EP1093382A2 - Use of an amphipathic compound for providing an adjuvant to a subunit vaccine - Google Patents

Use of an amphipathic compound for providing an adjuvant to a subunit vaccine

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Publication number
EP1093382A2
EP1093382A2 EP99929389A EP99929389A EP1093382A2 EP 1093382 A2 EP1093382 A2 EP 1093382A2 EP 99929389 A EP99929389 A EP 99929389A EP 99929389 A EP99929389 A EP 99929389A EP 1093382 A2 EP1093382 A2 EP 1093382A2
Authority
EP
European Patent Office
Prior art keywords
antigen
adjuvant
amphipathic compound
responders
use according
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99929389A
Other languages
German (de)
French (fr)
Inventor
Anne Darbouret
Florence Brunel
Jorge Ronco
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sanofi Pasteur Inc
Original Assignee
Aventis Pasteur SA
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Filing date
Publication date
Application filed by Aventis Pasteur SA filed Critical Aventis Pasteur SA
Publication of EP1093382A2 publication Critical patent/EP1093382A2/en
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/29—Hepatitis virus
    • A61K39/292—Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55505—Inorganic adjuvants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2730/00—Reverse transcribing DNA viruses
    • C12N2730/00011—Details
    • C12N2730/10011—Hepadnaviridae
    • C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
    • C12N2730/10134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the invention relates to the field of vaccine adjuvants.
  • the invention relates to the use of an amphipathic compound for the manufacture of a vaccine composition intended for non-responders.
  • the object of the invention is therefore to propose an improved vaccine as regards the rate of seroconversion which it makes it possible to obtain.
  • the invention proposes the use of an amphipathic compound for the preparation of a vaccine composition comprising at least one subunit antigen intended to be administered to target populations comprising individuals "Non-Responders" to said antigen .
  • the amphipathic compounds are derivatives of cholesterol linked to a quaternary ammonium or to a protonable amine by a carbamoyl bond.
  • These compounds, such as DC-chol can be found under basic form, in the form of salt, or, and this is the most frequent case, both under the 2 forms in equilibrium in a mixture, the shift of equilibrium towards one or the other dependent form the composition of the mixture and in particular its pH.
  • DC-chol which is particularly advantageous for the purposes of the invention is DC-chol which can be obtained from cholesteryl chloroformate and from N, N-dimethylethylenediamine, according to the method described in US Pat. No.
  • amphipathic compounds can be in the form of a dispersion in an aqueous or oily medium.
  • the vaccine composition which should be modified in order to reduce the number of individuals who are non-responders therein is a composition comprising at least one highly purified subunit antigen.
  • antigens are generally less immunogenic than less purified preparations obtained from whole germs, and can therefore lead to a higher proportion of Non-Responders.
  • the number of non-Responders to the hepatitis B antigen is reduced. It can be any hepatitis B antigen, and in particular an antigen containing the S and pre-S 2 regions , such as the antigen described in patent EP 0 273 811.
  • the vaccine composition according to the invention can be in liquid form or in lyophilized form.
  • the vaccine composition according to the invention can be a monovalent (that is to say intended to protect only against one disease) or plurivalent (protecting against several diseases) composition.
  • It can comprise, in addition to the adjuvant according to the invention, one or more other adjuvants intended, in a conventional manner, to increase the response of the immune system, whether it is a response of humoral type, of cellular type, or a mixture of the 2 types.
  • the vaccine composition according to the invention can also comprise all the ingredients usually present in vaccines: stabilizer, preservative, cryoprotective, etc.
  • This composition can be in liquid form or in the form of lyophilisate.
  • FIG. 1 represents the reaction for obtaining DC-chol.
  • Figures 2 and 3 and
  • Example 3 illustrate the results obtained in Example 2.
  • Example 1 Preparation of the immunization compositions.
  • a hepatitis B antigen suspension is prepared, according to the method described in patent EP 0 273 811, with the exception of the addition of aluminum hydroxide. It is therefore a suspension at 4 mg / l of antigen in 1 mM phosphate buffer at pH 6.8. This suspension is called Suspension A.
  • DC-chol powder obtained according to the mode of preparation described in Example 8 of patent application WO 96/40067 is available. This powder is suspended in 20 mM Tris / HCl buffer, 150 mM NaCl at Ph 6.8 under nitrogen at 4 ° C., maintaining stirring for 48 h. One then obtains a suspension at 2 g / l of DC-chol. By mixing volume to volume these 2 suspensions, a vaccine composition according to the invention is obtained which is distributed in doses of 0.5 ml each comprising 1 ⁇ g of antigen against hepatitis B and 0.5 mg of DC- chol.
  • a new immunization composition is prepared from suspension A to which an adjuvant C of the prior art is added in order to obtain, per 0.5 ml dose, a quantity of antigen of 1 ⁇ g and a quantity of adjuvant 0.5mg.
  • the nature and the dose of the adjuvant of the prior art were selected during comparative immunization tests against hepatitis B of BALB / C mice, by subcutaneous route.
  • the adjuvant selected is that which, during these tests has shown a strong capacity to increase the humoral response to the antigen considered, the results obtained with this adjuvant being clearly superior to those obtained with aluminum hydroxide or with DC-chol.
  • mice Each group of mice is immunized with a different vaccine composition:
  • a second group is immunized with doses of 0.5 ml each comprising the hepatitis B antigen and aluminum hydroxide.
  • a third group is immunized with doses of 0.5 ml each comprising the hepatitis B antigen and an adjuvant of the prior art.
  • a fourth group is immunized with doses of 0.5 ml of the vaccine composition according to the invention.
  • the immunization of the mice is carried out subcutaneously, one to two hours after the preparation of the immunizing compositions.
  • blood is taken for dosing, and a second injection is carried out under the same conditions as the first.
  • a second blood sample is taken three weeks after the second injection. All blood samples are coagulated, centrifuged; the collected serum is stored at -20 ° C until titration which is carried out by an ELISA technique.
  • Non-Responders in the group of mice having received the vaccine composition according to the invention while the use of the adjuvant of the prior art, yet capable of greatly increasing the immune response of mice already "Responders”, does not does not allow, even after the booster injection, to make all subjects "Responders”.
  • the immunization protocol is the same as that of Example 2.
  • results obtained show the effectiveness of the subject of the invention which makes it possible to increase the rate of seroconversion and therefore to reduce the number of "non-responder" subjects during immunization against a highly purified antigen.

