WO2018225785A1 - Composition de vaccin - Google Patents

Composition de vaccin Download PDF

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Publication number
WO2018225785A1
WO2018225785A1 PCT/JP2018/021696 JP2018021696W WO2018225785A1 WO 2018225785 A1 WO2018225785 A1 WO 2018225785A1 JP 2018021696 W JP2018021696 W JP 2018021696W WO 2018225785 A1 WO2018225785 A1 WO 2018225785A1
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Prior art keywords
vaccine composition
metal organic
organic structure
gas component
composition according
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PCT/JP2018/021696
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English (en)
Japanese (ja)
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浅利大介
加藤慎司
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株式会社Ａｔｏｍｉｓ
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Priority to JP2019523945A priority Critical patent/JPWO2018225785A1/ja
Publication of WO2018225785A1 publication Critical patent/WO2018225785A1/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
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants

Definitions

  • the present invention relates to a vaccine composition.
  • the object of the present invention is to provide an excellent vaccine composition.
  • a vaccine composition comprising an antigen for immunity induction and a gas component which is a gas at 25 ° C. and 100 kPa.
  • FIG. 1A is a CO adsorption profile of metal organic structure AP004 [MIL-100 (Fe)].
  • FIG. 1B is a NO adsorption profile of metal organic structure AP004 [MIL-100 (Fe)].
  • FIG. 2 is a NO adsorption profile of metal organic structure AP104 (BioMIL-3).
  • FIG. 3 is a view showing the measurement results of antigen-specific antibody titers in mouse serum.
  • FIG. 4A is a diagram showing the results of measurement of OVA-specific cytokine production.
  • FIG. 4B is a diagram showing the measurement results of OVA-specific cytokine production.
  • the vaccine composition according to the present invention contains an antigen for inducing immunity and a gas component that is a gas at 25 ° C. and 100 kPa (SATP).
  • a gas component that is a gas at 25 ° C. and 100 kPa (SATP).
  • Antigens include any substance that can induce an immune response. Although the said antigen is not specifically limited, For example, protein or a peptide is mentioned. In transdermal administration where the skin permeability of the antigen is required, it is preferable to use an antigen having a low molecular weight, for example, a peptide consisting of about 8 to 12 amino acids can be used. In addition, cancer antigen peptides, infectious pathogen-derived antigens, and the like can also be used as the antigen.
  • self antigens for example, antigens related to autoimmune diseases
  • endogenous antigens for example, antigens derived from cancer
  • foreign antigens for example, antigens related to allergies, and antigens derived from viruses and bacteria
  • Antigens related to autoimmune diseases include, for example, amyloid ⁇ and / or precursors thereof and fragment proteins and peptides thereof, which are considered to be the cause of Alzheimer's disease, ⁇ -synuclein and fragment proteins thereof which are considered to be the cause of Parkinson's disease And peptides, ⁇ -fodrin considered to be the cause of Sjogren's syndrome and its fragment proteins and peptides, thyroid hormone receptor and its fragment proteins and peptides considered to be the cause of Passow's disease, and causes of Guillain-Barre syndrome Gangliosides considered and fragmented proteins and peptides thereof, DNA and fragments thereof considered to cause systemic lupus erythematosus, cholesterol ester transfer protein considered to cause arteriosclerosis, apolipoprotein, and LDL and their fragment proteins and peptides, angiotensin I / II which is considered to cause hypertension and its fragment proteins and peptides, insulin, GAD and IL-1 ⁇ which are considered to
  • cancer-derived antigens include WT1, PR1, GPC3, HER-2, MAGE-A1, MAGE-A2, MAGE-A3, tyrosinase, gp100, CEA, hTRT, EGF receptor, mTERT, PRAME, PSMA, PSA- 1, cytochrome p450, NY-ESO-1, Survivine, MUC-1, MAGE-A10, and PAP-derived proteins and peptides.
  • Antigens related to allergies include, for example, allergens derived from trees (Acacia, alder tree, velvet blue-tailed potato, Japanese beech, birch, maple, mountain cedar, red cedar, boxwood, hinoki, American elm, Akinori, Togasawara, rubber, eucalyptus , Enoki, Hickory, American linden, Sugar maple, Mesquite, Casino cypress, Quercus, Olive, Pecan, Pepper, Pine, Squid, Russian olive, American sycamore, Sardine, Black walnut, Black willow, etc., Allergens derived from vegetation , Nagahagusa, Suzumenochahiki, Maize, Hirohoshi nokegusa, Seiban sorghum, Oats, Camouflage, Konukagusa, Barley, Rice, Hurghaya, Oawagaeri, Ayuyu, Akaza, Onamomi, Gishigi
