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WO2018030338A1 - Composition d'adjuvant - Google Patents

Composition d'adjuvant Download PDF

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Publication number
WO2018030338A1
WO2018030338A1 PCT/JP2017/028574 JP2017028574W WO2018030338A1 WO 2018030338 A1 WO2018030338 A1 WO 2018030338A1 JP 2017028574 W JP2017028574 W JP 2017028574W WO 2018030338 A1 WO2018030338 A1 WO 2018030338A1
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Prior art keywords
cpg
carbonate apatite
nanoparticles
administered
adjuvant composition
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English (en)
Japanese (ja)
Inventor
浩文 山本
吉岡　靖雄
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Research Foundation for Microbial Diseases of Osaka University BIKEN
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Research Foundation for Microbial Diseases of Osaka University BIKEN
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Priority to JP2018533448A priority Critical patent/JP7016317B2/ja
Publication of WO2018030338A1 publication Critical patent/WO2018030338A1/fr
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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
    • 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
    • A61K39/145—Orthomyxoviridae, e.g. influenza virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/52—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient

Definitions

  • the present invention relates to an adjuvant composition that is highly safe and can effectively improve protective immunity induced by a vaccine. Furthermore, the present invention relates to a vaccine preparation using the adjuvant composition.
  • the current vaccine development is shifting from live vaccines such as attenuated strains to development using pathogen-derived proteins and peptides as antigens with emphasis on safety.
  • proteins and peptides as antigens
  • stability in vivo is poor, it is difficult to migrate to lymph nodes where immunity is induced, and the rate of antigen migration to dendritic cells is extremely low. is there.
  • it is indispensable not only to induce antibody responses against pathogens, but also to induce cytotoxic T cells after delivering antigens into dendritic cells.
  • immunity induction is carried out using a peptide alone as an antigen, the antigen is decomposed by intracellular endosomes and cannot be induced to the cytoplasm, so that protective immunity cannot be effectively induced.
  • an adjuvant is used to increase the induction efficiency of protective immunity.
  • Adjuvants may also be used to efficiently induce protective immunity even in the case of live vaccines.
  • conventional adjuvants like proteins and peptides used as antigens, cannot control pharmacokinetics and are difficult to migrate to lymph nodes, and thus have a drawback that the induction of protective immunity cannot be effectively increased.
  • the conventional adjuvant causes unexpected side effects by moving to a non-target tissue other than lymph nodes. against the background of such conventional technology, it is desired to develop an adjuvant composition that is highly safe and can effectively improve the protective immunity induced by the vaccine.
  • Patent Document 1 discloses that carbonate apatite nanoparticles can efficiently deliver drugs such as nucleic acids and low-molecular compounds into cells. Patent Document 1 also discloses that carbonate apatite nanoparticles are efficiently immersed and accumulated in tumor tissue. Furthermore, Non-Patent Document 1 also discloses that carbonate apatite encapsulating siRNA is effective for treatment of solid cancer. However, there has been no report on a preparation using a combination of carbonate apatite nanoparticles and an adjuvant.
  • An object of the present invention is to provide an adjuvant composition that is highly safe and can effectively improve the protective immunity induced by a vaccine, and a vaccine preparation using the adjuvant composition.
  • Item 1 An adjuvant composition comprising CpG oligodeoxynucleotide and carbonate apatite particles.
  • Item 2. The adjuvant composition according to Item 1, wherein the CpG oligodeoxynucleotide is a K type.
  • Item 3. Item 3.
  • Item 4. Item 4. The adjuvant composition according to any one of Items 1 to 3, wherein the CpG oligodeoxynucleotide is complexed with the carbonate apatite particles.
  • a vaccine preparation comprising an antigen and the adjuvant composition according to any one of Items 1 to 4.
  • Item 6. The vaccine preparation according to Item 5, wherein the antigen is a protein or a peptide.
  • Item 7. Item 7. The vaccine preparation according to Item 5 or 6, wherein the antigen is a protein or peptide derived from influenza virus.
  • protective immunity induced by a vaccine can be effectively improved while ensuring high safety, more effective prevention or infection against viruses, bacteria, parasites and other infectious diseases It is possible to provide treatment strategies.
  • Example 1 it is the result of having measured the carbonate apatite (AP) particle
  • Example 1 it is the result of having observed the carbonate apatite (AP) particle
  • Example 2 after dendritic cells were cultured in a medium containing AP-CpG nanoparticles, the amount of cytokines (IFN- ⁇ , IL-12, and IFN- ⁇ ) in the supernatant was measured.
  • cytokines IFN- ⁇ , IL-12, and IFN- ⁇
  • Example 3 it is the result of measuring the amount of cytokines (IFN- ⁇ and IFN- ⁇ ) in the supernatant after culturing human peripheral blood mononuclear cells in a medium containing AP-CpG nanoparticles.
  • a is a graph plotting the fluorescence intensities of FSC-A and CpG ODN by FACS analysis in Example 4.
  • FIG. b is the result of measuring the amount of Ap-CpG nanoparticles taken up by dendritic cells in Example 4.
  • FIG. In Example 5 it is the result of measuring the uptake
  • Example 6 it is the result of measuring the amount of cytokines (IL-12, IFN- ⁇ and IL-6) in the supernatant after administering Ap-CpG nanoparticles to mice and culturing regional lymph node cells.
  • Example 7 it is the result of measuring the ovalbumin-specific antibody in plasma in the mouse administered with Ap-CpG nanoparticles and ovalbumin.
  • a is the result of measuring the hemagglutinin-specific antibody derived from influenza A strain H1N1 in plasma in Example 8 in mice administered with Ap-CpG nanoparticles and hemagglutinin derived from influenza A strain H1N1.
  • b is the result of measuring the survival rate and the body weight transition by intranasal administration of influenza virus to mice administered with Ap-CpG nanoparticles and hemagglutinin derived from influenza A strain H1N1 in Example 8.
  • a is the result of measuring the number of leukocytes and lymphocytes in blood in Example 9 in mice administered with Ap-CpG nanoparticles.
  • b is the result of measuring the weights of the spleen, lymph nodes, and liver in the mice administered with Ap-CpG nanoparticles in Example 9.
  • Example 10 after dendritic cells were cultured in a medium containing AP nanoparticles and CpG ODN, the amount of cytokines (IL-6, IL-12, and IFN- ⁇ ) in the supernatant was measured. .
