FR2824279A1 - CONCENTRATED W / O EMULSION - Google Patents

Abstract

The invention relates to a composition in the form of a water-in-oil type emulsion, comprising a fatty phase O and an aqueous phase W. The inventive composition is characterised in that the W/O weight ratio is greater than 1 and said composition is oily. The invention also relates to the use thereof as an adjuvant for vaccines and to a vaccine preparation method.

Description

a separator according to one of claims 1 to 11.

  - 1 - - The present invention relates to new adjuvants for vaccine compositions as well as compositions comprising at least one antigen, in particular an anti

  gene of viral, bacterial or parasitic origin and at least one adjuvant.

  The development of vaccines inactivated or containing purified antigens is becoming increasingly important, as it makes it possible to avoid undesirable side effects. However, improving the quality of antigens comes at the expense of their im

  munogène. It is for this reason that they are associated with immunity adjuvants.

  Immunity adjuvants are products that enhance the reactions of the immune system when administered in the presence of antigens of viral, bacterial or synthetic origin. They cause the massive appearance of macrophages at the injection site, then in the lymphatic nodules, increase the production of specific immunoglobu lines, antibodies, and stimulate many cells involved in

  immune defense mechanisms.

  These adjuvants are of various natures. They may for example consist of

liposomes or emulsions.

  Very effective Freund's adjuvants; they result from the association of a mineral oil and a mannitol ester containing or not containing a killed mycobacterium. The vaccines produced by mixing in equal parts a Freund's adjuvant with an aqueous antigenic medium, remain in fact the benchmarks worldwide for studies in the laboratory. They are in the form of water in oil emulsions (W / O), that is to say emulsions in which the continuous phase is oil. The active principle is generally localized in the aqueous phase which is often a buffered saline solution. This phase is in the form of drops separated from each other by an oily film. This formula

  tion allows to obtain amplified and prolonged blological responses over time.

  However emulsions of this type (W / O) are generally very viscous and are therefore difficult to inject. Elies often require the use of syringes with a large needle diameter and cause pain during injection and trauma

injection site.

  The aqueous phase content of the injectable W / O emulsions is around 30% to 40% by weight. The maximum encountered is 50%. However, this limit constitutes an obstacle in particular to the development of polyvalent vaccines, in which several antigens are associated and for which it would be preferable to obtain W / O emulsions having

  contents in aqueous phase greater than 50%.

  However, in a W / O emulsion with a low aqueous phase content, up to about 20%

  by weight, the viscosity of the emulsion is very close to the viscosity of the oil.

  The increase in the proportion in aqueous phase increases its viscosity. Thus - an emulsion at 20% by weight of water having a viscosity of 100 mPa.s, turns into a cream very difficult to inject when the water content is increased to 50

% in weight.

  Certain commercial oily adjuvants such as MONTANIDE_ ISA 70 allow to obtain W / O injectable emulsions containing approximately 30% by weight, of aqueous phase, 70% by weight of oily phase and having viscosities of the order of 50 to 100 mPa.s. Other adjuvants, such as MONTANIDE_ ISA 50, make it possible to obtain injectable W / O emulsions containing approximately 50% by weight of aqueous phase, 50% by weight of oily phase and having viscosities lower than 250 mPa.s. r

  The international patent application published under the number W0 99/20305 divul-

  gue the use of surfactants on mannitol oleate with mineral oils like MARCOL_ 52 to make emulsions, called fluids, of viscosities of the order of 500 mPa..s. The concept of fluid emulsion depends largely on the field of application. For injectable emulsions, the upper viscosity limit for a fluid emulsion is defined with respect to the viscosity of a reference W / O type emulsion, which contains 50% by weight of aqueous phase and 50% by weight of incomplete Freund's adjuvant (IFA); this viscosity is of the order of approximately 2,000 mPa.s. This sensation is considered to be very viscous. A W / O type emulsion will be called fluid if its viscosity is less than 4 times that of this reference emulsion, that is to say less than 500 mPa.s at 25 C

(BROOKFIELD LVT M 2 V 30).

  However, fluid emulsions are generally less stable than more viscous emulsions, since phase shifts are observed for only a few days,

after their preparation.

  The Applicant has therefore sought to develop water-in-oil type emulsions (W / O), concentrated in the aqueous phase which do not have the drawbacks set out above, that is to say which are fluid and stable. over time. The criterion of stability of the emuption being judged satisfied if it presents no major defect after 1

  months at the following three temperatures: 4 C, 25 C and 37 C.

