Composition comprising influenza vaccine

Abstract

The present invention provides a composition comprising a universal influenza vaccine antigen and a vaccine adjuvant.

Description

[Technical Field] This invention relates to a composition comprising a vaccine adjuvant and a universal influenza vaccine antigen. [Previous Technology] Component vaccines containing viral proteins or partial peptides exhibit superior safety compared to live or whole-particle inactivated vaccines. However, component vaccines tend to have lower immunogenicity. Therefore, to enhance the immunogenicity of epitopes and improve the immunogenicity of vaccines, methods combining adjuvants with antigens are being investigated. Adjuvants are additives that enhance humoral and / or cellular immune responses against antigens; currently used vaccine adjuvants include alum and saponins. Recent findings have shown that Toll-like receptors (TLRs) play a crucial role in activating innate immunity, one of the organism's defense mechanisms against microorganisms. Monophospholipid A (MPL) and oligodeoxynucleotides (CpG ODNs) containing unmethylated cytosine guanine (CpG) sequences exhibit immune activation via TLRs. It is generally believed that among the 13 known specific TLRs in the human body, 5 are associated with the recognition of bacterial components (TLR1 / 2 / 4 / 5 / 6), and the other 4 (TLR3 / 7 / 8 / 9) are also associated with the recognition of viral nucleic acids (see Non-Patent Literature 1). Furthermore, in the case of TLR7 and TLR8, low-molecular-weight compounds that mimic the single-stranded RNA of viruses as natural ligands are known to act as activators. For example, synthetic compounds such as pyrimidine compounds (see Patent Literature 1 and 2) and imidazoquinoline compounds (see Patent Literature 3) have been reported. By utilizing its activators, it activates TLR7 and / or TLR8, thereby activating dendritic cells (DCs) via a TLR and myeloid differentiation factor 88 (MyD88)-dependent signaling pathway. This results in enhanced expression of T cell co-stimulatory molecules (CD80, CD86, CD40) or the production of inflammatory cytokines including type I interferons (especially IFNα), TNFα, IL-6, or IL-12. Furthermore, regarding TLR7 and / or TLR8 activators, in addition to DC activation, they are also known to activate T cells or B cells, thereby stimulating NK cells to promote the production of IFNγ and other activities, with the expectation of possessing vaccine adjuvant activity. In fact, there are reports on the vaccine adjuvant activity of TLR7 and / or TLR8 activators such as resiquimod or imiquimod (see Non-Patent Literature 2). As a complex of TLR7 and / or 8 activators with other substances, known examples include: vaccine adjuvants formed by covalently linking fatty acids and imidazoquinoline compounds (see Patent Documents 4, 5, 6 and Non-Patent Document 4), or conjugate compounds of phospholipids and adenine compounds (see Patent Documents 7, 8, 9), conjugate compounds of fatty acids and adenine compounds (Non-Patent Document 5), conjugate compounds of fatty acid glycerides or phospholipids and adenine compounds formed by polyethylene glycol (see Patent Document 10), conjugate compounds of squalene and adenine compounds (see Patent Document 11), and conjugate compounds of squalene and pyrimidine compounds (see Patent Document 12). Current influenza hemagglutinin (hemagglutinin is sometimes referred to as "HA") vaccines, also known as influenza HA split vaccines, exert their protective effect against infection by inducing HA antibodies. HA antibodies bind to the portion of the viral membrane exposed on the outside of the hemagglutinin, known as the head region. However, this region exhibits the most structural variation among viral strains. As a result, there are cases where HA antibodies cannot bind to viruses with antigenic variants different from the vaccine strain, rendering the vaccine ineffective. In recent years, the industry has revealed that antibodies (stem antibodies) that bind to the stem region of hemagglutinin, which is less prone to antigenic variation, include infection defense antibodies (see Patent Document 13). To efficiently induce stem antibodies, HA stem proteins have been developed that successfully stabilize unstable stem regions by binding hemagglutinin to artificial variants or linkers, and human clinical trials have been conducted. However, the practical application of these HA stem proteins still faces challenges in improving immunogenicity and manufacturing processes. The report indicates that the infection defense efficacy of stem antibodies depends not only on their antigen-binding properties