WO2016089181A1 - 1 이상의 인플루엔자 바이러스 중화 결합 분자를 포함하는 애주번트 조성물 및 이를 포함하는 백신 조성물 - Google Patents
1 이상의 인플루엔자 바이러스 중화 결합 분자를 포함하는 애주번트 조성물 및 이를 포함하는 백신 조성물 Download PDFInfo
- Publication number
- WO2016089181A1 WO2016089181A1 PCT/KR2015/013279 KR2015013279W WO2016089181A1 WO 2016089181 A1 WO2016089181 A1 WO 2016089181A1 KR 2015013279 W KR2015013279 W KR 2015013279W WO 2016089181 A1 WO2016089181 A1 WO 2016089181A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- vaccine
- group
- adjuvant
- seq
- binding molecule
- Prior art date
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—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
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
Definitions
- the present invention relates to an adjuvant composition comprising at least one influenza virus neutralizing binding molecule and a vaccine composition comprising the same, more specifically, the use as an adjuvant to increase the efficacy of a vaccine by enhancing an immune response induced by the vaccine. It relates to an adjuvant composition comprising at least one human monoclonal antibody having neutralizing activity against influenza virus, and a vaccine composition comprising the same.
- Influenza virus is a disease caused by infection of the respiratory tract, which is common in winter, and is highly infectious and spreads easily to all age groups, especially those who are vulnerable (Treanor J, 2004, N Engl J). Med . 350 (3): 218-20).
- Enveloped virus belonging to the Orthomyxoviridae an influenza virus whose genome contains eight-segment negative-sense and single-strand RNA (ribonucleic acid) Influenza A virus is divided into several subtypes according to the major surface proteins HA (hemagglutinin) and NA (neuraminidase). To date, 17 HA and 10 NA are known (Cheung TK and Poon LL 2007, Ann NY Acad Sci. 1102: 1-25; Tong S, et al.
- Influenza viruses can infect birds, pigs, and humans depending on their type, and due to their genome consisting of RNA fragments, strains of viruses continue to develop due to combinations and mutations of various genes (Treanor J, 2004. N Engl J Med). 350 (3): 218-20). Because of this persistent mutation, it is difficult to obtain permanent immunity, so the most effective preventive method is to inoculate a vaccine against an influenza virus, which is expected to spread every year, to form a yearly immunity for a particular type. will be.
- influenza virus vaccine given annually is a trivalent or tetravalent vaccine, in which H1 and H3 subtypes of influenza A are mixed with one or two types of influenza B type HA.
- vaccines for various infectious diseases are added with a substance to increase immunogenicity, which is called an adjuvant.
- adjuvants approved for human use include MF59 with aluminum and oil-in-water emulsions consisting of aluminum hydroxide and aluminum phosphate, and AS04 with MPL and aluminum hydroxide, which are AS03 and TLR4 agonists. (Rappuoli R, 2011. Nature Reviews Immunology 11 , 11 (12): 865-72)
- the antibody against influenza A virus which the applicant has previously applied for, has shown neutralizing efficacy against various influenza subtypes.
- the antibody disclosed in Korean Patent Application No. 10-2011-0020061 is mainly derived from phylogenetic group 1 (H1, H2, H5, H9, etc.), the antibody disclosed in Korean Patent Application No. 10-2012-0107512 mainly showed neutralizing efficacy to phylogeny group 2 (H3, H7, etc.).
- a cocktail formulation that can prevent and cure both viruses belonging to Group 1 and Group 2, which are likely to be epidemic by mixing and administering two or more antibodies at the same time.
- the present inventors have completed the present invention by finding a new use of the previously developed antibody as an adjuvant by confirming that the effect of the vaccine can be increased by administering the influenza virus neutralizing antibody developed by the present inventor together with the vaccine.
- an object of the present invention is to provide an adjuvant composition comprising at least one influenza virus neutralizing binding molecule.
- Another object of the present invention is to provide a vaccine composition comprising the adjuvant composition and the target antigen.
- Another object of the present invention is to provide a preparation method comprising the adjuvant composition and the target antigen.
- Another object of the present invention is to provide a method of increasing the immune response to a target antigen by administering the adjuvant composition to a host.
- Another object of the present invention is to provide a method of immunizing by administering the vaccine composition to a host.
- Another object of the present invention is to provide a method for preparing an immunological product from an immunized host by administering the vaccine composition to a host.
- the present invention provides an adjuvant composition comprising at least one influenza virus neutralizing binding molecule.
- the binding molecule may bind to an epitope in the stem region of the hemagglutinin (HA) protein of influenza A virus.
- HA hemagglutinin
- the epitope of the binding molecule is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N
- It may include an adjuvant composition characterized in that at least one epitope selected from the group consisting of.
- the epitope of the binding molecule may comprise amino acid residues at positions 18, 38, 40, 291, 292 and 318 of the HA1 polypeptide.
- the epitope of the binding molecule may comprise amino acid residues at positions 18, 19, 20, 21, 41, 42, 45, 48, 49, 52 and 53 of the HA2 polypeptide.
- the epitope of the binding molecule comprises amino acid residues at positions 18, 38, 40, 291, 292 and 318 of the HA1 polypeptide and positions 18, 19, 20, 21 of the HA2 polypeptide And amino acid residues of 41, 42, 45, 48, 49, 52 and 53.
- the epitope of the binding molecule may comprise amino acid residues at positions 278 and 318 of the HA1 polypeptide.
- the epitope of the binding molecule may comprise positions 38, 39, 41, 42, 45, 48, 49, 52, and 53 amino acid residues of the HA2 polypeptide.
- the epitope of the binding molecule comprises an amino acid residue at the position of the HA1 polypeptide and / or HA2 polypeptide of an HA first monomer and HA1 of a second monomer adjacent to the first monomer. It may further comprise position 25, 32 and 33 amino acid residues of the polypeptide.
- the epitope of the binding molecule comprises amino acid residues at positions 278 and 318 of the HA1 polypeptide and positions 38, 39, 41, 42, 45, 48, 49, 52, and 53 of the HA2 polypeptide. Amino acid residues.
- the epitope of the binding molecule comprises an amino acid residue at the position of the HA1 polypeptide and HA2 polypeptide of an HA first monomer, and of the HA1 polypeptide of a second monomer adjacent to the first monomer. And may further comprise position 25, 32 and 33 amino acid residues.
- the epitope of the binding molecule comprises amino acid residues at positions 278 and 318 of the HA1 polypeptide and positions 38, 39, 41, 42, 45, 48, 49, 52, 53, Amino acid residues of 58 and 99;
- the epitope of the binding molecule comprises an amino acid residue at the position of the HA1 polypeptide and HA2 polypeptide of an HA first monomer, and of the HA1 polypeptide of a second monomer adjacent to the first monomer. Position 25, 27, 32, and 33 amino acid residues.
- the epitope of the binding molecule comprises amino acid residues at positions 54, 55, 278, 291 and 318 of the HA1 polypeptide, and positions 19, 20, 21, 38, 39, 41, Amino acid residues of 42, 45, 46, 48, 49, 52, 53, 56, 57 and 60.
- the epitope of the binding molecule comprises an amino acid residue at the position of the HA1 polypeptide and an HA2 polypeptide of an HA first monomer, and HA1 of an HA second monomer adjacent to the HA first monomer. Amino acid residues at positions 25, 32, 33, 310, 311, and 312 of the polypeptide.
- Amino acid position numbering of the epitopes is based on H3 HA numbering system.
