol or toluene. 
     
     
         32 . The process according to  claim 11 , wherein the dispersion aiding solvent is selected from the group consisting of an aromatic hydrocarbon, an aliphatic hydrocarbon, a halogenated hydrocarbon, a lower alcohol, an acetate, an ether and a ketone. 
     
     
         33 . The process according to  claim 27 , wherein the dispersion aiding solvent is selected from the group consisting of an aromatic hydrocarbon, an aliphatic hydrocarbon, a halogenated hydrocarbon, a lower alcohol, an acetate, an ether and a ketone.