and antibody quantity, but also, to a greater extent, on the subtype of IgG antibodies induced (see Non-Patent Literature 3). Specifically, it reveals the functional and cross-reactive nature of stem IgG2 antibody systems, and vaccines that strongly induce stem IgG2 antibodies are effective against a wide range of influenza infections. However, no effective method for enhancing the inducibility of stem IgG2 antibodies has yet been discovered. [Prior Art Literature] [Patent Literature] [Patent Document 1] International Publication No. 00 / 12487 [Patent Document 2] International Publication No. 2009 / 067081 [Patent Document 3] US Patent Publication No. 4689338 [Patent Document 4] International Publication No. 2005 / 001022 [Patent Document 5] International Publication No. 2005 / 018555 [Patent Document 6] International Publication No. 2012 / 024284 [Patent Document 7] International Publication No. 2010 / 048520 [Patent Document 8] International Publication No. 4689338 Patent Document No. 2011 / 017611 [Patent Document 9], International Publication No. 2011 / 139348 [Patent Document 10], International Publication No. 2010 / 093436 [Patent Document 11], International Publication No. 2017-061532 [Patent Document 12], International Publication No. 2017-056494 [Patent Document 13], Japanese Patent Publication No. 2016-516090 [Patent Document 14], International Publication No. 2016 / 109792 [Non-Patent Documents] [Non-Patent Literature 1] Iwasaki, A., Nat. Immunol. 2004, 5, 987 [Non-Patent Literature 2] Vaccine 2011, 29, 3341・MA Tomai et al, Exp. Rev. Vaccine, 6, 835 [Non-Patent Literature 3] DiLillo, DJ., Nat. Medicine 2014, 20, 143 [Non-Patent Literature 4] Vaccine. 2011 Jul 26;29(33):5434-42 [Non-Patent Literature 5] Bioconjugate Chem. 2009, 20, 1194-1200 [Non-Patent Literature 6] Curr Opin Virol. 2016, 17, 95-103 [Non-Patent Literature 7] Nat Med. 2015, 21, 1065-70 [Non-Patent Literature 8] Science. 2015, 349, 1301-6 [Summary of the Invention] [The problem the invention aims to solve] The problem this invention aims to solve is to discover an influenza HA split vaccine that effectively generates antibodies binding to the HA dry region of influenza viruses that are not prone to antigenic variation, and to provide a method for enhancing the inducibility of its IgG2 antibodies. [Technical Means for Solving the Problem] Through intensive research, the inventors discovered that by acidifying existing influenza HA split vaccines to expose the hemagglutinin dry region, the acid-treated influenza HA split vaccine can generate cross-reactive antibodies that bind to the LAH (long alpha helix) in the HA dry region, thereby functioning as a universal influenza vaccine antigen. Furthermore, they discovered that by adding a compound that enhances the physiological activity of TLR7 to the acid-treated influenza HA split vaccine, this compound functions as a vaccine adjuvant, increasing the IgG2 antibody induction capacity, thus completing this invention. That is, the present invention relates to the following. [Item 1] A composition comprising (1) and (2) of the following: (1) a universal influenza vaccine antigen; and (2) a vaccine adjuvant. [Item 2] The composition as described in Item 1, wherein the universal influenza vaccine antigen is an influenza HA split vaccine antigen capable of producing antibodies that bind to the LAH in the HA dry region. [Item 3] The composition as described in Item 2, wherein the influenza HA split vaccine antigen is in the shape of its HA dry region exposed to the outside. [Item 4] The composition as described in Item 1, wherein the universal influenza vaccine antigen is an influenza HA split vaccine antigen that enhances the antigenicity of the LAH in the HA dry region by means of the shape of the HA dry region being exposed to the outside, and can generate antibodies that bind to the LAH in the HA dry region. [Item 5] The composition described in any one of items 1 to 4, wherein the universal influenza vaccine antigen is manufactured by acid treatment of an influenza HA split vaccine. [Item 6] The composition described in any one of items 1 to 5, wherein the universal influenza vaccine antigen is manufactured by acid treatment of an influenza HA split vaccine that has not been treated with formalin. [Item 7] The composition described in any one of items 1 to 6, wherein the universal influenza vaccine antigen is manufactured by a manufacturing process including a step of acid treatment of an influenza HA split vaccine and a subsequent step of formalin treatment. [Item 8] The composition described in any one of items 1 to 7, wherein the universal influenza vaccine antigen is manufactured by subjecting