Adjuvant formulation comprising a submicron oil droplet emulsion

Abstract

An adjuvant composition, comprising a metabolizable oil and an emulsifying agent, wherein the oil and the detergent are present in the form of an oil-in-water emulsion having oil droplets substantially all of which are less than 1 micron in diameter. In preferred embodiments, the emulsifying agent is also an immunostimulating agent, such as a lipophilic muramyl peptide. Alternatively, an immunostimulating agent separate from the emulsifying agent can be used.

What is claimed:  
     
         1 - 35 . (canceled)  
     
     
         36 . A composition comprising: 
 (1) an adjuvant composition comprising: (a) 4-5% by volume of squalene; and (b) an emulsifying agent consisting essentially of polyoxyethylenesorbitan monooleate and sorbitan trioleate, wherein the total amount of polyoxyethylenesorbitan monooleate and sorbitan trioleate present is about 1% by weight (w/w); wherein said squalene and said emulsifying agent are present in the form of an oil-in-water emulsion having oil droplets substantially all of which are less than 1 micron in diameter and wherein said composition exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and    (2) an antigenic substance, wherein said antigenic substance is one or more antigenic substances selected from the group consisting of an antigenic substance from a virus, an antigenic substance from a bacterium and an antigenic substance from a parasite, wherein said adjuvant composition is capable of increasing the immune response to the antigenic substance, and further wherein said antigenic substance is not present in the internal phase of the adjuvant composition.    
     
     
         37 . The composition of  claim 36  wherein the antigenic substance comprises a viral subunit.  
     
     
         38 . The composition of  claim 36  wherein the antigenic substance is from a virus selected from the group consisting of hepatitis C virus (HCV), herpes simplex virus (HSV), human immunodeficiency virus (HIV), cytomegalovirus (CMV), influenza virus (flu), poliovirus, and rabies virus.  
     
     
         39 . The composition of  claim 38  wherein the antigenic substance is from herpes simplex virus (HSV).  
     
     
         40 . The composition of  claim 39  wherein the antigenic substance comprises HSV glycoprotein gD.  
     
     
         41 . The composition of  claim 38  wherein the antigenic substance is from human immunodeficiency virus (HIV).  
     
     
         42 . The composition of  claim 41  wherein the antigenic substance comprises HIV glycoprotein gp120.  
     
     
         43 . The composition of  claim 36  wherein the antigenic substance is from a bacterium.  
     
     
         44 . The composition of  claim 43  wherein the bacterium is selected from the group consisting of cholera, diphtheria, tetanus, and pertussis.  
     
     
         45 . The composition of  claim 36  wherein the antigenic substance is from a parasite.  
     
     
         46 . The composition of  claim 45  wherein the parasite comprises a malaria parasite.  
     
     
         47 . The composition of  claim 36  wherein the antigenic substance is used to immunize against meningitis.  
     
     
         48 . The composition of  claim 36  wherein the antigenic substance is selected from the group consisting of a protein, protein-polysaccharide, protein-lipopolysaccharide, polysaccharide, and lipopolysaccharide.  
     
     
         49 . The composition of  claim 36  wherein the antigenic substance comprises a hapten.  
     
     
         50 . The composition of  claim 36  further comprising an additional immunostimulatory agent.  
     
     
         51 . The composition of  claim 50  wherein the additional stimulatory agent is a muramyl peptide.  
     
     
         52 . The composition of  claim 36 , wherein said oil-in-water emulsion has oil droplets substantially all of which are less than 0.6 micron in diameter.  
     
     
         53 . The composition of  claim 36 , wherein said oil-in-water emulsion has oil droplets substantially all of which are less than 0.5 micron in diameter.  
     
     
         54 . The composition of  claim 52 , wherein said oil-in-water emulsion has oil droplets substantially all of which are 0.1 to 0.75 micron in diameter.  
     
     
         55 . The composition of  claim 38  wherein the antigenic substance is from influenza virus.  
     
     
         56 . The composition of  claim 55  wherein the antigenic substance comprises at least one influenza HA antigen.  
     
     
         57 . The composition of  claim 50  wherein the additional immunostimulatory agent is a component of a bacterial cell wall.  
     