Abstract

The invention concerns an amphipathic compound for preparing a vaccine composition comprising at least a subunit antigen to be administered to target populations comprising non-responders to said antigen. DC-chol is a particular amphiphatic compound.

Description

UTILISATION D'UN COMPOSE AMPHIPATHIQUE POUR ADJUVER UN VACCIN SOUS-UNITAIRE USE OF AN AMPHIPATHIC COMPOUND TO ADJUST A SUBUNIT VACCINE
L'invention concerne le domaine des adjuvants vaccinaux. En particulier, l'invention concerne l'utilisation d'un composé amphipathique pour la fabrication d'une composition vaccinale destinée à des sujets non-répondeurs.The invention relates to the field of vaccine adjuvants. In particular, the invention relates to the use of an amphipathic compound for the manufacture of a vaccine composition intended for non-responders.
Il est connu que, lorsqu'on vaccine une population d'individus contre une maladie, un certain nombre d'entre eux ne "répondent" pas à la vaccination, c'est-à-dire que leur système immunitaire ne semble pas réagir à l'antigène administré. Ce problème est plus ou moins important selon les maladies et les populations concernées, mais les fabricants de vaccins cherchent toujours à réduire, pour chacun des vaccins qu'ils mettent à disposition des médecins, le nombre de sujets susceptibles d'être des "Non-Répondeurs". Ce problème est particulièrement considéré pour les vaccins comprenant des antigènes purifiés tels que des vaccins sous-unitaires produits par génie génétique et notamment le vaccin contre l'hépatite ; or, si on connaît un certain nombre de composés amphipathiques, et notamment le 3 β-[N-(N',N'-diméthylaminoéthane)- carbamoyljcholesterol, communément appelé DC-chol, qui permettent d'accroître la réponse immunitaire de sujets déjà "répondeurs", et sont donc considérés comme de bons adjuvants vaccinaux, il est plus difficile de trouver des moyens pour surmonter le problème des Non-Répondeurs, d'autant que toutes les raisons pour lesquelles un individu est Non-Répondeur ne sont pas, pour l'instant, clairement identifiées. Les recherches effectuées dans ce domaine ont montré qu'il n'était pas possible d'extrapoler les résultats obtenus avec des adjuvants capables d'accroître la réponse immunitaire chez des sujets "Répondeurs" aux sujets "Non-Répondeurs", certains bons adjuvants chez les sujets répondeurs se révélant sans effet sur les sujets Non-Répondeurs.It is known that when a population of individuals is vaccinated against a disease, a certain number of them do not "respond" to vaccination, that is to say that their immune system does not seem to react to the antigen administered. This problem is more or less important according to the diseases and populations concerned, but the vaccine manufacturers are always seeking to reduce, for each of the vaccines they make available to doctors, the number of subjects likely to be "No- Responders ". This problem is particularly considered for vaccines comprising purified antigens such as subunit vaccines produced by genetic engineering and in particular the hepatitis vaccine; however, if a certain number of amphipathic compounds are known, and in particular 3 β- [N- (N ', N'-dimethylaminoethane) - carbamoyljcholesterol, commonly called DC-chol, which make it possible to increase the immune response of subjects already "responders", and are therefore considered good vaccine adjuvants, it is more difficult to find ways to overcome the problem of non-responders, especially since all the reasons why an individual is non-responder are not, for the time being, clearly identified. Research in this area has shown that it was not possible to extrapolate the results obtained with adjuvants capable of increasing the immune response in subjects "Responders" to subjects "Non-Responders", some good adjuvants in Responding subjects appear to have no effect on Non-Responding subjects.