  The subject of the invention is a composition comprising a fatty phase H and an aqueous phase E, in the form of a water-in-oil type emulsion and characterized

  in that the weight ratio W / O is greater than 1.

  According to a first particular aspect of the present invention, the weight ratio E

/ H is less than or equal to 3.

  The fatty phase constituting the composition which is the subject of the present invention, generally comprises one or more compounds chosen from oils of mineral, vegetable or animal origin, the alkyl esters of said oils, the alkyl esters of fatty acids or the ethers fatty acid alkyls, fatty acid esters and polyols or fatty alcohol ethers and polyols. As examples of oils of mineral origin, there are oils of petroleum origin, such as white mineral oils such as MARCOL_ 52 or DRAKEOL 6 VR. Examples of vegetable oils are peanut oil, olive oil, sesame oil, soybean oil, wheat germ oil, oil of grape pines, sunflower oil, castor oil, linseed oil, soybean oil, mas hulia, coconut oil, palm oil, oil nuts, hazelnut oil, rapeseed oil or squalane or olive squalene. As examples of oils of animal origin, there is oil

  of spermaceti, tallow oil, squalane or squalene extracted from fish livers.

  As examples of alkyl esters of oils, there are the ethyl, linear or branched propyl or butyl, linear or branched ethyl esters of said oils. As fatty acids suitable for the preparation of the esters mentioned above, there are more particularly, those comprising from 12 to 22 carbon atoms, such as for example, myristic acid, palmitic acid, olelic acid, Ricinoleic acid or

  Isostearic acid and advantageously a liquid fatty acid at 20 C.

  As examples of fatty acid esters or fatty acid ethers, there are the fatty acid alkyl esters, such as, ethyl oleate, methyl oleate, isopropyl myristate or octyl palmitate, esters of fatty acids and polyols or ethers of fatty alcohols and polyols, such as fatty acid monoglycerides, fatty acid diglycerides, fatty acid triglycerides, fatty acid esters with a polyglycerol or fatty acid and propylene glycol esters, and more particularly fatty acid esters with a hexol, such as for example sorbitol or mannitol, acid esters

  fatty with a hexcl anhydride, such as sorbitan or mannitane.

  In the context of the present invention, the fatty phase may comprise only one of the compounds mentioned above or else a mixture of several of the compounds mentioned above. The fatty phase, constituting the composition which is the subject of the present invention, also comprises one or more surfactants having an overall HLB number included

between 5 and 8.

  The fatty phase comprises approximately between 1% and 15% by weight, preferably in

  be 3% and 10% by weight of surfactants.

  - For the purposes of the present invention, the HLB number of a surfactant is calculated by the

  - formula HLB = 20 (1-lSlla), in which is Is represents the saponification index and the 'I'-

  acid dice of said surfactant. In the case of a mixture of surfactants, the HLB of the

  lange is the weighted sum of the H LB of each surfactant. These two clues, from sapo

  S nification and acid, are determined by methods described in the Pharmacopoeia eu-

  pean. A more particular subject of the invention is a composition as defined above in which the surfactant or the mixture of surfactants has a

  overall HLB number is greater than or equal to 6 and less than 8.

  The surfactants used are generally chosen from bodies

modified bold.

  The modified fatty substances used in the context of the present invention can

  be of vegetable or animal mineral origin. As modified fatty substances of mineral origin

  There are oils of petroleum origin.

  As modified fatty substances of vegetable origin, there are modified vegetable oils, for example, modified oils of peanut, olive, sesame, soybean, wheat germ, sunflower grape seeds , castor, flax, soy, mas, copra,

  palm, nuts, hazelnuts or rapeseed.

  As modified fatty substances of animal origin, there is for example squalane mo

  modified, modified squalene, modified spemmaceti oil or modified tallow oil.

  The term “modified fatty substances” denotes in particular the alkoxylated derivatives of

  fatty and more particularly the alkoxylated derivatives of oils or the alkoxylated derivatives of

  alkyl oils and more particularly, the ethoxylated and / or propoxylated derivatives of oils or the ethoxylated and / or propoxylated derivatives of the methyl, ethyl esters,

  linear or branched or butyl, linear or branched propyls, of said oils. Linen-

  vention more specifically relates to a composition as defined above.

  ment, in which the modified fatty substance is chosen from ethoxylated derivatives of oils

  having a number of EOs, between 1 and 10.