- the binding molecule In one embodiment of the invention, the binding molecule
- a light chain variable region comprising a CDR1 region of SEQ ID NO: 1, a CDR2 region of SEQ ID NO: 2, and a CDR3 region of SEQ ID NO: 3, and a CDR1 region of SEQ ID NO: 4, a CDR2 region of SEQ ID NO: 5, and SEQ ID NO: 6
- a binding molecule comprising a heavy chain variable region comprising a CDR3 region region;
- Binding molecule comprising a heavy chain variable region comprising a CDR3 region
- At least one binding molecule selected from the group consisting of.
- the CDRs of the variable regions were determined by conventional methods according to a system devised by Kabat et al. (Kabat et al., Sequences of Proteins of Immunological Interest (5 th ), National Institutes of Health, Bethesda, MD). (1991)].
- the CDR numbering used in the present invention used the Kabat method, but other binding molecules including CDRs determined according to other methods such as IMGT method, Chothia method and AbM method are also included in the present invention.
- the binding molecule In one embodiment of the invention, the binding molecule
- a binding molecule comprising a light chain comprising the polypeptide sequence of SEQ ID NO: 13 and a heavy chain comprising the polypeptide sequence of SEQ ID NO: 14;
- a binding molecule comprising a light chain comprising the polypeptide sequence of SEQ ID NO: 15 and a heavy chain comprising the polypeptide sequence of SEQ ID NO: 16
- At least one binding molecule selected from the group consisting of.
- the binding molecule comprises a binding molecule that binds to the Fc receptor on the cell surface.
- the present invention also provides a vaccine composition comprising the adjuvant composition and the target antigen.
- the target antigen may be a viral antigen, but is not limited thereto.
- the viral antigen is an influenza virus antigen.
- the influenza virus antigens include influenza A virus or influenza B virus antigens.
- the influenza virus antigen may be hemagglutinin (HA) or neuraminidase (NA), but is not limited thereto.
- the vaccine composition may be included in the weight ratio of the antigen and the adjuvant composition 1: 0.02 to 1: 200, preferably 1: 0.2 to 1:20 It is not limited to this.
- the weight ratio of antigen to adjuvant composition can be reduced or increased to modulate immunogenic activity.
- the present invention provides a vaccine composition to which an additional adjuvant composition is added in addition to the adjuvant composition.
- additional adjuvant compositions include, but are not limited to, Alum, metabolizable oils (eg squalene), tocols (eg alpha-tocopherol), sterols (eg cholesterol), saponins (Eg QS21), a ligand of Toll like receptor (eg poly (I: C), oligonucleotides with CpG motifs and / or LPS derivatives (eg 3D-MPL)).
- the present invention also provides a method for preparing a vaccine composition comprising the adjuvant composition and a target antigen.
- the vaccine composition may be, but is not limited to, an influenza virus vaccine composition.
- the present invention also provides a method for enhancing the immune response to a target antigen by administering the adjuvant composition to a host.
- the vaccine composition may be, but is not limited to, an influenza virus vaccine composition.
- the immune response may be a method characterized by being induced by a cell having an Fc receptor on the cell surface, but is not limited thereto.
- the present invention also provides a method for preventing a virus-derived disease comprising administering to a subject a vaccine composition comprising the adjuvant composition in an effective amount.
- the virus-derived disease may be an influenza virus-derived disease.
- the present invention also provides a method of immunizing by administering the vaccine composition to a host.
- the immunological product may be a T cell, a B cell, or an antibody.
- the immunological product may also be other types of cells having Fc receptors on the cell surface, such as B cells, such as neutrophils, macrophages, natural killer cells or dendritic cells.
- the "influenza A virus” described in the present invention is an enveloped virus belonging to the Orthomyxoviridae, which is negative-polar and single-stranded in eight segments.
- RNA ribonucleic acid
- A, B and C groups a genome of RNA (ribonucleic acid) as a genome, and is classified into A, B and C groups, and is divided into several subtypes according to major surface proteins HA (hemaggutinin) and NA (neuraminidase). To date, 17 HAs and 10 NAs are known.
- H1 subtypes described herein include H1N1, H1N2, H1N3, H1N4, H1N5, H1N6, H1N7, H1N8, H1N9 and H1N10.
- H2 subtypes described herein include H2N1, H2N2, H2N3, H2N4, H2N5, H2N6, H2N7, H2N8, H2N9 and H2N10.
- H5 subtypes described herein include H5N1, H5N2, H5N3, H5N4, H5N5, H5N6, H5N7, H5N8, H5N9 and H5N10.
- H9 subtypes described in the present invention include H9N1, H9N2, H9N3, H9N4, H9N5, H9N6, H9N7, H9N8, H9N9 and H9N10.
- H3 subtypes described in the present invention include H3N1, H3N2, H3N3, H3N4, H3N5, H3N6, H3N7, H3N8, H3N9 and H3N10.
- H7 subtypes described in the present invention include H7N1, H7N2, H7N3, H7N4, H7N5, H7N6, H7N7, H7N8, H7N9 and H7N10.
- Hemagglutinin (hereinafter referred to as” HA )” described in the present invention refers to the envelope glycoprotein of influenza virus. HA mediates influenza virus adsorption and penetration into host cells. There are 17 subtypes reported so far.
- NA neuroaminidase
- the term “neuraminidase (hereinafter referred to as” NA ”) described in the present invention refers to the envelope glycoprotein of influenza virus. NA plays an important role when influenza virus proliferates and then spreads. Ten subtypes have been reported so far.
- the "influenza vaccine” described in the present invention is the best way to prevent seasonal or pandemic influenza, and is largely divided into live and inactivated vaccines.
- Live vaccines have been developed and used live attenuated vaccines.
- Inactivated vaccines are whole virus vaccines using whole viruses incubated in embryonated eggs or cell cultures and inactivated with formalin, split vaccines in which viral envelopes are crushed with ethers, etc. And subunit vaccines with purified NA components.
- a vaccine containing one type of influenza B group together with H1 and H3 subtypes of influenza A group is said to be a trivalent vaccine and a vaccine containing two types of influenza B group is called a tetravalent vaccine.
- the "influenza vaccine” described in the present invention includes both trivalent and tetravalent seasonal, live and inactivated vaccines of pandemic.
- a “binding molecule” described herein binds to an intact immunoglobulin, a fusion protein or antigen having an Fc portion of an immunoglobulin, including a monoclonal antibody such as a chimeric, humanized or human monoclonal antibody.
- Enzymes, receptors which are capable of binding to variable domains or substrates comprising immunoglobulin fragments that compete with intact immunoglobulins for binding to monomeric HA or trimer HA of an immunoglobulin, such as influenza A virus It means protein. Regardless of the structure, the antigen-binding fragment binds to the same antigen recognized by intact immunoglobulins.
- An antigen-binding fragment comprises two or more continuations of the amino acid sequence of a binding molecule, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 30 contiguous amino acid residues, at least 35 contiguous amino acid residues, at least 40 contiguous amino acid residues.
- At least 50 contiguous amino acid residues at least 60 contiguous amino acid residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least 100 contiguous amino acid residues, at least 125 contiguous amino acid residues, Peptides or polypeptides comprising an amino acid sequence of at least 150 contiguous amino acid residues, at least 175 contiguous amino acid residues, at least 200 contiguous amino acid residues, or at least 250 contiguous amino acid residues.