  • Diseases affected by the infectious pathogen include, for example, adenovirus, herpes virus (eg, HSV-I, HSV-II, CMV, or VZV), poxvirus (eg, pressure ulcer or vaccinia, or contagious molluscum) Orthopoxvirus), picornavirus (eg rhinovirus or enterovirus), orthomyxovirus (eg influenza virus), paramyxovirus (eg parainfluenza virus, mumps virus, measles virus, respiratory organs) Endoplasmic reticulum virus (RSV)), coronavirus (eg SARS), papovavirus (eg papilloma virus such as those causing genital warts, vulgaris warts or plantar costus), hepadnavirus (eg hepatitis) B virus), Viral diseases, such as diseases resulting from viral infections such as Ravivirus (eg, hepatitis C virus or dengue virus) or retrovirus (eg, lentivirus
  • OVA ovalbumin
  • pneumococci pneumococci
  • influenza vaccine pneumococci
  • Cryj1 a major allergen of cedar pollen
  • HPV16 recombinant protein examples include ovalbumin (OVA), pneumococci, influenza vaccine, Cryj1 (a major allergen of cedar pollen), and HPV16 recombinant protein.
  • the content of the antigen in the vaccine composition is, for example, in the range of 1 ⁇ 10 ⁇ 7 to 1 ⁇ 10 ⁇ 1 mass%, preferably in the range of 1 ⁇ 10 ⁇ 6 to 1 ⁇ 10 ⁇ 2 mass%, More preferably, it is in the range of 2 ⁇ 10 ⁇ 6 to 2 ⁇ 10 ⁇ 3 mass%.
  • immune signaling substance means any substance used for transmitting an immune signal for inducing activation or differentiation of immune cells.
  • Immune signaling substances include, for example, lymphocytes (T cells, B cells, NK cells, etc.), monocytes (macrophages, Langerhans cells, dendritic cells, etc.), granulocytes (neutrophils, eosinophils, basophils). And / or factors that act on keratinocytes. Immune signaling substances are used for, for example, inducing differentiation of helper T cells, which are a type of lymphocyte, into various lineages such as Th1 cells, Th2 cells, Treg cells, Th17 cells, Tfh cells, and memory T cells. It may be a factor.
  • helper T cells which are a type of lymphocyte, into various lineages such as Th1 cells, Th2 cells, Treg cells, Th17 cells, Tfh cells, and memory T cells. It may be a factor.
  • the vaccine composition according to the present invention can be used, for example, for cancer vaccines and infectious disease vaccines.
  • the vaccine composition according to the present invention can be used, for example, for infectious disease vaccines and lifestyle-related disease vaccines.
  • the vaccine composition according to the present invention can be used, for example, for an allergy vaccine.
  • the vaccine composition according to the present invention can be used, for example, for an infectious disease vaccine.
  • the immune signaling substance induces Tfh cells the vaccine composition according to the present invention can be used, for example, for an infectious disease vaccine.
  • the vaccine composition according to the present invention can be used, for example, for an infectious disease vaccine or a cancer vaccine.
  • gas component described above is not particularly limited, and examples thereof include compounds shown in Table 1 below. These are non-limiting lists, and other gas molecules may be used. Of these, it is particularly preferable to use nitrogen monoxide, carbon monoxide, carbon dioxide, hydrogen sulfide or methane as the gas component.
  • the content of the gas component in the vaccine composition is, for example, in the range of 1 ⁇ 10 ⁇ 7 to 40% by mass, preferably in the range of 1 ⁇ 10 ⁇ 6 to 30% by mass, and more preferably 5 ⁇ 10 ⁇ It is within the range of 5 to 25% by mass.
  • the vaccine composition according to one embodiment of the present invention may further include a metal organic structure.
  • a metal organic structure When such a configuration is adopted, the performance of the vaccine composition can be improved by the action of the metal and the ligand in the metal organic structure.
  • the gas component by including at least a part of the gas component in the pores of the metal organic structure, the gas component can be more stably and quantitatively administered into the living body.
  • the metal organic structure is composed of a combination of a metal and a multidentate ligand.
  • multidentate ligand means a bidentate or higher ligand.
  • the type of the metal organic structure there is no particular limitation on the type of the metal organic structure. By appropriately combining the type and coordination number of the metal ion with the type and topology of the polydentate ligand, a metal organic structure having a desired structure can be produced.
  • the metal organic structure may be crystalline or amorphous.
