WO1998042374A1 - Immunostimulation induite par le chitosane - Google Patents

Immunostimulation induite par le chitosane Download PDF

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Publication number
WO1998042374A1
WO1998042374A1 PCT/US1997/005094 US9705094W WO9842374A1 WO 1998042374 A1 WO1998042374 A1 WO 1998042374A1 US 9705094 W US9705094 W US 9705094W WO 9842374 A1 WO9842374 A1 WO 9842374A1
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WO
WIPO (PCT)
Prior art keywords
solution
chitosan
antigen
oil
chitosan solution
Prior art date
Application number
PCT/US1997/005094
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English (en)
Inventor
Joseph S. Podolski
Mitzi L. Martinez
Original Assignee
Zonagen, Inc.
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Zonagen, Inc. filed Critical Zonagen, Inc.
Priority to PCT/US1997/005094 priority Critical patent/WO1998042374A1/fr
Priority to AU23478/97A priority patent/AU2347897A/en
Priority to JP10545633A priority patent/JP2000504350A/ja
Priority to CA002255867A priority patent/CA2255867C/fr
Priority to EP97916251A priority patent/EP0914154A1/fr
Publication of WO1998042374A1 publication Critical patent/WO1998042374A1/fr
Priority to AU19742/01A priority patent/AU771525B2/en

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    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • 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
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
    • 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
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55583—Polysaccharides

Definitions

  • the present invention relates generally to methods for potentiating an immune response in an animal, compositions to effect the potentiation, and methods to produce the compositions. More specifically, the invention provides methods comprising the use of an antigen/chitosan mixture or an antigen/chitosan/oil/surfactant emulsion to potentiate an immune response, antigen/chitosan mixtures or antigen/chitosan/oil/surfactant emulsions to effect potentiation, and methods to prepare the antigen/chitosan mixture or antigen/ chitosan/oil emulsion.
  • an adjuvant should potentiate long-lasting expression of functionally active antibodies, elicit cell-mediated immunity (CMI), and enhance production of memory T- and B-lymphocytes with highly specific immunoreactivity against an invading antigen.
  • CMI cell-mediated immunity
  • Antigens derived from closely related species are less competent in eliciting antibody production due to the fact that the host immune system is sometimes unable to clearly distinguish the foreign antigen from endogenous, or self antigens.
  • the dosage of the antigen, the purity of the antigen, and the frequency with which the antigen is administered are also factors which significantly contribute to the resulting antibody titer and specificity of the resulting antibodies.
  • Still other factors include the fonn, or complexity, of the antigen, and how the antigen is administered.
  • both the genetic makeup and overall physiological state of the immunized animal contribute to the extent to which an immune response is mounted. Of these factors, the form or complexity of the antigen is directly affected by immunization with an adjuvant.
  • helper T cells are required for B-cell antibody responses to most antigens.
  • an antigen is captured and processed by an antigen-presenting cell (APC), e.g. , circulating or tissue macrophages, and presented on the surface of the APC in association with a class II major histocompatibility (MHC) molecule.
  • APC antigen-presenting cell
  • MHC major histocompatibility
  • the antigen can interact with receptors on the surface of helper T cells thereby activating the particular subpopulation of cells to express and secrete any of a number of cytokines.
  • the nature of cytokine production depends on the subset of helper T cells activated, a result that can be modulated in part by the choice of adjuvant.
  • alum an aluminum salt adjuvant approved for clinical use in humans, has been reported to selectively activate T H 2 cells in mice [Grun and Maurer, Cell. Immunol. 121: 134-145 (1989)]
  • Freund's complete adjuvant (CFA) an emulsion of mineral oil with killed mycobacteria [Freund, et al. , Proc. Soc. Exp. Biol. Med. 37:509 (1937)]
  • CFA complete adjuvant
  • LPS lipopolysaccharide
  • Oil emulsions i.e. , Complete Freund's Adjuvant [CFA], Freund's incomplete adjuvant [FIA]
  • liposomes act through depot formation as does alum, thus allowing for slow release of antigen. Slow release of antigen permits extended exposure of the antigen to the immune system and also allows for initial immunization with a dosage of antigen that, if delivered at one time, would ordinarily be counterproductive to antibody formation.
  • alum [AlK(SO 4 ) 2 H 2 O]
  • Alum not only acts through T H 2 cell activation, depot formation and slow release of antigen following immunization [Edelman, Rev. Infect. Dis. 2:370-383 (1980); Warren, et al. , Ann. Rev. Immunol. 4:369-388 (1986)], but also through granuloma formation by attracting immunocompetent cells [White, et al. , J. Exp. Med. 702:73-82 (1955)] and activation of complement [Ramanathan, et al , Immunol. 57:881-888 (1979)].
  • alum is not without its negative side effects which include erythema, subcutaneous nodules, contact hypersensitivity, and granulomatous inflammation.
  • adjuvants which are widely employed outside of human application, are also the focus of continuing research to develop acceptable alternatives for use in humans. Included are the above mentioned oil emulsions (t ' .e. , CFA and FIA), bacterial products (i.e. , LPS, cholera toxin, mycobacterial components and whole killed Corynebacte ⁇ um parvum, Corynebacterium granulosum, and Bordetella pertussis, liposomes, immunostimulating complexes (ISCOMs), and naturally occurring and derivatized polysaccharides from other than bacterial sources.
  • oil emulsions i.e. , CFA and FIA
  • bacterial products i.e. , LPS, cholera toxin, mycobacterial components and whole killed Corynebacte ⁇ um parvum, Corynebacterium granulosum, and Bordetella pertussis, liposomes, immunostimulating complexes
  • LPS Lipopolysaccharide isolated from certain Gram-negative bacteria is one such polysaccharide even though the adjuvant properties of LPS are derived mainly from the lipid A region of the molecule, and not from the ⁇ -specific polysaccharide or core oligosaccharide regions of the molecule. LPS, which augments both humoral [Johnson, et al., J. Exp. Med.
  • Chitosan [j8-(l-4)-2-amino-2-deoxy-D-glucan] is a derivative of chitin and has been widely used in biomedical applications, due in part to is biodegradability by lysozyme and low toxicity in humans. These same properties have resulted in increased interest in chitosan as an immunopotentiating agent.
  • Matuhashi, et al in U.S. Patent No. 4,372,883, disclosed conjugation of soluble polysaccharides, including chitosan, to normally toxic antigens, conjugation thereby detoxifying the antigen and permitting its use as an immunogen.
  • Matuhashi et al. did not address the use of insoluble forms of chitosan, nor did Matuhashi compare the resulting serum antibody titer with that obtained from immunization with other known adjuvants.
  • Suzuki, et al. in U.S. Patent 4,971 ,956, disclosed the use of water soluble chitosan-oligomers as therapeutics for treatment of bacterial and fungal infections, as well as for the treatment of tumors.
  • Suzuki, et al discussed the difficulty in modifying chitosan to produce an appropriate water soluble form, disclosing that water-insoluble forms are impractical for therapeutic application.
  • Suzuki et al. does not disclose conjugation of an antigen to chitosan to effect enhanced immune response.
  • Mitsuhashi, et al. in U.S.
  • Patent 4,814, 169 disclosed the use of human protein conjugated to soluble polysaccharides, including chitosan, to generate antibodies against human protein in non-human animals.
  • Administration of the human protein/poly saccharide solution was by intravenous, intraperitoneal, or subcutaneous injection. Other routes, including oral and rectal administration, were not addressed in the disclosure.
  • the invention is directed to the use of chitosan formulations for potentiating an immune response in a host.
  • the present invention is directed to a method for potentiating an immune response comprising the steps of preparing a chitosan solution, incorporating an antigen into a phosphate buffer to form an antigen/phosphate buffer solution, lyophilizing the antigen/phosphate buffer solution to a lyophilized mixture, reconstituting the lyophilized mixture with the chitosan solution to form an antigen/chitosan mixture, and administering the mixture to an animal, including humans.
  • the antigen/chitosan mixture may be administered to the animal via oral, rectal, intravaginal routes as well as via intraperitoneal injection, intramuscular injection, or subcutaneous injection; administration may comprise a single route or a multiplicity of routes.
  • a composition which, comprises in combination lyophilized antigen/phosphate buffer and chitosan solution.
  • the antigen/chitosan mixture may be administered to the animal via oral, rectal, intravaginal routes as well as via intraperitoneal injection, intramuscular injection, or subcutaneous injection; administration may comprise a single route or a multiplicity of routes.
  • an immunogen comprising a lyophilized antigen/phosphate buffer and chitosan solution.
  • the antigen/chitosan mixture may be administered to the animal via oral, rectal, intravaginal routes as well as via intraperitoneal injection, intramuscular injection, or subcutaneous injection; administration may comprise a single route or a multiplicity of routes.
  • a method for preparing an immunogen comprising, preparing a chitosan solution, incorporating an antigen into a phosphate buffer to form an antigen/phosphate buffer solution, lyophilizing the antigen/phosphate buffer solution to a lyophilized mixture, and reconstituting the lyophilized mixture with the chitosan solution to form an antigen/chitosan mixture.
  • the present invention provides a method for potentiating an immune response comprising the steps of preparing a chitosan solution, preparing a sodium hydroxide solution, preparing an oil/ surfactant solution, wherein the oil can be metabolically degraded, mixing the chitosan solution with the sodium hydroxide solution, the oil/ surfactant solution, and the antigen to form an emulsion, and administering the emulsion to an animal.
  • the antigen may be, but is not limited to, a protein, carbohydrate, lipid, glycoprotein or combinations thereof.
  • the pH of the chitosan solution is about 5.0.
  • the emulsion may be administered to the animal via intraperitoneal injection, intramuscular injection, or subcutaneous injection.
  • the emulsion may also be administered alone, or in combination with any of a number of other adjuvants.
  • Immunization may comprise a single administration or a multiplicity of administrations.
  • the oil is squalene.
  • composition which, when administered to an animal, will potentiate an immune response, the composition comprising antigen, sodium hydroxide, oil, surfactant, and chitosan solution, wherein the oil can be metabolically degraded.
  • an immunogen comprising an antigen, sodium hydroxide solution, oil, surfactant, and a chitosan solution, wherein the oil can be metabolically degraded.
  • a method for preparing an immunogen comprising, of preparing a chitosan solution, preparing a sodium hydroxide solution, preparing an oil/ surfactant solution, wherein the oil can be metabolically degraded, mixing the chitosan solution with the sodium hydroxide solution, the oil/ surfactant solution, and the antigen to form an emulsion.
  • kits comprising a chitosan solution, a sodium hydroxide solution, and an oil/ surfactant solution.
  • compositions for immunopotentiation which comprise an antigen/chitosan mixture or an antigen/chitosan/oil/surfactant emulsion, as well as methods to prepare the antigen/chitosan mixture and the antigen/chitosan/oil/surfactant emulsion.
  • Example 1 demonstrates the preparation of antigen incorporated and lyophilized in phosphate buffer, which is subsequently reconstituted in a chitosan solution.
  • Example 2 provides a comparison of the ability of antigen incorporated into phosphate buffer and reconstituted in a chitosan solution to stimulate an immune response to that of a cu ⁇ ently available adjuvant.
  • Example 3 demonstrates the preparation of antigen incorporated in a chitosan/oil emulsion.
  • Examples 4 and 5 provide a comparison of the ability of different antigens incorporated into a chitosan/oil emulsion to stimulate an immune response to that of a cu ⁇ ently available adjuvant.
  • a 0.5 M phosphate buffer was prepared by diluting 15.6 ml of phosphoric acid (16 M; Mallinkrodt Chemical, Paris, KY) in 400 ml of deionized (18 mOhm: DI) water. The pH of the solution was adjusted to 7.3 with 10 N sodium hydroxide (Sigma Chemical Co. , St. Louis, MO). The total volume of the solution was adjusted to 500 ml by the addition of DI water.
  • a dilute chitosan solution was made by first preparing a 1 % chitosan in 2% acetic acid solution: 1 gm of chitosan (practical grade; Sigma Chemical Co. , St. Louis, MO) in 100 ml of 2% glacial acetic acid (Mallinkrodt Chemical, Paris, KY). The resulting 1 % chitosan in 2 % acetic acid solution was then diluted further by adding 7.4 ml of the solution to 2.6 ml of DI water to obtain a chitosan working solution. The pH of the final chitosan solution was between 6 and 7. 50 ⁇ L of a lOmg/ml ovalbumin (Sigma Chemical Co., St; Louis,
  • mice that were previously immunized (individually) with either a vaccine comprising 25 ⁇ g of ovalbumin with CFA (Sigma Chemical Co. , St. Louis, MO) or a vaccine comprising 25 ⁇ g of ovalbumin incorporated and lyophilized in phosphate buffer, and subsequently reconstituted in a chitosan solution (Test Group), as prepared in Example 1.
  • a vaccine comprising 25 ⁇ g of ovalbumin with CFA (Sigma Chemical Co. , St. Louis, MO) or a vaccine comprising 25 ⁇ g of ovalbumin incorporated and lyophilized in phosphate buffer, and subsequently reconstituted in a chitosan solution (Test Group), as prepared in Example 1.
  • mice Female Balb/c mice, 8 weeks of age, were immunized by a single intraperitoneal injection of the vaccine on day 0.
  • the ovalbumin CFA treament group contained 3 mice, while the test group (treated ovalbumin incorporated and lyophilized in phosphate buffer and reconstituted in a chitosan solution) contained 4 mice. Both experimental groups were bled on day 7, post-injection.
  • the CFA adjuvanted group was also bled on days 21 , 28, 35, 42, and 48 post- immunization.
  • the test group was also bled on days 26, 38, 38, 52, 70, 83, 102, 123, and 159 post-immunization.
  • Anti-ovalbumin serum antibody titers were determined by ELISA.
  • the test group animals developed a high antibody titer by day 26 (10,000). The high titer persisted past 83 days post-immunization, via a booster vaccination on day 42. Immediately following the booster vaccination, the titer increased to appproximately 64,000 (day 52) and persisted above 10,000 to approximately 123 days post-vaccination (original).
  • the Test Group values obtained were comparable to those of the standard adjuvant used by those of ordinary skill in the art, Complete Freund's Adjuvant.
  • the mean titer values in the test group animals were comparable to those seen with antigens cross-linked to chitosan with glutaraldehyde, which generally improves immunopotentiation over other commercially available adjuvants (PCT/US95/12189; WO 96/09805).
  • the present invention is a safe and comparable alternative adjuvant to both CFA and antigens cross-linked to chitosan via glutaraldehyde.
  • HlV-peptide- keyhole limpet hemocyamn conjugate (Example 4) or human zona pellucida B peptide-ovalbumin (Example 5) as antigens
  • any number of other antigens may be employed.
  • squalene those of ordinary skill in the art will appreciate that any oil that is readily metabolized by the recipient animal may be used (e.g. , corn, canola, peanut).
  • a 2% chitosan solution in 0.5 M sodium acetate was prepared by dissolving 4.1 g of sodium acetate (Sigma Chemical Co. , St. Louis, MO) in 50 ml of deionized (18 mOhm: DI) water with mixing. The pH of the solution was adjusted to 4.5 with approximately 7 ml of glacial acetic acid (Mallinkrodt Chemical, Paris, KY) and an additional 1.5 ml of glacial acetic acid was added to compensate for the effect of the addition of chitosan on the pH of the solution. The total volume of the solution was adjusted to 100 ml by the addition of DI water. 2 grams of chitosan (Sigma Chemical Co. , St.
  • the chitosan solution was then sterilized by autoclaving during a 25 minute cycle. The solution was cooled to room temperature in a biosafety cabinet. The chitosan solution was then clarified by centrifugation in an IEC clinical centrifuge (International Equipment Co. , Needham Hts. , MA) at setting 7 for 5 minutes. The supernatant was decanted from the pellet (insoluble chitosan/ chitin and contaminants). 87 to 90% (by weight) of the chitosan added was retained in the supernatant.
  • IEC clinical centrifuge International Equipment Co. , Needham Hts. , MA
  • a 50% sodium hydroxide solution was prepared by dissolving 50 gm of sodium hydroxide (Sigma Chemical Co. , St. Louis, MO) in 100 ml of deionized water, with mixing.
  • a squalene/ surfactant solution was prepared by combining 1500 ⁇ of squalene (2, 6, 10,15, 19,23-Hexamethyl-2, 6,10, 14,18,22- tetracosahexaene; Sigma Chemical Co. , St. Louis, MO) with 600 ⁇ L of the surfactant Pluronic ® L121 (BASF Corp. , Parsippany, NJ) and vortexed until homogeneous.
  • a chitosan/squalene/surfactant/antigen emulsion was prepared by adding approximately 420 ⁇ L of antigen (i.e. , HIV-peptide-keyhole limpet hemocyanin conjugate, Table 2; human zona pellucida B peptide-ovalbumin conjugates, Table 3) in water or urea to approximately 370 ⁇ L of 2 % chitosan in 0.5 M sodium acetate and vortexing.
  • the actual amount of antigen (i.e. , protein or peptide-carrier conjugate) used may range from 1 ⁇ g to several milligrams. 10 ⁇ L of the 50% sodium hydroxide were then added to the antigen/chitosan and the sample was vortexed.
  • Example 4 10 ⁇ L aliquots of the 50% sodium hydroxide were added until a stable cloudy precipitate formed. Approximately 140 ⁇ L of the previously prepared squalene/ surfactant solution was added to the above solutions of antigen & chitosan. The resulting solution was vortexed until a cloudy emulsion formed. Immediately prior to administration in the immunization studies as described in Examples 4 and 5, the resulting solution of chitosan/ squalene/ surfactant/antigen was mixed by vortexing or syringe aspiration. Example 4
  • mice were individually immunized with either a vaccine comprising various amounts of HIV-peptide-KLH conjugate [Saren et al. Vaccine Res. , 3:49-57; incorporated herein by reference] with the chitosan/ squalene/ surfactant emulsion or 20 ⁇ g of HIV-peptide-KLH conjugate with CFA.
  • mice Female, Balb/c mice, 8 weeks of age were immunized by a single 200 ⁇ L intraperitoneal injection of the vaccine on day 0.
  • a second immunization was given to Group 1 , at week 18 (126 days after the first immunization).
  • a second immunization was adminsitered to Groups 2 and 3 at week 24 (168 days after the first immunization).
  • the second immunization consisted of the unconjugated HIV peptide at the dosage indicated with the chitosan/ squalene/ surfactant emulsion in Groups 1-3.
  • the CFA group did not receive a second immunization.
  • the subject animals were bled on days 22, 35, 49, 63, 77, 91 , 119 (excluding Group 1), 140, and 149. Serum antibody titers were determined by ELISA.
  • Antigen human zona pellucida B peptide-ovalbumin conjugates
  • mice were individually immunized (intraperitoneal) with a vaccine comprising 6 different human zona pellucida B (ZPB) synthetic peptides [SEQ ID NOS. 1-6] adjuvanted with either the chitosan/ squalene/ surfactant emulsion or CFA.
  • ZPB human zona pellucida B
  • mice Female Balb/c mice, 8 weeks of female, were immunized by a 200 ⁇ L intraperitoneal injection of the vaccine (20 ⁇ g each of 6 different human ZPB synthetic peptides combined either with chitosan/squalene/surfactant emulsion (Group I) or CFA (Group II) on days 0 and 28. The Group ⁇ mice received CFA vaccine as the booster.
  • Serum antibody titers were determined by ELISA using plates coated with 1 ⁇ g per well of a mixture of the 6 peptides.
  • Antibody titers against full length purified recombinant human ZPB protein produced in Chinese hamster ovary cells [Harris et al. J. Seq. and Mapping, 4:361-393, 1994; incorporated herein by reference] were also determined by ELISA on plates ) coated with 50 ng of purified protein.
  • Peptide specific antibody titer s at day titers at day: Adjuvant 21 43 60 81 43 60 1:81
  • Val Ser Ser Lys Gly Pro Met lie Leu Leu Gin Ala Thr Lys Asp Pro 1 5 10 15

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)

Abstract

L'invention concerne des méthodes et des compositions permettant de renforcer une réponse immunitaire, dans lesquelles l'adjuvant immunostimulant est le chitosane. L'administration desdites compositions peut se faire par différentes voies.
PCT/US1997/005094 1997-03-25 1997-03-25 Immunostimulation induite par le chitosane WO1998042374A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
PCT/US1997/005094 WO1998042374A1 (fr) 1997-03-25 1997-03-25 Immunostimulation induite par le chitosane
AU23478/97A AU2347897A (en) 1997-03-25 1997-03-25 Chitosan induced immunopotentiation
JP10545633A JP2000504350A (ja) 1997-03-25 1997-03-25 キトサン誘導性免疫強化
CA002255867A CA2255867C (fr) 1997-03-25 1997-03-25 Immunostimulation induite par le chitosane
EP97916251A EP0914154A1 (fr) 1997-03-25 1997-03-25 Immunostimulation induite par le chitosane
AU19742/01A AU771525B2 (en) 1997-03-25 2001-02-13 Chitosan induced immunopotentiation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PCT/US1997/005094 WO1998042374A1 (fr) 1997-03-25 1997-03-25 Immunostimulation induite par le chitosane
CA002255867A CA2255867C (fr) 1997-03-25 1997-03-25 Immunostimulation induite par le chitosane

Publications (1)

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WO1998042374A1 true WO1998042374A1 (fr) 1998-10-01

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EP (1) EP0914154A1 (fr)
AU (1) AU2347897A (fr)
CA (1) CA2255867C (fr)
WO (1) WO1998042374A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001035994A2 (fr) * 1999-11-04 2001-05-25 Zonagen, Inc. Immunostimulation induite par le chitosane
EP2068917A2 (fr) * 2006-09-22 2009-06-17 Government of the USA, as Represented by the Secretary, Department of Health and Human Services Compositions et procédés pour améliorer la réponse immunitaire à l'aide des chitosanes
US7588774B2 (en) 2003-05-12 2009-09-15 Becton, Dickinson And Company Molecules enhancing dermal delivery of influenza vaccines
US20120164174A1 (en) * 2009-06-25 2012-06-28 Bioleaders Corporation Adjuvant composition containing poly-gamma-glutamic acid-chitosan nanoparticles

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0183556A2 (fr) * 1984-11-29 1986-06-04 IHARA CHEMICAL INDUSTRY Co., Ltd. Utilisation d'oligomères de chitine ou chitosane pour la fabrication d'un agent augmentant l'immunité contre les infections bactérielles et mycotiques et contre la croissance de tumeurs
WO1996009805A2 (fr) * 1994-09-23 1996-04-04 Zonagen, Inc. Immunostimulation induite par le chitosane
WO1996010421A1 (fr) * 1994-10-04 1996-04-11 Medeva Holdings B.V. Compositions vaccinales

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0183556A2 (fr) * 1984-11-29 1986-06-04 IHARA CHEMICAL INDUSTRY Co., Ltd. Utilisation d'oligomères de chitine ou chitosane pour la fabrication d'un agent augmentant l'immunité contre les infections bactérielles et mycotiques et contre la croissance de tumeurs
WO1996009805A2 (fr) * 1994-09-23 1996-04-04 Zonagen, Inc. Immunostimulation induite par le chitosane
WO1996010421A1 (fr) * 1994-10-04 1996-04-11 Medeva Holdings B.V. Compositions vaccinales

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MARCINKIEWICZ ET AL: "IMMUNOADJUVANT PROPERTIES OF CHITOSAN", ARCHIVUM IMMUNOLOGIAE ET THERAPIAE EXPERIMENTALIS, vol. 39, 1991, pages 127 - 132, XP002046373 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001035994A2 (fr) * 1999-11-04 2001-05-25 Zonagen, Inc. Immunostimulation induite par le chitosane
WO2001035994A3 (fr) * 1999-11-04 2002-06-27 Zonagen Inc Immunostimulation induite par le chitosane
US7588774B2 (en) 2003-05-12 2009-09-15 Becton, Dickinson And Company Molecules enhancing dermal delivery of influenza vaccines
EP2068917A2 (fr) * 2006-09-22 2009-06-17 Government of the USA, as Represented by the Secretary, Department of Health and Human Services Compositions et procédés pour améliorer la réponse immunitaire à l'aide des chitosanes
US20120164174A1 (en) * 2009-06-25 2012-06-28 Bioleaders Corporation Adjuvant composition containing poly-gamma-glutamic acid-chitosan nanoparticles
US9314520B2 (en) * 2009-06-25 2016-04-19 Bioleaders Corporation Adjuvant composition containing poly-gamma-glutamic acid-chitosan nanoparticles

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AU2347897A (en) 1998-10-20
CA2255867A1 (fr) 1998-10-01
EP0914154A1 (fr) 1999-05-12
CA2255867C (fr) 2008-02-05

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