  The term “modified fatty substances” also denotes esters of fatty acids and of polyols or ethers of fatty alcohols and of polyols, and more particularly, esters of fatty acids with a hexol, such as for example sorbitol or mannitol or esters of fatty acids with a hexol anhydride, such as sorbitan or mannitane, alkoxylated derivatives of fatty acid esters and of polyols or alkoxylated derivatives of fatty alcohol ethers and of polyols, like the triglycerides of alkoxylated fatty acids, the alkoxylated esters of polygly cerol of fatty acids, and more particularly the alkoxylated esters of fatty acids with a hexol, such as for example sorbitol or mannitol or the alkoxylated esters of acids fat

2824279

  with a hexol anhydride, such as sorbitan or mannitane having an EO number,

- between 1 and 20.

  The term “fatty acid and polyol esters” is used in the context of the present invention.

  vention, monoesters of fatty acids and polyois or polyesters of fatty acids and polyol such as for example diesters of fatty acids and polyols or triesters of fatty acids.

  fats and polyols. It is the same for the polyalkoxylated derivatives of said esters.

  The term “fatty acid and polyois ethers” is used in the context of the present invention.

  vention, monoethers of fatty acids and polyols or polyethers of fatty acids and polyol such as for example diethers of fatty acids and polyols or triethers of aci

  fats and polyols. The same is true for the polyalkoxylated derivatives of said ethers.

  The subject of the invention is more particularly, a composition as defined above, in which the modified fatty substances are chosen from the ethoxylated derivatives of fatty acid esters and of poiyols or the ethoxylated derivatives of fatty alcohol ethers and of polyols, and more particularly, the ethoxylated esters of fatty acids with a hexol, such as for example sorbitol or mannitol or the ethoxylated esters of fatty acids with a hexol anhydride, such as sorbitan or mannitane having a number of EOs,

between 5 and 10.

  As fatty acids suitable for the preparation of the modified fatty substances described above

  above, there are those comprising on average from 12 to 22 carbon atoms, such as for example, those comprising from 16 to 18 carbon atoms, such as oleic acid, acid

  ricinoleic or isostearic acid and advantageously liquid fatty acids at 20 C.

  The composition, as defined above, more particularly contains one or more

  several modified fatty substances as described above, derived from olelic acid.

  According to another particular aspect of the present invention, the surfactant or mixture of surfactants present in the fatty phase, essentially consists of one or more esters chosen from mannitane esters, sorbitan esters,

  alkoxylated mannitane esters or alkoxylated sorbitan esters.

  In a very particular way, the surfactant present in the fatty phase consists of a mixture of ma nnitane oleate and polyethoxylated mannane oleate, a mixture of sorbitan oleate and polyethoxylated sorbitan oleate, a mixture sorbitan oleate and polyethoxylated mannitane oleate or a mixture of mannitane oleate and polyethoxylated sorbitan oleate, said mixtures having an overall HLB number is

  greater than or equal to 6 and less than 8.

  The composition according to the invention can also comprise one or more conventional immune stimulating agents, such as Avridine_, N, Ndioctadecyl-N ', N'-bis (2

6 2824279

  hydroxyethyl) propanediamine, derivatives of MDP (muramyl dipeptide), in particular - threonyl-MDP, derivatives of mycolic acid or derivatives of Lipid A. The composition according to the invention optionally comprises one or more organic salts of cations water-soluble metals, such as for example calcium gluconate, manganese gluconate aluminum salicylate or aluminum acetate so-

  luble. When the adjuvant composition according to the invention comprises, a pharmaceutical salt

  only acceptable, it is at a concentration of 0.02 to 3000 mg / cm3, preferably

  rate 0.1 to 1000 mg / cm3, more preferably 0.1 to 150 mg / cm3.

  The composition according to the invention can also comprise one or more salts inso

  lubles generally used as immunity adjuvants, such as aluminum hydroxide

nium or calcium phosphate.

  According to another aspect of the present invention, that relates to a composition as defined above further comprising, at least one antigen or at least one in vivo generator of a compound comprising a sequence of amino acids and a

  non-zero quantity of a composition as defined above.

