Immunogenic composition in emulsion form comprising two dispersed phases, one comprising an antigen and the other comprising an immunostimulating agent

Patent metadata

Field	Value
Publication number	US-2016136288-A1
Application number	US-201514944825-A
Country	US
Kind code	A1
Filing date	Nov 18, 2015
Priority date	Feb 26, 2013
Publication date	May 19, 2016
Grant date	—

Abstract

The application relates to an immunogenic composition comprising a continuous aqueous phase and at least two dispersed phases 1 and 2 as droplets, wherein the dispersed phase 1 comprises a surfactant 1 bearing an antigen and the dispersed phase 2 comprises an immunostimulating agent 2, to its preparation method and its uses, notably for producing antibodies, as a drug, as a vaccine or in an immunization method.

First claim

What is claimed is:

1 . An immunogenic composition comprising a continuous aqueous phase and at least two dispersed phases 1 and 2 as droplets wherein:
the dispersed phase 1 comprises:
an amphiphilic lipid 1,
a solubilizing lipid 1 comprising at least one fatty acid glyceride,
a co-surfactant 1 comprising at least one chain consisting of alkylene oxide units,
a surfactant 1 bearing an antigen of the following formula (I):
(L 1 -X 1 -H 1 -Y 1 ) v -G-Z 1 -Ag   (I),

wherein:
L 1 represents a lipophilic group,
X 1 , Y 1 , Z 1 and G represent independently a binding group,
H 1 represents a hydrophilic group comprising a polyalkoxylated chain,
v is an integer from 1 to 8, and
Ag represents an antigen,
wherein the molar ratio of the surfactant 1 bearing an antigen of formula (I) over the sum of the co-surfactant 1 and of the surfactant 1 bearing an antigen of formula (I) is from 0.01% to 5%, and the dispersed phase 2 comprises:
an amphiphilic lipid 2,
a solubilizing lipid 2 comprising at least one fatty acid glyceride,
a co-surfactant 2 comprising at least one chain consisting of alkylene oxide units,
an immunostimulating agent 2,
with the proviso that the dispersed phase 2 is free of surfactant bearing an antigen of formula (I).

2 . The immunogenic composition according to claim 1 , wherein, in formula (I):
L 1 is selected from:
a group R or R—(C═O)—, wherein R represents a linear hydrocarbon chain comprising from 7 to 23 carbon atoms,
an ester or amide of fatty acids comprising from 8 to 24 carbon atoms and phosphatidylethanolamine, and
a poly(propylene oxide), and/or
X 1 , Y 1 and Z 1 are independently selected from:
a single bond,
a group Z selected from —O—, —NH—, —O(OC)—, —(CO)O—, —(CO)NH—, —NH(CO)—, —O—(CO)—O—, —NH—(CO)—O—, —O—(CO)—NH— and —NH—(CO)—NH,
a group Alk being an alkylene comprising from 1 to 8 carbon atoms optionally comprising a ring, and
a group Z-Alk, Alk-Z, Alk-Z-Alk or Z-Alk-Z wherein Alk and Z are as defined above and wherein both groups Z of the group Z-Alk-Z are identical or different, and/or
H 1 is selected from a poly(ethylene oxide) typically comprising from 3 to 500 ethylene oxide units, and/or G comprises at least one group G′ having one of the following formulae:

wherein A 102 represents CH or N, R 102 represents H or a linear hydrocarbon chain comprising from 1 to 6 carbon atoms, A 101 represents —O—, —NH—(CO)—or —O—(CO)—, R 100 represents H or a methyl, A 100 represents —O— or —NH— and R 101 represents H, Me or —OMe.

3 . The immunogenic composition according to claim 1 , wherein, in formula (I), the radical L 1 -X 1 —H 1 — consists in one of the groups of the following formulae:

wherein:
R 1 , R 2 , R 3 and R 4 represent independently a linear hydrocarbon chain comprising from 7 to 23 carbon atoms,
A 1 , A 2 , A 3 and A 4 represent O or NH,
m, n, o and p represent independently integers from 3 to 500, and
a represents an integer from 20 to 120,
M represents H or a cation.

4 . The immunogenic composition according to claim 1 , wherein the surfactant 1 of formula (I) has the following formula (I′):

wherein:
R 2 represents a linear hydrocarbon chain comprising from 7 to 23 carbon atoms,
A 2 represents O or NH, and
n represents an integer from 3 to 500,
Ag represents an antigen.