- Antigen-binding fragments described herein are in particular Fab, F (ab '), F (ab') 2, Fv, dAb, Fd, complementarity determining region (CDR) fragments, single-chain antibodies (scFv), 2 Bivalent single-chain antibodies, single-chain phage antibodies, diabodies, triabodies, tetrabodies, polypeptides containing one or more fragments of immunoglobulin sufficient to bind a particular antigen to the polypeptide, and the like It includes.
- the fragments may be produced synthetically or by enzymatic or chemical digestion of complete immunoglobulins or may be produced genetically by recombinant DNA techniques. Production methods are well known in the art.
- adjuvant refers to a substance or composition that is added to a vaccine or pharmaceutically active ingredients to increase and / or affect an immune response. Representatively means a carrier or auxiliary agent and / or other pharmaceutically active substance or composition for immunogen. Typically, the term “adjuvant” should be interpreted in a broad sense and capable of enhancing the immunogenicity of antigens incorporated into or administered with the adjuvant. It means a wide range of materials or strategies. Adjuvant may also be divided into, but not limited to, an immune potentiator, an antigen delivery system, or a combination thereof.
- immunological product refers to a protective immune mediator or cell, etc. arising from a host immunized by administration of an antigen and / or adjuvant composition, eg, activated T cells, B cells or It may mean an antibody, but is not limited thereto.
- an antigen and / or adjuvant composition eg, activated T cells, B cells or It may mean an antibody, but is not limited thereto.
- the term "pharmaceutically acceptable excipient” means an inert material that is combined into an active molecule, such as a drug, agent, or binding molecule, to produce an acceptable or convenient dosage form.
- Pharmaceutically acceptable excipients are nontoxic or are excipients that are acceptable to the recipient for their intended use, at least in the doses and concentrations in which the toxicity is used, and with other components of the formulation including drugs, preparations or binding powders. It is compatible.
- an effective amount refers to the amount of the binding molecule of the present invention that is effective in increasing the effectiveness of the vaccine when administered in combination with a vaccine of influenza A virus.
- composition comprising at least one influenza virus neutralizing binding molecule of the present invention has been found to increase the effectiveness of the vaccine, it can be used as an adjuvant to increase the immune response upon administration of the vaccine and to prevent disease caused by viruses. Very useful.
- FIG. 1 shows H1N1 vaccine composition intramuscularly in mice twice weekly (see Table 1), and serum was taken 13, 17 and 27 days after the first intramuscular injection to be specific for H1N1 influenza virus in serum.
- the antibody titer was confirmed by ELISA.
- FIG. 2 shows intramuscular injection of H1N1 vaccine composition into mice twice weekly (see Table 3), serum was collected 28 days after the first intramuscular injection, and specific antibody titers against H1N1 HA protein and H1N1 influenza virus in serum This is the result confirmed by ELISA.
- Figure 3 is inoculated with H1N1 influenza virus 10MLD 50 in the immunized mouse, and confirmed the survival rate and weight change of the immunized mouse.
- FIG. 4 shows H1N1 vaccine composition intramuscularly in mice twice weekly (see Table 5), and serum was collected 13, 20 and 27 days after the first intramuscular injection to be specific for H1N1 influenza virus in serum.
- the antibody titer was confirmed by ELISA.
- FIG. 6 shows that the H1N1 vaccine composition was intramuscularly injected into mice twice every two weeks (see Table 7), and serum was collected 13, 20, and 27 days after the first intramuscular injection to be specific for H1N1 influenza virus in serum.
- the antibody titer was confirmed by ELISA.
- Figure 7 is inoculated with H1N1 influenza virus 10MLD 50 in the immunized mouse, it is the result of confirming the survival rate and weight change of the immunized mouse.
- FIG. 8 shows intramuscular injection of H3N2 vaccine composition into mice twice weekly (see Table 9), serum was taken 13 and 27 days after the first intramuscular injection to determine specific antibody titers against H3N2 influenza virus in serum. The result confirmed by ELISA.
- FIG. 10 shows H1N1 vaccine composition intramuscularly injected to mice twice every two weeks (see Table 11), and serum was taken 13, 20 and 27 days after the first intramuscular injection to be specific for H1N1 influenza virus in serum.
- the antibody titer was confirmed by ELISA.
- FIG. 12 shows the ratio of B cells to immune cells in the furrow lymph nodes at 1, 3 and 7 days after the second intramuscular injection of the H1N1 vaccine composition into the mouse twice at intervals of 2 weeks (see Table 14). The result is confirmed.
- FIG. 13 shows H1N1 vaccine composition intramuscularly in mice twice weekly (see Table 15), and 4 weeks after the second intramuscular injection, H1N1 virus was inoculated with B cells in the spleen and body lymph nodes 1 and 3 days later. This is the result of checking the ratio of.
- FIG. 14 shows H1N1 vaccine composition intramuscularly in mice twice weekly (see Table 16), and serum was taken 13, 20, and 27 days after the first intramuscular injection to be specific for H1N1 influenza virus in serum.
- the antibody titer was confirmed by ELISA.
- H1N1 vaccine composition After the intramuscular injection of H1N1 vaccine composition into the mouse twice at intervals of 2 weeks as shown in Table 1 above, the immune response induced in each experimental group was confirmed. Serum was collected from each experimental group 13, 17 and 27 days after the first intramuscular injection, and the titer of the antibody was confirmed by ELISA.
- hemaglutinin inhibition assay As shown in Table 2 below, the effectiveness of the vaccine was evaluated by a hemaglutinin inhibition assay (HI assay, hemaglutinin inhibition assay) to confirm whether the therapeutic antibodies, CT120 antibody and CT149 antibody, as an adjuvant of the influenza virus vaccine.
- Serum was obtained from mice 13, 17 and 27 days after the administration of the vaccine and the adjuvant, and the antibody titers were able to inhibit the aggregation reaction between influenza virus and chicken erythrocytes.
- CT120 or CT149 When combined with influenza virus vaccine, CT120 or CT149 confirmed a similar or better immunogenic potency effect than Alum, an adjuvant. This confirmed that CT120 and CT149 were effective as an adjuvant of influenza virus vaccines.
- Example 2 H1N1 The vaccine as an antigen CT120 and CT149 Determination of Antigen Concentration to Determine the Effect of Adjuvant
- H1N1 vaccine (cell based) was administered alone from 0.01ug to 1ug or with Alum adjuvant, and then antibody titers were confirmed by ELISA and neutralizing antibody titers via HI. Standard is a trivalent vaccine on the market which was run for comparison.
- Group 11 Standard 0.3 ug - i.m. 5
- Group 12 H1N1 vaccine 0.05 ug - i.m. 5
- Group 13 H1N1 vaccine 0.05 ug Alum i.m. 5
- Group 14 H1N1 vaccine 0.01 ug - i.m. 5
- Group 15 H1N1 vaccine 0.01 ug Alum i.m. 5
- Group 16 Standard 0.03 ug - i.m. 5
- H1N1 vaccine composition After the intramuscular injection of H1N1 vaccine composition into the mouse twice at intervals of 2 weeks as shown in Table 3 above, the immune response induced in each experimental group was confirmed. Serum was collected from each experimental group 28 days after the first intramuscular injection, the antibody titers of HA protein and virus in serum were confirmed by ELISA, and the neutralizing antibody titers were confirmed by HI assay.
- the titer of the antibody was confirmed to increase in proportion to the amount of the administered antigen, and when the Alum adjuvant was added, it was confirmed that the antibody titer was generally higher. In addition, there was no significant difference in antibody titer against HA protein and antibody titer against H1N1 virus.