Le but de l'invention est donc de proposer un vaccin amélioré en ce qui concerne le taux de séroconversion qu'il permet d'obtenir. Pour cela, l'invention propose l'utilisation d'un composé amphipathique pour la préparation d'une composition vaccinale comprenant au moins un antigène sous-unitaire destinée à être administrée à des populations-cibles comprenant des individus "Non- Répondeurs" audit antigène.The object of the invention is therefore to propose an improved vaccine as regards the rate of seroconversion which it makes it possible to obtain. For this, the invention proposes the use of an amphipathic compound for the preparation of a vaccine composition comprising at least one subunit antigen intended to be administered to target populations comprising individuals "Non-Responders" to said antigen .
Selon l'invention, les composés amphipathiques sont des dérivés de cholestérol relié à un ammonium quaternaire ou à une aminé protonable par une liaison carbamoyle. Ces composés, tels le DC-chol peuvent se trouver sous forme basique, sous forme de sel, ou encore, et c'est le cas le plus fréquent, à la fois sous les 2 formes en équilibre dans un mélange, le déplacement de l'équilibre vers l'une ou l'autre forme dépendant de la composition du mélange et notamment de son pH. Un des composés amphipathiques particulièrement intéressant aux fins de l'invention est le DC-chol qui peut être obtenu à partir de cholestéryl chloroformate et de N, N-dimethylethylenediamine, selon la méthode décrite dans le brevet US 5283185 ou de façon préférée, selon la méthode décrite à l'exemple 8 de la demande de brevet WO 96/40067. Les 2 formes généralement en équilibre du DC-chol sont illustrées à la figure 1. Il est possible également d'utiliser un produit obtenu par réaction de cholestéryl chloroformate et de N,N,N triméthyléthylènediamine.According to the invention, the amphipathic compounds are derivatives of cholesterol linked to a quaternary ammonium or to a protonable amine by a carbamoyl bond. These compounds, such as DC-chol can be found under basic form, in the form of salt, or, and this is the most frequent case, both under the 2 forms in equilibrium in a mixture, the shift of equilibrium towards one or the other dependent form the composition of the mixture and in particular its pH. One of the amphipathic compounds which is particularly advantageous for the purposes of the invention is DC-chol which can be obtained from cholesteryl chloroformate and from N, N-dimethylethylenediamine, according to the method described in US Pat. No. 5,283,185 or, preferably, according to method described in Example 8 of patent application WO 96/40067. The 2 generally balanced forms of DC-chol are illustrated in Figure 1. It is also possible to use a product obtained by reaction of cholesteryl chloroformate and N, N, N trimethylethylenediamine.
Les composés amphipathiques peuvent se présenter sous forme de dispersion dans un milieu aqueux ou huileux.The amphipathic compounds can be in the form of a dispersion in an aqueous or oily medium.
La composition vaccinale qu'il convient de modifier afin de diminuer le nombre d'individus qui y sont Non-Répondeurs est une composition comprenant au moins un antigène sous-unitaire hautement purifié. En effet, de tels antigènes sont en général moins immunogènes que des préparations moins purifiées obtenues à partir de germes entiers, et peuvent donc conduire à une plus forte proportion de Non-Répondeurs. En particulier, selon l'invention, on réduit le nombre de Non-Répondeurs à l'antigène de l'hépatite B. Il peut s'agir de n'importe quel antigène de l'hépatite B, et notamment d'un antigène contenant les régions S et pré-S2, tel que l'antigène décrit dans le brevet EP 0 273 811.The vaccine composition which should be modified in order to reduce the number of individuals who are non-responders therein is a composition comprising at least one highly purified subunit antigen. Indeed, such antigens are generally less immunogenic than less purified preparations obtained from whole germs, and can therefore lead to a higher proportion of Non-Responders. In particular, according to the invention, the number of non-Responders to the hepatitis B antigen is reduced. It can be any hepatitis B antigen, and in particular an antigen containing the S and pre-S 2 regions , such as the antigen described in patent EP 0 273 811.