  • Example 11 it is the result of measuring ovalbumin-specific IgG1 and IgG2c in plasma in mice administered with Ap-CpG nanoparticles or AP nanoparticles and CpG ODN together with ovalbumin.
  • Example 12 it is the result of measuring hemagglutinin-specific IgG derived from influenza H1N1 strain in plasma in mice administered with Ap-CpG nanoparticles or AP nanoparticles and CpG ODN together with hemagglutinin from influenza A strain H1N1.
  • Example 12 it is the result of measuring hemagglutinin-specific IgG2c derived from influenza H1N1 strain in plasma in mice administered with Ap-CpG nanoparticles or AP nanoparticles and CpG ODN together with hemagglutinin from influenza A strain H1N1. .
  • a is the result of measuring hemagglutinin-specific antibodies derived from influenza A strain H1N1 in plasma in Example 13 in mice administered with AP nanoparticles, CpG ODN, and influenza A H1N1 strain-derived hemagglutinin.
  • b shows the results of measuring the survival rate and the body weight transition by intranasal administration of influenza virus to mice administered with AP nanoparticles, CpG ODN, and hemagglutinin derived from influenza A H1N1 strain in Example 13.
  • Reference Test Example 1 the results of measurement of ovalbumin-specific IgG1 and IgG2c in plasma in mice administered with AP-Poly (I; C) nanoparticles and ovalbumin.
  • Adjuvant Composition The adjuvant composition of the present invention is characterized by containing CpG oligodeoxynucleotide and carbonate apatite particles.
  • the adjuvant composition of the present invention will be described in detail.
  • the adjuvant composition of the present invention contains CpG oligodeoxynucleotide as an adjuvant.
  • CpG oligodeoxynucleotide as an adjuvant and using it together with carbonate apatite particles, it becomes possible to efficiently elicit protective immunity by a vaccine while ensuring high safety.
  • CpG oligodeoxynucleotide is an oligonucleotide having a CpG motif containing an unmethylated cytosine-guanine sequence.
  • CpG oligodeoxynucleotide may be abbreviated as CPG or CPG ODN.
  • the number of CpG motifs contained is not particularly limited, and may be one or more.
  • the number of bases of the CpG oligodeoxynucleotide is not particularly limited as long as it can function as an adjuvant.
  • the upper limit is 200 or less, preferably 100 or less, more preferably 50 or less, and still more preferably 40 or less. And most preferably 30 or less.
  • the lower limit is 5 or more, preferably 8 or more, more preferably 10 or more, and still more preferably 15 or more. More specifically, the number of bases of the CpG oligodeoxynucleotide is 5 to 200, preferably 8 to 50, more preferably 10 to 50, still more preferably 10 to 40, and most preferably 10 to 30. Is mentioned.
  • phosphodiester bond of the CpG oligodeoxynucleotide used in the present invention may be substituted with a phosphorothioate bond as necessary.
  • a phosphorothioate bond refers to a bond structure in which one of the oxygen atoms of the phosphate residue of the phosphodiester bond moiety is replaced with a sulfur atom. Formation of phosphorothioate bonds in CpG oligodeoxynucleotides can be performed according to conventionally known methods.
  • C type, D type, C type, and P type are known as CpG oligodeoxynucleotides depending on the base sequence.
  • any of these types of CpG oligodeoxynucleotides is used. May be.
  • C type or D type more preferably K type, particularly preferably K3 type CpG oligodeoxynucleotides are mentioned. .
  • K-type CpG oligodeoxynucleotides applied to humans include atcgactctcgagcgttctc (SEQ ID NO: 1, type K3), tcgtcgttttgtcgtttgtcgttt (SEQ ID NO: 2); K-type CpG applied to rodents (mouse, rat)
  • Examples of oligodeoxynucleotides are atcgactctcgagcgttctc (SEQ ID NO: 1, type K3), tccatgacgttcctgatgct (SEQ ID NO: 3), tccatgacgttcctgacgtt (SEQ ID NO: 4); Type), gggggacgatcgtcgggggg (SEQ ID NO: 6
  • CpG oligodeoxynucleotides are commercially available from Gene Design, InvivoGen, etc., and commercially available CpG oligodeoxynucleotides may be used in the present invention.
  • Carbonate apatite particles Carbonate apatite has a structure in which hydroxyl group of hydroxyapatite (Ca 10 (PO 4 ) 6 (OH) 2 ) is partially substituted with CO 3 , and has the general formula Ca 10-m X m (PO 4 ) 6 ( CO 3 ) 1-n Y n is a compound represented by
  • X is an element that can partially replace Ca in carbonate apatite, and examples thereof include Sr, Mn, and rare earth elements.
  • m is a positive number of usually 0 or more and 1 or less, preferably 0 or more and 0.1 or less, more preferably 0 or more and 0.01 or less, and still more preferably 0 or more and 0.001 or less.
  • Y is a group or element that can partially substitute CO 3 in carbonate apatite, and examples thereof include OH, F, and Cl.
  • n is usually a positive number from 0 to 0.1, preferably from 0 to 0.01, more preferably from 0 to 0.001, and even more preferably from 0 to 0.0001.
  • the average particle diameter of the carbonate apatite particles used in the present invention is not particularly limited as long as it is a size that can be administered into a living body and transferred into a dendritic cell that becomes a target cell. From the viewpoint of efficiently performing inward migration, it is preferably a nanoparticle, more specifically, usually 200 nm or less, preferably 50 nm or less, more preferably 1 to 40 nm, more preferably 1 to 20 nm, Particularly preferred is 5 to 10 nm.
  • the average particle diameter of the carbonate apatite means a volume average particle diameter measured using a zeta sizer when it is 100 nm or more, and when it is less than 100 nm, 50 particles are measured with an atomic force microscope (AFM). Mean value obtained by measuring the particle diameter of
  • the method for producing carbonate apatite having the average particle diameter described above is not particularly limited, and specifically, a step of preparing a dispersion in which carbonate apatite particles are dispersed in a pharmaceutically acceptable solvent, and the dispersion A method of undergoing a process of ultrasonic vibration treatment for the liquid is mentioned.
  • Carbonate apatite particles can be obtained according to a known method. For example, it can be obtained by preparing it in the presence of calcium ions, phosphate ions and bicarbonate ions in an aqueous solution.
  • concentration of each ion in the aqueous solution is not particularly limited as long as the carbonate apatite particles are formed, and can be appropriately set with reference to the following.
  • the calcium ion concentration in the aqueous solution is usually 0.1 to 1000 mM, preferably 0.5 to 100 mM, more preferably 1 to 10 mM.
  • the phosphate ion concentration in the aqueous solution is usually 0.1 to 1000 mM, preferably 0.5 to 100 mM, and more preferably 1 to 10 mM.
  • the concentration of hydrogen carbonate ions in the aqueous solution is usually 1.0 to 10,000 mM, preferably 5 to 1000 mM, and more preferably 10 to 100 mM.
  • the supply source of calcium ions, phosphate ions and hydrogen carbonate ions is not particularly limited as long as these ions can be supplied in an aqueous solution, and examples thereof include water-soluble salts of these ions.
  • CaCl 2 can be used as the calcium ion source
  • NaH 2 PO 4 .2H 2 O can be used as the phosphate ion source
  • NaHCO 3 can be used as the carbonate ion source.
  • each ion source is not particularly limited, and the aqueous solution may be prepared in any mixing order as long as carbonate apatite particles are obtained.
  • a first solution containing calcium ions is prepared, and a second solution containing phosphate ions and bicarbonate ions is prepared separately, and the first solution and the second solution are mixed.
  • An aqueous solution can be prepared.
  • the aqueous solution for preparing the carbonate apatite particles may contain components other than the above-described ion supply sources and other substances as long as the carbonate apatite particles are formed.
  • Ca or CO 3 in carbonate apatite may be partially substituted by adding fluorine ions, chlorine ions, Sr, Mn, or the like to the above composition in an aqueous solution.
  • the addition amount of fluorine ions, chlorine ions, Sr, and Mn is preferably within a range that does not significantly affect the particle size range of the carbonate apatite particles to be formed.
  • the aqueous solution for preparing the carbonate apatite particles may use water as a base, but various culture media and buffers for cell culture may be used.
  • Carbonate apatite particles can be obtained by adjusting the pH of the aqueous solution containing each of the above ions to a range of 6.0 to 9.0 and leaving (incubating) for a certain period of time.
  • the pH of the aqueous solution in forming the carbonate apatite particles is, for example, 7.0 to 8.5, preferably 7.1 to 8.5, more preferably 7.2 to 8.5, still more preferably 7.3 to 8.5, particularly preferably 7.4 to 8.5, and most preferably. 7.5 to 8.0.
  • the temperature condition of the aqueous solution in forming the carbonate apatite particles is not particularly limited as long as the carbonate apatite particles are formed, but is usually 10 ° C. or higher, preferably 25 to 80 ° C., more preferably 37 to 70 ° C. Can be mentioned.
  • the incubation time of the aqueous solution for forming the carbonate apatite particles is not particularly limited as long as the carbonate apatite particles are formed, but is usually 1 minute to 24 hours, preferably 10 minutes to 1 hour. The presence or absence of particle formation can be confirmed by observing under a microscope, for example.
  • the carbonate apatite particles often have an average particle diameter of more than 200 nm. Therefore, the carbonate apatite particles can be refined to reduce the average particle size to 200 nm or less (particularly 50 nm or less).
  • ultrasonic vibration treatment may be performed.
  • the ultrasonic vibration treatment is not a treatment in which an ultrasonic vibrator such as an ultrasonic crusher or a homogenizer used for so-called microbial cell crushing is directly brought into contact with a sample, but generally a precision instrument or a test.
  • This is a process using an ultrasonic cleaner in which an ultrasonic vibrator and a cleaning tank used for cleaning tubes and the like are integrated.
  • a liquid for example, water
  • a cleaning tank water tank
  • a container for example, made of plastic
  • the tube is floated and ultrasonic waves are applied to the dispersion through the liquid in the manner of washing the precision instrument. Thereby, the carbonate apatite particles can be refined simply and efficiently.
  • An apparatus that can be used for ultrasonic vibration treatment is capable of applying ultrasonic vibration to a container containing carbonate apatite particles indirectly through a solvent such as water, like the ultrasonic cleaner. If there is no particular limitation. From the viewpoint of versatility and ease of handling, it is preferable to use an ultrasonic cleaner equipped with an ultrasonic vibrator and a thermostatic bath.
  • the conditions for the above ultrasonic vibration treatment are not particularly limited as long as the particle diameter can be controlled within a predetermined range.
  • the temperature of the water tank can be appropriately selected from temperatures of 5 to 45 ° C., preferably 10 to 35 ° C., more preferably 20 to 30 ° C.
  • the high frequency output of the ultrasonic vibration treatment can be appropriately set, for example, in the range of 10 to 500 W, preferably 20 to 400 W, more preferably 30 to 300 W, and still more preferably 40 to 100 W.
  • the oscillation frequency is usually 10 to 60 Hz, preferably 20 to 50 Hz, and more preferably 30 to 40 Hz.
  • the ultrasonic vibration treatment time is, for example, 30 seconds to 30 minutes, preferably 1 to 20 minutes, and more preferably 3 to 10 minutes.
  • the type of container containing carbonate apatite particles used when performing ultrasonic vibration treatment is not limited as long as the particles can be refined to a predetermined particle diameter range, depending on the volume of the aqueous solution and the purpose of use. Can be selected as appropriate. For example, a plastic tube having a capacity of 1 to 1000 ml can be used.
  • the ultrasonic vibration treatment may be performed in the presence of albumin (that is, a state in which albumin is added to an aqueous solution containing carbonate apatite particles).
  • albumin that is, a state in which albumin is added to an aqueous solution containing carbonate apatite particles.
  • 0.1 to 500 mg / ml preferably 1 to 100 mg / ml, More preferably, about 1 to 10 mg / ml can be added.
  • albumin added to make carbonate apatite particles fine can be administered in vivo together with carbonate apatite particles in a state of being contained in the adjuvant composition of the present invention.
  • the CpG oligodeoxynucleotide may be present in a state of being complexed (encapsulated) with the carbonate apatite particles, or the CpG oligodeoxynucleotide and the carbonate apatite particles may be dispersed separately. There may be.
  • composite particles in which CpG oligodeoxynucleotides are combined with carbonate apatite particles are preferable.