- the protective immune effect against influenza virus was higher in the experimental group with the adjuvant compared to the experimental group administered with the antigen alone at all concentrations.
- the test group administered with the antigen alone was 80% of the 0.1ug test group. Survival rate was shown, and the experimental group administered with the adjuvant showed 100% survival rate.
- the 0.05ug experimental group showed 60% survival rate in the experimental group administered with antigen alone, and the experimental group administered with the adjuvant showed 100% survival rate.
- the HI titer was different in the 0.1ug experimental group, but the difference in the protective immune effect was small according to the presence or absence of an adjuvant.
- the final antigen concentration was determined to be 0.05 ug, which shows a large difference depending on the presence or absence of adjuvant. Animal experiments were conducted to confirm the effect of the adjuvant using this.
- Example 3 H1N1 The vaccine as an antigen CT120 and CT149 The effect of adjuvant
- the mouse form antibody was prepared by replacing the constant region of the Fc region with the IgG1 or IgG2a region of the mouse in human IgG1 form CT120, CT149.
- CT120 and CT120 were selected through the preliminary experiments (not prepared herein), the concentration showing the effect as an adjuvant.
- H1N1 vaccine cell based
- various concentrations of therapeutic antibodies were mixed with various concentrations of therapeutic antibodies and reacted at 37 ° C. for 1 hour
- the mice were injected intramuscularly twice every two weeks as shown in Table 5 below, and then 13 days from the first intramuscular injection.
- serum was collected from each experimental group to confirm antibody titer of H1N1 virus through serum ELISA and neutralizing antibody titer by HI assay.
- the antibody titer against H1N1 virus was generally higher in the experimental group using the mouse form CT120 (mIgG2a) as an adjuvant than the experimental group using the CT120 as an adjuvant.
- CT120 mouse form CT120
- the antibody titer was generally higher than the experimental group using Alum as an adjuvant, and the antibody titer was sharply increased in serum (D20 in FIG. 5) taken after the first 3 weeks of immunity. It was confirmed.
- the HI titer was increased up to 4 times according to the concentration in the experimental group using CT120 (mIgG2a) as an adjuvant as compared to the experimental group using the CT120 as an adjuvant.
- CT120 (mIgG2a) has a higher effect as an adjuvant than CT120, it was confirmed that the best effect when administered at a concentration of 0.5ug.
- the adjuvant effect of the CT149 was generally weak, and the antibody titer similar to the experimental group using the Alum adjuvant was shown in the experimental group using 0.5 ug of CT149 (mIgG2a).
- HI titer was not significantly increased in the experimental group using CT149 or CT149 (mIgG2a) as an adjuvant, and similar to the results of antibody titers measured by ELISA, 0.5 ug of CT149 (mIgG2a) was used.
- the experimental group (Group 10) showed the same HI titer as the experimental group (Group 11) using Alum adjuvant.
- CT120 (mIgG2a) showed the highest adjuvant effect against H1N1 vaccine. Among them, the most effective result was 0.5ug. In the case of CT149 (mIgG2a), an adjuvant effect was seen using 0.5ug, but it was lower than CT120 (mIgG2a).
- Example 4 H3N2 The vaccine as an antigen CT120 and CT149 The effect of adjuvant
- H3N2 vaccine (cell based) was produced using another influenza virus strain, Philippines / 2/82 (H3N2). Afterwards, animal experiments were conducted to confirm the adjuvant effects of CT120 and CT149 on the H3N2 vaccine.
- H3N2 vaccine was administered alone from 0.01ug to 1ug or with Alum adjuvant, and then antibody titer was confirmed by ELISA and neutralizing antibody titer was confirmed by HI.
- the mouse adapted Philippines / 2/82 (H3N2) virus was infected to confirm the change in survival rate and weight.
- H3N2 vaccine 0.05 ug - i.m. 5
- Group 11 H3N2 vaccine 0.05 ug Alum i.m. 5
- Group 12 H3N2 vaccine 0.01 ug - i.m. 5
- Group 13 H3N2 vaccine 0.01 ug Alum i.m. 5
- H3N2 vaccine composition After the intramuscular injection of H3N2 vaccine composition into the mouse twice at intervals of 2 weeks as shown in Table 9 above, the immune response induced in each experimental group was confirmed. Two weeks after each intramuscular injection, serum was collected from each experimental group, and antibody titers against H3N2 virus in serum were confirmed by ELISA, and neutralizing antibody titers were confirmed by HI assay.
- the titer of the antibody was increased in proportion to the amount of the administered antigen, and when the Alum adjuvant was added together, the titer of the antibody was generally higher.
- HI titer was higher in the experimental group to which the Alum was added as compared to the experimental group to which only the antigen was added.
- HI titers could not be identified in the experimental groups (groups 12 and 10) in which 0.01ug and 0.05ug of antigen were added, but the HI titers were 160 and 640 in the experimental groups in which Alum, an adjuvant, was added. It confirmed that it rises.
- H3N2 vaccine composition the difference in HI titer and survival rate according to the presence or absence of an adjuvant was shown at the antigen concentration (0.05ug) used in the H1N1 experiment performed in Example 3. Therefore, an animal experiment confirming the effect of the adjuvant of the therapeutic antibodies CT120 and CT149 using the H3N2 antigen concentration 0.05ug determined therefrom was carried out in the same manner as in Example 3, and the adjuvant of CT120 and CT149 using the H3N2 vaccine as an antigen. The effect as was confirmed.
- the hypothesis that the therapeutic antibody exhibits an adjuvant effect against the flu vaccine is that the immune response to the flu vaccine can be more efficiently generated by binding the Fc portion of the antibody to the Fc receptor present in immune cells.
- CT120 (mIgG2a) administration group As shown in Table 11, in the case of CT120 (mIgG2a) administration group, the experiment was conducted at 0.1ug and 0.5ug, concentrations showed the best effect as an adjuvant of the H1N1 vaccine in Example 3, CT120 (mIgG2a) F (ab ' 2)
- the amount of CT120 (mIgG2a) F (ab ') 2 is equal to the molar ratio between the group receiving the same amount as CT120 (mIgG2a) and the vaccine substance at 0.5ug, 0.1ug of CT120 (mIgG2a).
- mice After reacting the H1N1 vaccine with various concentrations of CT120 (mIgG2a) or CT120 (mIgG2a) F (ab ') 2 for 1 hour at 37 ° C, the mice were injected intramuscularly twice every two weeks as shown in Table 11 above. , The immune response induced in each experimental group was confirmed. Serum was collected from each experimental group 13, 20, and 27 days after the first intramuscular injection to identify antibody titers against H1N1 virus through serum ELISA and neutralizing antibody titers against H1N1 virus through HI.
- the antibody titer was relatively low when CT120 (mIgG2a) F (ab ') 2 was added, but the difference in antibody titer did not show a significant difference in all experimental groups.
- Group Ag Adjuvant Survival rate (%) Group 1 PBS - 0% Group 2 H1N1 vaccine 0.05 ug - 20% Group 3 PBS CT 120 ( mIgG2a ) F (ab ') 2 0.5 ug 0% Group 4 PBS CT 120 ( mIgG2a 0.5 ug 0% Group 5 H1N1 vaccine 0.05 ug Alum 80% Group 6 H1N1 vaccine 0.05 ug CT 120 ( mIgG2a 0.5 ug 100% Group 7 H1N1 vaccine 0.05 ug CT 120 ( mIgG2a ) 0.1 ug 90% Group 8 H1N1 vaccine 0.05 ug CT 120 ( mIgG2a ) F (ab ') 2 0.5 ug 50% Group 9 H1N1 vaccine 0.05 ug CT 120 ( mIgG2a ) F (ab ') 2 0.3 ug 30% Group 10 H1N1 vaccine 0.05 ug CT 120 ( mIgG
- CT120 (mIgG2a) F The experimental group (groups 8 to 11) to which ab ') 2 was added as an adjuvant showed a similar survival rate to the experimental group to which the antigen was administered alone (group 2).