     
         58 . An immunogenic composition comprising: 
 (1) an adjuvant composition comprising: (a) 1% to 12% by volume of oil comprising squalene; and (b) 0.02% to 2.5% by weight of an emulsifying agent comprising polyoxyethylenesorbitan monooleate; wherein said oil and said emulsifying agent are present in the form of an oil-in-water emulsion having oil droplets substantially all of which are less than 1 micron in diameter and wherein said composition exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and    (2) an antigenic substance, wherein said antigenic substance is one or more antigenic substances selected from the group consisting of an antigenic substance from a virus, an antigenic substance from a bacterium and an antigenic substance from a parasite, wherein said adjuvant composition is capable of increasing the immune response to the antigenic substance, and further wherein said antigenic substance is present in the external phase of the adjuvant composition.    
     
     
         59 . The composition of  claim 58  wherein the antigenic substance comprises a viral subunit.  
     
     
         60 . The composition of  claim 59  wherein the antigenic substance is from a virus selected from the group consisting of hepatitis C virus (HCV), herpes simplex virus (HSV), human immunodeficiency virus (HIV), cytomegalovirus (CMV), influenza virus (flu), poliovirus, and rabies virus.  
     
     
         61 . The composition of  claim 60  wherein the antigenic substance is from herpes simplex virus (HSV).  
     
     
         62 . The composition of  claim 61  wherein the antigenic substance comprises HSV glycoprotein gD.  
     
     
         63 . The composition of  claim 59  wherein the antigenic substance is from human immunodeficiency virus (HIV).  
     
     
         64 . The composition of  claim 63  wherein the antigenic substance comprises HIV glycoprotein gp120.  
     
     
         65 . The composition of  claim 59  wherein the antigenic substance is from influenza virus.  
     
     
         66 . The composition of  claim 65  wherein the antigenic substance comprises an influenza HA antigen.  
     
     
         67 . The composition of  claim 58  wherein the antigenic substance is from a bacterium.  
     
     
         68 . The composition of  claim 67  wherein the bacterium is selected from the group consisting of cholera, diphtheria, tetanus, and pertussis.  
     
     
         69 . The composition of  claim 58  wherein the antigenic substance is from a parasite.  
     
     
         70 . The composition of  claim 69  wherein the parasite comprises a malaria parasite.  
     
     
         71 . The composition of  claim 58  wherein the antigenic substance is used to immunize against meningitis.  
     
     
         72 . The composition of  claim 58  wherein the antigenic substance is selected from the group consisting of a protein, protein-polysaccharide, protein-lipopolysaccharide, polysaccharide, and lipopolysaccharide.  
     
     
         73 . The composition of  claim 58  wherein the antigenic substance comprises a hapten.  
     
     
         74 . The composition of  claim 58  further comprising an additional immunostimulatory agent.  
     
     
         75 . The composition of  claim 74  wherein the additional immunostimulatory agent is a muramyl peptide.  
     
     
         76 . The composition of  claim 58 , wherein said oil-in-water emulsion has oil droplets substantially all of which are less than 0.8 micron in diameter.  
     
     
         77 . The composition of  claim 58 , wherein said oil-in-water emulsion has oil droplets substantially all of which are less than 0.6 micron in diameter.  
     
     
         78 . The composition of  claim 58 , wherein said oil-in-water emulsion has oil droplets substantially all of which are 100 nm to 750 nm in diameter.  
     
     
         79 . The composition of  claim 78 , wherein said oil-in-water emulsion has oil droplets substantially all of which are 100 nm to 750 nm in diameter.  
     
     
         80 . The composition of  claim 58 , wherein squalene is present in the immunogenic composition in an amount of 1% to 4% by volume and the polyoxyethylenesorbitan monooleate is present in the immunogenic composition in an amount of about 1% by weight.  
     
     
         81 . The composition of  claim 79 , wherein the squalene is present in an amount of about 2.5% by volume.  
     
     
         82 . An immunogenic composition comprising: 
 (1) an adjuvant composition comprising: (a) 1% to 12% by volume of oil comprising squalene; and (b) 0.02% to 2.5% by weight of an emulsifying agent comprising polyoxyethylenesorbitan monooleate; wherein said oil and said emulsifying agent are present in the form of an oil-in-water emulsion having oil droplets substantially all of which are less than 0.6 micron in diameter and wherein said composition exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and    (2) an influenza HA antigen, wherein said adjuvant composition is capable of increasing the immune response to the influenza HA antigen, and further wherein said antigenic substance is present in the external phase of the adjuvant composition.    
     