  The term “antigen or at least one in vivo generator of a compound comprising an amino acid sequence” denotes either killed microorganisms, such as viruses, bacteria or parasites, or purified fractions of these microorganisms. , or living microorganisms whose pathogenic power has been reduced. As examples of viruses which can constitute an antigen according to the present invention, there is the rabies virus, herpes viruses, such as Aujeszky's disease virus, orthomixoviruses such as Influenzae, ies picornaviruses such as the vi, rus of the foot-and-mouth disease or the retroviruses

  such as HIV. As a microorganism of the bacterial type which can constitute an anti-

  gene according to the present invention, there may be mentioned E. Coli, and those of the genera Pasteurella, Furonculosis, Vibriosis, Staphylococcus and Streptococcus. Examples of parasites are those of the genera Trypanosoma, Plasmodium and Leishmania. Mention may also be made of recombinant viruses, in particular non-enveloped viruses, such as adenoviruses, vaccinia virus, Canarypox virus, herpes viruses or baculoviruses. A live, non-enveloped viral recombinant vector is also designated, the genome of which contains, inserted from

  preferably in a nonessential part for the replication of the corresponding enveloped virus

  laying, a sequence coding for an antigenic subunit inducing an antibody synthesis eVou a protective effect against the above enveloped virus or pathogenic microorganism; these antigenic subunits can be, for example, a protein, a glycoprotein, a peptide or a peptide fraction eVou which protects against infection by a living microorganism such as an envelope virus, a bacterium or a parasite. The exogenous gene inserted into the microorganism can, for example, come from an Au virus

jeszky or HIV.

  Mention may in particular be made of a recombinant plasmid consisting of a nucleotide sequence, into which an exogenous nucleotide sequence is inserted, originating from a microorganism or a pathogenic virus. The aim of this last nucleotide sequence is to allow the expression of a compound comprising an amino acid sequence, this compound itself having the aim of triggering an immune reaction in

a host organism.

  By generator "in vivo" of a compound comprising a sequence of amino acids born, we mean a whole biological product capable of expressing this compound in host organism into which said generator has been introduced in vivo. The compound comprising the amino acid sequence can be a protein, a peptide or a glycoprotein. These in vivo generators are generally obtained by processes derived from genetic engineering. More particularly, they can consist of living microorganisms, generally a virus, playing the role of a recombinant vector, into which a nuclectide sequence, in particular an exogenous gene, is inserted. These compounds are known as such and used in particular as a recombinant unitary vaccine. In this regard, reference may be made to the article by M. ELOIT et al., Joumal of virology (1990) 71, 2925-2431 and to the international patent applications published under the numbers WO-A- 91/00107 and WO -A-94/16681. The in vivo generators according to the invention can also consist of a recombinant plasmid comprising an exogenous nucleotide sequence, capable of ex

  priming in a host organism a compound comprising an amino acid sequence.

  Such recombinant plasmids and their mode of administration in a host organism were described in 1990, by LIN et al., Circulation 82: 2217,2221; COX et al., J. of VIROL, Sept. 1993, 67, 9, 5664-5667 and in the international application published under the number WO 95/25542. Depending on the nature of the nucleotide sequence included in the generator in vivo, the compound comprising the amino acid sequence which is expressed within the host organism, can: (i) be an antigen, and allow the triggering of a immune reaction, (ii) have a curative action vis-à-vis a disease, essentially a functional disease, which is triggered in the host organism. In this case, the general

  In vivo therapy allows treatment of the host, of the gene therapy type.

  By way of example, such a curative action may consist in a synthesis by the in vivo generator of cytokines, such as interleukins, in particular interleukin 2. These allow the triggering or the strengthening of an immune reaction aimed at

  selective elimination of cancer cells.

  The vaccine composition as defined above comprises an antigen concentration which depends on the nature of this antigen and on the nature of the subject treated. It is however particularly remarkable that an adjuvant according to the invention makes it possible to notably reduce the usual dose of antigen required. The adequate concentration of antigen can be determined in a conventional manner by a person skilled in the art. Generally, this dose is of the order of 0.1 g I cm3 to 1 g / cm3 more generally between 1 ug I cm3 and 100 mg I cm3. The concentration of said generator in vivo in the composition according to the invention again depends, in particular, on the nature of said generator and on the host in which it is administered. This concentration can be readily determined by those skilled in the art, based on routine experience. As an indication, when the in vivo generator is a recombinant microorganism, its concentration in the composition according to the invention generally between 102 and 1045 microorganisms / cm3 and preferably between 105 and 102 microorganisms I cm3. When the in vivo generator is a recombinant plasmid, its concentration in the composition according to the invention can be between 0.01 9 / dm3 and 100 91 dm3. The vaccine, as defined above, is prepared by mixing the adjuvant phase and the antigenic phase, possibly adding

  water or a pharmaceutically acceptable diluent medium.