  • “composite particles in which CpG oligodeoxynucleotides are complexed with carbonate apatite particles” means a state in which CpG oligodeoxynucleotides are adsorbed and supported on carbonate apatite particles by ionic bonds, hydrogen bonds, or the like. Point to.
  • the method for forming such composite particles is not particularly limited, but for example, in the aqueous solution for preparing carbonate apatite particles, by coexisting CpG oligodeoxynucleotide together with calcium ion, phosphate ion and hydrogen carbonate ion, Examples thereof include a method of simultaneously performing a step of forming carbonate apatite particles and a step of combining CpG oligodeoxynucleotide and carbonate apatite particles.
  • the method for forming the composite particles is the above-described method for producing carbonate apatite particles, in which CpG oligodeoxynucleotide is added to an aqueous solution for forming carbonate apatite particles in an amount of 0.1 to 1000 nM, preferably 0.5 to 600 nM. Preferably, it can be carried out by adding 1 to 400 nM and forming carbonate apatite particles according to the production method described above.
  • the ratio of CpG oligodeoxynucleotide to carbonate apatite particles is not particularly limited.
  • CpG oligodeoxynucleotide is 0.0001 to 10 ⁇ g, preferably 0.001 per 1 ⁇ g of calcium contained in carbonate apatite particles.
  • a ratio of ⁇ 1 ⁇ g, more preferably 0.01 ⁇ 1 ⁇ g can be mentioned.
  • the form of the adjuvant composition of the present invention is not particularly limited, but from the viewpoint of efficiently inducing protective immunity while suppressing the reaggregation of carbonate apatite particles and maintaining the state of the average particle diameter. It is preferably liquid.
  • the concentrations of CpG oligodeoxynucleotide and carbonate apatite may be set as appropriate so that the dose described later can be satisfied in consideration of the administration method and the like.
  • the concentration of CpG oligodeoxynucleotide is 0.2 to 20000 ⁇ g / ml, preferably 0.2 to 10,000 ⁇ g / ml, more preferably 0.2 to 1000 ⁇ g / ml.
  • carbonate apatite particles can be obtained by dissolving various ion supply substances in a solvent such as water, a medium, or a buffer. From the viewpoint of osmotic pressure, buffer capacity, sterility, etc., it is not necessarily suitable for administration to a living body (intravascular administration). For this reason, after the carbonate apatite particles are produced, the carbonate apatite particles are used by being dispersed in a solvent suitable for administration to a living body.
  • the carbonate apatite particles are usually separated from the solvent by centrifugation and recovered to remove the solvent. Replace operation is required. However, when such an operation is performed, the carbonate apatite particles are aggregated and the particles become enormous, so that the state changes to a state unsuitable for administration to a living body.
  • a solvent suitable for administration to a living body for example, physiological saline, etc.
  • the aggregated carbonate apatite particles are added to a solvent suitable for administration to the living body, and then the ultrasonic vibration treatment described above is performed, whereby the composite particles of CpG oligodeoxynucleotide and carbonate apatite particles are administered to the living body.
  • the adjuvant composition of the present invention is quickly administered after the carbonate apatite particles are dispersed in the form of fine particles by ultrasonic vibration treatment and before the particles aggregate.
  • administration within 1 minute, preferably within 30 seconds after the ultrasonic vibration treatment is suitable.
  • administration can be performed several minutes to several tens of minutes after the ultrasonic vibration treatment.
  • the adjuvant composition of the present invention is used for efficiently inducing protective immunity by vaccination.
  • the type of immunogen of the vaccine is not particularly limited as long as it can induce an immune response in vivo.
  • influenza virus avian influenza virus, parainfluenza virus, adenovirus, SARS virus AIDS virus, cytomegalovirus, hepatitis virus, Japanese encephalitis virus, measles virus, rubella virus, varicella-zoster virus, poliovirus, papilloma virus, herpes virus, mumps virus, rotavirus, cholera virus, rabies virus, Ebola hemorrhagic fever, Antigen proteins such as viruses that cause viral hemorrhagic fever such as Marburg disease, Lassa fever, Crimean-Congo hemorrhagic fever, antigenic peptides, or attenuated viruses that attenuate the pathogenicity of these viruses ; Diphtheria, tetanus, tuberculosis, pneumococci, meningococcus, staphylococci, Pse
  • an antigen protein or an antigen peptide is used as an immunogen for a vaccine, it may be produced by a gene recombination technique or may be produced by chemical synthesis.
  • an influenza vaccine decomposes and purifies viruses grown by cell culture techniques such as embryonated chicken eggs or Vero cells with ether or surfactant; or hemagglutinin, neuraminidase, nucleoprotein, matrix protein or a part thereof Etc. can be produced by genetic recombination techniques or chemical synthesis.
  • a preferred example of a vaccine that induces protective immunity with the adjuvant composition of the present invention is preferably an influenza vaccine, more preferably an influenza vaccine containing an influenza antigen protein or antigen peptide as an immunogen.
  • the subject of administration of the adjuvant composition of the present invention is not particularly limited as long as it is an animal to which the vaccine is administered.
  • animals for example, humans, monkeys, mice, rats, dogs, rabbits, cats, cows, horses, goats, etc. Mammals; birds such as chickens and ostriches.
  • the adjuvant composition of the present invention can efficiently induce protective immunity by administering at the same time as immunogen administration or before or after immunogen administration.
  • the adjuvant composition of the present invention may be administered in the state formulated as the same preparation (ie, vaccine preparation) mixed with the immunogen.
  • the immunogen and the adjuvant composition of the present invention may be prepared separately and formulated, and then administered after mixing at the time of use, or these may be administered separately sequentially.
  • the ratio of the adjuvant composition of the present invention to be administered and the immunogen may be appropriately set according to the type of immunogen used and the like.
  • the adjuvant composition of the present invention is CpG per 1 ⁇ g of immunogen.
  • the ratio is 0.0001 to 1 ⁇ g, preferably 0.001 to 0.6 ⁇ g, more preferably 0.01 to 0.3 ⁇ g, and still more preferably 0.01 to 0.2 ⁇ g in terms of oligodeoxynucleotide.
  • the administration form of the adjuvant composition of the present invention may be either parenteral administration or oral administration, but is preferably the same administration form as the immunogen.