an influenza HA split vaccine, which is a separate HA subtype, to acidic treatment. [Item 9] The composition described in any one of items 1 to 7, wherein the universal influenza vaccine antigen comprises two or more vaccine antigens that are manufactured by subjecting an influenza HA split vaccine as a separate HA subtype to acidic treatment. [Item 10] The composition of any one of items 1 to 9, wherein the vaccine adjuvant is a substance that enhances the physiological activity of TLRs. [Item 11] The composition of any one of items 1 to 10, wherein the vaccine adjuvant is a substance that enhances the physiological activity of one or more TLRs selected from the group consisting of TLR7, TLR8, and TLR9. [Item 12] The composition of any one of items 1 to 11, wherein the vaccine adjuvant is a substance that enhances the physiological activity of at least TLR7. [Item 13] The composition of any one of items 1 to 12, wherein the vaccine adjuvant is a substance that enhances the physiological activity of at least TLR8. [Item 14] The composition of any one of items 1 to 13, wherein the vaccine adjuvant is a substance that enhances the physiological activity of at least TLR9. [Item 15] The composition as described in Item 11 or 12, wherein the substance enhancing the physiological activity of TLR7 is a low molecular weight compound enhancing the physiological activity of TLR7, or a coupling compound formed by a low molecular weight compound enhancing the physiological activity of TLR7 and a lipid via a spacer group chemical bond. [Item 16] The composition as described in Item 15, wherein the low molecular weight compound enhancing the physiological activity of TLR7 or the low molecular weight compound enhancing the physiological activity of TLR7 contained in the coupling compound has a molecular weight of 200 to 600 and has an adenine skeleton, a pyrimidine skeleton, an imidazoquinoline skeleton, an imidazopyridine skeleton, or a quinazoline skeleton. [Item 17] The composition as described in Item 16, wherein the low molecular weight compound enhancing the physiological activity of TLR7 or the low molecular weight compound enhancing the physiological activity of TLR7 contained in the coupling compound has a pyrimidine skeleton, an adenine skeleton, or an imidazoquinoline skeleton. [Item 18] The composition described in any one of items 15 to 17, wherein the lipid is a lipid derived from squalene or squalane. [Item 19] The composition described in any one of items 15 to 18, wherein the compound that enhances the physiological activity of TLR7 is a low molecular weight compound having an adenine skeleton and a coupling compound formed by a spacer group chemically bonded to a lipid derived from squalene or squalane, or a pharmaceutically permissible salt thereof. [Item 20] The composition described in any one of items 15 to 18, wherein the compound that enhances the physiological activity of TLR7 is a low molecular weight compound having a pyrimidine skeleton and a coupling compound formed by a spacer group chemically bonded to a lipid derived from squalene or squalane, or a pharmaceutically permissible salt thereof. [Item 21] The composition described in any one of items 15 to 18 and 20, wherein the compound that enhances the physiological activity of TLR7 is of the following formula (1): [Chemical 1] [Figure] [In formula (1), X represents a methylene group, R1 represents a hydrogen atom or an alkyl group of 1 to 3 carbons that may be substituted with a hydroxyl group, R2 represents a hydrogen atom or an alkyl group of 1 to 6 carbons, R3 represents an alkyl group of 1 to 3 carbons, R4 represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group of 1 to 3 carbons or an alkoxy group of 1 to 3 carbons, Y1 represents a single bond or a methylene group, Y2 represents a single bond or -C(O)-, L represents a straight-chain extended alkyl group of 2 or 3 carbons. R5 and R6 independently represent hydrogen atoms or alkyl groups having 1 to 3 carbon atoms, or R5 may be together with R6 to form a substituted or unsubstituted 5 to 8 nitrogen-containing saturated heterocycle. In the case of substitution of the aforementioned 5 to 8 nitrogen-containing saturated heterocycle, it may be substituted by 1 to 4 substituents selected from the same or different hydroxyl and halogen atoms. m represents 0 or 1. [Chem. 2][Figure] all represent double bonds] represents the compound or a pharmaceutically permissible salt.[Item 22] The composition as described in any one of items 15 to 18, 20 and 21, wherein the coupling compound is (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenoyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.