- Example 5 it was confirmed that the Fc portion of the flu antibody plays an important role in the role of an adjuvant. Based on this, animal experiments were conducted to discover cells whose immune response is changed by binding to the Fc region among the immune cells expressing the Fc receptor. The amount of antibody was selected in Example 3 0.5ug of CT120 (mIgG2a) of the mouse form showing the best effect as an adjuvant of the H1N1 vaccine.
- mice After mixing the H1N1 vaccine and the adjuvant and reacted for 1 hour at 37 °C, as shown in Table 14 above, the mouse was injected into the muscle twice every two weeks. After that, one day, three days, and seven days later, each mouse was isolated from the spleen and the furrow lymph nodes, and various immune cell numbers and ratios were identified.
- CT120 adjuvant enhances the immune response of B cells through the Fc region.
- CT120 F (ab ') 2 group the difference of the control group with the PBS-administered mouse was not found. Therefore, the importance of the Fc region was confirmed once more. From this, CT120 immunoadjuvant was found to enhance immunogenicity of H1N1 vaccine substance by a mechanism different from that of Alum, a commercially available adjuvant.
- Example 6-1 it was confirmed that the CT120 adjuvant enhances B cell immunity. Based on this, animal experiments were conducted to determine whether CT120 immunoadjuvant could be involved in B cell defense immunity even in the presence of virus.
- B cells increased about 10% compared to the PBS control group in the experimental group administered with H1N1 vaccine and CT120.
- the experimental group administered with Alum in the vaccine also increased significantly compared to the PBS control group, but the percentage of B cells did not increase significantly compared to the experimental group containing CT120.
- the other groups contained B-1 cells (CD19 + B220-), immature B cells, the experimental group injected with CT120 in the H1N1 vaccine showed more mature B-2 cells (CD19). Only + B220 +). In the case of lymph nodes, the proportion of B cells did not increase rapidly as seen in the spleen, but it was confirmed that the increase was significant in three replicates.
- CT120 was confirmed to be effective as an adjuvant of the flu vaccine, and then the experiment was conducted to compare this effect with the effect of the adjuvant currently used in the flu vaccine.
- the adjuvant used in the current seasonal flu vaccine is MF59, which is used for Novadis Fluad.
- AddaVax TM InvivoGen, Catalog # vac-adx-10) having the same composition as MF59 was used.
- Example 3 0.5ug and 0.1ug of CT120 (mIgG2a) of a mouse form showing the best effect as an adjuvant of the H1N1 vaccine were selected.
- Addvax 100% means the amount of adjuvant to enter the HA content of 45ug of seasonal flu vaccine.
- H1N1 vaccine was mixed with CT120 (mIgG2a) or Addvax having the same composition as commercialized adjuvant and reacted at 37 ° C for 1 hour, the mice were injected intramuscularly twice every two weeks as shown in Table 16 above. Immune response induced in the experimental group was confirmed. Serum was collected from each experimental group 13, 20, and 27 days after the first intramuscular injection to identify antibody titers against H1N1 virus through serum ELISA and neutralizing antibody titers against H1N1 virus through HI.
- Group Ag Adjuvant HI titer Group 1 PBS - N.D.
- Group 2 H1N1 vaccine 0.05 ug - 20
- H1N1 vaccine 0.05 ug Alum 160
- H1N1 vaccine 0.05 ug CT 120 ( mIgG2a 0.5 ug 160
- H1N1 vaccine 0.05 ug CT 120 ( mIgG2a ) 0.1 ug 80
- Group 8 H1N1 vaccine 0.05 ug Addvax 100% 160
- Group 9 H1N1 vaccine 0.05 ug Addvax 10% 20
- the Addvax 100% test group (Group 8) showed HI titer similar to the experimental groups (Groups 6 and 7) in which 0.5ug and 0.1ug of CT120 (mIgG2a) was administered.
- The% administration groups (groups 10 and 9) had no similar or HI titer as the experimental group (group 2) administered with the H1N1 vaccine alone.
- Group Ag Adjuvant Survival rate (%) Group 1 PBS - 0 Group 2 H1N1 vaccine 0.05 ug - 20 Group 3 PBS CT 120 ( mIgG2a ) Fab 0.5 ug 0 Group 4 PBS Addvax 100% 0 Group 5 H1N1 vaccine 0.05 ug Alum 80 Group 6 H1N1 vaccine 0.05 ug CT 120 ( mIgG2a 0.5 ug 100 Group 7 H1N1 vaccine 0.05 ug CT 120 ( mIgG2a ) 0.1 ug 90 Group 8 H1N1 vaccine 0.05 ug Addvax 100% 90 Group 9 H1N1 vaccine 0.05 ug Addvax 10% 60 Group 10 H1N1 vaccine 0.05 ug Addvax One% 0
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Pulmonology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
Group | Antigen | Adjuvant | Route | Mouse # |
Group1 | PBS | - | i.m. | 4 |
Group2 | PBS | CT 120 10 ug | i.m. | 4 |
Group3 | PBS | CT 149 10 ug | i.m. | 4 |
Group4 | H1N1 split vaccine 0.2 ug | - | i.m. | 4 |
Group5 | H1N1 split vaccine 0.2 ug | CT 120 10 ug | i.m. | 4 |
Group6 | H1N1 split vaccine 0.2 ug | CT 120 5 ug | i.m. | 4 |
Group7 | H1N1 split vaccine 0.2 ug | CT120 1 ug | i.m. | 4 |
Group8 | H1N1 split vaccine 0.2 ug | CT120 0.5 ug | i.m. | 4 |
Group9 | H1N1 split vaccine 0.2 ug | CT149 10 ug | i.m. | 4 |
Group10 | H1N1 split vaccine 0.2 ug | CT149 5 ug | i.m. | 4 |
Group11 | H1N1 split vaccine 0.2 ug | CT149 1 ug | i.m. | 4 |
Group12 | H1N1 split vaccine 0.2 ug | CT149 0.5 ug | i.m. | 4 |
Group13 | H1N1 split vaccine 0.2 ug | Alum | i.m. | 4 |
Group | Antigen | Adjuvant | HI titer | ||
Day 13 | Day 17 | Day 27 | |||
Group1 | PBS | - | N.D. | N.D. | N.D. |
Group2 | PBS | CT 120 10 ug | N.D. | N.D. | N.D. |
Group3 | PBS | CT 149 10 ug | N.D. | N.D. | N.D. |
Group4 | H1N1 split vaccine 0.2 ug | - | N.D. | N.D. | 40 |