  The subject of the invention is preferably the emulsions of type E IH such as challenge

  previously denied, which are fluid.

  Fluid emulsion means emulsions whose viscosity, measured at 25 C with a BROOKFIELD LVT viscometer with rotating mobile (2 speed mobile 30 rpm) is

  less than 500 mPa.s (BROOKFIELD LVT M 2, V 30).

  The vaccine composition as described above can be used as a preventive or curative medicament. Depending on the nature of the antigen or generator in vivo, a composition according to the invention can be administered to fish, crustaceans such as shrimps, poultry, in particular, geese, turkeys, pigeons and chickens. , To canines such as dogs, felines such as cats, pigs, primates, bovines, ovids and horses. The composition according to the invention can also be ad min is trated to humans. The administration of the composition can be carried out in a conventional manner by the parenteral route, in particular by subcutaneous, intramuscular or intraperito injection.

  Neale. It can also be done orally or nasally.

  According to another aspect of the invention, the subject of the invention is a composition essentially consisting of 1% and 15% by weight, and preferably 3% and 10% by weight, of surfactants having an overall HLB number between 5 and 8, and from 85% to 9g%, and preferably from 90% to 97% by weight, of one or more compounds chosen from oils of mineral, vegetable or animal origin, the afkyl esters of said oils ,

9 2824279

  fatty acid alkyl esters or fatty acid alkyl ethers, fatty acid esters

  - fats and polyols or ethers of fatty alcohols and polyols.

  In the oily composition as defined above, the oil is more particular

  rely chosen from white mineral oils.

  In the oily composition as defined above, the surfactant or the mixture of surfactants preferably has an overall HLB number greater than or equal to

6 and less than 8.

  In the oily composition as defined above, the surfactant or the

  mixture of surfactants are chosen from modified fatty substances.

  The term “modified fatty substance” denotes in particular the alkoxylated derivatives of oils or the alkoxylated derivatives of alkyl esters of oils and more particularly, the ethoxylated derivatives or propoxylated oils or the ethoxylated and / or propoxylated derivatives of methyl, ethyl esters , linear or branched propyl or butyl, linear or branched, of said oils and more particularly from the ethoxylated derivatives of oils having a

  number of EOs, between 1 and 20, preferably between 5 and 10.

  The term “modified fatty substance” also denotes esters of fatty acids and of polyols or ethers of fatty alcohols and of polyols, and more particularly, esters of fatty acids with a hexol, such as for example sorbitol or mannitol or esters of fatty acids with a hexol anhydride, such as sorbitan or mannitane, alkoxylated derivatives of fatty acid esters and of polyols or ancoxylated derivatives of fatty alcohol ethers and of polyols, and more particularly, the alkoxylated esters of fatty acids with a hexot, such as for example sorbitol or mannitol or the aicoxylated esters of fatty acids with an anhydrous hexol, such as sorbitan or mannitane having a number d 'OE, between 1 and 20, more particularly, the ethoxylated derivatives of fatty acid esters and of polyols or the ethoxylated derivatives of ethers of fatty alcohols and of polyols, and very particularly, the ethoxylated esters of fatty acids with a hexol, such as for example pe sorbit ol or mannitol or the ethoxylated esters of fatty acids with a hexol anhydride, such as sorbitan or

  mannitane having an EO number between 5 and 10.

  In the above definition, the fatty acids suitable for the preparation of modified fatty substances preferably contain on average 12 to 22 carbon atoms.

  bone and more particularly 16 to 18 carbon atoms.

  According to a particular aspect of the present invention, in the oily composition as defined above, the surfactant or the mixture of surfactants are

derivatives of olelic acid.

  According to another particular aspect of the present invention, in the oily composition as defined above, the surfactant or surfactants are chosen from mannitane esters, sorbitan esters, alkoxylated mannitane esters

  - or the alkoxylated sorbitan esters.