  • the metal elements constituting the metal organic structure include arbitrary elements belonging to alkali metals (Group 1), alkaline earth metals (Group 2), and transition metals (Groups 3 to 12). Can be mentioned. Among these, from the viewpoint of biocompatibility, it is particularly preferable to use at least one metal element selected from the group consisting of calcium, magnesium, iron, zinc, aluminum, potassium, and sodium. However, even when a metal element other than these is used, there is no problem as long as biocompatibility as a metal organic structure is ensured.
  • the polydentate ligand constituting the metal organic structure is typically an organic ligand, and examples thereof include a carboxylate anion and a heterocyclic compound.
  • the carboxylic acid anion include dicarboxylic acid and tricarboxylic acid. Specific examples include anions of citric acid, malic acid, terephthalic acid, isophthalic acid, trimesic acid, and derivatives thereof.
  • the heterocyclic compound include bipyridine, imidazole, adenine, and derivatives thereof.
  • the ligand may be an amine compound, a sulfonate anion, or a phosphate anion.
  • the metal organic structure may further contain a monodentate ligand.
  • the combination of the metal and the ligand constituting the metal organic structure can be appropriately determined according to the function and the desired pore size.
  • the metal organic structure may contain 2 or more types of metal elements, and may contain 2 or more types of ligands.
  • the metal organic structure may be surface-modified with a polymer or the like.
  • metal organic structure for example, those listed in Table 1 of non-patent literature (Yabing He et al., Methane Storage in Metal-Organic Frameworks, Chem Soc Rev, 2014) can be used. . Or you may use the thing shown to the following Tables 2 thru
  • Particularly preferable metal organic structures include the following.
  • the content of the metal organic structure in the vaccine composition is, for example, in the range of 1 ⁇ 10 ⁇ 7 to 99.99999 mass%, preferably in the range of 1 ⁇ 10 ⁇ 6 to 99.99999999 mass%, and more Preferably, it is within the range of 5 ⁇ 10 ⁇ 6 to 99.99999 mass%.
  • the gas component is contained in the pores of the metal organic structure. Thereby, more stable and quantitative administration of the gas component becomes possible.
  • the other part of the gas component may be attached to the surface of the antigen or metal organic structure. Further, almost all of the gas component may be contained in the pores of the metal organic structure.
  • the metal organic structure When at least a part of the gas component is contained in the pores of the metal organic structure, the metal organic structure preferably has an irreversible adsorption / desorption profile. That is, the metal organic structure preferably has a larger amount of adsorption at the time of desorption than the amount of adsorption at the time of adsorption at the same pressure. In particular, it is preferable that the residual amount of adsorption of the metal organic structure is not zero when desorption from the pressurized state to the vacuum state is performed after the adsorption from the vacuum state to the pressurized state. In such a case, the gas component is easily retained in the pores of the metal organic structure even under low pressure conditions (for example, under atmospheric pressure).
  • the metal organic structure is configured to decompose in the living body and release at least a part of the gas component. It may be. Thereby, fine adjustments, such as a dosage of a gas component and a release rate, can be performed.
  • the method is not limited.
  • a metal organic structure solution or dispersion and a gas component solution or dispersion may be mixed.
  • the solid metal organic structure may be exposed to a gas component or a solution or dispersion thereof.
  • the vaccine composition according to one embodiment of the present invention may further contain a known adjuvant.
  • the vaccine composition may further contain an immunostimulatory agent such as a TLR ligand, an RLR ligand, an NLR ligand, and a cyclic dinucleotide.
  • the vaccine composition according to one embodiment of the present invention can be used, for example, in a state of being dissolved or dispersed in a solvent.
  • a solvent for example, physiological saline, phosphate buffered saline (PBS), glycerin, propylene glycol, polyethylene glycol, or fats and oils can be used.
  • the vaccine composition according to the present invention can be administered to a subject by any method.
  • subject refers to any animal, typically a mammal including humans, such as mice, rats, dogs, cats, rabbits, horses, which can induce an immune response upon administration of the vaccine composition in the practical stage. Meaning cow, sheep, pig, goat, monkey, chimpanzee, ferret, mole, etc. A particularly preferred subject is a human.