5 . The immunogenic composition according to claim 1 , wherein:
the amphiphilic lipid 1 and/or the amphiphilic lipid 2 is(are) a phospholipid, and/or the solubilizing lipid 1 and/or the solubilizing lipid 2 consist(s) of a mixture of saturated fatty acid glycerides including at least 10% by weight of C 12 fatty acids, at least 5% by weight of C 14 fatty acids, at least 5% by weight of C 16 fatty acids and at least 5% by weight of C18 fatty acids, and/or the co-surfactant 1 and/or the co-surfactant 2 is(are) selected from polyethyleneglycol/phosphatidyl-ethanolamine conjugate compounds, fatty acid and polyethyleneglycol ethers, fatty acid and polyethyleneglycol esters and block copolymers of ethylene oxide and propylene oxide, and the polyalkoxylated chain of the co-surfactant 1 and/or the co-surfactant 2 comprise(s) from 10 to 200 ethylene oxide/propylene oxide units.

6 . The immunogenic composition according to claim 1 , wherein the immunostimulating agent 2 is a “Toll-like receptor” (TLR) ligand.

7 . The immunogenic composition according to claim 1 , wherein:
the dispersed phase 1 comprises a biological targeting ligand 1 either grafted or not on the co-surfactant 1, and/or the dispersed phase 2 comprises a biological targeting ligand 2 either grafted or not on the co-surfactant 2, and/or the dispersed phase 1 comprises an agent 1 of interest selected from an optical agent or a physical agent, and/or the dispersed phase 2 comprises an agent 2 of interest selected from an optical agent a physical agent.

8 . The immunogenic composition according to claim 1 , wherein the dispersed phase 2 comprises:
oligodeoxynucleotide CpG as immunostimulating agent 2, a co-surfactant 2 comprising at least a poly (ethylene oxide) chain comprising at least 25 ethylene oxide units, a cationic surfactant 2, and optionally a helper lipid 2.

9 . The immunogenic composition according to claim 1 , wherein the dispersed phase 2 comprises:
from 0.1 to 10% by weight of monophosphoryl lipid A as immunostimulating agent 2 with respect to the weight of dispersed phase 2, and
from 0.1 to 40% by weight of co-surfactant 2 with respect to the weight of dispersed phase 2.

10 . A method for preparing the immunogenic composition as defined in claim 1 , comprising mixing an emulsion 1 comprising a continuous aqueous phase and at least a dispersed phase 1 as defined in claim 1 , and an emulsion 2 comprising a continuous aqueous phase and at least a dispersed phase 2 as defined in claim 1 .

11 . A drug or a vaccine comprising the immunogenic composition according to claim 1 .

12 . A method for inducing an immune response against an antigen, comprising the administration of the immunogenic composition according to claim 1 to a patient in need thereof.

13 . A method for treating or preventing an infection, a cancer, an inflammatory disease, an allergy, an age-related disease or a neurodegenerative disease comprising the administration of a pharmaceutically acceptable amount of the immunogenic composition according to claim 1 .

14 . The method according to claim 13 , wherein the infection is a viral, bacterial, parasite or prion type infection.

15 . A method for producing polyclonal antibodies, comprising the steps of:
(a) immunizing an animal with an immunogenic composition as defined according to claim 1 , so as to induce a humoral immune response against said antigen, and (b) harvesting the induced polyclonal antibodies directed against said antigen.

16 . A method for producing monoclonal antibodies, comprising the steps of:
(i) immunizing an animal with an immunogenic composition as defined according to any of claim 1 , (ii) recovering and isolating the B lymphocytes of the immunized animal in step (i), (iii) producing a hybridoma and cultivating said hybridoma in or

Assignees

• Commissariat à l'énergie atomique et aux énergies alternatives
• Inst Nat Sante Rech Med

Inventors

• Navarro Y Garcia Fabrice
• Bayon Emilie
• Marche Patrice
• Courant Thomas

Classifications

• A61K2039/55572

  Lipopolysaccharides; Lipid A; Monophosphoryl lipid A · CPC title

• A61K47/48215

  Human Necessities · mapped topic

• A61K2039/645

  Dendrimers; Multiple antigen peptides · CPC title

• C07K16/18

  against material from animals or humans · CPC title

• A61K2039/627

  characterised by the linker · CPC title