Water soluble adjuvant

Abstract

The present invention pertains to a compound useful as a vaccine adjuvant for a cancer vaccine, a method for producing said compound, a medicinal composition containing said compound, and a use of said compound as a vaccine adjuvant for a cancer vaccine.

Description

Water-soluble adjuvants

[0001] The present invention relates to compounds useful as vaccine adjuvants for vaccines (cancer vaccines or infectious disease vaccines), methods for producing the same, pharmaceutical compositions containing the compounds, and uses of the compounds as vaccine adjuvants for vaccines (cancer vaccines or infectious disease vaccines).

[0002] Cancer vaccine therapy generally aims to treat cancer by activating tumor-specific immune cells using proteins or peptides derived from tumor antigens. Therapies using tumor antigen peptides as antigens are called cancer peptide vaccine therapy. Generally, tumor antigen peptides alone have low immunogenicity, so vaccine adjuvants are used in combination with tumor antigens to induce cytotoxic T cells (CTLs), which are important for anti-tumor immunity. For example, because W / O emulsions have an aqueous phase as the internal phase, which makes it easy to retain antigenic peptides in the internal phase, it has been reported that the use of W / O emulsions as vaccine adjuvants has shown efficient induction of CTLs (Patent Document 1).

[0003] Examples of W / O emulsions used as vaccine adjuvants for tumor antigen peptides include an emulsion composition for dilution (Patent Document 1), Incomplete Freund's Adjuvant (IFA), and Montanide (registered trademark) (Non-Patent Documents 1 and 2). Complete Freund's Adjuvant (CFA), which is a W / O emulsion containing inactivated Mycobacterium tuberculosis, is also known. However, CFA is not approved for human administration due to its high toxicity (Non-Patent Document 2).

[0004] While the aim of improving vaccine activity has traditionally been to add inactivated bacterial cells to adjuvants, such as CFA, in recent years, vaccine adjuvants incorporating compounds with a clear mechanism of action have been developed. Toll-like receptor 7 (TLR7) has been reported to activate Th1 cells and enhance the cellular immunity required for antitumor activity (Non-Patent Document 3). Several easily manufactured low-molecular-weight compounds are known to act as ligands for TLR7, and in addition to the commercially available drug imiquimod, compounds with a pyrimidine backbone have been reported to act as TLR7 agonists (Patent Document 2).

[0005] A search is being conducted for compounds that have improved TLR7 agonists and physical properties suitable for use as adjuvants. For example, there have been reports of TLR7 agonists that have a phosphate group conjugated to them so that they can bind to insoluble metal particles, such as alum adjuvants (Patent Document 3, Non-Patent Documents 4 and 5).

[0006] International Publication WO2006 / 078059 International Publication WO2009 / 067081 International Publication WO2012 / 031140

[0007] J Immunother Cancer. 2016 Sep 20;4:56Semin Immunol. 2010 Jun;22(3):155-61.Vaccine. 2011 Apr 12;29(17):3341-55.Sci Transl Med. 2014 Nov 19;6(263):263ra160J. Med. Chem., 2016, 59 (12), pp 5868-5878

[0008] An object of the present invention is to provide a conjugated TLR7 agonist that enhances adjuvant activity.

[0009] The inventors conducted extensive research to search for TLR7 agonists that enhance adjuvant activity. As a result, they discovered that a TLR7 agonist having a pyrimidine backbone, conjugated with polyethylene glycol (PEG) and rendered water-soluble, exhibits excellent adjuvant activity, leading to the completion of the present invention. According to the present invention, there is provided a pyrimidine derivative represented by the following formula (1) (hereinafter, also referred to as the "compound of the present invention"):

[0010] That is, the present invention is as follows.

[0011] [Term 1] Formula (1): [wherein X is methylene, an oxygen atom, a sulfur atom, SO, SO ２ , or NR ５ (R ５ is a hydrogen atom or C １－６ R represents alkyl; １ is C １－６ alkyl, wherein the alkyl is selected from halogen, hydroxy, and C １－６ alkoxy), R ２ and R ３ are each independently a hydrogen atom or C １－６ Alkyl (the alkyl is not substituted with halogen, hydroxy, C １－６ alkoxy), R ４ is a hydrogen atom, halogen, hydroxy, C １－６ alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens), C １－６ represents alkoxy (which may be substituted with 1 to 3 of the same or different halogens) or cyano; L represents a linker; Y １ is -(CH ２ CH ２ O) ｍ -R ６ (R ６ is a hydrogen atom or C １－６and m represents an integer of 3 to 100.] or a pharmaceutically acceptable salt thereof.

[0012] [Item 2] The compound according to Item 1, or a pharmaceutically acceptable salt thereof, wherein X is methylene.

[0013] [Section 3] R １ But C １－３ Item 3. The compound according to Item 1 or 2, wherein the alkyl is optionally substituted with 1 to 3 of the same or different halogens, or a pharmaceutically acceptable salt thereof.

[0014] [Section 4] R １ Item 4. The compound according to Item 3, or a pharmaceutically acceptable salt thereof, wherein is methyl.

[0015] [Section 5] R ４ is a hydrogen atom, hydroxy, C １－３ Alkyl, or C １－３ Item 5. The compound according to any one of Items 1 to 4, which is alkoxy, or a pharmaceutically acceptable salt thereof.

[0016] [Section 6] R ４ Item 6. The compound according to Item 5, or a pharmaceutically acceptable salt thereof, wherein is a hydrogen atom, hydroxy, or methoxy.

[0017] [Section 7] R ２ But C １－６ Item 7. The compound according to any one of Items 1 to 6, wherein R is alkyl, or a pharmaceutically acceptable salt thereof.

[0018] [Section 8] R ３ is a hydrogen atom, or C １－３ Item 8. The compound according to any one of Items 1 to 7, wherein the alkyl is alkyl (which may be substituted with 1 to 3 hydroxyl groups), or a pharmaceutically acceptable salt thereof.

[0019] [Item 9] L is -O-, -NR Ｙ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR Ｙ -, -NR Ｙ C(O)-, -CH ２ NR Ｙ -, -CH２ O-, -OC(O)O-, -NR ７ C(O)O-, -OC(O)NR Ｙ -, -NR ７ C(O)NR Ｙ -, -OC(S)NR Ｙ - or -NR ７ C(S)NR Ｙ - (where R ７ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; R Ｙ represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or Y ２ (Y ２ is -(CH ２ CH ２ O) ｎ -R ８ (R ８ represents a hydrogen atom or alkyl having 1 to 6 carbon atoms, and n represents an integer of 3 to 100), or a pharmaceutically acceptable salt thereof.

[0020] [Item 10] L is —C(O)NR Ｙ -, -CH ２ NR Ｙ -, -C(O)O- or -CH ２ Item 10. The compound according to item 9, wherein R is O—, or a pharmaceutically acceptable salt thereof.

[0021] [Item 11] L is —C(O)NR Ｙ - or -CH ２ NR Ｙ Item 10. The compound according to Item 9, wherein - or a pharmaceutically acceptable salt thereof.

[0022] [Item 12] L is -CH ２ NR Ｙ - and R Ｙ is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or Y ２ Item 10. The compound according to Item 9, wherein:

[0023] [Item 13] L is -CH ２ NR Ｙ - and R ＹItem 10. The compound according to Item 9, or a pharmaceutically acceptable salt thereof, wherein is a hydrogen atom or alkyl having 1 to 6 carbon atoms.

[0024] [Section 14] Y １ But -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ Item 14. The compound according to any one of Items 1 to 13, wherein m is alkyl and m is an integer of 3 to 40, or a pharmaceutically acceptable salt thereof.

[0025] [Section 15] Y １ But -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ Item 15. The compound according to item 14, wherein m is alkyl and m is an integer of 3 to 20, or a pharmaceutically acceptable salt thereof.

[0026] [Term 16] Formula (2): or Equation (3): [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl (the alkyl may be substituted with 1 to 3 hydroxy groups); R ４ is a hydrogen atom, hydroxy, or methoxy; L is —CH ２ NR Ｙ -, -C(O)NR Ｙ -, -C(O)O- or -CH ２ O- and R Ｙ is a hydrogen atom, C １－６ Alkyl or Y ２ and Y １ is -(CH ２ CH ２ O) ｍ -R ６ and Y ２ is -(CH ２ CH ２ O)ｎ -R ８ and R ６ is a hydrogen atom or C １－６ alkyl, R ８ is a hydrogen atom or C １－６ and m and n are each independently an integer of 3 to 40.] or a pharmaceutically acceptable salt thereof.

[0027] [Term 17] Formula (2): or Equation (3): [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl (the alkyl may be substituted with 1 to 3 hydroxy groups); R ４ is a hydrogen atom, hydroxy, or methoxy; L is —CH ２ NR Ｙ - and R Ｙ is a hydrogen atom or C １－６ is alkyl, and Y １ is -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ and m is an integer of 3 to 20.] or a pharmaceutically acceptable salt thereof.

[0028] [Term 18] Formula (2): [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl, which may be substituted with one hydroxyl; R ４ is a hydrogen atom or methoxy, and L is —CH ２ NR Ｙ - and R Ｙ is a hydrogen atom or C １－６ is alkyl, and Y １is -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ and m is an integer of 3 to 40.] or a pharmaceutically acceptable salt thereof.

[0029] [Item 19] The compound according to Item 1 or a pharmaceutically acceptable salt thereof, selected from the following compound group: 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 1), 1-{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxyphenyl}-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 2), 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 3), 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-hydroxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 4), 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(20-hydroxy-3,6,9,12,15,18-hexaoxaicosan-1-yl)-3-methoxybenzamide (Example 5), 2,5,8,11-tetraoxatridecan-13-yl 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoate (Example 6), 5-{[2-methoxy-4-(2,5,8,11,14-pentaoxapentadecan-1-yl)phenyl]methyl}-6-methyl-N ４-pentylpyrimidine-2,4-diamine (Example 7), 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29-nonaoxa-2-azahentriacontan-31-ol (Example 8), 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71-tricosaoxa-2-azatriheptacontan-73-ol (Example 9), 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92,95,98,101,104,107-pentatriacontaoxa-2-azanonahexan-109-ol (Example 10), and 12-[(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3,6,9,15,18,21-hexaoxa-12-azatricosane-1,23-diol (Example 11).

[0030] [Item 20] The compound according to Item 1 or a pharmaceutically acceptable salt thereof, selected from the following compound group: 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 1), 1-{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxyphenyl}-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 2), 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 3), and 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-hydroxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol (Example 4).

[0031] [Item 21] A pharmaceutical composition comprising the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0032] [Item 22] The pharmaceutical composition according to Item 21, which is an emulsion formulation, an oily suspension formulation, a hydrogel formulation, or a lipid formulation.

[0033] [Item 23] The pharmaceutical composition according to Item 21, which is an emulsion formulation.

[0034] [Item 24] The pharmaceutical composition according to Item 23, wherein the emulsion formulation is a water-in-oil emulsion.

[0035] [Item 25] The pharmaceutical composition according to Item 24, wherein the emulsion formulation contains (1) ethyl oleate, octyldodecyl myristate, sorbitan monooleate, glyceryl monooleate, polyoxyethylene hydrogenated castor oil 20, glycerin, and sodium dihydrogen phosphate, or (2) Montanide ISA 51VG.

[0036] [Item 26] The pharmaceutical composition according to Item 21, which is a lipid formulation.

[0037] [Item 27] ​​The pharmaceutical composition according to Item 26, wherein the lipid formulation is a liposome formulation containing a phospholipid.

[0038] [Item 28] The pharmaceutical composition according to Item 26 or 27, wherein the lipid formulation is a liposome formulation containing sterols.

[0039] [Item 29] The pharmaceutical composition according to Item 28, wherein the sterol is cholesterol.

[0040] [Item 30] The pharmaceutical composition according to any one of Items 27 to 29, wherein the liposome preparation comprises one or more additives selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.

[0041] [Item 31] The pharmaceutical composition according to any one of Items 21 to 30, further comprising an antigen.

[0042] [Item 32] The pharmaceutical composition according to Item 31, wherein the antigen is a pathogen-derived antigen or a tumor antigen.

[0043] [Item 33] The pharmaceutical composition of Item 31, wherein the antigen is a tumor antigen.

[0044] [Item 34] The pharmaceutical composition of Item 33, wherein the tumor antigen is a tumor antigen peptide.

[0045] [Item 35] The tumor antigen peptide is selected from the group consisting of RMFPNAPYL (SEQ ID NO: 1), ALLPAVPSL (SEQ ID NO: 8), SLGEQQYSV (SEQ ID NO: 9), RVPGVAPTL (SEQ ID NO: 10), VLDFAPPGA (SEQ ID NO: 4), CMTWNQMNL (SEQ ID NO: 11), CYTWNQMNL (SEQ ID NO: 2), TYAGCLSQIF (SEQ ID NO: 18), and the formula (4): [wherein the bond between C and C represents a disulfide bond], and Formula (5): [wherein the bond between C and C represents a disulfide bond.] or a pharmaceutically acceptable salt thereof, and one or more peptides represented by an amino acid sequence selected from the group consisting of WAPVLDFAPPGASAYGSL (SEQ ID NO: 3), CWAPVLDFAPPGASAYGSL (SEQ ID NO: 12), WAPVLDFAPPGASAYGSLC (SEQ ID NO: 13), CNKRYFKLSHLQMHSRKHTG (SEQ ID NO: 14), CNKRYFKLSHLQMHSRKH (SEQ ID NO: 15), CNKRYFKLSHLQMHSRK (SEQ ID NO: 16), and KRYFKLSHLQMHSRKH (SEQ ID NO: 17), or a pharmaceutically acceptable salt thereof.

[0046] [Item 36] The tumor antigen peptide is selected from the group consisting of RMFPNAPYL (SEQ ID NO: 1), ALLPAVPSL (SEQ ID NO: 8), SLGEQQYSV (SEQ ID NO: 9), RVPGVAPTL (SEQ ID NO: 10), VLDFAPPGA (SEQ ID NO: 4), CMTWNQMNL (SEQ ID NO: 11), CYTWNQMNL (SEQ ID NO: 2), and the formula (4): [wherein the bond between C and C represents a disulfide bond], or a pharmaceutically acceptable salt thereof, and one or more peptides represented by amino acid sequences selected from the group consisting of WAPVLDFAPPGASAYGSL (SEQ ID NO: 3), CWAPVLDFAPPGASAYGSL (SEQ ID NO: 12), WAPVLDFAPPGASAYGSLC (SEQ ID NO: 13), CNKRYFKLSHLQMHSRKHTG (SEQ ID NO: 14), CNKRYFKLSHLQMHSRKH (SEQ ID NO: 15), CNKRYFKLSHLQMHSRK (SEQ ID NO: 16), and KRYFKLSHLQMHSRKH (SEQ ID NO: 17), or a pharmaceutically acceptable salt thereof.

[0047] [Item 37] The tumor antigen peptide is represented by formula (4): [wherein the bond between C and C represents a disulfide bond] or a pharmaceutically acceptable salt thereof, and a peptide represented by the amino acid sequence of SEQ ID NO: 3: WAPVLDFAPPGASAYGSL or a pharmaceutically acceptable salt thereof.

[0048] [Item 38] A vaccine adjuvant comprising the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0049] [Item 39] The vaccine adjuvant of Item 38, which is a vaccine adjuvant for a cancer vaccine.

[0050] [Item 40] The compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof, for use as a vaccine adjuvant.

[0051] [Item 41] The compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof, which is used as a vaccine adjuvant for a cancer vaccine.

[0052] [Item 42] A CTL inducer comprising the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0053] [Item 43] An immunostimulant comprising the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0054] [Item 44] A method for inducing CTLs in a mammal, which comprises administering to the mammal the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0055] [Item 45] A method for enhancing CTL induction in a mammal, which comprises administering to the mammal the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0056] [Item 46] A method for enhancing a specific immune response to an antigen in a mammal, which comprises administering to the mammal the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof.

[0057] [Item 47] Use of the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof for producing a vaccine adjuvant.

[0058] [Item 48] Use of the compound according to any one of Items 1 to 20 or a pharmaceutically acceptable salt thereof for producing a vaccine adjuvant for a cancer vaccine.

[0059] [Item 49] A kit comprising: a) a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound according to any one of items 1 to 20 or a pharmaceutically acceptable salt thereof; and b) an antigen or a pharmaceutical composition comprising the antigen.

[0060] [Item 50] A kit comprising: a) a compound represented by formula (1) of item 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof; and b) a tumor antigen or a pharmaceutical composition containing a tumor antigen.

[0061] [Item 51] A kit comprising: a) a compound represented by formula (1) of item 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof; and b) a pathogen-derived antigen or a pharmaceutical composition containing a pathogen-derived antigen.