[Item 23] The composition described in any one of items 15 to 18 and 20 to 22, wherein the compound that enhances the physiological activity of TLR7 is (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl]amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof. [Item 24] An influenza vaccine comprising the composition described in any one of items 1 to 23. [Item 25] A method for preventing influenza, comprising administering to a warm-blooded animal requiring prevention a preventively effective amount of the composition described in any one of items 1 to 23. [Item 26] A method for preventing influenza, comprising administering to a warm-blooded animal requiring prevention a preventively effective amount of a universal influenza vaccine antigen and a preventively effective amount of a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7). [Item 27] ​​A kit for the prevention of influenza, comprising a universal influenza vaccine antigen and a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7). [Item 28] A vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7) for use in combination with a universal influenza vaccine antigen to prevent influenza. [Item 29] A universal influenza vaccine antigen for use in combination with a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7) to prevent influenza. [Item 30] Use of a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7) for manufacturing a composition comprising a universal influenza vaccine antigen for the prevention of influenza. [Item 31] Use of a universal influenza vaccine antigen for manufacturing a composition comprising a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7) for the prevention of influenza. [Item 32] The method as described in Items 25 and 26, the kit as described in Item 27, the vaccine adjuvant as described in Item 28, or the antigen as described in Item 29, wherein the universal influenza vaccine antigen and the vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7) are administered simultaneously or separately. [Item 32-1] The method as described in Items 25 and 26, the kit as described in Item 27, the vaccine adjuvant as described in Item 28, or the antigen as described in Item 29, wherein the universal influenza vaccine antigen is administered before, after, or simultaneously with the administration of the vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7). [Item 33] A method for manufacturing a composition comprising an influenza HA split vaccine antigen and a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7), comprising the following steps: step a), wherein an influenza HA split vaccine antigen that generates antibodies binding to LAH in the HA dry region is generated by subjecting an untreated influenza HA split vaccine to acid treatment; and step b), wherein the vaccine antigen obtained in step a) is mixed with the vaccine adjuvant. [Item 34] A method for manufacturing a composition comprising an influenza HA split vaccine antigen and a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7), comprising the following steps: step a), wherein an influenza HA split vaccine is subjected to acid treatment; step b), wherein an influenza HA split vaccine antigen that generates antibodies binding to LAH in the HA dry region is subsequently subjected to formalin treatment; and step c), wherein the vaccine antigen obtained in step b) is mixed with the vaccine adjuvant. [Item 35] A method for manufacturing a kit comprising an influenza HA split vaccine antigen and a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7), comprising the following steps: step a), wherein an influenza HA split vaccine antigen that generates antibodies binding to LAH in the HA dry region is generated by subjecting an untreated influenza HA split vaccine to acid treatment; and step b), wherein the vaccine obtained in step a) is combined with the vaccine adjuvant to produce the kit. [Item 36] A method for manufacturing a kit comprising an influenza HA split vaccine antigen and a vaccine adjuvant (e.g., a substance that enhances the physiological activity of TLR7), comprising the following steps: step a), wherein the influenza HA split vaccine is subjected to acid treatment; step b), wherein the influenza HA split vaccine antigen is produced by subsequently subjected to formalin treatment to generate antibodies binding to the LAH in the HA dry region; and step c), wherein the vaccine obtained in step b) is combined with the vaccine adjuvant to produce the kit. [Effects of the Invention] According to the present invention, a universal influenza vaccine antigen is provided, and its vaccine activity can be further enhanced by adding a vaccine adjuvant.