  According to a very particular aspect of the present invention, the oily composition as defined above, contains as surfactant system, a mixture of mannitane oleate and ethoxylated mannitane oleate having a number of EOs, between 5 and 10. According to the last aspects of the present invention, the subject of the present invention is a process for the repair of a water-in-oil vaccine emulsion. as defined previously; comprising the addition in an EIH weight ratio greater than 1, of an aqueous medium containing at least one antigen or at least one generator in viYo of a compound comprising an amino acid sequence with the oily composition as defined above above, as well as the use of said oily composition as an adjuvant

  in injectable compositions such as vaccines.

  The process as defined above can be carried out cold, it is

  say at a temperature between û C and 30 C.

  The following examples illustrate the invention but do not limit it. Preparation of the compositions according to the invention.

  EXEMPTION 1 An emulsion is prepared according to the invention, by pouring an aqueous phase consisting of a saline solution of physiological saline, onto an oily phase, with stirring with a high shear mixer (SILVERSON L4RT). 200 g of emulsion are prepared for each test, the aqueous phase representing 65/0 of the weight of the emulsion. The fatty phase consists of: 90% by weight of MARCOLrM 52, 3% by weight of mannitane monooleate and

  7% by weight of polyethoxylated mannitane monooleate (EOI = 8).

  The mixture of mannitane monooleate and polyethoxylated mannitane monooleate (8

OE) has an HLB number equal to 6.2.

  The viscosity of the emulsion is checked after 24 hours of stabilization at 25 C

  (BROOKFIELD LVT M 2 V 30). It is equal to 430 mPa.s.

  The sample obtained is divided into three vials for evaluation of stabilities

  at the following three temperatures: 4 C, 25 C and 37 C.

  It is found that this emulsion is stable at the three temperatures tested. After 1

  After storage, the emulsion is not broken and the oil release is less than 5%.

11 2824279

Example 2:

  - An emulsion is prepared according to the invention as described in Example 1, in which the aqueous phase represents 65% by weight thereof, and in which the fatty phase constitutes 92% by weight of MARCOL_ 52, 4, 3% by weight of sorbitan trioleate and

  3.7% by weight of polyethoxylated sorbitan trioleate (OE number = 20).

  The mixture of sorbitan trioleate and polyethoxylated sorbitan trioleate (OE number

= 20) has an HLB number equal to 6.1.

  The viscosity of the emulsion is checked after 24 hours of stabilization at 25 C.

.

  It is equal to 134mPa.s. (BROOKFIELD LVT M 2 V 30).

  The sample obtained is divided into three vials for evaluation of stabilities

  at three temperatures (4 C / 25 C / 37 C).

  It is found that this emulsion is stable at the three temperatures tested After a

  months of storage, the emulsion is not broken and the release of oil is negligible.

Example 3:

  An emulsion according to the invention is prepared as described in Example 1, in which the aqueous phase represents 65% by weight thereof, and in which the fatty phase consists of: 92% by weight of MARCOL_ 52, 4% in weight of sorbitan monooleate and

  2.6% by weight of polyethoxylated mannitane monooleate (OE number = 8).

  The mixture of sorbitan monooleate and polyethoxylated mannitane monooleate

  (OE index = 8), has an HLB number equal to 7.6.

  The viscosity of the emulsion is checked after 24 hours of stabilization at 25 C.

  At Eile is equal to 360 mPa.s. (BROOKFIELD LVT M 2 V 30).

  The sample obtained is divided into three vials for evaluation of stabilities

  at three temperatures (4 C / 25 C / 37 C).

  It is found that this emulsion is stable at the three temperatures tested. After one month of storage, the emulsion is not broken and the oil release is less than 5%.