[0062] FIG. 1 shows the in vivo CTL induction ability against SEQ ID NO: 2 in Test Example 3, which was obtained by adding the compound synthesized in Example 1 or Reference Example 12 to a cocktail vaccine containing a compound represented by Formula 4, a peptide represented by SEQ ID NO: 3, and Montanide ISA 51 VG, as compared with HLA-A. ＊ 2 shows the results of an IFNγ ELISPOT assay using 24:02 transgenic mice. ＊3 shows the results of administering a cocktail vaccine containing a compound represented by SEQ ID NO: 4, a peptide represented by SEQ ID NO: 3, and Montanide ISA 51 VG, to which the compound synthesized in Example 1 was added, 7 days before and 7 days after transplantation of MCA-A24 / Kb-WT1 tumor cells into 24:02 transgenic mice, and measuring the tumor volume 27 days after transplantation. Fig. 3 shows the results of measuring the in vivo CTL induction ability against SEQ ID NO: 6 in Test Example 6, using a vaccine containing a mixture of the peptide represented by SEQ ID NO: 6 and Montanide ISA 51 VG, to which the compound synthesized in Example 1 was added, in terms of HLA-A ＊ 4 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 7, the vaccine was prepared by adding the compound synthesized in Example 1 to a vaccine containing a mixture of the peptide represented by SEQ ID NO: 5 and Montanide ISA 51 VG, and the in vivo CTL induction ability against SEQ ID NO: 5 was evaluated using HLA-A ＊ 5 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. Figure ... ＊ 6 shows the results of an IFNγ ELISPOT assay using 24:02 transgenic mice. In Test Example 9, a cocktail vaccine was prepared by mixing the compound represented by Formula 4, the peptide represented by SEQ ID NO: 3, and Montanide ISA 51 VG, to which the compound synthesized in Examples 8, 9, or 10, or Reference Example 12, was added. The results are shown in Fig. 6, which shows the in vivo CTL induction ability against SEQ ID NO: 1 in HLA-A ＊7 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. Figure 7 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 9, a cocktail vaccine was prepared by mixing the compound represented by Formula 4, the peptide represented by SEQ ID NO: 3, and Montanide ISA 51 VG, to which the compound synthesized in Examples 8, 9, or 10, or Reference Example 12, was added. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊ 8 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 10, a cocktail vaccine was prepared by mixing the compound represented by Formula 4, the peptide represented by SEQ ID NO: 3, and Montanide ISA 51 VG, to which the compound synthesized in Example 11 was added. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 9 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 10, a cocktail vaccine was prepared by mixing the compound represented by SEQ ID NO: 4, the peptide represented by SEQ ID NO: 3, and Montanide ISA 51 VG, to which the compound synthesized in Example 11 was added. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊ 10 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 11, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an emulsion composition 1 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 11 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 11, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an emulsion composition 1 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊12 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 12, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an emulsion composition 2 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 13 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 12, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an emulsion composition 2 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊ 14 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 13, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an emulsion composition 3 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 15 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 14, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an oily suspension of the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 16 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 14, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to an oily suspension of the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊17 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 15, a cocktail vaccine was prepared by hydrogel formulation of the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3, to which the compound synthesized in Example 1 was added. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 18 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 15, a cocktail vaccine was prepared by hydrogel formulation of the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3, to which the compound synthesized in Example 1 was added. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊ 19 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 16, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to a liposome preparation 1 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 20 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 17, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to a liposome preparation 2 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 21 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 17, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to a liposome preparation 2 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 4 was evaluated using HLA-A ＊22 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 18, a cocktail vaccine was prepared by adding the compound synthesized in Example 1 to a liposome preparation 3 containing the compound represented by Formula 4 and the peptide represented by SEQ ID NO: 3. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 23 shows the results of an IFNγ ELISPOT assay using 02:01 transgenic mice. In Test Example 19, a cocktail vaccine was prepared by mixing the peptide represented by SEQ ID NO: 1 and the peptide represented by SEQ ID NO: 17 with a pre-emulsified composition, to which the compound synthesized in Example 1 was added. The in vivo CTL induction ability against SEQ ID NO: 1 was evaluated using HLA-A ＊ 02:01 / HLA-DRB1 ＊ 24 shows the results of an IFNγ ELISPOT assay using 01:01 transgenic mice. In Test Example 19, a cocktail vaccine was prepared by mixing the peptide represented by SEQ ID NO: 1 and the peptide represented by SEQ ID NO: 17 with a pre-emulsified composition, to which the compound synthesized in Example 1 was added. The in vivo ability of the vaccine to induce helper T cells against SEQ ID NO: 17 was examined using HLA-A ＊ 02:01 / HLA-DRB1 ＊ 25 shows the results of an IFNγ ELISPOT assay using 01:01 transgenic mice. In Test Example 20, the compound represented by Formula 5 was mixed with a pre-emulsified composition to which the compound synthesized in Example 1 was added. The in vivo CTL induction ability against SEQ ID NO: 2 was evaluated using HLA-A ＊Figure 26 shows the results of an IFNγ ELISPOT assay using 24:02 transgenic mice. Figure 26 shows the results of measuring the spleen weight of mice administered with a vaccine in Test Examples 11 to 13, in which the compound represented by formula 4 and the peptide represented by sequence number 3 were combined in an emulsion composition and the compound synthesized in Example 1 was added. Figure 27 shows the results of measuring the spleen weight of mice administered with a vaccine in Test Examples 14 and 15, in which the compound represented by formula 4 and the peptide represented by sequence number 3 were combined in an oil suspension and a hydrogel formulation and the compound synthesized in Example 1 was added. Figure 28 shows the results of measuring the spleen weight of mice administered with a vaccine in Test Examples 16 to 18, in which the compound represented by formula 4 and the peptide represented by sequence number 3 were combined in a liposome formulation and the compound synthesized in Example 1 was added.

[0063] The terms used in this specification are explained below.

[0064] In this specification, the number of substituents in a group defined as "optionally substituted" or "substituted" is not particularly limited as long as substitution is possible. Furthermore, unless otherwise specified, the description of each group also applies when that group is a part or substituent of another group.

[0065] In the present specification, examples of "halogen" include fluorine, chlorine, bromine, and iodine. Preferred are fluorine and chlorine, and more preferred is fluorine.

[0066] "C １－６ "Alkyl" means a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms. １－４ alkyl", and more preferably "C １－３ "C alkyl" is an example. １－６Specific examples of "alkyl" include methyl, ethyl, propyl, 1-methylethyl, butyl, 2-methylpropyl, 1-methylpropyl, 1,1-dimethylethyl, pentyl, 3-methylbutyl, 2-methylbutyl, 2,2-dimethylpropyl, 1-ethylpropyl, 1,1-dimethylpropyl, hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, and 1-methylpentyl. １－４ Specific examples of "alkyl" include "C １－６ Specific examples of "alkyl" include those having 1 to 4 carbon atoms. １－３ Specific examples of "alkyl" include "C １－６ Specific examples of "alkyl" include those having 1 to 3 carbon atoms.

[0067] "C １－６ "Alkoxy" means "C １－６ It means "alkyloxy" and "C １－６ The "C alkyl" moiety is １－６ "C" is synonymous with "alkyl." １－６ The "alkoxy" is preferably "C １－４ Alkoxy" is preferred, and "C １－３ "Alkoxy" is an example. １－６ Specific examples of "alkoxy" include, for example, methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 2-methylpropoxy, 1-methylpropoxy, 1,1-dimethylethoxy, pentyloxy, 3-methylbutoxy, 2-methylbutoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, 1,1-dimethylpropoxy, hexyloxy, 4-methylpentyloxy, 3-methylpentyloxy, 2-methylpentyloxy, 1-methylpentyloxy, 3,3-dimethylbutoxy, 2,2-dimethylbutoxy, 1,1-dimethylbutoxy, and 1,2-dimethylbutoxy. １－４ Specific examples of "alkoxy" include "C １－６ Specific examples of "alkoxy" include those having 1 to 4 carbon atoms. １－３ Specific examples of "alkoxy" include "C １－６ Specific examples of "alkoxy" include those having 1 to 3 carbon atoms.

[0068] The term "linker" refers to a divalent group having two bonds within a functional group. Examples of the divalent group include C １－６ Alkylene, C ２－７ Alkenylene, C ２－７ Alkynylene, C ３－１０ Cycloalkylene, C ６－１０ Arylene, C ５－１０ Examples of the linker include heteroarylene, ether, amine, carbonyl, ester, amide, carbonate, carbamate, thiocarbamate, and thiourea. These examples may also be used in combination as a divalent group. Preferred examples of the linker include -O-, -NR Ｙ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR Ｙ -, -NR Ｙ C(O)-, -CH ２ NR Ｙ -, -CH ２ O-, -OC(O)O-, -NR ７ C(O)O-, -OC(O)NR Ｙ -, -NR ７ C(O)NR Ｙ -, -OC(S)NR Ｙ - or -NR ７ C(S)NR Ｙ - (wherein, R Ｙ , R ７ is as defined in item 9), and more preferably —C(O)NR Ｙ - or -CH ２ NR Ｙ -, and more preferably -CH ２ NR Ｙ Of the two bonds of the linker shown as specific examples here, the left bond is bonded to the benzene ring in the compound of formula (1), and the right bond is bonded to Y １ That is, the linker L is "-CH ２ NR Ｙ -", then the compound of formula (1) will have the following structure:

[0069] "C １－６"Alkylene" means a straight or branched chain saturated hydrocarbon having 1 to 6 carbon atoms. １－６ Specific examples of "alkylene" include methylene, ethylene, propylene, 1-methylethylene, butylene, 2-methylpropylene, 1-methylpropylene, 1,1-dimethylethylene, pentylene, 3-methylbutylene, 2-methylbutylene, 2,2-dimethylpropylene, 1-ethylpropylene, 1,1-dimethylpropylene, hexylene, 4-methylpentylene, and 3-methylpentylene, and preferably methylene or ethylene.

[0070] "C ２－７ "Alkenylene" means a straight or branched chain unsaturated hydrocarbon having 2 to 7 carbon atoms and containing 1 to 3 double bonds. ２－７ Specific examples of "alkenylene" include vinylene, propenylene, methylpropenylene, butenylene, methylbutenylene, pentenylene, hexenylene, and heptenylene, and preferred are vinylene and propenylene.

[0071] "C ２－７ "Alkynylene" means a straight or branched chain unsaturated hydrocarbon having 2 to 7 carbon atoms and containing one triple bond. ２－７ Specific examples of "alkynylene" include ethynylene, propynylene, methylpropynylene, butynylene, methylbutynylene, pentenylene, hexynylene, and heptynylene, and preferred are ethynylene and propynylene.

[0072] "C ３－１０ "Cycloalkylene" means a cyclic alkylene having 3 to 10 carbon atoms, and includes those having a partially bridged structure. ３－１０ Specific examples of "cycloalkylene" include cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, cyclooctylene, and adamantylene, and preferably cyclopropylene or cyclobutylene.

[0073] "C６－１０ "Arylene" means an aromatic hydrocarbon having 6 to 10 carbon atoms. ６－１０ Specific examples of "arylene" include phenylene, 1-naphthylene, and 2-naphthylene, and preferred is phenylene.

[0074] "C ５－１０ "Heteroarylene" means a monocyclic 5- to 7-membered aromatic heterocycle or a bicyclic 8- to 10-membered aromatic heterocycle containing 1 to 4 atoms independently selected from the group consisting of nitrogen, oxygen, and sulfur atoms. ５－１０ Specific examples of "heteroarylene" include pyridylene, pyridazinylene, isothiazolylene, pyrrolylene, furylene, thienylene, thiazolylene, imidazolylene, pyrimidinylene, thiadiazolylene, pyrazolylene, oxazolylene, isoxazolylene, pyrazinylene, triazolylene, imidazolidinylene, oxadiazolylene, triazolylene, tetrazolylene, indolylene, indazolylene, quinolylene, isoquinolylene, benzofuranylene, benzothienylene, benzoxazolylene, benzothiazolylene, benzisoxazolylene, benzisothiazolylene, benzotriazolylene, benzimidazolylene, and 6,11-dihydrodibenzo[b,e]thiepinylene. Preferred are pyridylene, pyrimidinylene, quinolylene, and isoquinolylene, and more preferred are pyridylene, furylene, and thienylene.

[0075] Among the compounds of the present invention represented by formula (1), X, Y １ , Y ２ , R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R Ｙ Preferred examples of L, m, and n are as follows, but the technical scope of the present invention is not limited to the range of compounds listed below.

[0076] X is preferably methylene, an oxygen atom, or NR ５ (R ５ is a hydrogen atom or C １－６ and more preferably, methylene.

[0077] Y １ As for -(CH ２ CH ２ O) ｍ -R ６ Examples include:

[0078] Y ２ As for -(CH ２ CH ２ O) ｎ -R ８ Examples include:

[0079] R １ Preferably, C １－６ alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens). More preferably, C １－６ Examples of alkyl include methyl, ethyl, propyl, 1-methylethyl, butyl, 2-methylpropyl, 1-methylpropyl, and 1,1-dimethylethyl, and more preferably, methyl.

[0080] R ２ is preferably (1) a hydrogen atom, or (2) C １－６ alkyl (the alkyl may be substituted with 1 to 3 substituents independently selected from halogen and hydroxy). More preferably, it is a hydrogen atom or C １－６ alkyl, more preferably C １－６ and even more preferably, C ３－４ Examples of alkyl include the following.

[0081] R ３ is preferably (1) a hydrogen atom, or (2) C １－６ alkyl (the alkyl may be substituted with 1 to 3 substituents independently selected from halogen and hydroxy);

[0082] R ３is more preferably (1) a hydrogen atom, or (2) C １－３ alkyl (the alkyl may be substituted with 1 to 3 hydroxy groups).

[0083] R ３ More preferably, (1) a hydrogen atom, or (2) C １－３ and alkyl (which may be substituted with one hydroxyl).

[0084] R ３ Even more preferably, (1) a hydrogen atom, or (2) C １－２ and alkyl (which may be substituted with one hydroxyl).

[0085] R ４ is preferably (1) a hydrogen atom, (2) a halogen, (3) a hydroxyl, or (4) C １－６ Alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens) (5) C １－６ (6) alkoxy (the alkoxy may be substituted with 1 to 3 of the same or different halogens); or (7) cyano.

[0086] R ４ More preferably, (1) a hydrogen atom, (2) a halogen, (3) a hydroxyl, or (4) C １－６ (5) alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens), or １－６ alkoxy (the alkoxy may be substituted with 1 to 3 of the same or different halogens).

[0087] R ４ More preferably, it is a hydrogen atom, hydroxy, C １－３ Alkyl, or C １－３ Examples include alkoxy.

[0088] R ４ More preferably, it is a hydrogen atom, hydroxy, or methoxy, even more preferably it is hydroxy or methoxy, and most preferably it is a hydrogen atom or methoxy.

[0089] R ５ is preferably a hydrogen atom or C １－３ More preferably, it is a hydrogen atom, methyl, ethyl, or propyl.

[0090] R ６ and R ８ are preferably each independently a hydrogen atom or C １－３ Examples of the alkyl include, more preferably, each independently includes a hydrogen atom, methyl, ethyl, or propyl, and even more preferably, includes a hydrogen atom or methyl.

[0091] R ７ is preferably a hydrogen atom or C １－３ More preferred are hydrogen atoms, methyl, ethyl, and propyl.

[0092] L is preferably (1)-O- (2)-NR Ｙ - (3)-C(O)- (4)-C(O)O- (5)-OC(O)- (6)-C(O)NR Ｙ - (7) -NR Ｙ C(O)-(8)-CH ２ NR Ｙ - (9) -CH ２ O- (10)-OC(O)O- (11)-NR ７ C(O)O- (12)-OC(O)NR Ｙ - (13)-NR ７ C(O)NR Ｙ - (14)-OC(S)NR Ｙ -, or (15)-NR ７ C(S)NR Ｙ - are listed.

[0093] L is more preferably (1)-O- (2)-NR Ｙ - (3)-C(O)- (4)-C(O)O- (5)-OC(O)- (6)-C(O)NR Ｙ - (7) -NR Ｙ C(O)-(8)-CH ２ NR Ｙ-, or (9) -CH ２ O- is an example.

[0094] More preferably, L is (1) —C(O)NR Ｙ -, (2)-CH ２ NR Ｙ -, (3) -C(O)O-, or (4) -CH ２ O- is an example.

[0095] Even more preferably, L is (1) —C(O)NR Ｙ -, or (2) -CH ２ NR Ｙ - are listed.

[0096] L is most preferably —CH ２ NR Ｙ - are some examples.

[0097] X, L, and R on the benzene ring ４ Preferred substitution positions include the following (1a) or (1aa):

[0098] R Ｙ is preferably a hydrogen atom, C １－６ Alkyl or Y ２ More preferably, it is a hydrogen atom or C １－６ Included are alkyl, and even more preferably, a hydrogen atom, methyl, ethyl, or propyl.

[0099] R Ｙ In another embodiment, preferably, a hydrogen atom, C １－６ Alkyl or Y ２ More preferably, a hydrogen atom, methyl, Y ２ Examples include:

[0100] Preferably, m and n are each independently an integer of 3 to 40, more preferably an integer of 4 to 40, and even more preferably an integer of 4 to 36.

[0101] In another embodiment, m and n each independently represent an integer of 3 to 40, preferably an integer of 3 to 20, and more preferably an integer of 5 to 20.

[0102] Preferred embodiments of the compound represented by formula (1) include the following compounds or pharmaceutically acceptable salts thereof:

[0103] Among the compounds represented by formula (1), the following (A) is a preferred embodiment: (A) X is methylene, an oxygen atom, a sulfur atom, SO, SO ２ , or NR ５ and R １ But C １－６ Alkyl (the alkyl is not substituted with halogen, hydroxy, C １－６ and R is optionally substituted with 1 to 5 substituents independently selected from the group consisting of alkoxy; ２ and R ３ are each independently: (1) a hydrogen atom, or (2) C １－６ Alkyl (the alkyl is not substituted with halogen, hydroxy, C １－６ and R is optionally substituted with 1 to 5 substituents independently selected from the group consisting of alkoxy; ４ (1) Hydrogen atom (2) Halogen (3) Hydroxy (4) C １－６ Alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens) (5) C １－６ (6) alkoxy (the alkoxy may be substituted with 1 to 3 of the same or different halogens), or (7) cyano; R ５ is (1) a hydrogen atom, or (2) C １－６ alkyl; R ６ and R ８ are each independently: (1) a hydrogen atom, or (2) C １－６ alkyl; R ７ is (1) a hydrogen atom, or (2) C １－６ alkyl; L is (1)-O- (2)-NR Ｙ - (3)-C(O)- (4)-C(O)O- (5)-OC(O)- (6)-C(O)NR Ｙ - (7) -NR Ｙ C(O)-(8)-CH ２ NRＹ - (9) -CH ２ O- (10)-OC(O)O- (11)-NR ７ C(O)O- (12)-OC(O)NR Ｙ - (13)-NR ７ C(O)NR Ｙ - (14)-OC(S)NR Ｙ -, or (15)-NR ７ C(S)NR Ｙ - and R Ｙ (1) Hydrogen atom (2) C １－６ alkyl, or (3) Y ２ and Y １ But -(CH ２ CH ２ O) ｍ -R ６ and Y ２ But -(CH ２ CH ２ O) ｎ -R ８ and m and n are each independently an integer of 3 to 100; or a pharmaceutically acceptable salt thereof.

[0104] Among the compounds represented by formula (1), the following (B) is a preferred embodiment: (B) X is methylene, an oxygen atom, or NR ５ and R １ But C １－６ alkyl, which may be substituted with 1 to 3 of the same or different halogens; R ２ is (1) a hydrogen atom, or (2) C １－６ alkyl; R ３ is (1) a hydrogen atom, or (2) C １－６ alkyl, which may be substituted with 1 to 3 hydroxy groups; R ４ (1) Hydrogen atom (2) Halogen (3) Hydroxy (4) C １－６ (5) alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens), or １－６alkoxy, which may be substituted with 1 to 3 of the same or different halogens; R ５ is (1) a hydrogen atom, or (2) C １－３ alkyl; R ６ and R ８ are each independently: (1) a hydrogen atom, or (2) C １－３ alkyl; L is (1)-O- (2)-NR Ｙ - (3)-C(O)- (4)-C(O)O- (5)-OC(O)- (6)-C(O)NR Ｙ - (7) -NR Ｙ C(O)-(8)-CH ２ NR Ｙ -, or (9) -CH ２ O-; R Ｙ (1) hydrogen atom, (2) C １－６ alkyl, or (3) Y ２ and Y １ But -(CH ２ CH ２ O) ｍ -R ６ and Y ２ But -(CH ２ CH ２ O) ｎ -R ８ and m and n are each independently an integer of 3 to 40; or a pharmaceutically acceptable salt thereof.

[0105] Among the compounds represented by formula (1), the following (C) is more preferred: (C) X is methylene; １ is methyl, ethyl, propyl, 1-methylethyl, butyl, 2-methylpropyl, 1-methylpropyl or 1,1-dimethylethyl; R ２ But C １－６ alkyl; R ３ is (1) a hydrogen atom, or (2) C １－３ alkyl, which may be substituted with one hydroxy; R ４ (1) Hydrogen atom (2) Hydroxy (3) C１－３ alkyl, or (4) C １－３ alkoxy; R ６ and R ８ are each independently a hydrogen atom, methyl, ethyl, or propyl; L is (1) —C(O)NR Ｙ -, (2)-CH ２ NR Ｙ -, (3) -C(O)O-, or (4) -CH ２ O-; R Ｙ is a hydrogen atom, methyl, ethyl, propyl or Y ２ and Y １ But -(CH ２ CH ２ O) ｍ -R ６ and Y ２ But -(CH ２ CH ２ O) ｎ -R ８ and m and n are each independently an integer of 3 to 40; or a pharmaceutically acceptable salt thereof.