  • the vaccine composition according to one embodiment of the present invention is configured to be administered onto the skin and / or mucous membrane, for example.
  • the vaccine composition may be any preparation usually used for transdermal administration, for example, liniments or lotions, external sprays such as aerosols, ointments, plasters, creams. , Gels, or patches such as tapes or poultices.
  • liniments or lotions external sprays such as aerosols, ointments, plasters, creams.
  • Gels or patches such as tapes or poultices.
  • the classification, definition, properties, production method and the like of these compositions are well known in the art, and refer to, for example, the Japanese Pharmacopoeia 16th edition.
  • the vaccine composition may be any formulation commonly used for mucosal administration, eg sublingual, nasal, buccal, rectal or vaginal administration, eg gel (jelly), cream, ointment.
  • Semi-solid preparations such as plasters and plasters, liquid preparations, powders, fine granules, granules, solid preparations such as films and tablets, orally disintegrating tablets, sprays for mucous membranes such as aerosols, inhalants, etc. It's okay.
  • the classification, definition, properties, production method and the like of these compositions are well known in the art, and refer to, for example, the Japanese Pharmacopoeia 16th edition.
  • the vaccine composition according to one embodiment of the present invention is configured to be administered, for example, by intradermal injection, subcutaneous injection, or intramuscular injection.
  • the pharmaceutical composition may be in a form that has a certain fluidity that can be administered by injection, such as a liquid, suspension, cream, and the like.
  • the classification, definition, properties, production method and the like of these compositions are well known in the art, and refer to, for example, the Japanese Pharmacopoeia 16th edition.
  • the vaccine composition may further contain an additive as necessary.
  • an additive for example, skin permeability enhancer, tonicity agent, antiseptic / disinfectant, antioxidant, Solubilizers, solubilizers, suspending agents, fillers, pH adjusters, stabilizers, absorption enhancers, release rate control agents, colorants, plasticizers, adhesives, etc., or combinations of two or more thereof Can be selected.
  • Example 1 NO nitrogen monoxide, Kyoto Teijin was bubbled into 100 mL of physiological saline (Otsuka raw food injection, Otsuka Pharmaceutical) at room temperature for 6 hours to prepare NO saturated physiological saline.
  • physiological saline Otsuka raw food injection, Otsuka Pharmaceutical
  • ZIF-8 Base Z1200, SIGMA-ALDRICH
  • OVA egg-derived albumin, Wako
  • Example 2 NO (nitrogen monoxide, Kyoto Teijin) was bubbled in 100 mL of physiological saline (Otsuka raw food injection, Otsuka Pharmaceutical) at room temperature for 6 hours to prepare NO saturated physiological saline. To 10 mL of the solution, 1 mg of OVA (egg-derived albumin, Wako) was added and mixed to obtain a sample solution.
  • physiological saline Otsuka raw food injection, Otsuka Pharmaceutical
  • OVA egg-derived albumin, Wako
  • NO nitrogen monoxide, Kyoto Teijin
  • physiological saline Otsuka raw food injection, Otsuka Pharmaceutical
  • ZIF-8 Base Z1200, SIGMA-ALDRICH
  • Example 6 A sample solution was prepared in the same manner as in Example 1 except that the antigen shown in Table 12 below was used.
  • Example 7 to 35 A sample solution was prepared in the same manner as in Example 1 except that the substances shown in Table 13 below were used as immune signaling substances.
  • Examples 36 to 145 A sample solution was prepared in the same manner as in Example 1 except that the metal organic structures shown in Tables 14 to 16 below were used. Abbreviations in Tables 14 to 16 are the same as those described in Tables 2 to 4, respectively.
  • sample stimulation PEC cells were dispensed at 1 ⁇ 10 6 cells / well in a 24-well plate, and each sample was added and incubated for 24 hours.
  • the serum collected from the mouse in advance was centrifuged at 3000 g for 10 minutes at 4 ° C., and the supernatant was collected. Using a solution obtained by diluting the blocking agent to 0.4 g / 100 mL with a phosphate buffer (Nacalai Tesque), serially dilute the above-mentioned supernatant or nasal wash 2 times, and add 50 ⁇ L each of the solutions. Left at room temperature for hours.
  • a phosphate buffer Nacalai Tesque
  • mice [Evaluation of humoral immunity using mice] Using the liquid prepared as described above, a mouse immunity test was conducted using a model animal for humoral immunity evaluation. 200 ⁇ L of an injection was administered subcutaneously to the back of a mouse (BALB / c mouse, female 7 weeks old) prepared in advance. One week after the administration, the same administration was again performed subcutaneously on the back of the mouse. Two weeks after the second administration, mouse serum was collected, and the serum OVA-specific IgG titer was measured by the ELISA method described above.