  • Specific examples of parenteral administration include intravenous administration, intramuscular administration, intraperitoneal administration, intradermal administration, subcutaneous administration, nasal administration, intraarticular administration, and mucosal administration.
  • parenteral administration is preferable, and intradermal administration, subcutaneous administration, or nasal administration is more preferable.
  • the dosage of the adjuvant composition of the present invention is appropriately set according to the type of vaccine to be used, the age and weight of the administration subject, the expected action, etc.
  • the administration animal is a human, it is usually 1
  • An amount of CpG oligodeoxynucleotide corresponding to 0.4 to 40000 ⁇ g / kg may be administered once to several times a day, preferably once a day (primary immunization).
  • the dose of the vaccine used together with the adjuvant composition of the present invention is also appropriately set according to the type of immunogen, the age and weight of the administration subject, etc.
  • An amount of immunogen corresponding to 0.4 to 40000 ⁇ g / kg may be administered once to several times a day, preferably once a day (primary immunization). Further, the adjuvant composition and vaccine of the present invention may be re-administered under the same conditions as the first immunization usually 2 to 6 weeks after the first immunization (boost immunization).
  • Vaccine Formulation The present invention further provides a vaccine formulation comprising the above adjuvant composition and an immunogen.
  • the vaccine preparation of the present invention may contain the adjuvant composition and the immunogen as the same preparation.
  • the adjuvant composition and the immunogen are separately formulated, and may be designed to be administered after being mixed at the time of use, or they may be separately administered sequentially. From the viewpoint of ease of use, preferably, the adjuvant composition and the immunogen are included as the same preparation.
  • the type of immunogen used in the vaccine preparation of the present invention, the dose and administration ratio of the immunogen and the adjuvant composition, the dosage form and administration method of the vaccine, etc. are the same as in the case of “1.
  • Adjuvant composition” above. is there.
  • the content of the adjuvant composition and the immunogen in the vaccine preparation of the present invention, the ratio thereof, and the like are appropriately set according to the dose and the administration ratio of the immunogen and the adjuvant composition.
  • OVA represents chicken ovalbumin
  • AP represents carbonate apatite
  • OVA + AP-CpG represents OVA and CpG oligos.
  • OVA + AP + CpG is a combination of OVA, CpG oligodeoxynucleotides, and carbonate apatite particles that are not complexed with CpG oligodeoxynucleotides.
  • HA- + AP-CpG represents a combination of OVA, CpG oligodeoxynucleotides, and carbonate apatite particles that are not complexed with CpG oligodeoxynucleotides.
  • other notations similar to this for example, “HA- + AP-CpG”, “HA + AP + CpG”, etc. have the same meaning.
  • Example 1 Production of Carbonate Apatite Particles Complexed with CpG Oligodeoxynucleotide
  • 0.37 g NaHCO 3 0.37 g NaHCO 3 , 90 ⁇ l NaH 2 PO 4 .2H 2 O (1M), and 180 ⁇ l CaCl 2 ( 1M) was added and dissolved in this order, and the pH was adjusted to 7.5 with 1N HCl. This was filtered through a 0.2 ⁇ m diameter filter.
  • Buffer A The solution thus obtained is hereinafter referred to as “Buffer A”.
  • FIG. 1 shows the results of measurement of carbonate apatite (AP) particles not complexed with CpG ODN and Ap-CpG nanoparticles obtained under the condition 4 shown in Table 1 with a zeta sizer.
  • AP carbonate apatite
  • FIG. 1 shows the results of measurement of carbonate apatite (AP) particles not complexed with CpG ODN and Ap-CpG nanoparticles obtained under the condition 4 shown in Table 1 with a zeta sizer.
  • AP carbonate apatite
  • FIG. 2 shows the results of observing the carbonate apatite (AP) particles not complexed with CpGNODN and the Ap-CpG particles obtained under the condition 4 shown in Table 1 by AFM.
  • the amount of CpG ODN was measured using (Qubit ssDNA Assay Kit, Thermo), and the ratio of CpG ODN contained in the carbonate apatite particles to the total amount of CpG ODN used for production (CpG ODN) The entrapment rate (%) was determined. Table 1 shows the results of the inclusion rate of the obtained CpG ⁇ ⁇ ODN.
  • Example 2 Analysis of the effect of Ap-CpG nanoparticles on the ability of dendritic cells to produce cytokines Bone marrow cells isolated from the femur of C57BL / 6JJmsSlc mice (hereinafter sometimes abbreviated as C57BL / 6 mice) Were cultured for 1 week at 37 ° C. using RPMI1640 medium containing 10% FCS and 1% antibiotics containing Flt3L (100 ng / mL) to induce dendritic cells.
  • the obtained dendritic cells were seeded at 5 ⁇ 10 5 cells / ml, and 24 hours later, AP-CpG nanoparticles obtained under the condition 4 or 5 of Example 1 were converted to 0.14 to 3.75 ⁇ g in terms of CpG ODN amount.
  • PRMI1640 medium containing / ml was added and cultured at 37 ° C. for 24 hours.
  • the supernatant after 24 hours of culture was collected, and the amount of cytokines (IFN- ⁇ , IL-12, and IFN- ⁇ ) in the supernatant was measured by ELISA.
  • CpG ODN K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO. 1, product name “K3 Et-Free”, product number “CN-65003” instead of AP-CpG nanoparticles.
  • K3 type oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO. 1, product name “K3 Et-
  • Example 3 Analysis of the effect of Ap-CpG nanoparticles on cytokine production ability of human peripheral blood mononuclear cells
  • CpG ODN K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO. 1, product name “K3 Et-Free”, product number “CN-65003” instead of AP-CpG nanoparticles. The same test was conducted when using Gene Design Co.).
  • Example 4 Verification of Ap-CpG nanoparticle uptake by mouse bone marrow-derived dendritic cells CpG ODN fluorescently labeled with Alexa 488 (K3 type, oligodeoxynucleotide comprising the nucleotide sequence shown in SEQ ID NO: 1, product name “K3 Et AP-CpG nanoparticles were obtained under the same conditions as in condition 4 of Example 1 except that “-Free”, product number “CN-65003” manufactured by Gene Design Co., Ltd.) was used.
  • dendritic cells were induced from mouse bone marrow cells under the conditions shown in Example 2.