[0106] Another embodiment of the compound represented by formula (1) includes the following (D): (D) a compound represented by formula (2) or (3) or a pharmaceutically acceptable salt thereof; [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl (the alkyl may be substituted with 1 to 3 hydroxy groups); R ４ is a hydrogen atom, hydroxy, or methoxy; L is —CH ２ NR Ｙ -, -C(O)NR Ｙ -, -C(O)O- or -CH ２ O- and R Ｙ is a hydrogen atom, methyl, ethyl, propyl or Y ２ and Y １ is -(CH ２ CH ２ O) ｍ -R ６and Y ２ But -(CH ２ CH ２ O) ｎ -R ８ and R ６ and R ８ are each independently a hydrogen atom, methyl, ethyl, or propyl, and m and n are each independently an integer of 3 to 40.

[0107] Another embodiment of the compound represented by formula (1) includes the following (E): (E) a compound represented by formula (2) or a pharmaceutically acceptable salt thereof; [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl, which may be substituted with one hydroxyl; R ４ is a hydrogen atom or methoxy, and L is —CH ２ NR Ｙ - and R Ｙ is a hydrogen atom, methyl, ethyl or propyl; Y １ is -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom, methyl, ethyl, or propyl, and m is an integer of 4 to 36.

[0108] The method for producing the compound of the present invention is described below. The compound of the present invention represented by formula (1) or a pharmaceutically acceptable salt thereof can be produced, for example, by the following production method. Production Method A-1: ​​In the compound of the present invention represented by formula (1) or a pharmaceutically acceptable salt thereof, -CR Ａ１ R Ａ２ NR Ｙ - or -CR Ａ１ R Ａ２ The compound (a1-2) having a linker represented by O- is produced by the following method. (In the formula, R １ , R ２ , R ３, R ４ , R ５ , X, and Y １ is as defined in Item 1, and R Ａ１ and R Ａ２ are each independently a hydrogen atom, C １－６ represents an alkyl group of the formula: LG ａ１ represents a leaving group, Nu ａ１ represents a nucleophile, L ａ１ represents the linker produced by this process)

[0109] In this step, the leaving group LG ａ１ and the nucleophile Nu ａ１ -Y １ In the presence or absence of a suitable base, compound (a1-1) is reacted with Nu ａ１ -Y １ The leaving group is not particularly limited, but preferably includes fluorine, chlorine, bromine, iodine, methanesulfonate, ethanesulfonate, p-toluenesulfonate, etc., and more preferably includes chlorine, bromine, and methanesulfonate. The nucleophile is not particularly limited, but preferably includes an amine (the amine is R defined in Item 9). Ｙ amines (which may be substituted with R defined in Item 9) and the like. Ｙ Examples of suitable solvents include an alcohol, which may be substituted with , and an alcohol. The base used in this step is selected from the bases exemplified below, and preferably sodium hydride or potassium hydride. The solvent used in this step is selected from the solvents exemplified below, and preferably DMF. The reaction time is usually about 5 minutes to about 48 hours, and preferably about 10 minutes to about 24 hours. The reaction temperature is usually about -78°C to about 100°C, and preferably about 0°C to about 100°C.

[0110] Production Method A-2 Among the compounds of the present invention represented by formula (1) or pharmaceutically acceptable salts thereof, —O—, —NR Ｙ -, -C(O)O-, -CH ２ NR Ｙ - or -CH２ The compound (a2-2) having a linker represented by O- is produced by the following method. (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , X, and Y １ is as defined in item 1, and LG ａ２ represents a leaving group, Nu ａ２ represents a nucleophile, L ａ２ represents the linker produced by this process)

[0111] In this step, the leaving group LG ａ２ and the nucleophile Nu ａ２ In the presence or absence of a suitable base, compound (a2-1) is reacted with LG ａ２ -Y １ This is a process for obtaining compound (a2-2) from ａ２ , Nu ａ２ and L ａ２ are the same as the leaving group, nucleophile, and linker described in Production Method A-1, respectively. Various reaction conditions are in accordance with those described in Production Method A-1.

[0112] The method for producing the compound of the present invention is described below. The compound of the present invention represented by formula (1) can be produced, for example, by the following production method. Production Method B-1 In the compound of the present invention represented by formula (1) or a pharmaceutically acceptable salt thereof, -CH ２ NR Ｙ The compound (b1-2) having a linker represented by - can be produced by the following method. (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , X, and Y １ is as defined in Item 1, and R Ｙ is as defined in item 9, and L ｂ１ represents the linker produced by this process)

[0113] This step is a reductive amination reaction between an aldehyde and an amine. In the presence of a suitable reducing agent, compound (b1-1) and R Ｙ -NH-Y １ This is a process for obtaining compound (b1-2) from the above. The reducing agent used in this process is not particularly limited, but preferred examples include sodium borohydride, triacetoxyborohydride, and picoline borane. The solvent used in this process is selected from the solvents exemplified below, and preferred examples include THF and chloroform. The reaction time is usually about 5 minutes to about 48 hours, and preferably about 10 minutes to about 24 hours. The reaction temperature is usually about −78° C. to about 100° C., and preferably about 0° C. to about 100° C.

[0114] Production Method B-2 Among the compounds of the present invention represented by formula (1) or pharmaceutically acceptable salts thereof, -NR Ｙ - or -CH ２ NR Ｙ The compound (b2-2) having a linker represented by - can be produced by the following method. (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , X, and Y １ is as defined in Item 1, and R Ｙ is as defined in item 9, and n ｂ is 0 or 1, and L ｂ２ represents the linker produced by this process)

[0115] This step is a reductive amination reaction between an aldehyde and an amine. １ This is a step of obtaining compound b2-2 from —CHO. Various reaction conditions are the same as those described in Production Method B-1.

[0116] The method for producing the compound of the present invention is described below. The compound of the present invention represented by formula (1) can be produced, for example, by the following production method. Production Method C-1 In the compound of the present invention represented by formula (1) or a pharmaceutically acceptable salt thereof, -O-, -NRＹ - or -NR Ｙ The compound (c1-2) having a linker represented by C(O)- is prepared by the following method. (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , X, and Y １ is as defined in item 1, and LG ｃ１ represents a leaving group, Nu ｃ１ represents a nucleophile, L ｃ１ represents the linker produced by this process)

[0117] In this step, the leaving group LG ｃ１ and the nucleophile Nu ｃ１ -Y １ In the presence of a suitable catalyst, and in the presence or absence of a suitable base, compound (c1-1) is reacted with a nucleophile, Nu ｃ１ -Y １ In this step, the catalyst may be a transition metal such as palladium, a salt thereof, a complex thereof, or a catalyst supported on a carrier such as a polymer. The leaving group in this step is not particularly limited, but preferred examples include boronic acid, boronic acid ester, halogen, trifluoromethanesulfonate, and more preferred examples include boronic acid, boronic acid ester, bromine atom, iodine atom, and trifluoromethanesulfonate. The nucleophile in this step is not particularly limited, but may be an amine (the amine is R defined in item 9). Ｙ Examples of suitable solvents include an alkyl group (which may be substituted with an alkyl group having 1 to 6 carbon atoms), an alcohol, an alkyl magnesium, an alkyl zinc, and an alkyl lithium, and more preferably an amine (which may be substituted with an alkyl group having 1 to 6 carbon atoms) and an alcohol. The solvent used in this step is selected from the solvents exemplified below, and preferably a dioxane-water mixed solvent. The reaction time is usually about 5 minutes to about 48 hours, and preferably about 10 minutes to about 24 hours. The reaction temperature is usually about −78° C. to about 100° C., and preferably about 0° C. to about 100° C.

[0118] The method for producing the compound of the present invention is described below: The compound of the present invention represented by formula (1) can be produced, for example, by the following production method.

[0119] Production Method D-1 Among the compounds of the present invention represented by formula (1) or pharmaceutically acceptable salts thereof, —C(O)O— or —C(O)NR Ｙ The compound (d1-2) having a linker is prepared by the following method. (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , X, and Y １ is as defined in Item 1, and Nu ｄ１ represents a nucleophile, L ｄ１ represents the linker produced by this process)

[0120] In this step, a compound (d1-1) having a carboxylic acid and Nu ｄ１ -Y １ In the presence of a suitable condensing agent, and in the presence or absence of a suitable base, compound (d1-1) is reacted with a nucleophile, Nu ｄ１ -Y １ In this step, the nucleophile is not particularly limited, but is preferably an amine (the amine is an amine having the R ＹExamples of suitable solvents include amines (which may be substituted with one alkyl group having 1 to 6 carbon atoms) and alcohols, and more preferably amines (which may be substituted with one alkyl group having 1 to 6 carbon atoms). The condensing agent used in this step is selected from condensing agents commonly used, and preferably HBTU, HATU, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (including hydrochloride). The base used in this step is selected from the bases exemplified below, and preferably tertiary alkylamines, and more preferably DIPEA and triethylamine. The solvent used in this step is selected from the solvents exemplified below, and preferably DMF, dichloromethane, chloroform, THF, and the like. The reaction time is usually about 5 minutes to about 48 hours, and preferably about 10 minutes to about 24 hours. The reaction temperature is usually about −78° C. to about 100° C., and preferably about 0° C. to about 100° C.

[0121] Production Method D-2 Among the compounds of the present invention represented by formula (1) or pharmaceutically acceptable salts thereof, —OC(O)— or —NR Ｙ The compound (d2-2) having a C(O)-linker is prepared by the following method. (In the formula R １ , R ２ , R ３ , R ４ , R ５ , X, and Y １ is as defined in Item 1, and Nu ｄ２ represents a nucleophile, L ｄ２ represents the linker produced by this process)

[0122] In this step, Y having a carboxylic acid １ -COOH and nucleophile Nu ｄ２ In the presence of a suitable condensing agent, in the presence or absence of a suitable base, a compound (d2-1) containing a nucleophile and an electrophilic agent Y １ This is a step of obtaining compound (d2-2) from —COOH. Various reaction conditions are the same as those described in step D-1.

[0123] The starting materials (a1-1, a2-1, b1-1, b2-1, c1-1, d1-1, d2-1) used in Production Methods A to D can be produced, for example, according to the method described in WO2009 / 067081.

[0124] The compound of the present invention represented by formula (1) can be produced, for example, by the following production method. Production Method E-1 (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , L, and Y １ is as defined in item 1, and LG ｅ１ and L.G. ｅ２ represents a leaving group, R ｅ is C １－６ represents an alkyl group)

[0125] The compound (e1-1) which is the starting material in step e-1 can be a commercially available product or can be produced by reacting the corresponding starting compound with the method described in Production Methods A to D.

[0126] Steps e-1 to e-4 are, for example, methods according to the production method described in WO2009 / 067081.

[0127] The base used in each step of each of the above-mentioned production methods should be appropriately selected depending on the type of reaction and raw material compound, and examples thereof include alkali bicarbonates such as sodium bicarbonate and potassium bicarbonate, alkali carbonates such as sodium carbonate and potassium carbonate, metal hydrides such as sodium hydride and potassium hydride, alkali metal hydroxides such as sodium hydroxide and potassium hydroxide, alkali metal alkoxides such as sodium methoxide and sodium t-butoxide, organometallic bases such as butyllithium and lithium diisopropylamide, and organic bases such as triethylamine, diisopropylethylamine, pyridine, 4-dimethylaminopyridine (DMAP), and 1,8-diazabicyclo[5.4.0]-7-undecene (DBU).

[0128] Condensing agents used in each step of the above-mentioned production methods include those described in Volume 22 of Jikken Kagaku Koza (edited by the Chemical Society of Japan, Maruzen). Examples include phosphate esters such as diethyl cyanophosphate and diphenylphosphoryl azide; carbodiimides such as 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (WSC.HCl) and dicyclohexylcarbodiimide (DCC); combinations of disulfides such as 2,2'-dipyridyl disulfide and phosphines such as triphenylphosphine; and N,N'-bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOPCl). Phosphorus halides; combinations of azodicarboxylic acid diesters such as diethyl azodicarboxylate and phosphines such as triphenylphosphine; 2-halo-1-lower alkylpyridinium halides such as 2-chloro-1-methylpyridinium iodide; 1,1'-carbonyldiimidazole (CDI); diphenylphosphoryl azide (DPPA); diethylphosphoryl cyanide (DEPC); 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium Examples of the tetrafluoroborates include tetrafluoroborate (TBTU) and 2-chloro-1,3-dimethylimidazolidinium tetrafluoroborate (CIB); and phosphates such as 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP), benzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (PYBOP), and 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU).

[0129] The solvent used in each step of each of the above-mentioned production methods should be selected appropriately depending on the type of reaction and raw material compounds, etc., and examples thereof include alcohols such as methanol, ethanol, and isopropanol; ketones such as acetone and methyl ketone; halogenated hydrocarbons such as methylene chloride and chloroform; ethers such as tetrahydrofuran (THF) and dioxane; aromatic hydrocarbons such as toluene and benzene; aliphatic hydrocarbons such as hexane and heptane; esters such as ethyl acetate and propyl acetate; amides such as N,N-dimethylformamide (DMF) and N-methyl-2-pyrrolidone; sulfoxides such as dimethyl sulfoxide (DMSO); nitriles such as acetonitrile; and water. These solvents can be used alone or in combination of two or more. Depending on the type of reaction, organic bases may also be used as solvents.

[0130] "Pharmaceutically acceptable salts" include acid addition salts and base addition salts. For example, acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, sulfate, hydroiodide, nitrate, and phosphate, and organic acid salts such as citrate, oxalate, phthalate, fumarate, maleate, succinate, malate, acetate, formate, propionate, benzoate, trifluoroacetate, methanesulfonate, benzenesulfonate, para-toluenesulfonate, and camphorsulfonate. Base addition salts include sodium salt, potassium salt, calcium salt, magnesium salt, barium salt, aluminum salt, and the like. or organic base salts of trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, tromethamine [tris(hydroxymethyl)methylamine], tert-butylamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylamine, and the like; and further include amino acid salts of basic or acidic amino acids such as arginine, lysine, ornithine, aspartic acid, or glutamic acid. Suitable salts and pharmaceutically acceptable salts of the starting compounds and target compounds are conventional non-toxic salts, including acid addition salts such as organic acid salts (e.g., acetate, trifluoroacetate, maleate, fumarate, citrate, tartrate, methanesulfonate, benzenesulfonate, formate, or para-toluenesulfonate) and inorganic acid salts (e.g., hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, or phosphate); salts with amino acids (e.g., arginine, aspartic acid, or glutamic acid); metal salts such as alkali metal salts (e.g., sodium salt, potassium salt, or alkaline earth metal salts (e.g., calcium salt, or magnesium salt); ammonium salts; or organic base salts (e.g., trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt, or N,N'-dibenzylethylenediamine salt), which can be appropriately selected by those skilled in the art.

[0131] When it is desired to obtain a salt of the compound of the present invention, if the compound of the present invention is obtained in the form of a salt, it may be purified as it is, or if it is obtained in the free form, it may be dissolved or suspended in an appropriate organic solvent and an acid or base may be added to form a salt by a conventional method. In addition, the compound of the present invention and its pharmaceutically acceptable salts may exist in the form of a solvate with water or various solvents, and these adducts are also encompassed by the present invention.

[0132] In addition, one or more H of the compound represented by formula (1) ２ Deuterium conversion products converted to H(D) are also included in the compounds represented by formula (1).

[0133] The present invention also includes a compound represented by formula (1) or a pharmaceutically acceptable salt thereof. It also includes solvates thereof, such as hydrates or ethanol solvates. Furthermore, the present invention also includes all tautomers, all existing stereoisomers, and all crystalline forms of compound (1) of the present invention.

[0134] Among the compounds (1) of the present invention, there may exist optical isomers based on optically active centers, atropisomers based on axial or planar chirality resulting from constraints on intramolecular rotation, other stereoisomers, tautomers, geometric isomers, and the like, and all possible isomers and mixtures thereof, including these, are included in the scope of the present invention.

[0135] A mixture of optical isomers of the compound of the present invention can be produced by a conventional method. Examples of production methods include a method using a starting material having an asymmetric center or a method introducing asymmetry at an intermediate stage. For example, in the case of optical isomers, optical isomers can be obtained by using an optically active starting material or by performing optical resolution at an appropriate stage in the production process. Examples of optical resolution methods include a diastereomeric method in which, when the compound represented by formula (1) or an intermediate thereof has a basic functional group, a salt is formed using an optically active acid (e.g., monocarboxylic acid such as mandelic acid, N-benzyloxyalanine, lactic acid, dicarboxylic acid such as tartaric acid, o-diisopropylidenetartaric acid, malic acid, or sulfonic acid such as camphorsulfonic acid or bromocamphorsulfonic acid) in an inert solvent (e.g., alcoholic solvents such as methanol, ethanol, 2-propanol, etc.; ether solvents such as diethyl ether, ester solvents such as ethyl acetate, hydrocarbon solvents such as toluene, aprotic solvents such as acetonitrile, or a mixed solvent of two or more of the above solvents). When the compound of the present invention represented by formula (1) or an intermediate thereof has an acidic functional group such as a carboxyl group, optical resolution can also be carried out by forming a salt using an optically active amine (e.g., an organic amine such as 1-phenylethylamine, quinine, quinidine, cinchonidine, cinchonine, or strychnine).

[0136] The compound of the present invention represented by formula (1) or an intermediate thereof can be separated and purified by methods known to those skilled in the art. Examples of such methods include extraction, distribution, reprecipitation, column chromatography (e.g., silica gel column chromatography, ion exchange column chromatography, or preparative liquid chromatography), and recrystallization. Examples of recrystallization solvents that can be used include alcoholic solvents such as methanol, ethanol, or 2-propanol; etheric solvents such as diethyl ether; ester solvents such as ethyl acetate; aromatic hydrocarbon solvents such as benzene or toluene; ketone solvents such as acetone; halogenated solvents such as dichloromethane or chloroform; hydrocarbon solvents such as hexane; aprotic solvents such as dimethylformamide or acetonitrile; water; or a mixture of these solvents. Other purification methods that can be used include those described in Volume 1 of "Experimental Chemistry Lectures" (edited by the Chemical Society of Japan, Maruzen). The molecular structure of the compound of the present invention can be easily determined by spectroscopic techniques such as nuclear magnetic resonance, infrared absorption, and circular dichroism spectroscopy, as well as mass spectrometry, with reference to the structures derived from the respective starting compounds.

[0137] Furthermore, the intermediates or final products in the above production methods can be converted into other compounds included in the present invention by appropriately converting their functional groups, particularly by extending various side chains using amino groups, hydroxyl groups, carbonyl groups, halogen groups, etc., as a stepping stone, and by carrying out the above-mentioned protection and deprotection as necessary. Functional group conversion and side chain extension can be carried out by commonly used general methods (see, for example, Comprehensive Organic Transformations, R.C. Larock, John Wiley & Sons Inc. (1999)).