  • ELISPOT method Splenocytes (3 ⁇ 10 6 cells / well) and antigenic peptide (100 ⁇ M) or antigenic protein (100 ⁇ g / mL) are placed in a well of an ELISPOT plate (R & D Systems) on which an anti-mouse IFN- ⁇ antibody is immobilized, together with a culture solution. The cells were co-cultured at 37 ° C. under 5% CO 2 for 20 hours, and the number of IFN- ⁇ producing cell spots (spot number / 3 ⁇ 10 6 cells) was measured by ELISPOT method.
  • mice [Cellular immunity evaluation using mice] Using the solution prepared as described above, a mouse immunity test was conducted using a model animal for cellular immunity evaluation. 200 ⁇ L of an injection was administered subcutaneously to the back of a mouse (C57BL / 6 mouse, female 7 week old) prepared in advance. One week after the administration, the same administration was again performed subcutaneously on the back of the mouse. One week after the second administration, mouse spleen was collected, and OVA antigen-specific CTL was measured by the ELISPOT method described above.
  • FIG. 1A is a CO adsorption profile of AP004 [MIL-100 (Fe)].
  • FIG. 1B is a NO adsorption profile of AP004 [MIL-100 (Fe)].
  • FIG. 2 is a NO adsorption profile of AP104 (BioMIL-3). In these examples, the adsorption / desorption profile was irreversible.
  • the adsorption amount at the time of desorption was larger than the adsorption amount at the time of adsorption. Further, the amount of residual adsorption when the desorption from the pressurized state to the vacuum state was performed after the adsorption from the vacuum state to the pressurized state was not zero.
  • mice serum and spleen cells were collected, and serum OVA-specific IgG antibody and IgG2a antibody were measured by ELISA.
  • spleen cells were used to simultaneously evaluate the production amounts of OVA-specific IFN- ⁇ and IL-4.
  • the specific evaluation method is as follows.
  • an antigen As an antigen, an OVA-containing solution (100 ⁇ g / mL) diluted with a carbonate buffer was prepared. 100 ⁇ L of this was added to a 96-well plate for ELISA and allowed to stand overnight.
  • Wells were washed three times with a preliminarily prepared washing solution (PBS containing Tween 20).
  • a blocking solution obtained by diluting a blocking agent (Block Ace, Dainippon Sumitomo Pharma Co., Ltd.) to 4 g / 100 mL with purified water was added 200 ⁇ L at a time, and the mixture was allowed to stand at room temperature for 2 hours. Thereafter, the wells were washed three times with a washing solution.
  • the serum collected from the mouse in advance was centrifuged at 3000 g for 10 minutes at 4 ° C., and the supernatant was collected.
  • a solution obtained by diluting the blocking agent with a phosphate buffer (Nacalai Tesque) to 0.4 g / 100 mL the above supernatant was serially diluted two times. 50 ⁇ L of each of the obtained diluted solutions was added and left at room temperature for 2 hours.
  • an HRP-labeled anti-mouse IgG antibody Goat-anti mouse IgG Fc HRP, BETHYL
  • an HRP-labeled anti-mouse IgG2a antibody in a solution obtained by diluting the blocking agent with phosphate buffer (Nacalai Tesque) to 0.4 g / 100 mL
  • Goat-anti mouse IgG2a Fc HRP, BETHYL was diluted 10,000 times. 100 ⁇ L of this diluted solution was added and left at room temperature for 1 hour.
  • TMB solution ELISA POD TMB kit, Nacalai Tesque
  • IgG antibody titer or IgG2a antibody titer in mouse serum was determined by Log2.
  • the immune characteristics could be controlled. Moreover, the immune characteristics could be further changed by combining the gas component and the metal organic structure.
  • FIGS. 4A and 4B These results are shown in FIGS. 4A and 4B.
  • the immune characteristics could be controlled by using the gas component.
  • the immune characteristics could be further changed by combining the gas component and the metal organic structure.

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Abstract

L'invention a pour objet de fournir une excellente composition de vaccin. La composition de vaccin de l'invention contient un antigène destiné à induire une immunité, et un composant gazeux liquide à 25°C et 100kPa.
PCT/JP2018/021696 2017-06-06 2018-06-06 Composition de vaccin WO2018225785A1 (fr)

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CN112592487A (zh) * 2020-12-11 2021-04-02 成都理工大学 腺嘌呤及柠檬酸与Zn2+构筑的MOFs材料及其制备方法

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CN112022836A (zh) * 2020-09-02 2020-12-04 山东大学 一种无需冷藏储存的金属有机框架纳米疫苗的制备方法
CN112592487A (zh) * 2020-12-11 2021-04-02 成都理工大学 腺嘌呤及柠檬酸与Zn2+构筑的MOFs材料及其制备方法

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