Implementation Method

Claims

1. A composition comprising (1) and (2) of the following: (1) an influenza vaccine antigen that generates antibodies binding to the LAH in the HA dry region; and (2) a vaccine adjuvant; wherein the influenza vaccine antigen is manufactured by acid treatment of an influenza HA split vaccine, wherein the acid treatment is performed at pH 3.0 to 6.5; and the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

2. The composition of claim 1, wherein the influenza vaccine antigen has an exposed HA dry region.

3. The composition of claim 1, wherein the influenza vaccine antigen is an influenza HA split vaccine antigen that enhances the antigenicity of the LAH in the HA dry region exposed to the outside and can generate antibodies that bind to the LAH in the HA dry region.

4. The composition of claim 1, wherein the influenza vaccine antigen is manufactured by acid treatment of an untreated influenza HA split vaccine.

5. The composition of claim 1, wherein the influenza vaccine antigen is manufactured by a manufacturing process including a step of acidifying an influenza HA split vaccine and a subsequent step of formalinizing the influenza HA split vaccine.

6. The composition of claim 1, wherein the influenza vaccine antigen is manufactured by subjecting an influenza HA split vaccine, which is a separate HA subtype, to acidic treatment.

7. The composition of claim 1, wherein the influenza vaccine antigen comprises two or more vaccine antigens comprising influenza HA split vaccine antigens manufactured by subjecting an influenza HA split vaccine as a separate HA subtype to acidic treatment.

8. The composition of any one of claims 1 to 7, wherein the acid treatment is performed at pH 4.0 to 6.

0.

9. The composition of any one of claims 1 to 7, wherein the acid treatment is performed at pH 4.4 to 5.

8.

10. The composition of any one of claims 1 to 7, wherein the compound is (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl]amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

11. The composition of claims 1 to 7, wherein the compound is (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; or a pharmaceutically permissible salt thereof.

12. An influenza vaccine comprising a composition of any one of claims 1 to 11.

13. Use of a composition of any one of claims 1 to 11 for manufacturing a vaccine for preventing influenza in warm-blooded animals.

14. Use of an influenza vaccine antigen and vaccine adjuvant for manufacturing a composition for preventing influenza in warm-blooded animals, wherein the influenza vaccine antigen produces antibodies that bind to the LAH region of the HA dry area; wherein the influenza vaccine antigen is manufactured by subjecting an influenza HA split vaccine to acid treatment, wherein the acid treatment is performed at pH 3.0 to 6.5; and the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

15. A kit for preventing influenza, comprising an influenza vaccine antigen and a vaccine adjuvant, wherein the influenza vaccine antigen produces antibodies that bind to the LAH region of the HA dry area; wherein the influenza vaccine antigen is manufactured by acidifying an influenza HA split vaccine, wherein the acidification is performed at pH 3.0 to 6.5; and the vaccine adjuvant is selected from the following compound: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

16. An influenza vaccine antigen for use in combination with a vaccine adjuvant to prevent influenza, wherein the influenza vaccine antigen produces antibodies that bind to the LAH region of the HA dry area; wherein the influenza vaccine antigen is manufactured by subjecting an influenza HA split vaccine to acidic treatment, wherein the acidic treatment is performed at pH 3.0 to 6.5; and wherein the vaccine adjuvant is selected from the following compound: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