[0138] The temperature for salt formation is selected from the range of −50°C to the boiling point of the solvent, preferably from the range of 0°C to the boiling point, and more preferably from room temperature to the boiling point of the solvent. To improve optical purity, it is desirable to first raise the temperature to near the boiling point of the solvent. When filtering out the precipitated salt, cooling can be performed as needed to improve the yield. The amount of optically active acid or amine used is preferably in the range of about 0.5 to about 2.0 equivalents relative to the substrate, preferably around 1 equivalent. If necessary, the crystals can be recrystallized in an inert solvent (e.g., alcoholic solvents such as methanol, ethanol, and 2-propanol; etheric solvents such as diethyl ether; esteric solvents such as ethyl acetate; hydrocarbon solvents such as toluene; aprotic solvents such as acetonitrile; or a mixed solvent of two or more of the above solvents) to obtain a highly pure optically active salt. Furthermore, if necessary, the optically resolved salt can be treated with an acid or base by a conventional method to obtain a free form.

[0139] Of the raw materials and intermediates in each of the production methods explained above, those for which the production method is not particularly described are commercially available compounds or can be synthesized from commercially available compounds by methods known to those skilled in the art or methods similar thereto.

[0140] The present invention provides compounds of formula (1) as defined above or pharmaceutically acceptable salts thereof, which are useful as vaccine adjuvants, preferably as vaccine adjuvants for cancer vaccines.

[0141] The present invention also provides a pharmaceutical composition containing the compound represented by formula (1) defined above or a pharmaceutically acceptable salt thereof in combination with a pharmaceutically acceptable diluent or carrier (hereinafter referred to as the pharmaceutical composition of the present invention).

[0142] The compound of the present invention or a pharmaceutically acceptable salt thereof can be used as an adjuvant for maintaining or enhancing the immunostimulatory activity of an active ingredient having immunostimulatory activity. Specifically, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of inducing or enhancing antigen-specific antibodies, specifically antigen-specific IgG, more specifically Th1-type antigen-specific IgG (e.g., IgG2c). Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of increasing cytotoxic T cells (CTLs). Alternatively, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of inducing CTLs in mammals or enhancing CTL induction in mammals. Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of increasing CD4-positive (i.e., MHC class II-restricted) and / or CD8-positive (i.e., MHC class I-restricted) T cells. Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of increasing antigen-specific T cells. Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of increasing memory T cells, specifically CD8-positive effector memory T cells. Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof has the characteristic that, when administered to a mammal, the effect of increasing CTLs is higher than when a compound not containing the same molar number of PEG structures is administered. Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof has the activity of activating immunocompetent cells. The pharmaceutical composition of the present invention may contain a tumor antigen. As the tumor antigen, a tumor antigen protein or a tumor antigen peptide derived from the tumor antigen protein can be used. As the tumor antigen peptide, the antigen peptides described below can be preferably used, with more preferred examples including tumor antigen peptides derived from NY-ESO-1, MAGE-3, WT1, OR7C1, or Her2 / neu, and even more preferred examples including tumor antigen peptides derived from WT1. Furthermore, peptides derived from neoantigens generated as a result of genetic abnormalities in tumors can also be used together with the compound of the present invention or a pharmaceutically acceptable salt thereof.Furthermore, a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof and a tumor antigen has an effect of inhibiting the growth of a tumor expressing the antigen, or a preventive effect of suppressing the occurrence of a tumor expressing the antigen. Therefore, the compound of the present invention or a pharmaceutically acceptable salt thereof is useful as a drug for treating or preventing cancer when used as a pharmaceutical composition in combination with the tumor antigen shown below.

[0143] The tumor antigen peptide is not particularly limited, and peptides described in International Publication WO2014 / 157692 or International Publication WO2014 / 157704A1 can be used. One embodiment of the tumor antigen peptide includes, for example, a tumor antigen peptide which is a peptide consisting of the following amino acid sequence or a pharmaceutically acceptable salt thereof: RMFPNAPYL (SEQ ID NO: 1), ALLPAVPSL (SEQ ID NO: 8), SLGEQQYSV (SEQ ID NO: 9), RVPGVAPTL (SEQ ID NO: 10), VLDFAPPGA (SEQ ID NO: 4), CMTWNQMNL (SEQ ID NO: 11), CYTWNQMNL (SEQ ID NO: 2), WAPVLDFAPPGASAYGSL (SEQ ID NO: 3), CWAPVLDFAPPGASAYGSL (SEQ ID NO: 12), WAPVLDFAPPGASAYGSLC (SEQ ID NO: 13), CNKRYFKLSHLQMHSRKHTG (SEQ ID NO: 14), CNKRYFKLSHLQMHSRKH (SEQ ID NO: 15), CNKRYFKLSHLQMHSRK (SEQ ID NO: 16), KRYFKLSHLQMHSRKH (SEQ ID NO: 17) or TYAGCLSQIF (SEQ ID NO: 18). [wherein the bond between C and C represents a disulfide bond] and Formula (5): [wherein the bond between C and C represents a disulfide bond] or a pharmaceutically acceptable salt thereof can also be used as the tumor antigen peptide of the present invention.

[0144] Tumor antigen peptides can be synthesized according to methods commonly used in peptide chemistry, including those described in literature (Peptide Synthesis, Interscience, New York, 1966; The Proteins, Vol. 2, Academic Press Inc., New York, 1976).

[0145] In another embodiment, the pharmaceutical composition of the present invention may contain an antigen. Examples of such antigens include pathogen-derived antigens, such as proteins or partial peptides derived from pathogens such as viruses or bacteria. Furthermore, complexes of these antigens with carriers are also included in the category of antigens herein. Examples of such complexes include those in which an antigen (including, but not limited to, a protein or a peptide) is chemically crosslinked to a carrier protein via a linker well known to those skilled in the art, and those in which the antigen is contained in a virus-like particle (VLP). Therefore, the compound of the present invention or a pharmaceutically acceptable salt thereof, when used in combination with the antigen, is useful as a drug for treating or preventing infectious diseases caused by viruses, bacteria, or the like.

[0146] The pharmaceutical composition of the present invention can be administered, for example, parenterally, specifically via intravascular (e.g., intravenous), subcutaneous, intradermal, intramuscular, intratumoral, lymph node, or transdermal administration.

[0147] In one embodiment, the pharmaceutical composition of the present invention may contain a compound represented by formula (1) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier.

[0148] The pharmaceutical composition of the present invention may be in the form of a liquid or the like.

[0149] The liquid preparations of the present invention include aqueous solution preparations or aqueous suspensions, oily solution preparations or oily suspensions, hydrogel preparations, lipid preparations, and emulsion preparations. Examples of aqueous solution preparations or aqueous suspensions include preparations in which an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof is dissolved or dispersed in water. Examples of oily solution preparations or oily suspensions include preparations in which an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof is dissolved or dispersed in an oily component. Examples of hydrogel preparations include preparations in which an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof is dissolved or dispersed in water to impart viscosity. Examples of lipid preparations include liposome preparations containing an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof. Examples of emulsion preparations include preparations containing an aqueous solution and an oily composition containing an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof. Other embodiments of the liquid preparation of the present invention include aqueous solution preparations or aqueous suspensions in which a tumor antigen and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof is dissolved or dispersed in water; oily solution preparations or oily suspensions in which a tumor antigen and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof is dissolved or dispersed in an oily component; and emulsion preparations containing an aqueous solution and an oily composition. Examples of additives used in the aqueous solution preparations or aqueous suspensions of the present invention include purified water, water for injection, buffers, pH adjusters, stabilizers, isotonicity agents, solubilizers, and solubilizers. Examples of additives used in the oily solution formulation or oily suspension formulation of the present invention include buffers, pH adjusters, stabilizers, isotonicity agents, animal and vegetable oils and fats, hydrocarbons, fatty acids, fatty acid esters, solubilizers, and solubilizing agents.Examples of additives used in the hydrogel formulation of the present invention include purified water, water for injection, buffers, pH adjusters, stabilizers, isotonicity agents, solubilizers, solubilizers, thickeners, etc. Examples of additives used in the liposome formulation of the present invention include purified water, water for injection, buffers, pH adjusters, stabilizers, isotonicity agents, solubilizers, solubilizers, lipids, etc.

[0150] The emulsion formulation of the present invention may be an oil-in-water emulsion (also referred to as an O / W emulsion), a water-in-oil emulsion (also referred to as a W / O emulsion), a water-in-oil-in-water emulsion (also referred to as a W / O / W emulsion), or an oil-in-water-in-oil emulsion (also referred to as an O / W / O emulsion). A preferred emulsion formulation of the present invention is a water-in-oil emulsion (W / O emulsion). The emulsion formulation of the present invention can be produced by emulsifying an aqueous phase and an oil phase using a known method. The antigen (tumor antigen or pathogen-derived antigen) and / or the compound represented by Formula (1) or a pharmaceutically acceptable salt thereof may be contained in either or both of the oil and aqueous phases of the emulsion. Examples of additives used in the emulsion formulation of the present invention include water, buffers, pH adjusters, stabilizers, isotonicity agents, animal and vegetable oils and fats, hydrocarbons, fatty acids, fatty acid esters, glycerin fatty acid esters, hydrophilic surfactants, and lipophilic surfactants.Here, examples of water include purified water and water for injection, buffers include phosphates and organic acid salts, pH adjusters include hydrochloric acid and sodium hydroxide, stabilizers include glycerin, propylene glycol, sulfites, and the like, isotonic agents include table salt, glucose, sucrose, mannitol, and the like, animal and vegetable oils include olive oil, soybean oil, and cod liver oil, hydrocarbons include liquid paraffin, squalene, squalane, and the like, fatty acids include oleic acid and myristic acid, and fatty acid esters include oleic acid. Examples of surfactants that can be used include ethyl myristate, octyldodecyl myristate, cetyl 2-ethylhexanoate, and isopropyl myristate. Examples of glycerin fatty acid esters include medium-chain fatty acid triglycerides, medium-chain fatty acid diglycerides, and medium-chain fatty acid monoglycerides. Examples of hydrophilic surfactants include polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid esters, and polysorbates. Examples of lipophilic surfactants include glycerin monooleate, glycerin dioleate, sorbitan monooleate (Span 80 (registered trademark)), sorbitan sesquioleate, sorbitan dioleate, sorbitan trioleate (Span 85 (registered trademark)), PEG-30 dipolyhydroxystearate, and plant-derived surfactants (saponin, etc.).

[0151] Specific examples of the additive composition for the emulsion formulation of the present invention include, but are not limited to, the emulsifying composition for dilution described in International Publication WO 2006 / 078059, Montanide ISA 51 VG (Seppic), Montanide ISA 720 VG (Seppic), Incomplete Freund's Adjuvant (IFA), etc. Examples of the W / O emulsion formulation of the present invention include a formulation containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof, ethyl oleate, octyldodecyl myristate, sorbitan monooleate, glycerin monooleate, polyoxyethylene hydrogenated castor oil 20, glycerin, and sodium dihydrogen phosphate; or a formulation containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof, and Montanide ISA 51 VG.

[0152] In the liposome preparation of the present invention, the term "liposome" refers to a microvesicle having an internal phase composed of a lipid multilayer, such as a bilayer membrane (lipid bilayer) of amphiphilic lipid molecules. Here, the lipid multilayer is preferably a lipid bilayer.

[0153] The liposome preparation of the present invention contains amphipathic lipid molecules. The amphipathic lipid molecules preferably contain one or more types of "phospholipids." Examples of "phospholipids" include phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, and sphingomyelin. Preferred examples of "phospholipids" include phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, sphingomyelin, and phosphatidylserine. More preferred examples of "phospholipids" include phosphatidylcholine, sphingomyelin, and phosphatidylserine. The fatty acid residue in the "phospholipid" is not particularly limited, but examples include saturated or unsaturated fatty acid residues having 14 to 18 carbon atoms, and specific examples include acyl groups derived from fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid. Also usable are naturally occurring phospholipids such as egg yolk lecithin and soybean lecithin, and hydrogenated egg yolk lecithin and hydrogenated soybean lecithin (also called hydrogenated soybean phospholipid or hydrogenated soybean phosphatidylcholine) in which the unsaturated fatty acid residues of these phospholipids have been hydrogenated. The amount (molar fraction) of phospholipid relative to the total liposome membrane components is not particularly limited, but is preferably 30 to 80%, more preferably 40 to 70%.

[0154] The liposome encapsulating the compound of the present invention may contain sterols. Examples of sterols include cholesterol, β-sitosterol, stigmasterol, campesterol, brassica sterol, ergosterol, and fucosterol. A preferred example of the sterol is cholesterol. The amount (molar fraction) of sterols relative to the total liposome membrane components is not particularly limited, but is preferably 0 to 60%, more preferably 10 to 50%, and even more preferably 30 to 50%.

[0155] The liposome encapsulating the compound of the present invention may contain a polymer-modified lipid. A polymer-modified lipid means a lipid modified with a polymer. The polymer-modified lipid is represented by "lipid-polymer". The polymer portion of the polymer-modified lipid is preferably a hydrophilic polymer, and more preferably a hydrophilic polymer in which the end of the polymer not bound to the lipid is alkoxylated. More preferred examples of the polymer portion of the polymer-modified lipid include hydrophilic polymers in which the end of the polymer not bound to the lipid is methoxylated, ethoxylated, or propoxylated. Most preferred examples of the polymer portion of the polymer-modified lipid include hydrophilic polymers in which the end of the polymer not bound to the lipid is methoxylated. The polymer portion of the polymer-modified lipid is not particularly limited, but may include, for example, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, and propoxypolyvinylpyrrolidone. The polymer portion of the polymer-modified lipid is preferably polyethylene glycol, methoxypolyethylene glycol, methoxypolypropylene glycol, ethoxypolyethylene glycol, ethoxypolypropylene glycol, propoxypolyethylene glycol, and propoxypolypropylene glycol. The polymer portion of the polymer-modified lipid is more preferably polyethylene glycol, methoxypolyethylene glycol, ethoxypolyethylene glycol, ethoxypolypropylene glycol, and propoxypolyethylene glycol. The polymer portion of the polymer-modified lipid is even more preferably polyethylene glycol and methoxypolyethylene glycol. The polymer portion of the polymer-modified lipid is most preferably methoxypolyethylene glycol.The molecular weight of the polymer portion of the polymer-modified lipid is not particularly limited, and examples thereof include 100 to 10,000 daltons, preferably 500 to 8,000 daltons, more preferably 1,000 to 7,000 daltons, even more preferably 1,500 to 5,000 daltons, and most preferably 1,500 to 3,000 daltons. The lipid portion of the polymer-modified lipid is not particularly limited, and examples thereof include phosphatidylethanolamine and diacylglycerol. The lipid portion of the polymer-modified lipid is preferably a phosphatidylethanolamine having a saturated or unsaturated fatty acid residue having 14 to 18 carbon atoms, and a diacylglycerol having a saturated or unsaturated fatty acid residue having 14 to 18 carbon atoms, more preferably a phosphatidylethanolamine having a saturated fatty acid residue having 14 to 18 carbon atoms, and a diacylglycerol having a saturated fatty acid residue having 14 to 18 carbon atoms, and even more preferably a phosphatidylethanolamine having a palmitoyl group or a stearoyl group, and a diacylglycerol having a palmitoyl group or a stearoyl group. The lipid portion of the polymer-modified lipid is most preferably distearoylphosphatidylethanolamine. The amount (molar fraction) of the polymer-modified lipid relative to the total liposome membrane components is not particularly limited, but is preferably 0 to 20%, more preferably 1 to 10%, and even more preferably 2 to 6%.

[0156] Liposomes encapsulating the compounds of the present invention may contain pharmaceutically acceptable additives. Examples of additives include inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers. Inorganic acids include phosphoric acid, hydrochloric acid, and sulfuric acid. Inorganic acid salts include sodium hydrogen phosphate, sodium chloride, ammonium sulfate, and magnesium sulfate. Organic acids include citric acid, acetic acid, succinic acid, and tartaric acid. Organic acid salts include sodium citrate, sodium acetate, disodium succinate, and sodium tartrate. Sugars include glucose, sucrose, mannitol, sorbitol, and trehalose. Buffers include L-arginine, L-histidine, trometamol (trishydroxymethylaminomethane, Tris), and salts thereof. Antioxidants include, for example, sodium sulfite, L-cysteine, sodium thioglycolate, sodium thiosulfate, ascorbic acid, and tocopherol. Polymers include, for example, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, and sodium carboxymethylcellulose.

[0157] The oil suspension formulation of the present invention can contain an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof in the oil component in either or both of a dissolved and dispersed state. Examples of additives used in the oil suspension formulation of the present invention include buffers, pH adjusters, stabilizers, isotonicity agents, animal and vegetable oils and fats, hydrocarbons, fatty acids, fatty acid esters, solubilizers, and solubilizers. Examples of buffering agents include phosphates and organic acid salts, examples of pH adjusting agents include hydrochloric acid and sodium hydroxide, examples of stabilizers include glycerin, propylene glycol, and sulfites, examples of isotonic agents include table salt, glucose, sucrose, and mannitol, examples of animal and vegetable oils and fats include olive oil, soybean oil, and cod liver oil, examples of hydrocarbons include liquid paraffin, squalene, and squalane, examples of fatty acids include oleic acid and myristic acid, examples of fatty acid esters include ethyl oleate, octyldodecyl myristate, cetyl 2-ethylhexanoate, isopropyl myristate, sucrose fatty acid esters, glycerin fatty acid esters, sorbitan fatty acid esters, and propylene glycol fatty acid esters, and examples of solubilizing agents or solubilizing aids include glycerin, propylene glycol, macrogols, and ethanol.

[0158] The hydrogel formulation of the present invention may be, for example, a formulation in which an antigen (tumor antigen or pathogen-derived antigen) and / or a compound represented by formula (1) or a pharmaceutically acceptable salt thereof is dissolved or dispersed in water to impart viscosity. Examples of additives used in the hydrogel formulation of the present invention include purified water, water for injection, buffers, pH adjusters, stabilizers, isotonicity agents, solubilizers, solubilizers, thickeners, etc. Examples of buffers include phosphates and organic acid salts. Examples of pH adjusters include hydrochloric acid and sodium hydroxide. Examples of stabilizers include glycerin, propylene glycol, sulfites, etc. Examples of isotonicity agents include table salt, glucose, sucrose, mannitol, etc. Examples of solubilizers or solubilizers include glycerin, propylene glycol, macrogols, ethanol, etc. Examples of thickeners include carmellose sodium, poloxamers, povidone, etc.

[0159] The compound represented by formula (1) or a pharmaceutically acceptable salt thereof described herein, or the pharmaceutical composition of the present invention can be used in combination with other drugs (also referred to as concomitant drugs in this specification) in addition to the above-mentioned tumor antigens.

[0160] In certain embodiments, the compound represented by formula (1) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the present invention can be used in combination with an "immunomodulator" in addition to the tumor antigen. As used herein, the term "immunomodulator" refers to any agent that controls the transmission of costimulatory signals by interacting with molecules involved in the transmission of costimulatory signals on antigen-presenting cells and / or T cells during T cell activation by antigen-presenting cells, or that directly or indirectly controls the function of molecules involved in the establishment of immune tolerance (immunosuppression) in the immune system. Because tumor antigen peptides are drugs that increase tumor-reactive CTLs within tumors, their use in combination with an immunomodulator may reduce the dosage of the immunomodulator and potentially alleviate adverse events. In other words, the combined use of a WT1 antigen peptide and an immunomodulator can provide patients with a therapy that combines greater efficacy and safety.