Group5 | H1N1 split vaccine 0.2 ug | CT 120 10 ug | N.D. | N.D. | 40 |
Group6 | H1N1 split vaccine 0.2 ug | CT 120 5 ug | N.D. | N.D. | 80 |
Group7 | H1N1 split vaccine 0.2 ug | CT120 1 ug | N.D. | N.D. | 40 |
Group8 | H1N1 split vaccine 0.2 ug | CT120 0.5 ug | N.D. | N.D. | 40 |
Group9 | H1N1 split vaccine 0.2 ug | CT149 10 ug | N.D. | N.D. | 80 |
Group10 | H1N1 split vaccine 0.2 ug | CT149 5 ug | N.D. | N.D. | 40 |
Group11 | H1N1 split vaccine 0.2 ug | CT149 1 ug | N.D. | 40 | 160 |
Group12 | H1N1 split vaccine 0.2 ug | CT149 0.5 ug | N.D. | N.D. | 160 |
Group13 | H1N1 split vaccine 0.2 ug | Alum | N.D. | 20 | 80 |
Group | Ag | Adjuvant | Route | Mouse # |
Group 1 | PBS | - | i.m. | 5 |
Group 2 | H1N1 vaccine 1 ug | - | i.m. | 5 |
Group 3 | H1N1 vaccine 1 ug | Alum | i.m. | 5 |
Group 4 | Standard 3 ug | - | i.m. | 5 |
Group 5 | H1N1 vaccine 0.5 ug | - | i.m. | 5 |
Group 6 | H1N1 vaccine 0.5 ug | Alum | i.m. | 5 |
Group 7 | H1N1 vaccine 0.2 ug | - | i.m. | 5 |
Group 8 | H1N1 vaccine 0.2 ug | Alum | i.m. | 5 |
Group 9 | H1N1 vaccine 0.1 ug | - | i.m. | 5 |
Group 10 | H1N1 vaccine 0.1 ug | Alum | i.m. | 5 |
Group 11 | Standard 0.3 ug | - | i.m. | 5 |
Group 12 | H1N1 vaccine 0.05 ug | - | i.m. | 5 |
Group 13 | H1N1 vaccine 0.05 ug | Alum | i.m. | 5 |
Group 14 | H1N1 vaccine 0.01 ug | - | i.m. | 5 |
Group 15 | H1N1 vaccine 0.01 ug | Alum | i.m. | 5 |
Group 16 | Standard 0.03 ug | - | i.m. | 5 |
Group | HI titer | |
Group 1 | PBS | N.D. |
Group 2 | H1N1 1 ug | 160 |
Group 3 | H1N1 1 ug + alum | 1280 |
Group 4 | Standard 3 ug | 80 |
Group 5 | H1N1 0.5 ug | 160 |
Group 6 | H1N1 0.5 ug + alum | 320 |
Group 7 | H1N1 0.2 ug | 80 |
Group 8 | H1N1 0.2 ug + alum | 160 |
Group 9 | H1N1 0.1 ug | 20 |
Group 10 | H1N1 0.1 ug + alum | 160 |
Group 11 | Standard 0.3 ug | N.D. |
Group 12 | H1N1 0.05 ug | 20 |
Group 13 | H1N1 0.05 ug + alum | 80 |
Group 14 | H1N1 0.01 ug | N.D. |
Group 15 | H1N1 0.01 ug + alum | N.D. |
Group 16 | Standard 0.03 ug | N.D. |
Group | Ag | Adjuvant | Route | Mouse # |
Group 1 | PBS | - | i.m. | 10 |
Group 2 | PBS | CT 120 0.5 ug | i.m. | 10 |
Group 3 | PBS | CT 120 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 4 | H1N1 vaccine 0.05 ug | - | i.m. | 10 |
Group 5 | H1N1 vaccine 0.05 ug | CT 120 0.05 ug | i.m. | 10 |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 0.1 ug | i.m. | 10 |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 0.5 ug | i.m. | 10 |
Group 8 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.05 ug | i.m. | 10 |
Group 9 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | i.m. | 10 |
Group 10 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 11 | H1N1 vaccine 0.05 ug | Alum | i.m. | 10 |
Group 12 | Standard 0.15 ug | - | i.m. | 10 |
Ag | Adjuvant | HI titer | |
G1 | PBS | - | N.D. |
G2 | PBS | CT 120 0.5 ug | N.D. |
G3 | PBS | CT 120 ( mIgG2a ) 0.5 ug | N.D. |
G4 | H1N1 vaccine 0.05 ug | - | 20 |
G5 | H1N1 vaccine 0.05 ug | CT 120 0.05 ug | 40 |
G6 | H1N1 vaccine 0.05 ug | CT 120 0.1 ug | 20 |
G7 | H1N1 vaccine 0.05 ug | CT 120 0.5 ug | 20 |
G8 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.05 ug | 40 |
G9 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | 80 |
G10 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | 160 |
G11 | H1N1 vaccine 0.05 ug | Alum | 80 |
G12 | Standard 0.15 ug | - | N.D. |
Group | Ag | Adjuvant | Route | Mouse # |
Group 1 | PBS | - | i.m. | 10 |
Group 2 | PBS | CT 149 0.5 ug | i.m. | 10 |
Group 3 | PBS | CT 149 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 4 | H1N1 vaccine 0.05 ug | - | i.m. | 10 |
Group 5 | H1N1 vaccine 0.05 ug | CT 149 0.01 ug | i.m. | 10 |
Group 6 | H1N1 vaccine 0.05 ug | CT 149 0.05 ug | i.m. | 10 |
Group 7 | H1N1 vaccine 0.05 ug | CT 149 0.5 ug | i.m. | 10 |
Group 8 | H1N1 vaccine 0.05 ug | CT 149 ( mIgG2a ) 0.01 ug | i.m. | 10 |
Group 9 | H1N1 vaccine 0.05 ug | CT 149 ( mIgG2a ) 0.05 ug | i.m. | 10 |
Group 10 | H1N1 vaccine 0.05 ug | CT 149 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 11 | H1N1 vaccine 0.05 ug | Alum | i.m. | 10 |
Group 12 | Standard 0.15 ug | - | i.m. | 10 |
Ag | Adjuvant | HI titer | |
G1 | PBS | - | N.D. |
G2 | PBS | CT 149 0.5 ug | N.D. |
G3 | PBS | CT 149 ( mIgG2a ) 0.5 ug | N.D. |
G4 | H1N1 vaccine 0.05 ug | - | 20 |
G5 | H1N1 vaccine 0.05 ug | CT 149 0.01 ug | 20 |
G6 | H1N1 vaccine 0.05 ug | CT 149 0.05 ug | 40 |
G7 | H1N1 vaccine 0.05 ug | CT 149 0.5 ug | 20 |
G8 | H1N1 vaccine 0.05 ug | CT 149 ( mIgG2a ) 0.01 ug | 20 |
G9 | H1N1 vaccine 0.05 ug | CT 149 ( mIgG2a ) 0.05 ug | 40 |
G10 | H1N1 vaccine 0.05 ug | CT 149 ( mIgG2a ) 0.5 ug | 80 |
G11 | H1N1 vaccine 0.05 ug | Alum | 80 |
G12 | Standard 0.15 ug | - | N.D. |
Group | Ag | Adjuvant | Route | Mouse # |
Group 1 | PBS | - | i.m. | 5 |
Group 2 | H3N2 vaccine 1 ug | - | i.m. | 5 |
Group 3 | H3N2 vaccine 1 ug | Alum | i.m. | 5 |
Group 4 | H3N2 vaccine 0.5 ug | - | i.m. | 5 |
Group 5 | H3N2 vaccine 0.5 ug | Alum | i.m. | 5 |
Group 6 | H3N2 vaccine 0.2 ug | - | i.m. | 5 |
Group 7 | H3N2 vaccine 0.2 ug | Alum | i.m. | 5 |
Group 8 | H3N2 vaccine 0.1 ug | - | i.m. | 5 |
Group 9 | H3N2 vaccine 0.1 ug | Alum | i.m. | 5 |
Group 10 | H3N2 vaccine 0.05 ug | - | i.m. | 5 |
Group 11 | H3N2 vaccine 0.05 ug | Alum | i.m. | 5 |
Group 12 | H3N2 vaccine 0.01 ug | - | i.m. | 5 |
Group 13 | H3N2 vaccine 0.01 ug | Alum | i.m. | 5 |
Group | Ag | Adjuvant | HI titer |
Group 1 | PBS | - | N.D. |
Group 2 | H3N2 vaccine 1 ug | - | 160 |
Group 3 | H3N2 vaccine 1 ug | Alum | 320 |
Group 4 | H3N2 vaccine 0.5 ug | - | 160 |
Group 5 | H3N2 vaccine 0.5 ug | Alum | 320 |
Group 6 | H3N2 vaccine 0.2 ug | - | 80 |