     
         83 . A composition comprising: 
 (1) an immunostimulating amount of an adjuvant composition comprising: (a) 1-12% by volume of squalene; and (b) an emulsifying agent consisting essentially of polyoxyethylenesorbitan monooleate and sorbitan trioleate, wherein the total amount of polyoxyethylenesorbitan monooleate and sorbitan trioleate present is 0.02-2.5% by weight (w/w); wherein said squalene and said emulsifying agent are present in the form of an oil-in-water emulsion having oil droplets substantially all of which are less than 1 micron in diameter and wherein said composition exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and    (2) an antigenic substance, wherein said antigenic substance is one or more antigenic substances selected from the group consisting of an antigenic substance from a virus, an antigenic substance from a bacterium and an antigenic substance from a parasite, wherein said adjuvant composition is capable of increasing the immune response to the antigenic substance, and further wherein said antigenic substance is present in the external phase of the adjuvant composition.    
     
     
         84 . A composition produced by the method comprising: 
 mixing (1) an immunostimulating amount of an oil-in-water emulsion adjuvant composition comprising: (a) 4-5% by volume of squalene; and (b) 0.02-2.5% by weight of an emulsifying agent consisting essentially of polyoxyethylenesorbitan monooleate, wherein the oil-in-water emulsion has droplets substantially all of which are less than 0.5 micron in diameter, and wherein said composition exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and    (2) an influenza antigen.    
     
     
         85 . The composition of  claim 82 , wherein the influenza antigen comprises HA from two A strains and one B strain.  
     
     
         86 . The composition of  claim 84 , wherein the influenza antigen comprises HA from two A strains and one B strain.  
     
     
         87 . The composition of  claim 82 , wherein the adjuvant composition includes a phosphate buffer.  
     
     
         88 . The composition of  claim 84 , wherein the adjuvant composition includes a phosphate buffer.  
     
     
         89 . The composition of  claim 87 , wherein the phosphate buffer has a pH between 6.0-8.0.  
     
     
         90 . The composition of  claim 88 , wherein the phosphate buffer has a pH between 6.0-8.0.  
     
     
         91 . A composition comprising: 
 (1) an adjuvant composition comprising: (a) 1-12% by volume of squalene; and (b) an emulsifying agent comprising a sorbitan fatty acid ether, wherein the total amount of sorbitan fatty acid ether present is 0.02-2.5% by weight (w/w); wherein said squalene and said emulsifying agent are present in the form of an oil-in-water emulsion having oil droplets substantially all of which are less than 0.5 microns in diameter and wherein said composition exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and    (2) an antigenic substance, wherein said antigenic substance is one or more antigenic substances selected from the group consisting of an antigenic substance from a virus, an antigenic substance from a bacterium and an antigenic substance from a parasite, wherein said adjuvant composition is capable of increasing the immune response to the antigenic substance, and further wherein said antigenic substance is present in the external phase of the adjuvant composition.    
     
     
         92 . The composition of  claim 91 , wherein the sorbitan fatty acid ether is selected from the group consisting of: sorbitan monolaurate, sorbitan monopalmitate, sorbitan monoestearate, sorbitan monooleate, sorbitan sesquioleate, and sorbitan trioleate.  
     
     
         93 . The composition of  claim 92 , wherein the antigenic substance is an influenza antigen.  
     
     
         94 . A composition produced by the method comprising: 
 mixing (1) an oil-in-water emulsion adjuvant composition comprising: (a) 4-5% by volume of squalene; and (b) 0.02-2.5% by weight of an emulsifying agent comprising a sorbitan fatty acid ether, wherein the oil-in-water emulsion has droplets substantially all of which are less than 0.5 microns in diameter, and wherein said emulsion exists in the absence of any polyoxypropylene-polyoxyethylene block copolymer; and (2) an influenza antigen.    
     
     
         95 . The composition of  claim 94 , wherein the sorbitan fatty acid ether is selected from the group consisting of: sorbitan monolaurate, sorbitan monopalmitate, sorbitan monoestearate, sorbitan monooleate, sorbitan sesquioleate, and sorbitan trioleate.  
     
     
         96 . The composition of  claim 92 , further comprising mannitol.