Pour apprécier l'efficacité de l'objet de l'invention, on utilise des modèles animaux. On compare ainsi, en parallèle, les résultats obtenus avec les composés selon l'invention et ceux obtenus avec les adjuvants classiques de l'art antérieur, sur un ensemble de souris non-syngéniques OF 1 qui, à l'instar de la population humaine, est composé à la fois de sujets Répondeurs et de sujets Non-Répondeurs. La comparaison du nombre de sujets séro-convertis dans chacun des groupes permet d'apprécier l'intérêt de l'objet de l'invention.To assess the effectiveness of the object of the invention, animal models are used. Thus, in parallel, the results obtained with the compounds according to the invention and those obtained with the conventional adjuvants of the prior art are compared, on a set of non-syngeneic OF 1 mice which, like the human population. , is composed of both Responder and Non-Responder subjects. The comparison of the number of sero-converted subjects in each of the groups makes it possible to appreciate the interest of the subject of the invention.
Un autre test peut être effectué sur des souris appartenant à une souche décrite comme étant Non-Répondeuse à l'antigène considéré, telles que les souris B10.MH2f pour l'antigène de l'hépatite B. La comparaison des résultats obtenus dans chacun des groupes permet d'apprécier l'efficacité de l'objet de l'invention. La composition vaccinale selon l'invention peut se présenter sous forme liquide ou sous forme lyophilisée.Another test can be carried out on mice belonging to a strain described as being non-responder to the antigen considered, such as the B10.MH 2f mice for the hepatitis B antigen. The comparison of the results obtained in each groups allows to assess the effectiveness of the object of the invention. The vaccine composition according to the invention can be in liquid form or in lyophilized form.
La composition vaccinale selon l'invention peut être une composition monovalente (c'est-à-dire destinée à ne protéger que contre une maladie) ou plurivalente (protégeant contre plusieurs maladies).The vaccine composition according to the invention can be a monovalent (that is to say intended to protect only against one disease) or plurivalent (protecting against several diseases) composition.
Elle peut comprendre, outre l'adjuvant selon l'invention, un ou plusieurs autres adjuvants destinés, de façon classique, à accroître la réponse du système immunitaire, qu'il s'agisse d'une réponse de type humorale, de type cellulaire, ou d'un mélange des 2 types.It can comprise, in addition to the adjuvant according to the invention, one or more other adjuvants intended, in a conventional manner, to increase the response of the immune system, whether it is a response of humoral type, of cellular type, or a mixture of the 2 types.
De façon classique, la composition vaccinale selon l'invention peut comprendre, en outre, tous les ingrédients habituellement présents dans les vaccins : stabilisant, conservateur, cryoprotecteur... Cette composition peut se présenter sous forme liquide ou sous forme de lyophilisât.Conventionally, the vaccine composition according to the invention can also comprise all the ingredients usually present in vaccines: stabilizer, preservative, cryoprotective, etc. This composition can be in liquid form or in the form of lyophilisate.
Les exemples qui suivent illustrent un mode de réalisation de l'invention.The examples which follow illustrate an embodiment of the invention.
La figure 1 représente la réaction d'obtention du DC-chol. Les figures 2 etFIG. 1 represents the reaction for obtaining DC-chol. Figures 2 and
3 illustrent les résultats obtenus à l'exemple 2.3 illustrate the results obtained in Example 2.
Exemple 1 : Préparation des compositions d'immunisation.Example 1: Preparation of the immunization compositions.
On prépare, une suspension d'antigène de l'hépatite B, selon la méthode décrite dans le brevet EP 0 273 811 , à l'exception de l'addition d'hydroxyde d'aluminium. Il s'agit donc d'une suspension à 4mg/l d'antigène en tampon phosphate 1 mM à ph 6,8. Cette suspension est appelée Suspension A.A hepatitis B antigen suspension is prepared, according to the method described in patent EP 0 273 811, with the exception of the addition of aluminum hydroxide. It is therefore a suspension at 4 mg / l of antigen in 1 mM phosphate buffer at pH 6.8. This suspension is called Suspension A.