Group 7 | H3N2 vaccine 0.2 ug | Alum | 640 |
Group 8 | H3N2 vaccine 0.1 ug | - | 80 |
Group 9 | H3N2 vaccine 0.1 ug | Alum | 320 |
Group 10 | H3N2 vaccine 0.05 ug | - | N.D. |
Group 11 | H3N2 vaccine 0.05 ug | Alum | 640 |
Group 12 | H3N2 vaccine 0.01 ug | - | N.D. |
Group 13 | H3N2 vaccine 0.01 ug | Alum | 160 |
Group | Ag | Adjuvant | Route | Mouse # |
Group 1 | PBS | - | i.m. | 10 |
Group 2 | H1N1 vaccine 0.05 ug | - | i.m. | 10 |
Group 3 | PBS | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | i.m. | 10 |
Group 4 | PBS | CT 120 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 5 | H1N1 vaccine 0.05 ug | Alum | i.m. | 10 |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | i.m. | 10 |
Group 8 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | i.m. | 10 |
Group 9 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.3 ug | i.m. | 10 |
Group 10 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.1 ug | i.m. | 10 |
Group 11 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.06 ug | i.m. | 10 |
Group | Ag | Adjuvant | Route | HI titer |
Group 1 | PBS | - | i.m. | N.D. |
Group 2 | H1N1 vaccine 0.05 ug | - | i.m. | 20 |
Group 3 | PBS | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | i.m. | N.D. |
Group 4 | PBS | CT 120 ( mIgG2a ) 0.5 ug | i.m. | N.D. |
Group 5 | H1N1 vaccine 0.05 ug | Alum | i.m. | 160 |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | i.m. | 160 |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | i.m. | 80 |
Group 8 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | i.m. | 40 |
Group 9 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.3 ug | i.m. | 20 |
Group 10 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.1 ug | i.m. | 20 |
Group 11 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.06 ug | i.m. | 40 |
Group | Ag | Adjuvant | Survival rate(%) |
Group 1 | PBS | - | 0% |
Group 2 | H1N1 vaccine 0.05 ug | - | 20% |
Group 3 | PBS | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | 0% |
Group 4 | PBS | CT 120 ( mIgG2a ) 0.5 ug | 0% |
Group 5 | H1N1 vaccine 0.05 ug | Alum | 80% |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | 100% |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | 90% |
Group 8 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | 50% |
Group 9 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.3 ug | 30% |
Group 10 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.1 ug | 30% |
Group 11 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.06 ug | 30% |
Group | Ag | Adjuvant | Route | Mouse# |
Group1 | PBS | - | i.m | 3 |
Group2 | H1N1 vaccine 0.05 ug | - | i.m | 3 |
Group3 | - | CT 120 ( mIgG2a ) 0.5 ug | i.m | 3 |
Group4 | H1N1 vaccine 0.05 ug | Alum | i.m | 3 |
Group5 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | i.m | 3 |
Group6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) F(ab')2 0.5 ug | i.m | 3 |
Group | Ag | Adjuvant | Route | Mouse# |
Group1 | PBS | - | i.m | 3 |
Group2 | H1N1 vaccine 0.05 ug | - | i.m | 3 |
Group3 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | i.m | 3 |
Group4 | H1N1 vaccine 0.05 ug | Alum | i.m | 3 |
Group | Ag | Adjuvant | Route | Mouse # |
Group 1 | PBS | - | i.m. | 10 |
Group 2 | H1N1 vaccine 0.05 ug | - | i.m. | 10 |
Group 3 | PBS | CT 120 ( mIgG2a ) Fab 0.5 ug | i.m. | 10 |
Group 4 | PBS | Addvax 100% | i.m. | 10 |
Group 5 | H1N1 vaccine 0.05 ug | Alum | i.m. | 10 |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | i.m. | 10 |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | i.m. | 10 |
Group 8 | H1N1 vaccine 0.05 ug | Addvax 100% | i.m. | 10 |
Group 9 | H1N1 vaccine 0.05 ug | Addvax 10% | i.m. | 10 |
Group 10 | H1N1 vaccine 0.05 ug | Addvax 1% | i.m. | 10 |
Group | Ag | Adjuvant | HI titer |
Group 1 | PBS | - | N.D. |
Group 2 | H1N1 vaccine 0.05 ug | - | 20 |
Group 3 | PBS | CT 120 ( mIgG2a ) Fab 0.5 ug | N.D. |
Group 4 | PBS | Addvax 100% | N.D. |
Group 5 | H1N1 vaccine 0.05 ug | Alum | 160 |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | 160 |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | 80 |
Group 8 | H1N1 vaccine 0.05 ug | Addvax 100% | 160 |
Group 9 | H1N1 vaccine 0.05 ug | Addvax 10% | 20 |
Group 10 | H1N1 vaccine 0.05 ug | Addvax 1% | N.D. |
Group | Ag | Adjuvant | Survival rate(%) |
Group 1 | PBS | - | 0 |
Group 2 | H1N1 vaccine 0.05 ug | - | 20 |
Group 3 | PBS | CT 120 ( mIgG2a ) Fab 0.5 ug | 0 |
Group 4 | PBS | Addvax 100% | 0 |
Group 5 | H1N1 vaccine 0.05 ug | Alum | 80 |
Group 6 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.5 ug | 100 |
Group 7 | H1N1 vaccine 0.05 ug | CT 120 ( mIgG2a ) 0.1 ug | 90 |
Group 8 | H1N1 vaccine 0.05 ug | Addvax 100% | 90 |
Group 9 | H1N1 vaccine 0.05 ug | Addvax 10% | 60 |
Group 10 | H1N1 vaccine 0.05 ug | Addvax 1% | 0 |
Claims (11)
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US15/533,269 US20180008702A1 (en) | 2014-12-05 | 2015-05-12 | Adjuvant composition containing at least one influenza virus neutralizing and binding molecule and vaccine composition containing same |
EP15866215.5A EP3228323B1 (en) | 2014-12-05 | 2015-12-05 | Adjuvant composition containing at least one influenza virus neutralizing and binding molecule and vaccine composition containing same |
JP2017530090A JP6496409B2 (ja) | 2014-12-05 | 2015-12-05 | 1以上のインフルエンザウイルス中和結合分子を含むアジュバント組成物およびこれを含むワクチン組成物 |
ES15866215T ES2902702T3 (es) | 2014-12-05 | 2015-12-05 | Composición adyuvante que contiene por lo menos una molécula neutralizante y de unión al virus de la gripe, y composición de vacuna que la contiene |
CN201580065445.0A CN106999570B (zh) | 2014-12-05 | 2015-12-05 | 含至少一种流感病毒的中和性结合分子的佐剂组合物及含该佐剂组合物的疫苗组合物 |