17. Use of a vaccine adjuvant for manufacturing a composition comprising an influenza vaccine antigen for the prevention of influenza, wherein the influenza vaccine antigen generates antibodies that bind to the LAH region of the HA dry area; wherein the influenza vaccine antigen is manufactured by subjecting an influenza HA split vaccine to acid treatment, wherein the acid treatment is performed at pH 3.0 to 6.5; and wherein the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

18. Use of an influenza vaccine antigen for manufacturing a composition comprising a vaccine adjuvant for the prevention of influenza, wherein the influenza vaccine antigen produces antibodies that bind to the LAH region of the HA dry region; wherein the influenza vaccine antigen is manufactured by subjecting an influenza HA split vaccine to acidic treatment, wherein the acidic treatment is performed at pH 3.0 to 6.5; and wherein the vaccine adjuvant is selected from the following compound: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

19. As in request item 15, in which the influenza vaccine antigen and vaccine adjuvant are administered simultaneously or separately.

20. The antigens in claim 16, wherein the influenza vaccine antigens and vaccine adjuvants are administered simultaneously or separately.

21. A method for manufacturing a composition comprising an influenza HA split vaccine antigen and a vaccine adjuvant, comprising the following steps: step a), wherein an influenza HA split vaccine antigen generating antibodies binding to the LAH region of the HA dry region is generated by subjecting an untreated influenza HA split vaccine to acid treatment, wherein the acid treatment is performed at pH 3.0–6.5; and step b), wherein the vaccine antigen obtained in step a) is mixed with a vaccine adjuvant; wherein the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

22. A method for manufacturing a composition comprising an influenza HA split vaccine antigen and a vaccine adjuvant, comprising the following steps: step a), wherein the influenza HA split vaccine is subjected to an acidic treatment, wherein the acidic treatment is performed at pH 3.0 to 6.5; step b), wherein the influenza HA split vaccine antigen is obtained by subsequently performing formalin treatment to produce antibodies binding to the LAH in the HA dry region; and step c), wherein the vaccine antigen obtained in step b) is mixed with a vaccine adjuvant; wherein the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

23. A method for manufacturing a kit comprising an influenza HA split vaccine antigen and a vaccine adjuvant, comprising the following steps: step a), which involves subjecting an untreated influenza HA split vaccine to acid treatment to generate an influenza HA split vaccine antigen that produces antibodies binding to the LAH region of the HA dry region, wherein the acid treatment is performed at pH 3.0–6.5; and step b), which involves combining the vaccine antigen obtained in step a) with a vaccine adjuvant to prepare the kit; wherein the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

24. A method for manufacturing a kit comprising an influenza HA split vaccine antigen and a vaccine adjuvant, comprising the following steps: step a), wherein the influenza HA split vaccine is subjected to an acidic treatment, wherein the acidic treatment is performed at pH 3.0–6.5; step b), wherein the influenza HA split vaccine antigen is obtained by subsequently performing formalin treatment to produce antibodies that bind to the LAH in the HA dry region; and step c), wherein the vaccine antigen obtained in step b) is combined with a vaccine adjuvant to prepare the kit; wherein the vaccine adjuvant is selected from the following compounds: (4E,8E,12E,16E,20E)-N-{2-[{4-[(2-amino-4-{[(3S)-1-hydroxyhexane-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]benzyl}(methyl)amino]ethyl}-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenylamine; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; (4E,8E,12E,16E,20E)-1-(4-{4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoyl}piperidin-1-yl)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaen-1-one; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexenyl]amino}ethyl)-3-methoxybenzamide; 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(2-{[(4E,8E,12E,16E,20E)-4,8,12,17,21,25-hexamethylhexacarbon-4,8,12,16,20,24-hexaenyl](methyl)amino}ethyl)-3-methoxybenzamide; or a pharmaceutically permissible salt thereof.

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