On dispose de poudre de DC-chol obtenue selon le mode de préparation décrit à l'exemple 8 de la demande de brevet WO 96/40067. Cette poudre est mise en suspension dans du tampon Tris/HCI 20 mM, NaCI 150mM à Ph 6,8 sous azote à 4°C, en maintenant l'agitation pendant 48 h. On obtient alors une suspension à 2g/l de DC-chol. En mélangeant volume à volume ces 2 suspensions, on obtient une composition vaccinale selon l'invention que l'on répartit par doses de 0,5 ml comprenant chacune 1 μg d'antigène contre l'hépatite B et 0,5 mg de DC-chol.DC-chol powder obtained according to the mode of preparation described in Example 8 of patent application WO 96/40067 is available. This powder is suspended in 20 mM Tris / HCl buffer, 150 mM NaCl at Ph 6.8 under nitrogen at 4 ° C., maintaining stirring for 48 h. One then obtains a suspension at 2 g / l of DC-chol. By mixing volume to volume these 2 suspensions, a vaccine composition according to the invention is obtained which is distributed in doses of 0.5 ml each comprising 1 μg of antigen against hepatitis B and 0.5 mg of DC- chol.
En ajoutant à la suspension A de l'hydroxyde d'aluminium (Alhydrogel® fourni par la Société SUPERFOS BIOSECTOR), on prépare une nouvelle composition immunisante comprenant, cette fois, 1 μg d'antigène et 1 mg d'hydroxyde d'aluminium par dose de 0,5ml.By adding aluminum hydroxide (Alhydrogel® supplied by the company SUPERFOS BIOSECTOR) to suspension A, a new immunizing composition is prepared, this time comprising 1 μg of antigen and 1 mg of aluminum hydroxide per 0.5ml dose.
Une nouvelle composition d'immunisation est préparée à partir de la suspension A à laquelle on ajoute un adjuvant C de l'art antérieur afin d'obtenir par dose de 0,5ml une quantité d'antigène de 1 μg et une quantité d'adjuvant de 0,5mg. La nature et la dose de l'adjuvant de l'art antérieur ont été sélectionnés lors d'essais comparatifs d'immunisation contre l'hépatite B de souris BALB/C, par voie sous-cutanée. L'adjuvant retenu est celui qui, lors de ces essais à montré une forte capacité d'accroître la réponse humorale à l'antigène considéré, les résultats obtenus avec cet adjuvant étant nettement supérieurs à ceux obtenus avec l'hydroxyde d'aluminium ou avec le DC-chol.A new immunization composition is prepared from suspension A to which an adjuvant C of the prior art is added in order to obtain, per 0.5 ml dose, a quantity of antigen of 1 μg and a quantity of adjuvant 0.5mg. The nature and the dose of the adjuvant of the prior art were selected during comparative immunization tests against hepatitis B of BALB / C mice, by subcutaneous route. The adjuvant selected is that which, during these tests has shown a strong capacity to increase the humoral response to the antigen considered, the results obtained with this adjuvant being clearly superior to those obtained with aluminum hydroxide or with DC-chol.
Exemple 2 : Immunisation de souris OF1EXAMPLE 2 Immunization of OF1 Mice
On dispose de 4 groupes de 10 souris femelles OF1 non-syngéniques âgées de 6 à 8 semaines qui représentent une population hétérogène, reflétant la variabilité de la population humaine.There are 4 groups of 10 non-syngeneic female OF1 mice aged 6 to 8 weeks which represent a heterogeneous population, reflecting the variability of the human population.
Chaque groupe de souris est immunisé avec une composition vaccinale différente :Each group of mice is immunized with a different vaccine composition:
• un 1er groupe est immunisé avec des doses de 0,5 ml de la suspension A mentionnée à l'exemple 1 , et contenant donc uniquement l'antigène de l'hépatite B.• a 1st group is immunized with doses of 0.5 ml of suspension A mentioned in example 1, and therefore containing only the hepatitis B antigen.
• un second groupe est immunisé avec des doses de 0,5 ml comprenant chacune l'antigène de l'hépatite B et de l'hydroxyde d'aluminium..• a second group is immunized with doses of 0.5 ml each comprising the hepatitis B antigen and aluminum hydroxide.
• un troisième groupe est immunisé avec des doses de 0,5ml comprenant chacune l'antigène de l'hépatite B et un adjuvant de l'art antérieur.• a third group is immunized with doses of 0.5 ml each comprising the hepatitis B antigen and an adjuvant of the prior art.
• un quatrième groupe est immunisé avec des doses de 0 ,5ml de la composition vaccinale selon l'invention. L'immunisation des souris est effectuée par voie sous-cutanée, une à deux heures après la préparation des compositions immunisantes. Trois semaines après la première immunisation, on prélève du sang pour dosage, et on procède à une seconde injection dans les mêmes conditions que la première. Un second prélèvement de sang est réalisé trois semaines après la seconde injection. Tous les échantillons de sang sont coagulés, centrifugés ; le sérum récolté est stocké à -20°C jusqu'au titrage qui est réalisé par une technique ELISA.• a fourth group is immunized with doses of 0.5 ml of the vaccine composition according to the invention. The immunization of the mice is carried out subcutaneously, one to two hours after the preparation of the immunizing compositions. Three weeks after the first immunization, blood is taken for dosing, and a second injection is carried out under the same conditions as the first. A second blood sample is taken three weeks after the second injection. All blood samples are coagulated, centrifuged; the collected serum is stored at -20 ° C until titration which is carried out by an ELISA technique.
Les résultats obtenus pour chacun des groupes de souris sont illustrés sur la figure 2 qui indique, en unité ELISA arbitraires les moyennes géométriques des titres en anticorps (GMT) obtenues avec chacune des compositions testées après la 1ère injection, et sur la figure 3 qui indique les moyennes GMT obtenues après l'injection de rappel.The results obtained for each of the groups of mice are shown in Figure 2 which shows, in the geometric mean arbitrary ELISA unit titre (GMT) achieved with each of the compositions tested after the 1st injection, and Figure 3 which indicates the GMT averages obtained after the booster injection.
On remarque ainsi que, dès la première injection, il n'y a pas de sujetWe notice that, from the first injection, there is no subject
Non-Répondeurs dans le groupe de souris ayant reçu la composition vaccinale selon l'invention, alors que l'utilisation de l'adjuvant de l'art antérieur, pourtant capable d'accroître fortement la réponse immunitaire des souris déjà "Répondeurs", ne permet pas, même après l'injection de rappel, de rendre tous les sujets "Répondeurs".Non-Responders in the group of mice having received the vaccine composition according to the invention, while the use of the adjuvant of the prior art, yet capable of greatly increasing the immune response of mice already "Responders", does not does not allow, even after the booster injection, to make all subjects "Responders".
Exemple 3 : Immunisation de souris B10.MH2f EXAMPLE 3 Immunization of B10.MH 2f Mice
On dispose des 4 préparations d'immunisation décrites à l'exemple 2 que l'on administre cette fois à 4 groupes de 8 souris B10.M qui sont des souris décrites pour être "Non-Répondeurs" à l'antigène de l'hépatite B.We have the 4 immunization preparations described in Example 2 which are administered this time to 4 groups of 8 B10.M mice which are mice described to be "Non-Responders" to the hepatitis antigen B.
Le protocole d'immunisation est le même que celui de l'exemple 2.The immunization protocol is the same as that of Example 2.
Les résultats obtenus sont indiqués dans le tableau 1 ci-après ; il s'agit du nombre de souris séroconverties, après l'injection de rappel, dans chacun des groupes testés.The results obtained are shown in Table 1 below; this is the number of seroconverted mice, after the booster injection, in each of the groups tested.
Pour ce test, on considère qu'il y a séroconversion lorsque les taux d'anticorps induits sont détectables lors du dosage ELISA.For this test, it is considered that there is seroconversion when the levels of antibodies induced are detectable during the ELISA assay.
Tableau 1Table 1
Les résultats obtenus montrent l'efficacité de l'objet de l'invention qui permet d'accroître le taux de séroconversion et donc de réduire le nombre de sujet "Non-Répondeurs" lors de l'immunisation contre un antigène hautement purifié. The results obtained show the effectiveness of the subject of the invention which makes it possible to increase the rate of seroconversion and therefore to reduce the number of "non-responder" subjects during immunization against a highly purified antigen.

Claims

REVENDICATIONS
1. Utilisation d'un composé amphipathique pour la préparation d'une composition vaccinale comprenant au moins un antigène sous-unitaire destinée à être administrée à des populations-cibles comprenant des individus "Non- Répondeurs" audit antigène.1. Use of an amphipathic compound for the preparation of a vaccine composition comprising at least one subunit antigen intended to be administered to target populations comprising individuals "Non-Responders" to said antigen.
2. Utilisation selon la revendication 1 caractérisée en ce que ledit antigène est un antigène pour la vaccination contre l'hépatite B.2. Use according to claim 1 characterized in that said antigen is an antigen for vaccination against hepatitis B.
3. Utilisation selon une des revendications précédentes, caractérisée en ce que ledit composé amphipathique est du DC-chol.3. Use according to one of the preceding claims, characterized in that said amphipathic compound is DC-chol.
4. Utilisation selon une des revendications précédentes, caractérisée en ce que la composition vaccinale est une composition plurivalente.4. Use according to one of the preceding claims, characterized in that the vaccine composition is a plurivalent composition.
5. Utilisation selon une des revendications précédentes, caractérisée en ce que le composé amphipathique est présent sous forme de dispersion dans un milieu aqueux ou huileux.5. Use according to one of the preceding claims, characterized in that the amphipathic compound is present in the form of a dispersion in an aqueous or oily medium.
6. Utilisation selon une des revendications précédentes, caractérisée en ce que le composé amphipathique est présent dans une formulation lyophilisée.6. Use according to one of the preceding claims, characterized in that the amphipathic compound is present in a lyophilized formulation.
7. Utilisation selon une des revendications précédentes, caractérisée en ce qu'elle comprend en outre au moins un autre adjuvant vaccinal. 7. Use according to one of the preceding claims, characterized in that it further comprises at least one other vaccine adjuvant.
EP99929389A 1998-07-03 1999-07-02 Use of an amphipathic compound for providing an adjuvant to a subunit vaccine Withdrawn EP1093382A2 (en)

Applications Claiming Priority (3)

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FR9808700A FR2781160B1 (en) 1998-07-03 1998-07-03 USE OF AN AMPHIPATHIC COMPOUND TO ADJUST A SUBUNIT VACCINE
FR9808700 1998-07-03
PCT/FR1999/001604 WO2000001345A2 (en) 1998-07-03 1999-07-02 Use of an amphipathic compound for providing an adjuvant to a subunit vaccine

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US20100209452A1 (en) * 2007-10-03 2010-08-19 Immunovaccine Technologies, Inc Compositions comprising an antigen, an amphipathic compound and a hydrophobic carrier, and uses thereof
ES2524699T3 (en) * 2008-06-05 2014-12-11 Immunovaccine Technologies Inc. Compositions comprising liposomes, an antigen, a polynucleotide and a vehicle comprising a continuous phase of a hydrophobic substance
AU2012321022B2 (en) 2011-10-06 2017-03-23 Immunovaccine Technologies Inc. Liposome compositions comprising an adjuvant that activates or increases the activity of TLR2 and uses thereof

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WO1995004523A1 (en) * 1993-08-06 1995-02-16 Opperbas Holding B.V. A method for preparation of vesicles loaded with biological structures, biopolymers and/or oligomers
FR2726764B1 (en) * 1994-11-14 1997-01-31 Pasteur Merieux Serums Vacc ADJUVANT FOR VACCINE COMPOSITION
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AU4621799A (en) 2000-01-24
US6472159B1 (en) 2002-10-29
FR2781160A1 (en) 2000-01-21

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