  • the obtained dendritic cells were seeded at 1 ⁇ 10 7 cells / ml, and 24 hours later, RPMI1640 medium containing 1.25 ⁇ g / ml of fluorescently labeled AP-CpG nanoparticles in terms of CpG ODN amount was added, and 37 Incubate at 180 ° C. for 180 minutes. During the culture, dendritic cells were collected over time, the fluorescence intensity of the dendritic cells was measured by FACS analysis, and the ratio of the dendritic cells (K3-positive dendritic cells) incorporated with CpG ODN was determined.
  • K3 positive dendritic cells those with a fluorescence intensity of about 1 ⁇ 10 4 were classified as K3 low positive, and those with a K3 fluorescence intensity of over 3 ⁇ 10 4 were classified as K3 high positive.
  • CpG ODN fluorescently labeled with Alexa 488 instead of AP-CpG nanoparticles K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product
  • the number “CN-65003” manufactured by Gene Design Co., Ltd.
  • FIG. 5a shows a plot of the fluorescence intensity of FSC-A and CpGFACODN by FACS analysis.
  • FIG. 5b shows the results of determining the ratio of dendritic cells (K3-positive dendritic cells) that incorporated CpG ODN when AP-CpG nanoparticles and CpG ODN were added. From this result, it was confirmed that AP-CpG nanoparticles were efficiently taken up by dendritic cells as compared to CpG ODN alone.
  • Example 5 Analysis of Ap-CpG nanoparticle uptake mechanism by mouse bone marrow-derived dendritic cells Under the conditions shown in Example 4 above, fluorescently labeled AP-CpG nanoparticles and mouse dendritic cells were prepared.
  • Mouse dendritic cells are seeded at 5 ⁇ 10 6 cells / ml, and 24 hours later, scavenger receptor inhibitor poly I is added at 0, 4, or 20 ⁇ g / ml, or endocytosis inhibitor cytochalasin D RPMI1640 medium containing 0, 0.8, 4, or 20 ⁇ M was added and incubated at 37 ° C. for 60 minutes.
  • scavenger receptor inhibitor poly I was 0, 4, or 20 ⁇ g / ml, or endocytosis.
  • RPMI1640 medium containing 0, 0.8, 4, or 20 ⁇ M of cytochalasin D, an inhibitor was added and incubated at 37 ° C. for 60 minutes. After culturing, the dendritic cells were collected, the fluorescence intensity of the dendritic cells was measured by FACS analysis, and the proportion of dendritic cells (K3-positive dendritic cells) into which CpG ODN was incorporated was determined. The ratio of K3 positive dendritic cells under each condition was calculated with the ratio of K3 positive dendritic cells when cultured in a medium without poly I or cytochalasin D added as 100%.
  • Example 6 Analysis of the effect of Ap-CpG nanoparticles on the in vivo cytokine production capacity of mouse regional lymph node cells 6 to 8 weeks old C57BL / 6 mice were obtained under condition 4 of Example 1.
  • AP-CpG nanoparticles (8 ⁇ g / mouse in terms of CpG ODN amount) were administered to the ears, and regional lymph nodes were collected 24 hours later. Cells in the lymph nodes were collected, PRMI1640 medium was added, and cultured at 37 ° C. for 8 hours. The supernatant after 8 hours of culture was collected, and the amount of cytokine (IL-12, IFN- ⁇ and IL-6) in the supernatant was measured by ELISA.
  • CpG ODN K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free” was used instead of AP-CpG nanoparticles.
  • Product number“ CN-65003 ”manufactured by Gene Design Co., Ltd. was administered at 40 or 8 ⁇ g / mouse in the same manner.
  • Example 7 Verification of protective immunity induction effect of Ap-CpG nanoparticles 6- to 8-week-old C57BL / 6 mice were treated with AP-CpG nanoparticles obtained in condition 4 of Example 1 (in terms of CpG ODN amount). 10 ⁇ g / mouse) and OVA (100 ⁇ g / mouse) mixture were administered to the ridge (day 0), and 14 days later, the same amount of AP-CpG nanoparticles and OVA were again administered to the ridge (day 14). . One week after the final administration, blood was collected, and OVA-specific IgG, IgG1, and IgG2c in plasma were measured by ELISA.
  • the ratio of antigen-specific cytotoxic T cells (CD8 + CD44 + Tetramer + cells) in the regional lymph nodes was measured by FACS.
  • regional lymph nodes were collected from the mice, cultured in RPMI1640 medium containing 5 ⁇ g / ml of OVA-derived peptide for 24 hours, and then the IFN- ⁇ concentration in the culture supernatant was measured by ELISA.
  • CpG ODN K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”,
  • product number “CN-65003” manufactured by Gene Design Co., Ltd.
  • Al hydroxide Alum
  • FIG. 8a shows the result of measuring the OVA-specific antibody in plasma diluted 100,000 times.
  • FIG. 8b shows the measurement results of each antibody when the dilution ratio of plasma is changed.
  • FIG. 8c shows the ratio of antigen-specific cytotoxic T cells (CD8 + CD44 + Tetramer + cells) in the regional lymph nodes, and when the lymphocytes isolated from the lymph nodes were restimulated with the antigen.
  • the measurement result of the produced IFN- ⁇ concentration is shown.
  • Example 8 Verification of protective immunity induction effect of Ap-CpG nanoparticles 6-week-old C57BL / 6 mice were treated with AP-CpG nanoparticles obtained in condition 4 of Example 1 (in terms of CpG ODN amount). 10 ⁇ g / mouse) and hemagglutinin (HA) derived from influenza A strain H1N1 (0.5 ⁇ g / mouse) were administered to the ridge (day 0), and 14 days later, the same amount of AP-CpG nanoparticles and HA was administered to the ridge (day 14). One week after the final administration, blood was collected, and HA-specific IgG, IgG1, and IgG2c in plasma were measured by ELISA.
  • HA hemagglutinin
  • FIG. 9a shows the results of measuring HA-specific antibodies in plasma.
  • FIG. 9b shows the measurement results of the survival rate and weight transition of mice. From these results, it was revealed that AP-CpG nanoparticles induce the same level of protection against influenza infection as 5 times the amount of CpG ODN.