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Claims (18)

  1. Composition in the form of an emulsion of the water-in-oil type, comprising a fatty phase H and an aqueous phase E, characterized in that the weight ratio W / O is greater than 1. 2. Composition as defined in claim 1, in which the ratio W / O is less than or equal to 3.   3. Composition as defined in any one of claims 1 or 2,   in which the fatty phase constituting the composition comprises one or more compounds chosen from oils of mineral, vegetable or animal origin, the alkyl esters of said oils, the alkyl esters of fatty acids or the alkyl ethers of fatty acids, esters of fatty acids and polyols or ethers of fatty alcohols and polyols. 4. Composition as defined in claim 2, in which the oil is   chosen from white mineral oils.   5. Composition as defined in any one of Claims 1 to 4,   in which the fatty phase also comprises one or more surfactants having   a global HLB number between 5 and 8.   6. Composition as defined in any one of claims 1 or 5,   in which the fatty phase comprises approximately between 1% and 15% by weight and preferably   between 3% and 10% by weight of surfactants.   7. Composition as defined in any one of claims 5 or 6,   wherein the surfactant or mixture of surfactants has an HLB number   overall greater than or equal to 6 and less than 8.   8. Composition tç !! e quç dfiniç à! 'Unç que! Conquç of claims 5 to 7;   in which the surfactant or the mixture of surfactants are chosen from modified fatty substances.   9. Composition as defined in claim 8, in which the modified fatty substances are chosen from alkoxylated derivatives of oils or alkoxylated derivatives of alkyl esters of oils and more particularly, ethoxylated derivatives or propoxylated oils or the ethoxylated and / or propoxylated derivatives of the methyl, ethyl, linear or branched or butyl, linear or branched propyl esters, of said oils and more particularly from the ethoxylated derivatives of oils having a number of EOs, between 1   and 20, preferably between 5 and 10.   10. Composition as defined in claim 8, in which the modified fatty substances are chosen from esters of fatty acids and of polyols or ethers of al 13 2824279   fatty cools and poyols, and more particularly, esters of sra acids with a hexol, such as for example sorbitol or mannitol or esters of fatty acids with an anhydrous hexol, such as sorbitan or mannitane , alkoxylated derivatives of fatty acid esters and polyols or alkoxylated derivatives of fatty alcohol ethers and polyols, and more particularly, alkoxylated esters of fatty acids with a hexol, such as for example sorbitol or mannitol or alkoxylated esters of fatty acids with a hexol anhydride,   such as sorbitan or mannitane having an EO number between 1 and 20.   11. Composition as defined in claim 10, in which the modified fatty substances are chosen from ethoxylated derivatives of fatty acid esters and of polyols or ethoxylated derivatives of ethers of fatty alcohols and of polyols, and more in particular, the ethoxylated esters of fatty acids with a hexol, such as for example sorbitol or man nitol or the ethoxylated esters of fatty acids with a hexol anhydride, such as sorbitan   or mannitane having an EO number between 5 and 10.   12. Composition as defined in one of claims 10 or 11, in the   which fatty acids suitable for the preparation of modified fatty substances, contain on average from 12 to 22 carbon atoms and more particularly 16 to 18 carbon atoms.   13. Composition as defined in any one of claims 5 to 12,   wherein the surfactant or mixture of surfactants are derivatives of Oleic acid.   14. Composition as defined in any one of claims 5 to 13,   in which the surfactant or surfactants are chosen from mannitane esters, sorbitan esters, alkoxylated mannitane esters or esters of alkoxylated sorbitan.   15. omission tç !! ç quç dPfinite to any of claims 7 to 14,   in which the surfactant present in the fatty phase consists of a mixture of mannitane oleate and polyethoxylated mannitane oleate, a mixture of sorbitan oleate and polyethoxylated sorbitan oleate, a mixture of sorbitan oleate and polyethoxylated mannitane oleate or a mixture of mannitane oleate and oleate polyethoxylated sorbitan.   16. Composition as defined in any one of Claims 1 to 15,   further comprising at least one antigen or at least one in vivo generator of a com   posed comprising an amino acid sequence.   17. Composition as defined in one of claims 1 to 16, characterized in that it is fluid. At there 2824279   18. Process for the preparation of a composition as defined in any one   conch of claim 16 or 17, comprising addition in a weight ratio E I   H greater than 1, of an aqueous medium containing at least one antigen or at least one   in viYo generator of a compound and comprising an amino acid sequence at a   position essentially consisting of 1% and 15% by weight, and preferably 3% and% by weight, of surfactants having an overall HLB number of between 5 and 8, and from 85% to 99%, and preferably of 90% to 97% by weight, of one or more compounds   chosen from oils of mineral, vegetable or animal origin, the alkyl esters of   so-called oils, alkyl esters of fatty acids or alkyl ethers of fatty acids,   esters of fatty acids and polyols or ethers of fatty alcohols and polyols, and the emulsifier