US16/130,405 US10610587B2 (en) | 2014-12-05 | 2018-09-13 | Adjuvant composition containing at least one influenza virus neutralizing and binding molecule and vaccine composition containing same |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR20140173469 | 2014-12-05 | ||
KR10-2014-0173469 | 2014-12-05 | ||
KR10-2015-0113515 | 2015-08-11 | ||
KR1020150113515A KR20160068636A (ko) | 2014-12-05 | 2015-08-11 | 1 이상의 인플루엔자 바이러스 중화 결합 분자를 포함하는 애주번트 조성물 및 이를 포함하는 백신 조성물 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US15/533,269 A-371-Of-International US20180008702A1 (en) | 2014-12-05 | 2015-05-12 | Adjuvant composition containing at least one influenza virus neutralizing and binding molecule and vaccine composition containing same |
US16/130,405 Division US10610587B2 (en) | 2014-12-05 | 2018-09-13 | Adjuvant composition containing at least one influenza virus neutralizing and binding molecule and vaccine composition containing same |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2016089181A1 true WO2016089181A1 (ko) | 2016-06-09 |
Family
ID=56092041
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2015/013279 WO2016089181A1 (ko) | 2014-12-05 | 2015-12-05 | 1 이상의 인플루엔자 바이러스 중화 결합 분자를 포함하는 애주번트 조성물 및 이를 포함하는 백신 조성물 |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2016089181A1 (ko) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20040067687A (ko) * | 2003-01-24 | 2004-07-30 | 한국생명공학연구원 | A형 간염 바이러스 중화 인간 단일클론항체 |
US20100040635A1 (en) * | 2008-03-28 | 2010-02-18 | Sea Lane Biotechnologies | Neutralizing antibodies to influenza viruses |
KR20110047193A (ko) * | 2008-07-25 | 2011-05-06 | 인스티튜트 포 리서치 인 바이오메드슨 | 중화 항-인플루엔자 a 바이러스 항체 및 이의 용도 |
KR20110102198A (ko) * | 2010-03-08 | 2011-09-16 | (주)셀트리온 | 인간 b 세포에서 생산된 인플루엔자 a 바이러스 중화 활성을 가지는 인간 단일클론 항체 |
KR20130035916A (ko) * | 2011-09-30 | 2013-04-09 | (주)셀트리온 | 인간 b 세포에서 생산된 인플루엔자 a 바이러스 중화 활성을 가지는 결합 분자 |
KR20140119641A (ko) * | 2013-03-29 | 2014-10-10 | (주)셀트리온 | 2 이상의 인플루엔자 a 바이러스 중화 결합 분자를 포함하는 조성물 |
-
2015
- 2015-12-05 WO PCT/KR2015/013279 patent/WO2016089181A1/ko active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20040067687A (ko) * | 2003-01-24 | 2004-07-30 | 한국생명공학연구원 | A형 간염 바이러스 중화 인간 단일클론항체 |
US20100040635A1 (en) * | 2008-03-28 | 2010-02-18 | Sea Lane Biotechnologies | Neutralizing antibodies to influenza viruses |
KR20110047193A (ko) * | 2008-07-25 | 2011-05-06 | 인스티튜트 포 리서치 인 바이오메드슨 | 중화 항-인플루엔자 a 바이러스 항체 및 이의 용도 |
KR20110102198A (ko) * | 2010-03-08 | 2011-09-16 | (주)셀트리온 | 인간 b 세포에서 생산된 인플루엔자 a 바이러스 중화 활성을 가지는 인간 단일클론 항체 |
KR20130035916A (ko) * | 2011-09-30 | 2013-04-09 | (주)셀트리온 | 인간 b 세포에서 생산된 인플루엔자 a 바이러스 중화 활성을 가지는 결합 분자 |
KR20140119641A (ko) * | 2013-03-29 | 2014-10-10 | (주)셀트리온 | 2 이상의 인플루엔자 a 바이러스 중화 결합 분자를 포함하는 조성물 |
Non-Patent Citations (1)
Title |
---|
See also references of EP3228323A4 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Krammer et al. | Universal influenza virus vaccines that target the conserved hemagglutinin stalk and conserved sites in the head domain | |
Nachbagauer et al. | Age dependence and isotype specificity of influenza virus hemagglutinin stalk-reactive antibodies in humans | |
WO2014158001A1 (ko) | 2 이상의 인플루엔자 a 바이러스 중화 결합 분자를 포함하는 조성물 | |
WO2013048153A2 (ko) | 인간 b 세포에서 생산된 인플루엔자 a 바이러스 중화 활성을 가지는 결합 분자 | |
US10676520B2 (en) | Antibodies useful in passive influenza immunization | |
WO2011111966A2 (en) | Human monoclonal antibodies derived from human b cells and having neutralizing activity against influenza a viruses | |
EP2793945B1 (en) | Antibodies useful in passive influenza immunization | |
IL241552B2 (en) | Preparation and methods based on neutralizing antibodies administered intranasally for increased therapeutic efficacy | |
AU2022271473A1 (en) | Neutralizing anti-influenza binding molecules and uses thereof | |
US10610587B2 (en) | Adjuvant composition containing at least one influenza virus neutralizing and binding molecule and vaccine composition containing same | |
US20190345230A1 (en) | Broad-spectrum monoclonal anti-flu b antibody and uses thereof | |
WO2017095140A1 (ko) | 면역글로불린 fc가 융합된 인터루킨-7 융합 단백질을 포함하는 인플루엔자 바이러스 감염의 예방 또는 치료용 약학적 조성물 | |
WO2016089181A1 (ko) | 1 이상의 인플루엔자 바이러스 중화 결합 분자를 포함하는 애주번트 조성물 및 이를 포함하는 백신 조성물 | |
WO2015147611A1 (en) | Influenza a virus-specific monoclonal antibody and method for treating and diagnosing influenza infection using the same | |
Li et al. | Rapid isolation of a potent human antibody against H7N9 influenza virus from an infected patient | |
Wang et al. | Generation, characterization, and protective ability of mouse monoclonal antibodies against the HA of A (H1N1) influenza virus | |
WO2018030777A1 (ko) | 항-인플루엔자 바이러스 항체의 안정한 액체 약제학적 제제 | |
WO2021215855A1 (ko) | 삼량체를 형성하는 인플루엔자 바이러스 표면 단백질 유래 재조합 혈구응집소 단백질 및 이의 용도 | |
WO2023167487A1 (ko) | 사스-코로나바이러스-2 감염증 예방용 재조합 발현 벡터 및 그 응용 | |
WO2021145720A1 (ko) | 바이러스 감염병 예방 또는 치료용 항원 조성물 | |
JP2022535523A (ja) | Pan-ノイラミニダーゼ阻害抗体 | |
KR20130128751A (ko) | 신종 인플루엔자 바이러스 ha 단백질 특이적 항체 | |
US20170306001A1 (en) | Chimerization and characterization of a monoclonal antibody with potent neutralizing activity across multiple influenza a h5n1 clades | |
NZ626716B2 (en) | Antibodies useful in passive influenza immunization |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 15866215 Country of ref document: EP Kind code of ref document: A1 |
|
REEP | Request for entry into the european phase |
Ref document number: 2015866215 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2017530090 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 15533269 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |