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WO2003007869A2 - Compositions immunogenes contenant des antigenes, des vecteurs geniques et des microspheres biodegradables chargees d'adjuvants - Google Patents

Compositions immunogenes contenant des antigenes, des vecteurs geniques et des microspheres biodegradables chargees d'adjuvants Download PDF

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Publication number
WO2003007869A2
WO2003007869A2 PCT/BR2002/000099 BR0200099W WO03007869A2 WO 2003007869 A2 WO2003007869 A2 WO 2003007869A2 BR 0200099 W BR0200099 W BR 0200099W WO 03007869 A2 WO03007869 A2 WO 03007869A2
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Prior art keywords
antigen
gene
pharmaceutical composition
immune response
microspheres
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WO2003007869A3 (fr
Inventor
Célio LOPES SILVA
José Maciel Junior RODRIGUES
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Universidade Federal de Minas Gerais
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Universidade Federal de Minas Gerais
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Priority to CA002453959A priority Critical patent/CA2453959A1/fr
Priority to EP02744957A priority patent/EP1420823A2/fr
Priority to AU2002317056A priority patent/AU2002317056A1/en
Publication of WO2003007869A2 publication Critical patent/WO2003007869A2/fr
Anticipated expiration legal-status Critical
Publication of WO2003007869A3 publication Critical patent/WO2003007869A3/fr
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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
    • A61K9/1647—Polyesters, e.g. poly(lactide-co-glycolide)
    • 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
    • A61K39/02—Bacterial antigens
    • A61K39/04—Mycobacterium, e.g. Mycobacterium tuberculosis
    • 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
    • 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
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55583—Polysaccharides
    • 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
    • A61K2039/55588—Adjuvants of undefined constitution
    • A61K2039/55594—Adjuvants of undefined constitution from bacteria

Definitions

  • the present invention is related to processes of production of pharmaceutical compositions which are able to stimulate a specific immune response, cellular and/or humoral, in a host immunized with an antigen or a gene vector that codifies the antigen of 'interest.
  • Such compositions involve biodegradable microspheres based on a copolymer derived from polyesters, capable of encapsulating and controlling the release of antigens, gene vectors, and adjuvants that stimulate mediators of the immune response.
  • the term "adjuvant" refers to a substance that does not share immune epitopes with the antigen of interest itself, but is able to stimulate the immune response against the antigen of interest .
  • Patent US 4,340,588 describes the use of trehalose dimycolate as adjuvant in a vaccine composition containing antigens of Brucella abortus (BA, 45/20 strain) .
  • impure natural mixtures of trehalose dimycolate were obtained from several sources and were denominated "cord factor". These factors usually present 90% dimycolate and 10% monomycolate, as well as a small amount of exogenous material, which contains from 95 to 99% trehalose dimycolate after appropriate purification by chromatographic techniques.
  • Aqueous suspensions containing an insoluble fraction of mycobacteria cellular wall were proposed (US 5,759,554).
  • Aqueous suspensions are prepared by the following steps: (i) rupture of the bacteria and, subsequent centrifugation; (ii) elimination of protein from the fraction based on cellular wall (cellular debris obtained in the centrifugation step) employing proteolytic enzymes; (iii) treatment with detergents and washing of fraction resulting from step (ii) , (iv) freeze-drying and (v) suspension of the freeze-dried fraction in an aqueous solution, such as saline solution.
  • aqueous solution such as saline solution.
  • the stimulation of the immune system through the employment of a suspension based on mycobacterial fractions stimulates the human or animal organism to neutralize an infectious agent or delay the growth of cancerous cells.
  • this procedure does not allow a controlled adjuvant release, which could result in optimization of the immune response.
  • Processes that target the antigen, gene vectors and/or adjuvant have also been described in the literature, as in patent US 5,643,605.
  • the use of adjuvant and/or microencapsulated antigens is described for therapeutic or prophylactic purposes.
  • the composition presented under patent US 5,643,605 contains microspheres of PLGA (D-L-lactide-co-glycolide)
  • the adjuvant used was saponin (QS21) or mura ildipeptide (MDP) , which are able to stimulate an immune response where: (i) the volume of the aqueous adjuvant incorporated into the polymer is equal or lower than 1 mL for 3 grams of polymer; (ii) the molar ratio between monomers of lactic and glycolic acid can vary .from 100:0 to 0:100; (iii) the inherent viscosity of PLGA 5 polymer varies from 0.1 to 1.2 dL/g; (iv) the diameters of
  • the microspheres vary from 20 to 100 ⁇ m, and (v) the adjuvant is released from microspheres according to a triphasic model in which, during the first phase, less than 30% of the adjuvant is released in a period of 1 day;
  • Patent US 6,048,551 describes a method of preparation of microspheres containing gene vectors which, are employed in the treatment of tumors.
  • the multiple emulsion method was used.
  • the gene vector was dissolved in an aqueous phase and encapsulated, associated or not to antiviral drugs that were also dissolved in the aqueous phase.
  • the diameter of the particles ranged between 1 and 200 ⁇ m.
  • Patent US 6,309,569 claims a method of encapsulating DNA in a polymer microparticle where the excess of organic solvent is extracted by adding the (water-in-oil) -in-water emulsion to an aqueous phase to extract the solvent, thereby forming polymer microparticles of a size up to 10
  • This invention presents an immunogenic composition involving, at least, adjuvant and/or protein; and/or plasmids containing protein-expressing genes, entrapped into polymeric microspheres with an average diameter smaller than 20 ⁇ m, preferentially between 1 and 10 ⁇ m. Therefore, the composition contains, besides the adjuvant, an antigen or a gene vector that codifies the antigen (preferentially from parasites and pathogenic agents), which include antigens of extra and intracellular bacteria, eucariotic organisms such as fungi and protozoa, among others.
  • the adjuvants are, preferentially, those which stimulate citokyne production.
  • the plasmid carrying the gene of Mycobacterium proteins as, for instance, the hsp65 gene (heat shock protein of 65 kDa from
  • the purified or recombinant protein may be encapsulated in polymeric microspheres derived from lactic acid and copolymers of lactic and glycolic acid
  • the trehalose dimycolate may be encapsulated in microspheres of PLGA.
  • the system which includes trehalose dimycolate and/or the plasmid containing the gene of hsp65 protein, or still the protein itself, can also be applied in the prevention and therapy of tuberculosis, when the disease is already installed in the human or animal host.
  • microencapsulation of a gene vector, antigen and/or - adjuvant according to the present invention can be obtained by the following protocols:.
  • Protocol A (i) dissolution of the polymer derived from lactic and glycolic acids (ratio in % of weight between 100:0 and 0:100) in an organic solvent, which is non miscible or partially miscible in water in a concentration between 0.2 and 10%, as for example, ethyl acetate, methylene chloride, chloroform, among others; (ii) dissolution of the lipophilic antigen and/or adjuvant in the solution (i) , that contains the polymer, (iii) addition of the solution .obtained in step(ii) to a polyol solution, for example, polyvinyl alcohol (1 to 10%) , aiming the formation of an oil-in-water emulsion, (iv) stirring of this emulsion under an appropriated speed (200 to 1000 rpm) , in order to ' obtain microspheres with an average
  • diameter smaller than 10 ⁇ preferentially between 1 and 5
  • the suspension of microspheres obtained may be, alternatively, collected by centrifugation or filtration, washed with an aqueous solvent, then freeze-dried.
  • Protocol B (i) dissolution of the polymer derived from lactic and glycolic acids (ratio in % of weight between 100:0 and 0:100) in an organic solvent in a concentration between 0.2 and 10%, for example, methylene chloride, ethyl acetate, chloroform, among others, with or without a lipophilic adjuvant such as trehalose dimycolate; (ii) dissolution of the gene vector, antigen and/or adjuvant of hydrophilic nature, in 0.1 to 1 mL of an aqueous phase, (iii) addition of the solution obtained in (ii) to the polymeric solution obtained in (i) , followed by agitation under a speed ranging between 500 to 15,000 rpm, leading to the formation of a water-in-oil emulsion (or dispersion) , (iv) addition of this primary emulsion (or dispersion), after adequate period of agitation (1 to 10 minutes) , to an emulsifying aque
  • Microspheres are prepared following the emulsion and solvent evaporation technique (Lewis, D. H. 1990. Controlled release of bioactive agents from lactide/glicolide polymers. In Chasin M & Langer R. Biodegradable polymers as drug delivery systems. Marcel Dekker, New York, 1-43) .
  • Five mg of trehalose dimycolate (SIGMA) and 125 mg, of PLGA (50 : 50) biodegradable (Resomer RG 505, MW 78,000, Boehringer Ingelheim, Germany) were diluted in 30 mL of methylene chloride. This organic phase was mixed with 100 mL of an aqueous phase containing 3% of
  • polyvinyl alcohol (Mowiol® 40-88, SIGMA Aldrich Chemicals) as surfactant, aiming the formation of an oil-in-water stable emulsion.
  • a mechanical agitation 800 rpm was applied for a 6-hour period at room temperature.
  • Microspheres were collected by centrifugation at 10.000 x g, washed 3 times with sterile water or saline, freeze-dried and stored at 4°C. Presence of trehalose dimycolate in microspheres was determined by thin-layer chromatography after dissolution of the particles in methylene chloride. Diameter . of the particles was determined by laser difractometry in a CILAS 1064 Liquid apparatus (Cilas, France) showing average
  • E. coli, DH5 ⁇ strain, transformed with the gene vectors pCDNA3 or pCDNA3modified (pCDNA3m) , and with the plasmids containing the hsp65 gene (pCD AS-hap-f- ⁇ pCDNA3modified-hsp65) are cultured in LB BROTH BASE medium
  • pCDNA3 vector (Invitrogen®, Carlsbad, CA, USA), which is previously digested with Bam HI and Not I (Gibco BRL,
  • Plasmids are purified by anion exchange resin (Concert High Purity Maxiprep System, GIBCO BRL) . Evaluation of plasmid
  • E. coli, BL21 strain, transformed with pIL-161 plasmid containing the M. leprae hsp65 gene are cultured in 500 mL of LB BROTH BASE liquid medium (GIBCO BRL, Scotland) containing ampicillin (SIGMA) in the concentration of 100
  • Microspheres are obtained by multiple emulsion followed by solvent evaporation method: the aqueous phase (0.3 L) containing only the plasmid vector that does not codify the hsp65 protein (pCDNA3 or PcDNA3modified) ; or the vector containing the gene that codifies the hsp65 protein
  • plasmid For preparation of microspheres containing plasmid, 4 mg of plasmid are dissolved into 0.3 mL of aqueous phase and, in case of recombinant hsp65, 1 mg of protein is dissolved into 0.3 mL of aqueous phase. This emulsion is added to an external aqueous phase (100 mL) ,
  • Microspheres are collected by centrifugation at 10.000 x g in a Himac CR21 centrifuge (Hitachi, rotor R20A2), washed 3
  • Microspheres are obtained by multiple emulsion and solvent evaporation method: the aqueous phase (0.3 L) containing pCDNA3, PcDNA3modified, pCDNA3-hsp65, PcDNA3modified-hsp65 or recombinant hsp65 ( 4 mg for plasmid or 1 mg for protein) , is emulsified in 30 mL of methylene chloride containing 0 . 5 mg of trehalose dimycolate and 400 mg of PLGA 50 : 50 (Resomer RG 505 da Boehringer Ingelheim) under strong agitation ( 8 , 000 rpm) ,
  • Microspheres are collected by centrifugation at 10. 000 x g in a Himac CR21 centrifuge (Hitachi, rotor R20A2 ) , washed 3 times with sterile water, freeze-dried and
  • Particle diameter is evaluated by laser difractometry in a CILAS 1064 Liquid apparatus (Cilas , France ) .
  • the average particle diameter should be between 1
  • EXAMPLE 7 Evaluation of hsp65 expression in eucariotic cells transfected with plasmid-loaded microspheres J774 cells (tumoral lineage cells originally from BALB/c mice) and J774-hsp65 (J774 cells transfected with retroviral vector [pZIPNeoSV(x) ] carrying the M. Leprae hsp65 gene) are cultured in 1640 RPMI medium (SIGMA) supplemented with 2 % of heat inactivated fetal calf serum,
  • J774 cells receive 1 mL of RPMI medium and are used as negative control for hsp65 expression;
  • J774-hsp65 cells receive 1 mL of RPMI medium and are used as positive control for hsp65 expression;
  • C) J774 cells receive 1 mL of RPMI medium containing pCDNA3modified-hsp65-loaded microspheres (10 microspheres/cell) . Particles are ressuspended in medium and the concentration of this suspension is adjusted to 5xl0 6 particles/mL, counting the particles in a Neubauer chamber;
  • J774 cells receive 1 mL of RPMI medium containing
  • pCDNA3modified-hsp65 complexed with Lipofectin® (GIBCO BRL) Twenty-four hours after incubation, the culture supernatants are removed and replaced with 1 L of RPMI medium. Medium is substituted every 48 hours.
  • the cells are washed 3 times in PBS solution containing 1% of bovine serum albumin (BSA) (SIGMA). They are fixed in 4% paraformaldehyde solution for 30 minutes at room temperature and then rewashed . All the washes are performed 3 times. After that, endogenous peroxidase blockage- is carried out by incubation with PBS containing 3% of H 2 0 2 for 30 minutes at room temperature. After washing, the cells are incubated with blocking buffer (3 % of rabbit serum, 1% of BSA and 0.01% of Triton X 100) for 1 hour at room temperature, in order to block nonspecific linkages. Following that, anti-hsp65 antibody is added and
  • mice 6 to 8 weeks old, were obtained from the animal facilities at the School of Medicine of Ribeirao Preto, University of Sao Paulo, and maintained in standard laboratory conditions. All procedures were carried out according to the Ethic Committee recommendations.
  • mice were immunized with different microsphere formulations by intramuscular or intratracheal route.
  • Microsphere doses varied from 100 to 500 mg/kg and the schedules were of single dose for intratracheal route and 1 or 3 doses for intramuscular route.
  • Mice were either challenged or killed, 30 or 60 days after administration of the microspheres, and the immune response was evaluated.
  • mice received, by intratracheal route, a dose of 10 colony-forming units (CFU) of M. tuberculosis H37Rv strain, under anesthesia
  • EXAMPLE 9 Cytokine measurement Cytokine levels produced by lung and spleen cells from mice immunized with microspheres are measured by ELISA. Total lung tissue is homogenized for 5 minutes in an Ultraturrax T 25 IKA (Labortechnik, Germany) at 4°C. Homogenized tissue is centrifuged at 10,000 x g for 15
  • IL-4 (BVD4-1D11, BVD6-24G2), IFN- ⁇ (R4-6A2, XMG1.2) and IL-12 (C15.6, C17.8), as well as recombinant cytokines, may be purchased from Pharmingen (San Diego, CA) .
  • IL-4 (BVD4-1D11, BVD6-24G2)
  • IFN- ⁇ R4-6A2, XMG1.2
  • IL-12 C15.6, C17.8
  • recombinant cytokines may be purchased from Pharmingen (San Diego, CA) .
  • TNF- ⁇ , IL-10, IL-6, IL-4, IFN- ⁇ and IL-12 are determined in supernatant by ELISA technique, as previously described (Haagmans, B. L., A. J. van den Eertwegh, E. Claassen, M. C. Horzinek, and V. E. Schijns. 1994. Tumor necrosis factor-alpha production during cytomegalpvirus infection in immunosuppressed rats. J. Gen. Virol. 75:779-787), according to manufacturer instructions (Pharmingen, San Diego, CA) . For each assay, a standard curve is built with
  • EXAMPLE 10 Nitric oxide measurement Cells from bronchoalveolar lavage recovered from mice injected with microspheres containing trehalose dimycolate are cultured in RPMI medium containing 10% of fetal calf serum (GIBCO-BRL, Grand Island, NY), 10 mM Hepes, 20 mM sodium bicarbonate, penicillin, and streptomycin at 100 ⁇ g/ml (GIBCO-BRL) .
  • nitric oxide Production of nitric oxide is evaluated by measuring nitrite (N0 2 ⁇ ) production in cell culture supernatant by Greiss method (Stuehr, D.J., and C. F. Nathan. 1989. Nitric oxide. A macrophage product responsible for cystostasis and respiratory inhibition in tumor target cells. J. Exp. Med. 169:1543-1555) . The standard curve is constructed using a serial dilution of a
  • Interferon-gamma (IFN- ⁇ ) levels were significant after in vitro stimulation with recombinant hsp65 in mice immunized with recombinant hsp65-loaded microspheres. Mice immunized with recombinant hsp65 (r-hsp65) in PBS or unloaded
  • microspheres did not produce significant levels of IFN- ⁇ .
  • IFN- ⁇ levels produced after in vitro stimulation with, concanavalin A (ConA) were used as positive control.
  • Interleukin 12 (IL-12) levels were significant in mice which received r-hsp65 in PBS or loaded into microspheres. This result confirms the possibility of administering the formulations resulting from the present invention by a route other than the parenteral one.
  • IFN- ⁇ production pg/mL
  • the formulations object of the present invention were also able to elicit specific immune response.
  • BALB/c mice were immunized with a single dose of 5 mg of microspheres containing plasmid/mouse by intramuscular route. Sixty days after immunization, there was evidence of specific immune
  • Table IV IL-12 production (pg/mL) in spleen cells culture supernatant from BALB/c mice immunized by intramuscular injection with plasmid and r-hsp65 loaded into microspheres, 60 days after immunization
  • Table V IL-6 production (pg/mL) in spleen cells culture supernatant from BALB/c mice immunized by intramuscular injection with plasmid and r-hsp65 encapsulated into microspheres , 60 days after immunization
  • Tables VI and VII show that, after intratracheal administration, the formulation obtained in this invention was able to elicit the production of several cytokines by lung cells, as well as the activation of alveolar acrophages as determined by nitric oxide production.
  • Table VI illustrates the results of cytokine production by lung cells from mice injected with microspheres containing TDM and controls (PBS, control lipid CtLip and trehalose dibehenate DBT) . The animals were injected by intratracheal route. Sixty days later, their lungs were removed and homogenized for measurement of cytokine in supernatant by ELISA. Data are representative of one experiment, repeated 3 times.
  • Table VII illustrates the results of nitric oxide production by cells from bronchoalveolar lavage from mice injected with microspheres containing TDM and controls (PBS, control lipid, and trehalose dibehenate) . Around 10 cells from bronchoalveolar lavage were obtained in each
  • Table VII Evaluation of nitric oxide levels in broncoalveolar lavage cells culture supernatant from BALB/c mice treated by intratracheal route.
  • Table VIII illustrates the IFN- ⁇ levels produced in spleen cell culture supernatant from BALB/c mice immunized with microspheres containing plasmid plus TDM and controls
  • Table IX illustrates the levels of antibodies in serum from BALB/c mice immunized with microspheres containing, plasmid plus TDM and controls, 30 days after immunization.
  • Tables X, XI and XII illustrate cytokine levels produced in lung homogenate from BALB/c mice challenged with M. tuberculosis H37Rv strain after injection of microspheres containing plasmid plus TDM and controls.
  • Table X IL-10 production (pg/mL) in lung homogenate from BALB/c mice immunized and challenged with M. tuberculosis
  • Table XI IL-12 production (pg/mL) in lung homogenate from BALB/c mice immunized and challenged with M. tuberculosis
  • Table XIII illustrates the ability of different microsphere formulations containing DNA or protein plus TDM, to confer protection in BALB/c mice against challenge with virulent strain of M. Tuberculosis .

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Abstract

L'invention concerne des compositions capables de stimuler une réponse immune chez un hôte auquel on a inoculé un antigène d'intérêt ou le vecteur génique qui le codifie. Cette composition comporte des microsphères à base de copolymères dérivés d'acide lactique et d'acide glycolique enrobant des antigènes, des vecteurs géniques et/ou des adjuvants présentant des propriétés immunostimulatrices obtenues à partir de fractions mycobactériennes. L'invention se caractérise en ce que l'on utilise du tréhalose dimycolate associé à une protéine de choc thermique ou au plasmide renfermant le gène le codifiant, enrobé dans des microsphères dont les dimensions n'excèdent pas 10 νm, capable d'induire des anticorps spécifiques et des réponses immunes cellulaires, ainsi que la sécrétion de cytokines impliquées dans la protection contre des maladies infectieuses et cancers, après administration d'une ou de plusieurs doses.
PCT/BR2002/000099 2001-07-17 2002-07-17 Compositions immunogenes contenant des antigenes, des vecteurs geniques et des microspheres biodegradables chargees d'adjuvants Ceased WO2003007869A2 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
CA002453959A CA2453959A1 (fr) 2001-07-17 2002-07-17 Compositions immunogenes contenant des antigenes, des vecteurs geniques et des microspheres biodegradables chargees d'adjuvants
EP02744957A EP1420823A2 (fr) 2001-07-17 2002-07-17 Compositions immunogenes contenant des antigenes, des vecteurs geniques et des microspheres biodegradables chargees d'adjuvants
AU2002317056A AU2002317056A1 (en) 2001-07-17 2002-07-17 Immunogenic compositions containing antigens, gene vectors and adjuvants-loaded biodegradable microspheres

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
BRPI0103887 2001-07-17
BR0103887-7A BR0103887C1 (pt) 2001-07-17 2001-07-17 Composições imunogênicas contendo microesferas biodegradáveis encapsulando antìgenos, vetores gênicos e adjuvantes
BRC10103887 2002-07-04

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WO2003007869A3 WO2003007869A3 (fr) 2004-03-18

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007001448A2 (fr) 2004-11-04 2007-01-04 Massachusetts Institute Of Technology Particules polymeres revetues a diffusion regulee comme vecteurs efficaces d'administration par voie orale de produits biopharmaceutiques
CN1317560C (zh) * 2005-10-19 2007-05-23 华中科技大学 一种提高量子点编码微球的编码稳定性的方法
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WO2008098165A2 (fr) 2007-02-09 2008-08-14 Massachusetts Institute Of Technology Bioréacteur oscillant pour la culture de cellules
JP2010523595A (ja) 2007-04-04 2010-07-15 マサチューセッツ インスティテュート オブ テクノロジー ポリ（アミノ酸）ターゲッティング部分
CN101861165A (zh) 2007-10-12 2010-10-13 麻省理工学院 疫苗纳米技术
CN101376023B (zh) * 2008-10-10 2011-12-28 山西海森生物制品有限公司 一种禽用疫苗缓释剂及其制备与接种方法
US8277812B2 (en) 2008-10-12 2012-10-02 Massachusetts Institute Of Technology Immunonanotherapeutics that provide IgG humoral response without T-cell antigen
US8343497B2 (en) 2008-10-12 2013-01-01 The Brigham And Women's Hospital, Inc. Targeting of antigen presenting cells with immunonanotherapeutics
US8591905B2 (en) 2008-10-12 2013-11-26 The Brigham And Women's Hospital, Inc. Nicotine immunonanotherapeutics
US8343498B2 (en) 2008-10-12 2013-01-01 Massachusetts Institute Of Technology Adjuvant incorporation in immunonanotherapeutics
EP2661253B1 (fr) * 2011-01-04 2017-04-19 Archivel Farma, SL Formulation de liposomes pour le traitement ou la prévention de la tuberculose
WO2013104943A1 (fr) 2012-01-12 2013-07-18 Archivel Farma, S.L. Vaccin mycobacterium tuberculosis-complex (mtb-c) contre l'asthme
EP3142852B1 (fr) 2014-05-14 2021-12-01 Dicar, Inc. Couvercle de protection comportant des renforts à motifs

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH01207237A (ja) * 1988-02-15 1989-08-21 Sawai Seiyaku Kk 腫瘍壊死因子誘起剤
US6048551A (en) * 1997-03-27 2000-04-11 Hilfinger; John M. Microsphere encapsulation of gene transfer vectors

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014046983A1 (fr) * 2012-09-21 2014-03-27 Intensity Therapeutic Procédé de traitement du cancer
US9636406B2 (en) 2012-09-21 2017-05-02 Intensity Therapeutics, Inc. Method of treating cancer
US10888618B2 (en) 2012-09-21 2021-01-12 Intensity Therapeutics, Inc. Method of treating cancer

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CN1549728A (zh) 2004-11-24
BR0103887C1 (pt) 2005-11-08
WO2003007869A3 (fr) 2004-03-18

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