  • Example 9 Verification of safety of Ap-CpG nanoparticles 7-week-old C57BL / 6 mice were treated with AP-CpG nanoparticles obtained in condition 4 of Example 1 (10 ⁇ g / mouse in terms of CpG ODN amount). After administration to the ridge, blood was collected 24 hours later, and the number of leukocytes and lymphocytes in the blood was counted. In addition, 7-week-old C57BL / 6 mice were subjected to AP-CpG nanoparticles (10 ⁇ g / mouse / times in terms of CpG ODN amount) obtained in Condition 4 of Example 1 three times every other day. After 24 hours from the last administration, the spleen, lymph node, and liver weights of the mice were measured.
  • CpG ODN K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN -65003 ”(manufactured by Gene Design) (10 or 50 ⁇ g / mouse / time), and carbonate apatite (AP) particles not combined with CpG ODN were administered in the same manner.
  • FIG. 10a shows the results of measuring the number of white blood cells and lymphocytes in the blood
  • FIG. 10b shows the results of measuring the weights of the spleen, lymph nodes, and liver. From these results, it was clarified that CpGNODN was improved in safety by being enclosed in carbonate apatite.
  • Example 10 Evaluation of the ability to induce cytokine production in the absence of complexation of carbonate apatite particles and CpG oligodeoxynucleotides Bone marrow cells isolated from the femur of C57BL / 6J mice were 10% containing Flt3L (100 ng / mL) Dendritic cells were induced by culturing for 1 week at 37 ° C. using FCS and RPMI1640 medium containing 1% antibiotics.
  • the obtained dendritic cells were seeded at 5 ⁇ 10 5 cells / ml, and 24 hours later, carbonate apatite (AP) nanoparticles obtained under the condition 0 of Example 1 (about 1.25 ⁇ g / ml in terms of calcium amount) ) And CpG ODN (K3 type, oligodeoxynucleotide consisting of the nucleotide sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN-65003”, Gene Design Co., Ltd.) (3.75 ⁇ g / ml, 11.25 PRMI1640 medium containing .mu.g / ml, 33.75 .mu.g / ml) was added and cultured at 37.degree.
  • K3 type oligodeoxynucleotide consisting of the nucleotide sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN-65003”, Gene Design Co., Ltd.
  • cytokines IL-6, IL-12, and IFN- ⁇
  • AP is used alone instead of the combination of AP nanoparticles and CpG, and CpG ODN (K3 type, oligodeoxynucleotide comprising the nucleotide sequence shown in SEQ ID NO: 1, product name “K3 Et -Free ", product number” CN-65003 "(manufactured by Gene Design Co., Ltd.) were also used in the same manner.
  • Example 11 Verification of effect of induction of protective immunity by administration in a state in which carbonate apatite particles and CpG oligodeoxynucleotides are not complexed 6-week-old C57BL / 6 mice were obtained under condition 0 of Example 1
  • Carbonate apatite (AP) nanoparticles (about 125 ⁇ g / mouse in terms of calcium content)
  • CpG ODN (K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN -65003 ”(manufactured by Gene Design) (10 ⁇ g / mouse) and OVA (100 ⁇ g / mouse) were administered to the ridge (day 0), and 14 days later, the same amount of AP nanoparticles, CpG ODN And a mixed solution of OVA were administered to the ridge (day 14).
  • a mixed solution of AP-CpG nanoparticles (10 ⁇ g / mouse in terms of CpG ODN amount) and OVA (100 ⁇ g / mouse) obtained under condition 4 of Example 1 was added to C57BL / 6 mice aged 6 to 8 weeks. It was administered to the ridge (day 0), and 14 days later, the same amounts of AP-CpG nanoparticles and OVA were again administered to the ridge (day 14). One week after the final administration, blood was collected, and OVA-specific IgG1 and IgG2c in plasma were measured by ELISA.
  • OVA 100 ⁇ g / mouse
  • CpG ODN K3 type, oligodeoxynucleotide comprising the nucleotide sequence shown in SEQ ID NO: 1, product name “K3 Et -Free ”, product number“ CN-65003 ”, manufactured by Gene Design
  • OVA 100 ⁇ g / mouse
  • Alum 500 ⁇ g / mouse in terms of aluminum
  • Example 12 Verification of effect of induction of protective immunity by administration in a state in which carbonate apatite particles and CpG oligodeoxynucleotides are not complexed 6- to 8-week-old C57BL / 6 mice were obtained under condition 0 of Example 1
  • Carbonate apatite (AP) nanoparticles (about 125 ⁇ g / mouse in terms of calcium content)
  • CpG ODN (K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN -65003 "(manufactured by Gene Design) (10 ⁇ g / mouse) and hemagglutinin (HA) (0.5 ⁇ g / mouse) from influenza
  • HA hemagglutinin
  • HA 10 or 50 ⁇ g / mouse is administered, HA (0.5 ⁇ g / mouse) and Alum (aluminum content)
  • HA 10 or 50 ⁇ g / mouse
  • Alum aluminum content
  • FIGS. FIG. 13 shows the result of measuring HA-specific IgG in plasma
  • FIG. 14 shows the result of measuring HA-specific IgG2c in plasma.
  • the dilution rate in FIGS. 13 and 12 is the dilution rate of plasma at the time of antibody measurement. From this result, it was clarified that AP promotes induction of antigen-specific antibody production only by mixing with CpG.
  • Example 13 Verification of the induction effect of protective immunity by administration in a state in which carbonate apatite particles and CpG oligodeoxynucleotides are not complexed. C57BL / 6 mice aged 6 to 8 weeks were obtained under condition 0 of Example 1.
  • Carbonate apatite (AP) nanoparticles (about 125 ⁇ g / mouse in terms of calcium content), CpG ODN (K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN -65003 "(manufactured by Gene Design) (10 ⁇ g / mouse) and hemagglutinin (HA) derived from influenza A strain H1N1 (HA) (0.5 ⁇ g / mouse) was administered to the ear (day 0), and 14 days later Again, the same amount of AP nanoparticles, CpG ODN (K3 type, oligodeoxynucleotide consisting of the base sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN-65003”, manufactured by Gene Design) and The HA mixture was administered to the ear (day 1).
  • CpG ODN K3 type, oligodeoxynucleot
  • HA-specific IgG, IgG1, and IgG2c in plasma were measured by ELISA.
  • HA 0.5 ⁇ g / mouse
  • AP about 100 ⁇ g / mouse in terms of calcium
  • HA 0.5 ⁇ g / mouse
  • CpG ODN K3 type, oligodeoxynucleotide consisting of the nucleotide sequence shown in SEQ ID NO: 1, product name “K3 Et-Free”, product number “CN-65003”, manufactured by Gene Design
  • FIG. 15a shows the measurement results of HA-specific IgG in plasma.
  • FIG. 15 b shows the measurement results of the survival rate and weight transition of mice. From this result, it was clarified that the induction of antigen-specific antibody production was strongly promoted only by mixing AP, CpG and HA. In addition, it was revealed that the influenza infection protective ability was strongly induced in the mixed group of AP, CpG and HA.
  • Reference Test Example 1 Verification of protective immunity induction effect of carbonate apatite particles combined with Poly (I; C) as an adjuvant Instead of using CpG ODN, except that Poly (I; C) was used, Carbonate apatite (AP-Poly (I; C)) nanoparticles in which Poly (I; C) was complexed were obtained under the same conditions as in Condition 5 of Example 1.
  • AP-Poly (I; C) nanoparticles obtained above 0.1 or 1 ⁇ g / mouse in terms of Poly (I; C) amount
  • OVA 100 ⁇ g / mouse
  • the obtained results are shown in FIG.
  • the dilution rate in FIG. 16 is the dilution rate of plasma at the time of antibody measurement. From these results, it was found that even when AP-Poly (I; C) was used as an adjuvant and used in combination with carbonate apatite particles, effective induction of protective immunity was not observed.

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Abstract

L'objectif de la présente invention est de fournir : une composition d'adjuvant qui est hautement sûre et peut efficacement améliorer une immunité protectrice induite par un vaccin; et une préparation de vaccin utilisant la composition d'adjuvant. Un oligodésoxynucléotide CpG est choisi parmi des adjuvants, et l'oligodésoxynucléotide CpG est utilisé en combinaison avec carbonate-apatite. La composition d'adjuvant est hautement sûre et peut efficacement améliorer une immunité protectrice induite par un vaccin.
PCT/JP2017/028574 2016-08-08 2017-08-07 Composition d'adjuvant Ceased WO2018030338A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2022523226A (ja) * 2019-03-04 2022-04-21 エムペグ エルエイ リミテッド ライアビリティ カンパニー 増強された生理活性を有する多量体オリゴヌクレオチド
WO2022158440A1 (fr) 2021-01-19 2022-07-28 浩文 山本 Composition d'administration de médicament, son procédé de production et son utilisation
EP4262758A4 (fr) * 2020-12-16 2024-11-06 Dynavax Technologies Corporation Procédé de quantification d'oligonucléotides contenant une cpg dans des formulations comprenant de l'alun
US12378551B2 (en) 2017-02-06 2025-08-05 Mpeg La, L.L.C. Multimeric oligonucleotides having decreased kidney clearance

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006509838A (ja) * 2002-11-12 2006-03-23 財団法人理工学振興会 細胞導入剤、細胞導入方法、細胞導入剤の製造方法、細胞導入剤製造用組成物、および、細胞導入剤製造用キット
JP2009514841A (ja) * 2005-11-04 2009-04-09 ノバルティス ヴァクシンズ アンド ダイアグノスティクス エスアールエル 粒子状アジュバントと免疫増強物質との組合せを含むインフルエンザワクチン
WO2017007027A1 (fr) * 2015-07-09 2017-01-12 国立研究開発法人物質・材料研究機構 Complexe oligonucléotidique immunostimulant
JP2017088601A (ja) * 2015-11-09 2017-05-25 医療法人 医潤会 生体投与を目的とした薬物導入剤および製造方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006509838A (ja) * 2002-11-12 2006-03-23 財団法人理工学振興会 細胞導入剤、細胞導入方法、細胞導入剤の製造方法、細胞導入剤製造用組成物、および、細胞導入剤製造用キット
JP2009514841A (ja) * 2005-11-04 2009-04-09 ノバルティス ヴァクシンズ アンド ダイアグノスティクス エスアールエル 粒子状アジュバントと免疫増強物質との組合せを含むインフルエンザワクチン
WO2017007027A1 (fr) * 2015-07-09 2017-01-12 国立研究開発法人物質・材料研究機構 Complexe oligonucléotidique immunostimulant
JP2017088601A (ja) * 2015-11-09 2017-05-25 医療法人 医潤会 生体投与を目的とした薬物導入剤および製造方法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
KNUSCHKE, T. ET AL.: "Immunization with Biodegradable Nanoparticles Efficiently Induces Cellular Immunity and Protects against Influenza Virus Infection", THE JOURNAL OF IMMUNOLOGY, vol. 190, no. 12, 2013, XP055464014, ISSN: 0022-1767 *
XU, Z. ET AL.: "Multifunctional nanoparticles co-delivering Trp2 peptide and CpG adjuvant induce potent cytotoxic T-lymphocyte response against melanoma and its lung metastasis", JOURNAL OF CONTROLLED RELEASE, vol. 172, 2013, pages 259 - 265, XP028772929, ISSN: 0168-3659 *
YOSHITOMO HONDA ET AL.: "Synergistic immunostimulating effect of CpG DNA/calcium phosphate complex", THE JOURNAL OF THE JAPANESE SOCIETY FOR DENTAL MATERIALS AND DEVICES, vol. 28, 2009, ISSN: 0286-5858 *
ZHENG, J. ET AL.: "Electro-deposited calcium phosphate compounds on graphene sheets: Blossoming flowers", MATERIALS LETTERS, vol. 179, 9 May 2016 (2016-05-09), pages 122 - 125, XP029562443, ISSN: 0167-577X *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12378551B2 (en) 2017-02-06 2025-08-05 Mpeg La, L.L.C. Multimeric oligonucleotides having decreased kidney clearance
JP2022523226A (ja) * 2019-03-04 2022-04-21 エムペグ エルエイ リミテッド ライアビリティ カンパニー 増強された生理活性を有する多量体オリゴヌクレオチド
EP4262758A4 (fr) * 2020-12-16 2024-11-06 Dynavax Technologies Corporation Procédé de quantification d'oligonucléotides contenant une cpg dans des formulations comprenant de l'alun
WO2022158440A1 (fr) 2021-01-19 2022-07-28 浩文 山本 Composition d'administration de médicament, son procédé de production et son utilisation

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