[0161] An "immunomodulator" may be an agent selected from, but is not limited to, an antibody, a nucleic acid, a protein, a peptide, and a small molecule compound. In the description of an "immunomodulator," the term "antibody" also includes antibody fragments. Examples of antibody fragments include antibody heavy and light chain variable regions (VH and VL), F(ab')2, Fab', Fab, Fv, Fd, sdFv, and scFV. In the description of an "immunomodulator," a protein refers to any protein other than an antibody. "Immunomodulators" include, for example, immune checkpoint inhibitors, costimulatory molecule agonists, immune activators, and small molecule inhibitors.

[0162] "Immune checkpoint inhibitors" inhibit the immunosuppressive effects of cancer cells and antigen-presenting cells. Examples of immune checkpoint inhibitors include, but are not limited to, drugs against molecules selected from the group consisting of: (1) CTLA-4 (ipilimumab, tremelimumab, etc.); (2) PD-1 (nivolumab, pembrolizumab, AMP-224, AMP-514 (MEDI0680), pidilizumab (CT-011), etc.); (3) LAG-3 (IMP-321, BMS-986016, etc.); (4) BTLA; (5) KIR (IPH2101, etc.); (6) TIM-3 (LY3321367, CA-327, etc.); (7) PD-L1 (durvalumab (MEDI 4736), MPDL3280A, BMS-936559, avelumab (MSB0010718C), BMS-1001, BMS-1116, CA-170, CA-327, etc.); (8) PD-L2; (9) B7-H3 (MGA-271, etc.); (10) B7-H4; (11) HVEM; (1 2) GAL9; (13) CD160; (14) VISTA (e.g., ombatilimab (JNJ-61610588), HMBD-002, CA-170); (15) BTNL2; (16) TIGIT; (17) PVR; (18) BTN1A1; (19) BTN2A2; (20) BTN3A2 (Nat Rev Drug Discov. 2013; 12: 130-146; Nikkei Medical Cancer Review 2014; 9; Nat Rev Immunol. 2014; 14: 559-69); (21) CSF1-R; (22) VSIG-3; (23) CD112; (24) CD112R, and (25) CD96.

[0163] "Costimulatory molecule agonists" activate T cells by transmitting auxiliary signals mediated by costimulatory molecules on T cells or antigen-presenting cells, thereby attenuating the immunosuppressive effects of cancer cells or antigen-presenting cells. Examples of costimulatory molecule agonists include, but are not limited to, agents directed against molecules selected from the following group: (1) 4-1BB (2) 4-1BB-L; (3) OX40 (4) OX40-L; (5) GITR; (6) CD28; (7) CD40; (8) CD40-L (9) ICOS; (10) ICOS-L; (11) LIGHT; (12) CD27; and (13) DNAM-1.

[0164] An "immunostimulator" efficiently stimulates killer T cells in lymph nodes by directly or indirectly activating immune cells such as T cells and dendritic cells. Examples of immune activators include, but are not limited to, Toll-like receptor (TLR) agonists, stimulator of interferon genes (STING) agonists, cytokines, and agents against heat shock proteins (HSPs).

[0165] Examples of "Toll-like receptor (TLR) agonists" include, but are not limited to, TLR1 / 2 agonists, TLR2 agonists, TLR3 agonists (PolyI:C and the like), TLR4 agonists (S-type lipopolysaccharide, paclitaxel, lipid A, monophosphoryl lipid A and the like), TLR5 agonists (flangerine and the like), TLR6 / 2 agonists (MALP-2 and the like), TLR7 agonists, TLR7 / 8 agonists (gardiquimod, imiquimod, loxoribine, resiquimod (R848) and the like), TLR7 / 9 agonists (hydroxychloroquine sulfate and the like), TLR8 agonists (motolimod (VTX-2337) and the like), TLR9 agonists (CpG-ODN and the like), and TLR11 agonists (profilin).

[0166] Examples of "cytokines" include, but are not limited to, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, interferon (INF)-α, INF-β, INF-γ, SCF, GM-CSF, G-CSF, M-CSF, erythropoietin, thrombopoietin, MIP (macrophage inflammatory protein), and MCP (monocyte chemoattractant protein).

[0167] "Heat shock proteins (HSPs)" include, but are not limited to, HSP70, HSP90, HSP90α, HSP90β, HSP105, HSP72, and HSP40. Drugs against HSPs include HSP inhibitors. For example, HSP90 inhibitors include, but are not limited to, tanespimycin (17-AAG), luminespib (AUY-922, NVP-AUY922), alvespimycin (17-DMAG) hydrochloride, ganetespib (STA-9090), BIIB021, onarespib (AT13387), geldanamycin, and NVP-BEP80. 0, SNX-2112 (PF-04928473), PF-4929113 (SNX-5422), KW-2478, XL888, VER155008, VER-50589, CH5138303, VER-49009, NMS-E973, PU-H71, HSP990 (NVP-HSP990), or KNK437.

[0168] Examples of "small molecule inhibitors" include, but are not limited to, histone deacetylation inhibitors, histone demethylation inhibitors, histone acetyltransferase inhibitors, histone methyltransferase inhibitors, DNA methyltransferase inhibitors, anthracycline antibiotics, platinum agents, MAPK inhibitors, β-catenin inhibitors, STAT3 inhibitors, NF-kB inhibitors, JAK inhibitors, mTOR inhibitors, IDO inhibitors, COX-2 inhibitors, CXCR4 inhibitors, and arginase inhibitors.

[0169] Examples of "histone deacetylation inhibitors" include, but are not limited to, vorinostat (SAHA, MK0683), entinostat (MS-275), panobinostat (LBH589), trichostatin A (TSA), mocetinostat (MGCD0103), BG45, BRD73954, belinostat (PXD101), romidepsin (FK228, decipeptide), 4SC-202, HPOB, LMK-235, CAY10603, tasquinimod, TMP269, and Nexturastat. A, Rocilinostat (ACY-1215), RGFP966, RG2833 (RGFP109), Scriptaid, Tubastatin A, Pracinostat (SB939), CUDC-101, M344, PCI-34051, Dacinostat (LAQ824), Tubastatin A hydrochloride, Avexinostat (PCI-24781), CUDC-907, AR-42, Sodium phenylbutyrate, Resminostat, Tubacin, Xinostat (JNJ-26481585) dihydrochloride, MC1568, Givinostat (ITF2357), Droxinostat, Chidamide (C S055, HBI-8000), CHR-2485, CHR-3996, DAC-060, FRM-0334 (EVP-0334), MGCD-290, CXD-101 (AZD-9468), CG200745, arginine butyrate, sulforaphane, SHP-141, CUDC-907, YM753 (OBP-801), sodium valproate, apicidin, and CI994 (Tacedinaline).

[0170] Examples of "histone demethylation inhibitors" include, but are not limited to, GSK J4 HCl, OG-L002, JIB-04, IOX1, SP2509, ORY-1001 (RG-6016), GSK J1, ML324, GSK-LSD1 2HCl, etc.

[0171] "Histone acetyltransferase inhibitors" include, but are not limited to, for example, C646, MG149, Remodelin, and Anacardiic Acid.

[0172] Examples of "histone methyltransferase inhibitors" include, but are not limited to, pinometostat (EPZ5676), EPZ005678, GSK343, BIX01294, tazemetostat (EPZ6438), 3-deazaneplanocin A (DZNeP) HCl, UNC1999, MM-102, SGC0946, entacapone, EPZ015666, UNC0379, EI1, MI-2 (Menin-MLL Inhibitor), MI-3 (Menin-MLL Inhibitor), PFI-2, GSK126, EPZ04777, BRD4770, GSK-2816126, and UNC0631.

[0173] "DNA methyltransferase inhibitors" include, but are not limited to, decitabine, azatidine, RG108, thioguanine, zebularine, SGI-110, CC-486, SGI-1027, lomeguatrib, and procainamide hydrochloride.

[0174] "Anthracycline antibiotics" inhibit DNA unwinding by intercalating between DNA strands. Examples of anthracycline antibiotics include, but are not limited to, doxorubicin, liposomal doxorubicin, daunorubicin, pirarubicin, epirubicin, idarubicin, aclarubicin, amrubicin, aloin, and mitoxatrone.

[0175] "Platinum agents" are not particularly limited, but examples include cisplatin, carboplatin, miboplatin, nedaplatin, satraplatin (JM-126), oxaliplatin (ELOXATIN), triplatin tetranitrate, or DDS preparations thereof.

[0176] Examples of "MAPK inhibitors" include, but are not limited to, SB203580, dramapimod (BIRB796), SB202190 (FHPI), LY2228820, VX-702, SB239063, pexmetinib (ARRY-614), PH-797804, VX-745, and TAK-715.

[0177] Examples of "β-catenin inhibitors" include, but are not limited to, XAV-939, ICG-001, IWR-1-endo, Wnt-C59 (C59), LGK-974, KY02111, IWP-2, IWP-L6, WIKI4, and FH535.

[0178] Examples of "STAT3 inhibitors" include, but are not limited to, S3I-201, Stattic, niclosamide, nifuroxazide, napabucasin (BBI608), cryptotanshinone, HO-3867, WHI-P154, FLLL32, STA-21, WP1066, and SH-4-54.

[0179] Examples of "NF-kB inhibitors" include, but are not limited to, QNZ (EVP4593), sodium 4-aminosalicylate, JSH-23, phenethyl caffeate, sodium salicylate, andrographolide, and SC75741.

[0180] Examples of "JAK inhibitors" include, but are not limited to, ruxolitinib (INCB018424), tofacitinib (CP-690550) citrate, AZD1480, fedratinib (SAR302503, TG101348), AT9283, tyrphostin B42 (AG-490), momelotinib (CYT387), tofacitinib (CP-690550, tasocitinib), WP1066, TG101209, gandotinib (LY2784544), and NVP-BSK805. 2HCl, baricitinib (LY3009104, INCB02850), AZ960, CEP-33779, pacritinib (SB1518), WHI-P154, XL019, S-ruxolitinib (INCB018424), ZM39923 HCl, decernotinib (VX-509), cerdulatinib (PRT062070, PRT2070), filgotinib (GLPG0634), FLLL32, peficitinib (ASP015K, JNJ-54781532), GLPG0634 analogue, Go6976 or Curcumol.

[0181] Examples of "mTOR inhibitors" include, but are not limited to, sirolimus (rapamycin), deforolimus (AP23573, MK-8669), everolimus (RAD-001), temsirolimus (CCI-779, NSC683864), zotarolimus (ABT-578), and biolimus A9 (umirolimus), AZD8055, KU-0063794, voxtalisib (XL765, SAR245409), MHY1485, dactolisib (BEZ235, NVP-BEZ235), PI-103, torkinib (PP242), and the like.

[0182] "IDO inhibitors" include, but are not limited to, NLG919, INCB024360 analogs, indoximod (NLG-8189), and Epacadostat (INCB024360).

[0183] Examples of "COX2 inhibitors" include, but are not limited to, valdecoxib, rofecoxib, carprofen, celecoxib, lumiracoxib, tolfenamic acid, nimesulide, niflumic acid, asaraldehyde, lornoxicam, meclofenamate sodium, amfenac sodium hydrate, diclofenac sodium, ketoprofen, ketorolac, naproxen sodium, indomethacin, ibuprofen, aspirin, mefenamic acid, bromfenac sodium, oxaprozin, zaltoprofen, and nepafenac.

[0184] Examples of "CXCR4 inhibitors" include, but are not limited to, WZ811, Plerixafor (AMD3100), and Plerixafor 8HCl (AMD3100 8HCl).

[0185] The compounds represented by formula (1) described herein, and pharmaceutically acceptable salts thereof, and compositions thereof can be used in combination with one or more drugs selected from the group consisting of "hormonal therapeutic agents," "immunotherapeutic agents," "biological preparations," "cell growth factors," "cell growth factor inhibitors," "cell growth factor receptor inhibitors," "radiotherapeutic agents," "adjuvants," or "chemotherapeutic agents." For example, the peptides, compounds, and pharmaceutically acceptable salts thereof, as well as combinations thereof, can be used in combination with 1 to 5, 1 to 3, or 1 drug selected from the above group.

[0186] Examples of "hormonal therapy agents" include corticosteroid drugs (for example, steroidal anti-inflammatory drugs, estrogen preparations, progesterone preparations, androgen preparations, etc.), antiestrogens, estrogen regulators, estrogen synthesis inhibitors, antiandrogens, androgen regulators, androgen synthesis inhibitors, LH-RH agonist preparations, LH-RH antagonist preparations, aromatase inhibitors, steroid lactonase inhibitors, birth control pill preparations, retinoids, and agents that slow the metabolism of retinoids.

[0187] Examples of "hormonal therapeutic agents" include fosfestrol, diethylstilbestrol, fluoxymesterol, chlorotrianisene, methyltestosterone, medroxyprogesterone acetate, megestrol acetate, chlormadinone acetate, cyproterone acetate, danazol, allylestrenol, gestrinone, mepartricin, raloxifene, ormeloxifene, levormeloxifene, tamoxifen citrate, toremifene citrate, iodoxifene, birth control pills, mepitiostane, testololactone, aminoglutethimide, and goserelin acetate. , buserelin, leuprorelin, leuprolide, droloxifene, epitiostanol, ethinylestradiol sulfonate, estramustine, fadrozole hydrochloride, anastrozole, terorazole, ketoconazole, letrozole, exemestane, vorozole, formestane, exemestane, flutamide, bicalutamide, nilutamide, enzalutamide, mifepristone, finasteride, dexamethasone, prednisolone, betamethasone, triamcinolone, abiraterone, liarozole, bexarotene, or DN101.

[0188] Examples of "immunotherapeutic agents" include picibanil, krestin, sizofiran, lentinan, ubenimex, interferon (IL)-α, interferon (IL)-β, interferon (IL)-γ, interleukin, macrophage colony-stimulating factor, granulocyte colony-stimulating factor, erythropoietin, lymphotoxin, BCG vaccine, Corynebacterium parvum, levamisole, polysaccharide K, procodazole, anti-CTLA4 antibody, anti-PD-1 antibody, or TLR agonist (e.g., TLR7 agonist, TLR8 agonist, TLR9 agonist).

[0189] Examples of "biological agents" include, but are not limited to, interleukin-2 (Aldesleukin), interferon-α, interferon-β, interferon-γ, erythropoietin (EPO), granulocyte colony-stimulating factor (filgrastin), granulocyte-macrophage colony-stimulating factor (sargramostim), IL13-PE38QQR, bacillus calmette-guerin, levamisole, octreotide, CPG7909, Provenge, GVAX, Myvax, Favld, lenalidomide, trastuzumab, rituximab, gemtuzumab ozogamicin, alemtuzumab, endostatin, ibritumomab tiuxetan, tositumomab, and cetaxel. Cimab, zanolimumab, ofatumumab, HGS-ETR1, pertuzumab, M200, SGN-30, matuzumab, adecatumab, denosumab, zalutumumab, MDX-060, nimotuzumab, MORAb-003, Vitaxin, MDX-101, MDX-010, DPC4 antibody, NF-1 antibody, NF-2 antibody, Rb antibody, p53 Examples of antibodies include antibodies, WT1 antibodies, BRCA1 antibodies, BRCA2 antibodies, ganglioside (GM2), prostate-specific antigen (PSA), α-fetoprotein (AFP), carcinoembryonic antigen (CEA), melanoma-associated antigens (MART-1, gap100, MAGE1,3 tyrosine), papilloma virus E6 and E7 fragments, or DDS formulations thereof.

[0190] The cell growth factors in the above-mentioned "cell growth factors," "cell growth factor inhibitors," and "cell growth factor receptor inhibitors" may be any substance that promotes cell growth, and examples thereof include peptides with a molecular weight of 20,000 or less that exert their effects at low concentrations by binding to receptors.

[0191] The "cell growth factor" is not particularly limited, but examples thereof include epidermal growth factor (EGF), insulin-like growth factor (IGF (e.g., insulin, IGF-1, IGF-2, etc.)), transforming growth factor (TGF (e.g., TGF-alpha, TGF-beta)), nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), colony stimulating factor (CSF (e.g., granulocyte-colony stimulating factor (G-CSF)), granulocyte-macrophage-colony stimulating factor (GM-CSF)), platelet-derived growth factor (PGF), and the like. Examples of growth factors include PDGF, erythropoietin (EPO), fibroblast growth factors (FGFs (e.g., acidic FGF, basic FGF, KGK (Keratinocyte Growth Factor), FGF-10, etc.)), hepatocyte growth factor (HGF), heregulin, and angiopoietin. The cell growth factor is synonymous with growth factor.

[0192] Examples of "cell growth factor inhibitors" include, but are not limited to, epidermal growth factor inhibitors (EGF inhibitors), insulin-like growth factor inhibitors (IGF inhibitors), nerve growth factor inhibitors (NGF inhibitors), brain-derived neurotrophic factor inhibitors (NGF inhibitors), vascular endothelial growth factor inhibitors (VEGF inhibitors), colony-stimulating factor inhibitors (CSF inhibitors), platelet-derived growth factor inhibitors (PDGF inhibitors), erythropoietin inhibitors (EPO inhibitors), fibroblast growth factor inhibitors (FGF inhibitors), hepatocyte growth factor inhibitors (HGF inhibitors), heregulin inhibitors, and angiopoietin inhibitors. Note that cell growth factor inhibitors are synonymous with growth factor inhibitors.

[0193] Examples of "cell growth factor receptor inhibitors" include, but are not limited to, epidermal growth factor receptor inhibitors (EGFR inhibitors), insulin-like growth factor receptor inhibitors (IGFR inhibitors), nerve growth factor receptor inhibitors (NGFR inhibitors), brain-derived neurotrophic factor receptor inhibitors (NGFR inhibitors), vascular endothelial growth factor inhibitors (VEGF inhibitors), colony-stimulating factor inhibitors (CSF inhibitors), platelet-derived growth factor receptor inhibitors (PDGFR inhibitors), erythropoietin receptor inhibitors (EPOR inhibitors), fibroblast growth factor receptor inhibitors (FGFR inhibitors), hepatocyte growth factor receptor inhibitors (HGFR inhibitors), heregulin receptor inhibitors, and angiopoietin receptor inhibitors. Note that cell growth factor receptor inhibitor is synonymous with growth factor receptor inhibitor.

[0194] "Radiotherapeutic agents" include, but are not limited to, radioactive substances and radiosensitizers.

[0195] "Adjuvants" are used to suppress side effects and vomiting caused by anticancer drugs, and include, but are not limited to, for example, aprepitant, ondansetron, lorazepam, dexamethasone, diphenhydramine, ranitidine, cimetidine, ranitidine, famotidine, cimetidine, Procrit, epoetin alfa, filgrastim, oprelvekin, leucovorin, and granulocyte-macrophage colony-stimulating factor (GM-CSF).

[0196] Examples of "chemotherapeutic agents" that can be used include, but are not limited to, alkylating agents, platinum compounds, antimetabolites, topoisomerase inhibitors, DNA intercalators, antimitotic agents, anticancer antibiotics, plant-derived anticancer agents, epigenomic drugs, immunomodulatory agents, molecularly targeted therapeutic agents, angiogenesis inhibitors, and other chemotherapeutic agents. Representative examples are described below.

[0197] Examples of "alkylating agents" include, but are not limited to, nitrogen mustard, nitrogen mustard-N-oxide hydrochloride, chlorambucil, cyclophosphamide, ifosfamide, thiotepa, carboquone, improsulfan tosylate, busulfan, nimustine hydrochloride, mitobronitol, melphalan, dacarbazine, procarbazine, ranimustine, estramustine sodium phosphate, triethylenemelamine, carmustine, lomustine, streptozodin, pipobroman, etoglucide, and altretamine. , ambamustine, dibrospidium hydrochloride, fotemustine, prednimustine, bendamustine, uramustine, semustine, pumitepa, ribomustine, temozolomide, treosulfan, trofosfamide, zinostatin stimalamer, adozelesin, systustine, bizelesin, mecloethamine, uracil mustard, streptozocin, trabectedin, becaterin, chlormethine, mannosulfan, triaziquone, procarbacine, canfosfamide, nitrosoureas, and DDS preparations thereof.

[0198] "Platinum preparations" are not particularly limited, but examples include cisplatin, carboplatin, miboplatin, nedaplatin, satraplatin, oxaliplatin, triplatin tetranitrate, and DDS preparations thereof.

[0199] "Antimetabolites" include, but are not limited to, antifolates, pyrimidine metabolism inhibitors, purine metabolism inhibitors, ribonucleotide reductase inhibitors, and nucleotide analogs.

[0200] Examples of "antimetabolites" include, but are not limited to, mercaptopurine, 6-mercaptopurine riboside, thioinosine, methotrexate, pemetrexed, eocitabine, enocitabine, cytarabine, cytarabine ocfosfate, ancitabine hydrochloride, 5-FU drugs (e.g., fluorouracil, Calzoner, Bennan, Ruconal, Lunavon, tegafur, tegafur-uracil, tegafur-gimeracil-oteracil potassium (TS-1), UFT, doxifluridine, carmofur, galocitabine, emitefur, capecitabine, etc.), aminopterin, nelarabine, leukoporin calcium, tabloid, butosin, folinate calcium, levofolinate calcium, cladribine, emitefur, fludara iben, gemcitabine, hydroxycarbamide, pentostatin, piritrexim, idoxuridine, mitoguazone, tiazofurin, ambamustine, bendamustine, floxuridine, nelarabine, leucovorin, hydroxyurea, thioguanine, asparaginase, bortezomib, raltitrexed, clofarabine, enocitabine, sapacitabine, azacitidine , sulfadiazine, sulfamethoxazole, trimethoprim, liproxstatin-1, D4476, xanthohumol, Epacadostat (INCB024360), Vidofludimus, P7C3, GMX1778 (CHS828), NCT-501, SW033291, Ro61-8048, and DDS formulations thereof.

[0201] Examples of "topoisomerase inhibitors" include, but are not limited to, doxorubicin, daunorubicin, epirubicin, idarubicin, anthracenedione, mitoxantrone, mitomycin C, bleomycin, dactinomycin, plicatomycin, irinotecan, camptothecin, rubitecan, belotecan, etoposide, teniposide, topotecan, amsacrine, and DDS preparations thereof.

[0202] "DNA intercalators" are not particularly limited, but examples include proflavine, doxorubicin (adriamycin), daunorubicin, dactinomycin, thalidomide, and DDS preparations thereof.

[0203] Examples of "antimitotic agents" include, but are not limited to, paclitaxel, paclitaxel derivatives (e.g., DHA-paclitaxel, polyglutamated paclitaxel, nab-paclitaxel, paclitaxel micelles, 7α-glucosyloxyacetylpaclitaxel, BMS-275183, etc.), docetaxel, vinorlevin, vincristine, vinblastine, vindesine, vinzolidine, etoposide, teniposide, ixabepilone, larotaxel, ortataxel, tesetaxel, ispinesib, colchicine, vinflunine, and DDS formulations thereof.

[0204] Examples of "anticancer antibiotics" include, but are not limited to, actinomycin D, actinomycin C, mitomycin C, chromomycin A3, mithramycin A, bleomycin hydrochloride, bleomycin sulfate, peplomycin sulfate, daunorubicin hydrochloride, doxorubicin hydrochloride, aclarubicin hydrochloride, pirarubicin hydrochloride, epirubicin hydrochloride, alumubicin hydrochloride, neocarzinostatin, zinostatin stimalamer, mithramycin, sarkomycin, carzinophilin, mitotane, zorubicin hydrochloride, mitoxantrone hydrochloride, idarubicin hydrochloride, liposomal doxavircin, and DDS preparations thereof.

[0205] Examples of "plant-derived anticancer agents" include, but are not limited to, irinotecan, nogitecan, etoposide, etoposide phosphate, eribulin, sobuzoxane, vinblastine sulfate, vincristine sulfate, vindesine sulfate, teniposide, paclitaxel, paclitaxel injection, docetaxel, DJ-927, vinorelbine, topotecan, and DDS formulations thereof.

[0206] Examples of "epigenomic drugs" include, but are not limited to, DNA methylation inhibitors, histone deacetylase (HDAC) inhibitors, DNA methyltransferase (DNMT) inhibitors, histone deacetylase activators, histone demethylase inhibitors, and methylated nucleotides.

[0207] Examples of "epigenomic drugs" include, but are not limited to, vorinostat, belinostat, mocetinostat (MGCD0103), entinostat (SNDX-275), romidepsin, azacitidine, decitabine, GSK2879552 2H1, SGC707, ORY-1001 (RG-6016), PFI-4, SirReal2, GSK2801, CPI-360, GSK503, AMI-1, CPI-169, and DDS formulations thereof.

[0208] "Immunomodulators" are not particularly limited, but include, for example, thalidomide, lenalidomide, pomalidomide, and DDS preparations thereof.

[0209] The "molecularly targeted therapeutic agent" may be a low molecular weight compound or an antibody. Examples of the "molecularly targeted therapeutic agent" include, but are not limited to, kinase inhibitors, proteasome inhibitors, monoclonal antibodies, mTOR inhibitors, TNF inhibitors, and T cell inhibitors.

[0210] Examples of "kinase inhibitors" include, but are not limited to, tyrosine kinase inhibitors, serine / threonine kinase inhibitors, Raf kinase inhibitors, CDK (cyclin-dependent kinase) inhibitors, and MEK (mitogen-activated protein kinase) inhibitors.

[0211] Specifically, examples of "kinase inhibitors" include, but are not limited to, imatinib, gefitinib, erlotinib, afatinib, dasatinib, bosutinib, vandetanib, sunitinib, axitinib, pazopanib, lenvatinib, lapatinib, nintedanib, nilotinib, crizotinib, ceritinib, alectinib, ruxolitinib, tofacitinib, ibrutinib, sorafenib, vemurafenib, dabrafenib, palbociclib, trametinib, regorafenib, cedibanib, lestaurtinib, Examples of such anti-cancer drugs include vandetinib, vatalanib, seliciclib, tivantinib, canertinib, pelitinib, tesevatinib, cediranib, motesanib, midostaurin, foretinib, cabozanteinib, selumetinib, neratinib, volasertib, saracatinib, enzastaurin, tandutinib, semaxanib, alvocidib, ICR-62, AEE788, PD0325901, PD153035, TK787, amcasertib (BBI503), E6201, E7050, and DDS formulations thereof.

[0212] "Proteasome inhibitors" are not particularly limited, but examples include bortezomib, carfilzomib, and DDS formulations thereof.

[0213] The "monoclonal antibody" is not particularly limited, and examples thereof include an anti-CD22 antibody, an anti-CD20 antibody, an anti-CD25 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD5 antibody, an anti-CD52 antibody, an anti-epidermal growth factor receptor antibody (EGFR antibody), an anti-vascular endothelial growth factor antibody (VEGF antibody), an anti-TNF-α antibody, an anti-IL-1 receptor antibody, an anti-IL-2 receptor antibody, an anti-IL-5 receptor antibody, an anti-IL-6 receptor antibody, an anti-HER2 antibody, an anti-IgE antibody, an anti-IgG antibody, an anti-respiratory syncytial virus antibody, an anti-CCR4 antibody, an anti-CTLA-4 (cytotoxic T-lymphocyte-associated antigen 4, CD152) antibody, an anti-PD-1 antibody, an anti-RANKL (receptor activator of nuclear factor κB ligand) antibody, an anti-c-Met antibody, and an anti-CXCR4 antibody.

[0214] Specifically, the "monoclonal antibody" is not particularly limited, but examples thereof include ibritumomab tiuxetan, rituximab, cetuximab, infliximab, basiliximab, and brentuximab. Examples include vedotin, tocilizumab, trastuzumab, bevacizumab, omalizumab, mepolizumab, gemtuzumab, ozogamicin, palivizumab, ranibizumab, certolizumab, ocrelizumab, mogamulizumab, eculizumab, pertuzumab, alemtuzumab, inotuzumab, panitumumab, ofatumumab, golimumab, adalimumab, ramucirumab, nivolumab, anakinra, denosumab, ipilimumab, pembrolizumab, matuzumab, farletuzumab, MORAb-004, MORA-b009, and DDS formulations thereof.

[0215] Examples of "mTOR inhibitors" include, but are not limited to, everolimus (RAD001), rapamycin (sirolimus), AZD8055, temsirolimus (CCI-779, NSC683864), KU-0063794, voxtalisib (XL-765, SAR245409), MHY1485, dactolisib (BEZ235), PI-103, torkinib (PP242), ridaforolimus (deforolimus, MK-8669), INK-128 (MLN0128), torin1, omipalisib (GSK2126458, GSK458), OSI- 027, PF-04691502, apitolisib (GDC-0980, RG7422), GSK1059615, gedatolisib (PF-05212384, PKI-587), WYE-132, PP121, WYE-354, AZD2014, Torin2, WYE-687, CH5132799, WAY-600, ETP-46464, GDC-0349, XL388, zotarolimus (ABT-578), tacrolimus (FK506), BGT226 (NVP-BGT226), Palomid 529 (P529), chrysophanic acid, and DDS formulations thereof.

[0216] "TNF inhibitors" include, but are not limited to, etanercept, lenalidomide (CC-5013), pomalidomide, thalidomide, necrostatin-1, and QNZ (EVP4593).

[0217] "T cell inhibitors" are not particularly limited, but examples include abatacept and the like.

[0218] Examples of "angiogenesis inhibitors" include, but are not limited to, CM101, IFN-α, IL-12, platelet factor-4, suramin, semaxanib, thrombospondin, VEGFR antagonists, angiogenesis inhibitory steroids plus heparin, cartilage-derived angiogenesis inhibitory factor, matrix metalloproteinase inhibitors, batimastat, marimastat, angiostatin, endostatin, 2-methoxyestradiol, tecogalan, thrombospondin, αVβ3 inhibitors, linomide, ADH-1, E7820, and DDS formulations thereof.

[0219] "Other chemotherapeutic agents" include, but are not limited to, for example, fistenalide, sobuzoxane, obatoclax, efaproxiral, tipifarnib, lonafarnib, etc.

[0220] The pharmaceutical composition of the present invention may further contain other additives, such as surfactants, antioxidants, preservatives, and soothing agents.

[0221] The compound of formula (1) or a pharmaceutically acceptable salt thereof can be administered simultaneously with or at a later time than the antigenic substance (immunogen). The dose is usually 5 to 5,000 mg / m 2(body surface area), i.e., a unit dose in the range of about 0.1 ng / kg to 100 mg / kg, which usually provides an effective dose as a vaccine adjuvant. Unit dosage forms such as injections usually contain, for example, 1 ng to 250 mg of active ingredient. Preferably, a daily dose in the range of 1 ng to 50 mg / kg is used. However, this daily dose may need to be varied depending on the host being treated, the specific route of administration, and the severity of the disease being treated. Therefore, the optimal dose may be determined by the practitioner treating the individual patient or warm-blooded animal.

[0222] As used herein, "treatment" is used in a sense that includes alleviating, in whole or in part, any, some or all of the symptoms of a disease, or arresting or slowing the progression of the pathological condition.

[0223] As used herein, "prevention" is used to mean both primary prevention of a disease (preventing the onset of a disease) and secondary prevention (preventing recurrence in patients whose symptoms have been alleviated or whose illness has been cured after the onset of the disease, i.e., preventing recurrence).

[0224] The compound of the present invention or a pharmaceutically acceptable salt thereof has immune adjuvant activity in vitro or in vivo, and is therefore useful as a vaccine adjuvant for maintaining or enhancing the immunogenicity of an antigen (tumor antigen or pathogen-derived antigen).

[0225] The compound of the present invention or a pharmaceutically acceptable salt thereof has adjuvant activity for cellular immunity in vitro or in vivo, and is therefore useful as a vaccine adjuvant for maintaining or enhancing the immunogenicity of tumor antigens.

[0226] The compound of the present invention or a pharmaceutically acceptable salt thereof can be used to maintain or improve the activity of an immunopotentiator (immunostimulator), i.e., a substance that induces an antigen (tumor antigen or pathogen-derived antigen)-specific immune response, which is a therapeutic or preventive agent for a disease. A pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof and a substance that enhances a tumor antigen-specific immune response or a pathogen-specific immune response (also referred to as a tumor antigen or a pathogen-derived antigen) is also an aspect of the present invention. The tumor antigen includes, but is not limited to, an antigen protein or an antigen peptide (partial peptide) derived from the antigen protein, a tumor antigen protein or a tumor antigen peptide (partial peptide) derived from the tumor antigen protein, and complexes of these with a carrier.

[0227] In a specific embodiment of the present invention, the compound of the present invention or a pharmaceutically acceptable salt thereof can be administered in combination with a tumor antigen protein or tumor antigen peptide for cancer immunotherapy to treat or prevent cancer, such as leukemia, myelodysplastic syndrome, multiple myeloma, malignant lymphoma, gastric cancer, colon cancer, lung cancer, breast cancer, germ cell cancer, liver cancer, skin cancer, bladder cancer, prostate cancer, uterine cancer, cervical cancer, ovarian cancer, brain tumor, bone cancer, pancreatic cancer, head and neck cancer, cutaneous or intraorbital malignant melanoma, rectal cancer, anal cancer, testicular cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, parenchymal carcinoma, and the like. These include fibrosarcoma, urethral cancer, penile cancer, chronic or acute leukemia including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, and chronic lymphocytic leukemia, childhood solid tumors, lymphocytic lymphoma, kidney or ureter cancer, renal pelvis carcinoma, central nervous system (CNS) tumors, primary CNS lymphoma, tumor angiogenesis, spinal tumors, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, squamous cell carcinoma, squamous cell carcinoma, T-cell lymphoma, glioblastoma multiforme, malignant melanoma, non-small cell lung cancer, renal cell carcinoma, and asbestos-induced cancer. Here, treatment or prevention of cancer includes prevention of metastatic disease and tumor recurrence, as well as prevention and treatment of paraneoplastic syndromes.

[0228] In a specific embodiment of the present invention, the compound of the present invention or a pharmaceutically acceptable salt thereof can be administered in combination with an active ingredient of a vaccine for preventing infectious diseases to prevent various infectious diseases, for example, viral diseases such as genital warts, common warts, plantar warts, hepatitis B, hepatitis C, herpes simplex virus, molluscum contagiosum, smallpox, human immunodeficiency virus (HIV), human papillomavirus (HPV), respiratory syncytial virus, norovirus, cytomegalovirus (CMV), varicella-zoster virus (VZV), rhinovirus, adenovirus, coronavirus, influenza, and parainfluenza; bacterial diseases such as tuberculosis, Mycobacterium avium, and leprosy; fungal diseases, chlamydia, candida, aspergillus, cryptococcal meningitis, pneumocystis carinii, cryptosporidiosis, histoplasmosis, toxoplasmosis, malaria, trypanosoma infection, and leishmaniasis. The active ingredients of infectious disease preventive vaccines are not particularly limited, and include substances derived from microorganisms / pathogens such as bacteria, fungi, protozoa, and viruses that cause infectious diseases, such as antigenic proteins, antigenic peptides (partial peptides) derived from such proteins, polysaccharides, lipids, and combinations thereof, or combinations of substances derived from the microorganisms / pathogens and carriers.

[0229] Viral antigen peptides derived from viral antigens are not particularly limited, and examples thereof include influenza matrix protein peptide 58-66 (Jager E et al., Int. J. Cancer 67:54(1996)), HPV16 E7 peptide 86-93 (van Driel WJ et al., Eur. J. Cancer 35:946(1999)), HPV E7 peptide 12-20 (Scheibenbogen C et al., J. Immunother 23:275(2000)), HPV16 E7 peptide 11-20 (Smith JWI et al., J. Clin. Oncol. 21:1562(2003)), HSV2 gD (Berman PW et al., Science 227:1490(1985)), CMV gB (Frey SE et al., Infect Dis. 180:1700(1999)), Gonczol E. et al., Exp. Opin. Biol. Ther. 1:401(2001)), CMV pp65 (Rosa CL et al., Blood 100:3681(2002), Gonczol E. et al., Exp. Opin. Biol. Ther. 1:401(2001)), and the like.

[0230] The carrier used in the present invention is a substance that chemically and / or physically binds an antigenic protein or antigenic peptide, and examples thereof include proteins and lipids, and are not particularly limited. Examples of such carriers include CRM197 (Vaccine. 2013 Oct 1;31(42):4827-33), KLH (Cancer Immunol Immunother. 2003 Oct;52(10):608-16), virus-like particles (PLoS ONE 5(3): e9809), and liposomes (J Liposome Res. 2004;14(3-4):175-89).

[0231] The antigen protein can be prepared by cloning cDNA encoding the antigen protein and expressing it in host cells according to basic texts such as Molecular Cloning 2nd Ed., Cold Spring Harbor Laboratory Press (1989).

[0232] Antigen peptides can be synthesized according to methods commonly used in peptide chemistry, including those described in literature (Peptide Synthesis, Interscience, New York, 1966; The Proteins, Vol. 2, Academic Press Inc., New York, 1976).

[0233] In one aspect of the present invention, there is provided a kit containing: a) a compound represented by formula (1) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the compound represented by formula (1) or a pharmaceutically acceptable salt thereof; b) an antigen (tumor antigen or pathogen-derived antigen) or a pharmaceutical composition containing an antigen (tumor antigen or pathogen-derived antigen). The antigen is not particularly limited as long as it is an antigen that can be used as an active ingredient in a vaccine, and examples thereof include the above-mentioned antigen protein or an antigen peptide (partial peptide) derived from the antigen protein, as well as complexes of these with a carrier.

[0234] In one aspect of the present invention, there is provided a kit containing: a) a compound represented by formula (1) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof; b) a tumor antigen or a pharmaceutical composition containing a tumor antigen. Here, the tumor antigen is not particularly limited as long as it is a tumor antigen that can be used as an active ingredient in a cancer vaccine, and examples thereof include the above-mentioned tumor antigen proteins or tumor antigen peptides (partial peptides) derived from the tumor antigen proteins, as well as complexes of these with carriers.

[0235] In one aspect of the present invention, there is provided a kit containing: a) a compound represented by formula (1) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof; b) a pathogen-derived antigen or a pharmaceutical composition containing a pathogen-derived antigen. Here, the pathogen-derived antigen is not particularly limited as long as it is a pathogen-derived antigen that can be used as an active ingredient in an infectious disease vaccine, and examples thereof include the above-mentioned pathogen-derived antigen protein or a pathogen-derived antigen peptide (partial peptide) derived from the pathogen-derived antigen protein, as well as a complex of these with a carrier.

[0236] One aspect of the present invention provides the use of a compound of formula (1), or a pharmaceutically acceptable salt thereof, for the manufacture of a vaccine adjuvant. Also provided is the use of a compound of formula (I), as defined above, or a pharmaceutically acceptable salt thereof, as a vaccine adjuvant in the manufacture of a vaccine for the treatment of cancer or an infectious disease.

[0237] One aspect of the present invention provides the use of a compound of formula (1), or a pharmaceutically acceptable salt thereof, for the manufacture of a vaccine adjuvant for a cancer vaccine. Also provided is the use of a compound of formula (I), as defined above, or a pharmaceutically acceptable salt thereof, as a vaccine adjuvant in the manufacture of a cancer vaccine for the treatment of cancer.

[0238] One aspect of the present invention provides use of a compound represented by formula (1) or a pharmaceutically acceptable salt thereof for the manufacture of a vaccine adjuvant for an infectious disease vaccine. Also, one aspect of the present invention provides use of a compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof as a vaccine adjuvant in the manufacture of an infectious disease vaccine for the treatment of infectious diseases.

[0239] Furthermore, as one aspect of the present invention, there is provided a method for treating, preventing the progression of, or preventing cancer or an infectious disease, which comprises the step of administering to a patient a compound represented by formula (I) defined above, or a pharmaceutically acceptable salt thereof, together with an antigen (tumor antigen or pathogen-derived antigen).

[0240] Furthermore, as one aspect of the present invention, there is provided a method for treating, preventing the progression of, or preventing cancer, which comprises the step of administering to a patient a compound represented by formula (I) as defined above, or a pharmaceutically acceptable salt thereof, together with a tumor antigen.

[0241] Furthermore, as one aspect of the present invention, there is provided a method for treating, preventing the progression of, or preventing an infectious disease, which comprises the step of administering to a patient a compound represented by formula (I) defined above, or a pharmaceutically acceptable salt thereof, together with an antigen derived from a pathogen.

[0242] The present invention will be specifically explained below with reference to examples, but the present invention is not limited to these examples in any way.

[0243] Fmoc: 9-fluorenylmethyloxycarbonyl Boc: tert-butoxycarbonyl Alko: p-alkoxybenzyl alcohol PEG: polyethylene glycol tBu: tert-butyl HBTU: O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate DIPEA: N,N-diisopropylethylamine DMF: N,N-dimethylformamide TFA: trifluoroacetic acid TIS: triisopropylsilane THF: tetrahydrofuran TBS: tert-butyldimethylsilyl group TBDPS: tert-butyldiphenylsilyl group

[0244] The measurement conditions for high performance liquid chromatography mass spectrometry (LCMS) are as follows.

[0245] LCMS Condition A MS detector: LCMS-IT-TOF HPLC: Shimadzu Nexera X2 LC 30AD Column: Kinetex 1.7μ C18 100A New column 50×2.1 mm Flow rate: 1.2 ml / min Measurement wavelength: 254 / 220 nm Mobile phase: Solution A; 0.1% formic acid aqueous solution Solution B; acetonitrile Time program: Step Time (min) 1 0.01-1.40 Solution A:Solution B = 90:10 to 5:95 2 1.40-1.60 Solution A:Solution B = 5:95 3 1.61-2.00 Solution A:Solution B = 99:1

[0246] LCMS Condition B MS detector: ACQUITY (registered trademark) SQdetector (Waters) HPLC: ACQUITY (registered trademark) system Column: Waters ACQUITY HPLC (registered trademark) BEH C18 (1.7 μm, 2.1 mm × 30 mm) Flow rate: 0.8 ml / min Measurement wavelength: 254 / 220 nm Mobile phase: Solution A: 0.06% formic acid / acetonitrile, Solution B: 0.06% formic acid aqueous solution Time program: 0.0-1.30 Solution A: Solution B = 2:98 to 96:4 Column temperature: 25°C

[0247] Reference Example 1: Synthesis of a peptide consisting of the amino acid sequence: RMFPNAPYL (Arg-Met-Phe-Pro-Asn-Ala-Pro-Tyr-Leu) (SEQ ID NO: 1) Starting from 1.00 g of Fmoc-Lys(Boc)-Alko-PEG resin (Watanabe Chemical Industry Co., Ltd.; 0.23 mmol / g, 0.23 mmol), a peptide chain was assembled by solid-phase synthesis using the Fmoc / tBu method. A CS336X peptide synthesizer (CS Bio) was used for solid-phase synthesis, and deprotection of the Fmoc group was carried out by treatment with a 20% piperidine solution in DMF for 5 minutes and 20 minutes. Coupling of the protected amino acid was carried out by reacting with a DMF solution of 1.05 mmol of the protected amino acid, 1 mmol of HBTU, and 2 mmol of DIPEA for 1 hour. The resulting resin was washed with DMF and ether and then dried under reduced pressure to obtain a peptide resin. To this peptide resin, 10 mL of a mixture of TFA / water / TIS (volume ratio: 94 / 2.5 / 2.5) was added, and the mixture was shaken at room temperature for 2 hours. After filtering off the resin, the reaction mixture was concentrated under reduced pressure. The reaction mixture was ice-cooled, and diethyl ether (50 mL) was added. The resulting precipitate was collected by filtration, washed with ether, and dried under reduced pressure to obtain the crude peptide. The resulting crude peptide was dissolved in a mixture of 20% aqueous acetic acid and acetonitrile (volume ratio: 1 / 1) and purified under the conditions shown below to obtain 0.16 g of trifluoroacetate of RMFPNAPYL (Arg-Met-Phe-Pro-Asn-Ala-Pro-Tyr-Leu) (SEQ ID NO: 1). An acetate salt was prepared from the resulting trifluoroacetate according to a standard method and used for evaluation. Mass spectrometry: m / z = 554.73 [M+2H] ＋2, retention time: 0.82 min (LCMS condition A) Purification conditions HPLC system: Gilson high-throughput HPLC preparative system Column: YMC ODS-A 3 cmφ×25 cm, 10 μm Eluent 1: 0.1% TFA water Eluent 2: 0.035% TFA acetonitrile Flow rate: 20 mL / min Gradient method:

[0248] The peptides shown in Table 1 were obtained as trifluoroacetates using the corresponding starting materials according to the method described in Reference Example 1. Since this compound is not a compound of the present invention, it is used as a Reference Example. Note that the compound in Reference Example 3 was converted to an acetate salt according to a standard method and used for the subsequent evaluation.

[0249] According to the method described in WO 2014 / 157692, the compound shown in Table 2 was obtained as a trifluoroacetate salt (in the formula, the bond between C and C represents a disulfide bond). This compound is not a compound of the present invention, and therefore is used as a reference example.

[0250]

[0251] Reference Example 9 Synthesis of N-2,2,3,3-pentamethyl-4,7,10,13,16-pentaoxa-3-silaoctadecane-18-amine

[0252] Process 1 To a solution of the known compound 14-amino-3,6,9,12-tetraoxatetradecan-1-ol (1.60 g) in THF (25 mL), triethylamine (4.7 mL) and ethyl trifluoroacetate (2.4 mL) were added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (mobile phase: chloroform / methanol) to give 2,2,2-trifluoro-N-(14-hydroxy-3,6,9,12-tetraoxatetradecan-1-yl)acetamide (1.00 g). m / z = 334 [M+H] + , Rt = 0.507 (LCMS condition B)

[0253] Process 2 To a solution of the compound obtained in Step 1 of Reference Example 9 (3.91 g) in DMF (20 mL), triethylamine (4.90 mL) and tert-butyldimethylchlorosilane (3.54 g) were added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was diluted with ethyl acetate, washed with water and saturated brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (mobile phase: hexane / ethyl acetate) to give 2,2,2-trifluoro-N-(2,2,3,3-tetramethyl-4,7,10,13,16-pentaoxa-3-silaoctadecane-18-yl)acetamide (3.70 g). m / z = 448 [M+H] + , Rt = 1.153 (LCMS condition B)

[0254] Process 3 Cesium carbonate (6.46 g) and iodomethane (1.6 g) were added to a DMF (20 mL) solution of the compound (4.44 g) obtained in Step 2 of Reference Example 9, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate, washed with water and saturated brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (mobile phase: hexane / ethyl acetate) to give 2,2,2-trifluoro-N-methyl-N-(2,2,3,3-tetramethyl-4,7,10,13,16-pentaoxa-3-silaoctadecane-18-yl)acetamide (3.31 g). m / z = 463 [M+H] + , Rt = 1.210 (LCMS condition B)

[0255] Process 4 Potassium carbonate (70 mg) was added to a solution of the compound obtained in Step 3 of Reference Example 9 (117 mg) in methanol (5 mL), and the mixture was stirred at room temperature for 5 hours. The reaction mixture was concentrated, and the resulting crude product was purified by silica gel chromatography (mobile phase: chloroform / methanol) to give N-2,2,3,3-pentamethyl-4,7,10,13,16-pentaoxa-3-silaoctadecane-18-amine (67 mg). m / z = 366 [M+H + ], Rt = 0.718 (LCMS condition B)

[0256] Reference Example 10 Synthesis of N,2,2-trimethyl-3,3-diphenyl-4,7,10,13,16-pentaoxa-3-silaoctadecane-18-amine The title compound was obtained according to the method of Reference Example 9. m / z = 490 [M+H] + , Rt = 0.953 (LCMS condition B)

[0257] Reference Example 11 Synthesis of (3S)-3-{[2-amino-5-({2-methoxy-4-[(methylamino)methyl]phenyl}methyl)-6-methylpyrimidin-4-yl]amino}hexan-1-ol The title compound was synthesized according to the method described in WO2012 / 066336. m / z = 194 [M+2H] +2 , Rt = 0.552 (LCMS condition B)

[0258] Reference Example 12 5-({2-methoxy-4-[(methylamino)methyl]phenyl}methyl)-6-methyl-N ４ Synthesis of -pentylpyrimidine-2,4-diamine The known compound 5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-6-methyl-N ４ The title compound was obtained from -pentylpyrimidine-2,4-diamine by the same reaction and treatment as in Example 1. m / z = 179 [M+2H] +2 , Rt = 0.475 (LCMS condition B)

[0259] Reference Example 13: Synthesis of 3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-methoxybenzaldehyde The title compound was obtained from the known compound methyl 3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-methoxybenzoate by a reaction and treatment method according to WO2017 / 061532. m / z = 179 [M+2H] +2 , Rt = 0.475 (LCMS condition B)

[0260] Reference Example 14: Synthesis of 3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-hydroxybenzaldehyde A solution of boron tribromide in dichloromethane (1.0 M, 0.8 mL) was added to a solution of Reference Example 13 (104 mg) in dichloromethane (5 mL) under ice cooling, and the mixture was stirred at room temperature for 6 hours. Saturated aqueous sodium bicarbonate was added to the reaction mixture, followed by extraction with chloroform. The organic layer was dried over sodium sulfate, filtered, and then concentrated. The resulting crude product was purified by silica gel chromatography (mobile phase: chloroform / methanol) to give the title compound (54 mg). m / z = 329 [M+H] + , Rt = 0.748 (LCMS condition B)

[0261] Reference Example 15 5-({2-methoxy-5-[(methylamino)methyl]phenyl}methyl)-6-methyl-N ４ Synthesis of -pentylpyrimidine-2,4-diamine The title compound was obtained by carrying out the reaction and treatment in accordance with the methods of Reference Example 13 to Example 3. m / z = 179 [M+2H] +2 , Rt = 0.599 (LCMS condition B)

[0262] Reference Example 16: Synthesis of 2-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-[(methylamino)methyl]phenol The title compound was obtained by carrying out the reaction and treatment in accordance with the methods of Reference Example 14 to Example 3. m / z = 173 [M+2H] +2 , Rt = 0.562 (LCMS condition B)

[0263] Reference Example 17 N,2,2,3,3-pentamethyl-4,7,10,13,16,19,22,25,28,31-decaoxa-3-silatritriacontan-33-amine The title compound was obtained according to the method of Reference Example 9. m / z = 587 [M+H] + , Rt = 0.865 (LCMS condition B)

[0264] Reference Example 18 N,2,2,3,3-pentamethyl-4,7,10,13,16,19,22,25,28,31,34,37,40,46,49,52,55,58,61,64,67,70,73-tetracosaoxa-3-silapentaheptacontan-75-amine The title compound was obtained according to the method of Reference Example 9. m / z = 402 [M+3H] +3 , Rt = 0.946 (LCMS condition B)

[0265] Reference Example 19 N,2,2,3,3-pentamethyl-4,7,10,13,16,19,22,25,28,31,34,37,40,46,49,52,55,58,61,64,67,70,73,76,79,82,85,88,91,94,97,100,103,106,109-hexatriacontaoxa-3-silahendecahectan-111-amine The title compound was obtained according to the method of Reference Example 9. m / z = 866 [M+2H] +2 , Rt = 0.948 (LCMS condition B)

[0266] Example 1 Synthesis of 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol trifluoroacetate The known compound 5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-6-methyl-N ４ To a solution of 1,4-pentylpyrimidine-2,4-diamine (75 mg) in acetonitrile (3 mL) were added Reference Example 9 (76 mg), potassium carbonate (65 mg), and potassium iodide (67 mg), and the mixture was stirred at 60° C. for 8 hours. The reaction solution was concentrated and then purified by reversed-phase HPLC in the same manner as in Reference Example 1 to give the title compound (96 mg). m / z = 290 [M+2H] +2 , Rt = 0.623 (LCMS condition B) 1H-NMR (CDCl3): δ 7.34 (s, 1H), 6.92 (d, J = 8.0 Hz, 1H), 6.96 (d, J = 8.0 Hz, 1H), 6.08 (t, J = 5.6 Hz, 1H), 3.96 (s, 3H), 3.88 (m, 2H), 3.70 (m, 2H), 3.64-3.55 (m, 18H), 3.47 (s, 1H), 3.34 (dd, J = 6.8, 12 Hz, 2H), 2.81 (s, 3H), 2.47 (s, 3H), 1.49-1.42 (m, 2H), 1.30-1.23 (m, 2H), 1.21-1.13 (m, 2H), 0.85 (t, J = 7.2 Hz, 3H)

[0267] Example 2 Synthesis of 1-{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxyphenyl}-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol trifluoroacetate The title compound was obtained from the known compound (3S)-3-[(2-amino-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-6-methylpyrimidin-4-yl)amino]hexan-1-ol by the reaction and treatment described in Example 1. m / z = 305 [M+2H] +2 , Rt = 0.527 (LCMS condition B)

[0268] Example 3 Synthesis of 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol trifluoroacetate To a solution of Reference Example 13 (64.3 mg) in THF (5 mL) were added Reference Example 9 (101 mg), acetic acid (5.4 μL), and sodium triacetoxyborohydride (199 mg), and the mixture was stirred at room temperature for 24 hours. Water was added to the reaction solution, which was neutralized with saturated aqueous sodium bicarbonate and then extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over sodium sulfate, filtered, and concentrated. The crude product was purified by HPLC to obtain the target TBDPS-protected compound. The obtained protected compound was dissolved in methanol and stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by reverse-phase HPLC to obtain the title compound (26 mg). m / z = 290 [M+2H] +2 , Rt = 0.661 (LCMS condition B) 1 H-NMR (CDCl3): δ 7.32 (dd, J = 1.6, 8.4 Hz, 1H), 7.13 (d, J = 1.6 Hz, 1H), 6.87 (d, J = 8.4 Hz, 1H), 6.24 (t, J = 4.8 Hz, 1H), 4.26 (m, 2H), 3.86 (s, 3H), 3.81 (m, 2H), 3.64-3.49 (m, 18H), 3.29 (dd, J = 6.8, 12.8 Hz, 2H), 2.70 (s, 3H), 2.43 (s, 3H), 1.44-1.36 (m, 2H), 1.24-1.17 (m, 2H), 1.13-1.07 (m, 2H), 0.80 (t, J = 7.2 Hz, 3H)

[0269] Example 4 Synthesis of 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-hydroxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol trifluoroacetate To a solution of Reference Example 14 (30 mg) in THF (5 mL) were added Reference Example 9 (49 mg), acetic acid (2.6 μL), and sodium triacetoxyborohydride (97 mg), and the mixture was stirred at room temperature for 24 hours. Water was added to the reaction solution, which was neutralized with saturated aqueous sodium bicarbonate and then extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over sodium sulfate, filtered, and concentrated. The crude product was purified by HPLC to obtain the target TBDPS-protected compound. The obtained protected compound was dissolved in methanol and stirred at room temperature for 12 hours. The reaction solution was concentrated and purified by reverse-phase HPLC to obtain the title compound (13 mg). m / z = 290 [M+2H] +2 , Rt = 0.661 (LCMS condition B) 1 H-NMR (CDCl3): δ 7.30 (t, J = 5.2 Hz, 1H), 7.07 (d, J = 2.0 Hz, 1H), 6.97 (dd, J = 1.6, 8.0 Hz, 1H), 6.89 (d, J = 8.4 Hz, 1H), 4.10 (m, 2H), 3.70 (t, J = 5.6 Hz, 2H), 3.63 (d, J = 5.6 Hz, 2H), 3.55-3.49 (m, 18H), 3.26 (dd, J = 7.2, 13.2 Hz, 2H), 2.68 (s, 3H), 2.43 (s, 3H), 1.47-1.40 (m, 2H), 1.24-1.15 (m, 2H), 1.13-1.05 (m, 2H), 0.76 (t, J = 6.8 Hz, 3H)

[0270] Example 5 Synthesis of 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(20-hydroxy-3,6,9,12,15,18-hexaoxaicosan-1-yl)-3-methoxybenzamide Diisopropylethylamine (25.9 mg), 20-amino-3,6,9,12,15,18-hexaoxaicosan-1-ol (31.3 mg), and HATU (33.5 mg) were added to a solution of 4-[(2-amino-4-{[(2S)-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoic acid (30 mg), a known compound, in DMF (1 mL), and the mixture was stirred at room temperature for 24 hours. Saturated aqueous sodium bicarbonate was added to the reaction solution, and the mixture was extracted with a chloroform / methanol (10:1) mixture. The extract was then dried over magnesium sulfate, filtered, and concentrated. The resulting crude product was purified by aminosilica gel column chromatography (chloroform / methanol as the mobile phase) to give the title compound (28.1 mg). m / z = 683 [M+H] + , Rt = 0.558 (LCMS condition B) 1 H-NMR (CDCl3): δ 7.49 (1H, s), 7.21 (1H, d, J = 7.9 Hz), 7.05 (1H, s),6.95 (1H, d, J = 7.9 Hz), 4.77 (1H, d, J = 6.7 Hz), 4.68 (2H, s), 4.01-3.92 (1H, m), 3.94 (3H, s), 3.75-3.68 (4H, m), 3.67-3.51 (26H, m), 3.42-3.35 (1H, m), 2.28 (3H, s), 1.45-1.36 (1H, m), 1.29-1.20 (1H, m), 1.16-1.06 (2H, m), 0.80 (3H, t, J = 7.3 Hz).

[0271] Example 6 Synthesis of 2,5,8,11-tetraoxatridecan-13-yl 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoate To a solution of the known compound 4-[(2-amino-4-{[(2S)-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoic acid (30 mg) in THF (1.5 mL), 2,5,8,11-tetraoxatridecan-13-ol (501 mg), diisopropylethylamine (36.2 mg), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (46.1 mg) were added, and the mixture was stirred at 60°C for 10 hours. The reaction solution was purified by silica gel column chromatography (mobile phase: chloroform / methanol) to obtain the title compound (16.2 mg). m / z = 565 [M+H] + , Rt = 0.665 (LCMS condition B) 1 H-NMR (CDCl3):δ 7.56 (1H, d, J = 7.9 Hz), 7.51 (1H, s), 6.99 (1H, d, J = 7.9 Hz), 6.12 (1H, d, J = 7.9 Hz), 4.41 (2H, t, J = 4.6 Hz), 4.20-4.10 (1H, m), 3.93 (3H, s), 3.86-3.41 (18H, m), 3.29 (3H, s), 3.02 (1H, q, J = 7.5 Hz), 2.44 (3H, s), 1.50-1.22 (2H, m), 1.12-0.91 (2H, m), 0.74 (3H, t, J = 7.0 Hz).

[0272] Example 7 5-{[2-methoxy-4-(2,5,8,11,14-pentaoxapentadecan-1-yl)phenyl]methyl}-6-methyl-N ４ Synthesis of -pentylpyrimidine-2,4-diamine Sodium hydride (7.2 mg, content >55%) was added to 2,5,8,11-tetraoxatridecan-13-ol and stirred at room temperature for 1 hour. After that, the known compound 5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-6-methyl-N ４To the mixture was added 20 mg of pentylpyrimidine-2,4-diamine and the mixture was stirred at 60°C for 3 hours. After adding water, the mixture was extracted with chloroform, dried over magnesium sulfate, filtered, and concentrated. The resulting crude product was purified by silica gel column chromatography (mobile phase: chloroform / methanol) to give the title compound (6.7 mg). m / z = 536 [M+H] + , Rt = 0.850 (LCMS condition B) 1 H-NMR (CDCl3):δ 6.88 (1H, s), 6.84 (1H, d, J = 7.3 Hz), 6.78 (1H, d, J = 7.3 Hz), 5.80-5.40 (2H, br), 4.46 (2H, s), 3.86 (3H, s), 3.67-3.46 (19H, m), 3.30 (3H, s), 3.24 (2H, td, J = 7.0, 5.5 Hz), 2.35 (3H, s), 1.36 (2H, m), 1.23-1.05 (4H, m), 0.78 (3H, t, J = 7.0 Hz).

[0273] Example 8 Synthesis of 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29-nonaoxa-2-azahentriacontan-31-ol The known compound 5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-6-methyl-N ４Acetonitrile (1 mL) was added to a mixture of 2,4-pentylpyrimidine-2,4-diamine (40 mg), Reference Example 17 (64.6 mg), potassium iodide (36.6 mg), and potassium carbonate (30.5 mg), and the mixture was stirred at 80°C for 2 hours. The reaction solution was diluted with chloroform, and then insoluble matter was filtered off and the mixture was concentrated. Chloroform (1 mL) and TFA (0.1 mL) were then added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was neutralized with aqueous sodium carbonate, extracted with chloroform, dried over magnesium sulfate, filtered, and concentrated. The resulting crude product was purified by aminosilica gel column chromatography (mobile phase: chloroform / methanol) to give the title compound (28.4 mg). m / z = 400 [M+2H] +2 , Rt = 0.583 (LCMS condition B) 1 H-NMR (CDCl3):δ 6.86 (1H, s), 6.81 (1H, d, J = 7.9 Hz), 6.73 (1H, d, J = 7.9 Hz), 6.00-5.00 (2H, br), 3.85 (3H, s), 3.68 (2H, t, J = 4.6 Hz), 3.63-3.50 (40H, m), 3.44 (2H, s), 3.24 (2H, q, J = 6.5 Hz), 2.54 (2H, t, J = 6.1 Hz), 2.32 (3H, s), 2.19 (3H, s), 1.41-1.34 (2H, m), 1.23-1.16 (2H, m), 1.13-1.05 (2H, m), 0.78 (3H, t, J = 7.3 Hz).

[0274] Example 9 Synthesis of 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71-tricosaoxa-2-azatriheptacontan-73-ol The reaction was carried out in the same manner as in Example 8 using Reference Example 18 (83 mg) to give the title compound (31.6 mg). m / z = 708 [M+2H] +2, Rt = 0.672 (LCMS condition B) 1 H-NMR (CDCl3):δ 6.83 (1H, s), 6.79 (1H, d, J = 7.9 Hz), 6.69 (1H, d, J = 7.3 Hz), 4.79 (1H, s), 4.56 (2H, s), 3.83 (3H, s), 3.76-3.40 (99H, m), 3.21 (2H, dd, J = 12.5, 7.0 Hz), 2.53 (2H, t, J = 6.1 Hz), 2.24 (3H, s), 2.18 (3H, s), 1.39-1.32 (2H, m), 1.22-1.14 (2H, m), 1.12-1.04 (2H, m), 0.77 (3H, t, J = 7.0 Hz).

[0275] Example 101 Synthesis of (4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92,95,98,101,104,107-pentatriacontaoxa-2-azanonahexan-109-ol The reaction was carried out in the same manner as in Example 8 using Reference Example 19 (95 mg) to give the title compound (26.3 mg). m / z = 487 [M+4H] +4 , Rt = 0.693 (LCMS condition B) 1H-NMR (CDCl3):δ 6.83 (1H, s), 6.79 (1H, d, J = 7.9 Hz), 6.69 (1H, d, J = 9.2 Hz), 4.81 (1H, s), 4.60 (2H, s), 3.83 (3H, s), 3.76-3.38 (147H, m), 3.21 (2H, dd, J = 12.5, 7.0 Hz), 2.53 (2H, t, J = 5.8 Hz), 2.24 (3H, s), 2.18 (3H, s), 1.39-1.32 (2H, m), 1.21-1.14 (2H, m), 1.12-1.06 (2H, m), 0.77 (3H, t, J = 7.3 Hz).

[0276] Example 11 Synthesis of 12-[(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3,6,9,15,18,21-hexaoxa-12-azatricosane-1,23-diol The title compound (13.1 mg) was obtained by reacting 3,6,9,15,18,21-hexaoxa-12-azatricosane-1,23-diol (40.7 mg) in the same manner as in Example 1. m / z = 349 [M+2H] +2 , Rt = 0.554 (LCMS condition B) 1 H-NMR (CDCl3):δ 6.85 (1H, s), 6.79 (1H, d, J = 7.9 Hz), 6.73 (1H, d, J = 7.9 Hz), 4.85 (1H, t, J = 5.5 Hz), 4.57 (2H, s), 3.83 (3H, s), 3.68-3.42 (34H, m), 3.21 (2H, td, J = 7.2, 5.3 Hz), 2.67 (4H, t, J = 6.1 Hz), 2.24 (3H, s), 1.41-1.33 (2H, m), 1.23-1.09 (4H, m), 0.78 (3H, t, J = 7.0 Hz).

[0277] Test Example 1 Human TLR7 reporter gene assay TLR7 / NF-κB / SEAPorter ＴＭThe HEK293 cell line (Imgenex Corporation) is a stable co-transfected cell line expressing full-length human TLR7 and a secreted alkaline phosphatase (SEAP) reporter gene under the transcriptional control of the NF-κB response element. TLR7 expression in this cell line was examined by flow cytometry. Stably expressing transformants were selected using the antibiotics blasticidin and geneticin. TLR signaling leads to NF-κB translocation, and promoter activation results in SEAP gene expression. The cells were incubated with the compounds synthesized in the Examples and Reference Examples for 24 hours at 37°C in the presence of 0.1% (v / v) dimethyl sulfoxide (DMSO), and TLR7-specific activation was assessed by measuring the level of SEAP produced. The degree of human TLR7 activation by the compounds of the present invention was assessed using a human TLR7 reporter gene assay, and the concentration of compound that produced half-maximal SEAP levels (EC ５０ ) are shown in Tables 3 and 4.

[0278] Test Example 2 Mouse TLR7 reporter gene assay HEK-Blue ＴＭ The mTLR7 cell line (Invivogen) is a stable co-transfected cell line expressing full-length mouse TLR7 and a secreted SEAP reporter gene under the transcriptional control of the NF-κB response element. TLR7 expression in this cell line was tested by RT-PCR. Stably expressing transformants were selected using the antibiotics blasticidin and zeocin. TLR signaling leads to NF-κB translocation, and promoter activation results in SEAP gene expression. TLR7-specific activation was assessed by incubating the cells with the compounds synthesized in the Examples and Reference Examples in the presence of 0.1% (v / v) DMSO at 37°C for 20-24 hours, and then measuring the level of SEAP produced. The extent of mouse TLR7 activation by the compounds of the present invention was assessed using a mouse TLR7 reporter gene assay, and the concentration of compound that produces half-maximal levels of SEAP (EC ５０ ) are shown in Tables 5 and 6.

[0279] The results of Test Examples 1 and 2 suggest that both the example compounds of the present invention and the reference example compounds act as agonists of human and mouse TLR7.

[0280] Test Example 3 HLA-A ＊ Evaluation of in vivo CTL induction ability using 24:02 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1 and the compound synthesized in Reference Example 12, a cocktail vaccine was prepared by mixing the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 with Montanide ISA 51 VG (hereinafter referred to as "cocktail vaccine b"), to which the compound synthesized in Example 1 was added, and a vaccine was also prepared by adding the compound synthesized in Reference Example 12 to cocktail vaccine b, and these were used to prepare HLA-A ＊ The compound was administered to 24:02 transgenic mice, and its adjuvant activity was evaluated by an antigen-specific cytotoxic T cell (CTL) induction test. CYTWNQMNL (SEQ ID NO: 2) contained in the compound represented by formula 4 binds to HLA-A. ＊ 24:02-restricted WT1 protein-derived antigen peptide.

[0281] HLA-A ＊ 24:02 Transgenic mice (C57BL / 6CrHLA-A2402 / K ｂ ) is a human MHC, HLA-A ＊ 24:02 and mouse MHC H-2K ｂ This mouse expresses chimeric HLA. ＊ It is possible to induce CTLs in 24:02-positive humans using peptides capable of inducing CTLs (Int J Cancer. 2002; 100: 565-70).

[0282] Whether or not CTLs against the peptide of interest (SEQ ID NO: 2) were induced by administration of cocktail vaccine b was determined by measuring whether IFNγ was produced when splenocytes derived from the above mice were restimulated with the peptide (SEQ ID NO: 2). Furthermore, whether or not the compound synthesized in Example 1 or Reference Example 12 exhibits adjuvant activity in vivo was determined by comparing the number of CTLs induced by administration of cocktail vaccine b with the number of CTLs induced by administration of a vaccine prepared by adding the compound synthesized in Example 1 or Reference Example 12 to cocktail vaccine b, and determining whether or not there was an increase in the number of CTLs.

[0283] Specifically, the compound represented by formula 4 and the peptide represented by SEQ ID NO:3 were dissolved in DMSO and then mixed with water for injection to achieve diluted concentrations of 6 mg / mL for the compound represented by formula 4 and 4.5 mg / mL for the peptide represented by SEQ ID NO:3. This peptide dilution was mixed with an equal volume of Montanide ISA 51 VG to form an emulsion, preparing cocktail vaccine b, which was administered intradermally at the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO:3. Alternatively, a vaccine prepared by adding the compound synthesized in Example 1 to the peptide dilution used in preparing cocktail vaccine b was administered intradermally at the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4, 225 μg / mouse of the peptide represented by SEQ ID NO:3, and 100 ng / mouse of the compound synthesized in Example 1. Alternatively, a vaccine prepared by adding the compound represented by Reference Example 12 to the peptide dilution solution used to prepare cocktail vaccine b was administered intradermally to the base of the tail of mice so that the compound represented by formula 4 was 300 μg / mouse, the peptide represented by SEQ ID NO: 3 was 225 μg / mouse, and the compound synthesized in Reference Example 12 was 69 ng / mouse. Administration was carried out twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２After euthanasia by gas, the spleens were removed and splenocytes were prepared. IFNγ production was measured using an IFNγ ELISPOT assay kit. The day before splenocyte preparation, the ELISPOT plate was treated with anti-mouse IFNγ antibody, and on the day of preparation, it was blocked with RPMI1640 medium containing 10% fetal bovine serum (FBS). ＊ 2.5 × 10 splenocytes from 24:02 transgenic mice ５ The cells were seeded onto a blocked ELISPOT plate at 1000x1000 cells / well. The peptide (SEQ ID NO: 2) was added to the splenocytes at a final concentration of 10 μg / mL in the presence of 0.1% (v / v). The peptide-added splenocytes were incubated at 37°C, 5% CO ２ After incubation, the supernatant was removed, and the number of spots on the colored ELISPOT plate was counted using an Immuno Spot Analyzer (CTL).

[0284] HLA-A ＊ The results of the IFNγ ELISPOT assay using 24:02 transgenic mice are shown in Figure 1. In Figure 1, the vertical axis indicates the number of cells in the inoculated cells that produced IFNγ in response to stimulation, and the horizontal axis indicates the vaccine administered to the mice. The black and white bars in Figure 1 indicate HLA-A ＊ The figure shows the results of culturing splenocytes derived from 24:02 transgenic mice in the presence and absence of the peptide represented by SEQ ID NO: 2. That is, the difference between the values ​​of the black and white bars indicates the number of IFNγ-producing cells specific to the peptide represented by SEQ ID NO: 2, i.e., CTLs, induced in vivo by administration of the vaccine. The values ​​of the white bars are hardly observed in Figure 1. This indicates that the splenocytes of the mice hardly reacted in the absence of the peptide of interest. As a result of this test, HLA-A ＊Induction of CTLs reactive to the peptide represented by SEQ ID NO: 2 was confirmed in 24:02 transgenic mice. Furthermore, the number of CTLs reactive to the peptide represented by SEQ ID NO: 2 was observed to be greater when the compound synthesized in Example 1 or the compound synthesized in Reference Example 12 was added to cocktail vaccine b. Furthermore, the increase in the number of CTLs was greater when the compound synthesized in Example 1 was added to cocktail vaccine b compared to when the compound synthesized in Reference Example 12 was added to cocktail vaccine b.

[0285] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of induced CTLs, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo. Furthermore, it was shown that the effect of the compound synthesized in Example 1 in increasing CTLs is higher than that of the compound synthesized in Reference Example 12, which does not contain a PEG structure.

[0286] Test Example 4 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1 and the compound synthesized in Reference Example 12, a cocktail vaccine (hereinafter referred to as "cocktail vaccine a") was prepared by mixing the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 with a pre-emulsified composition, and the compound synthesized in Example 1 or the compound synthesized in Reference Example 12 was added to the cocktail vaccine. ＊ The adjuvant activity was evaluated by an antigen-specific CTL induction test. (In the formula, the bond between C and C represents a disulfide bond.) RMFPNAPYL (SEQ ID NO: 1) contained in formula 4 and VLDFAPPGA (SEQ ID NO: 4) contained in the peptide represented by SEQ ID NO: 3 are HLA-A ＊ 02:01-restricted WT1 protein-derived antigen peptide.

[0287] HLA-A ＊The 02:01 transgenic mouse (C57BL / 6CrHLA-A2.1DR1) lacks mouse MHC and contains human MHC, HLA-A. ＊ 02:01 and mouse MHC H-2D ｂ Chimeric HLA and HLA-DRB1 ＊ These mice express HLA-A 01:01. ＊ It is possible to induce CTLs in O2:01-positive humans using peptides capable of inducing CTLs (Eur J Immunol. 2, 004; 34: 3, 060-9).

[0288] Whether or not CTLs against the antigen peptide (SEQ ID NO: 1 or SEQ ID NO: 4) were induced by administration of cocktail vaccine a was determined by measuring whether IFNγ was produced when splenocytes derived from the above mice were restimulated with the peptide of interest. Furthermore, whether or not the compound synthesized in Example 1 or Reference Example 12 exhibits adjuvant activity in vivo was determined by comparing the number of CTLs induced by administration of cocktail vaccine a with the number of CTLs induced by administration of a vaccine prepared by adding the compound synthesized in Example 1 or Reference Example 12 to cocktail vaccine a, and determining whether or not the number of CTLs increased.

[0289] Specifically, in vivo adjuvant activity was evaluated as follows. 0.312 g of sodium dihydrogen phosphate dihydrate was dissolved in 80 g of water for injection. 14.0 g of ethyl oleate, 14.0 g of octyldodecyl myristate, 2.0 g of sorbitan monooleate, 2.8 g of glycerin monooleate, 200.4 g of polyoxyethylene hydrogenated castor oil, and 0.4 g of glycerin were mixed. 2.354 mL (equivalent to 2.077 g) of this mixture was placed in a test tube, and 0.396 mL (equivalent to 0.396 g) of sodium dihydrogen phosphate aqueous solution was gradually added while stirring with a mixer (Ultra Turrax T10, IKA Corporation, or Touch Mixer MT-51, Yamato Scientific) to emulsify the mixture to prepare a pre-emulsified composition. The amount of the solution prepared was adjusted as needed. The compound represented by formula 4 and the peptide represented by SEQ ID NO: 3 were dissolved in DMSO and then mixed with water for injection to achieve a diluted concentration of 3 mg / mL for the compound represented by formula 4 and 2.25 mg / mL for the peptide represented by SEQ ID NO: 3. This peptide dilution was mixed with an equal amount of the above-mentioned pre-emulsified composition to form an emulsion, which was then administered intradermally to the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO: 3. Alternatively, a vaccine prepared by adding the compound synthesized in Example 1 to the peptide dilution used in preparing cocktail vaccine a was administered intradermally to the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4, 225 μg / mouse of the peptide represented by SEQ ID NO: 3, and 32.5 ng / mouse of the compound synthesized in Example 1. Alternatively, a vaccine prepared by adding the compound synthesized in Reference Example 12 to the peptide dilution solution used to prepare cocktail vaccine a was administered intradermally to the base of the tail of mice at a dose of 300 μg / mouse of the compound represented by formula 4, 225 μg / mouse of the peptide represented by SEQ ID NO: 3, and 22.5 ng / mouse of the compound synthesized in Reference Example 12. After one week, the mice were treated with CO ２After euthanasia by gas, the spleens were removed and splenocytes were prepared. IFNγ production was measured using an IFNγ ELISPOT assay kit. The day before preparation of splenocytes, the ELISPOT plate was treated with anti-mouse IFNγ antibody, and on the day of preparation, the plate was blocked with RPMI 1640 medium containing 10% FBS. 1.25 × 10 prepared splenocytes were cultured. ５ The cells were seeded onto a blocked ELISPOT plate at 1000x1000 cells / well. The peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added to the splenocytes at a final concentration of 10 μg / mL in the presence of 0.1% (v / v) DMSO. The peptide-added splenocytes were incubated at 37°C, 5% CO ２ The cells were cultured overnight under 5% CO₂ at 27°C for in vitro peptide restimulation. After incubation, the supernatant was removed, and the ELISPOT plate was developed according to the attached protocol. The number of developed spots was counted using an ImmunoSpot Analyzer.

[0290] As a result of this test, the HLA-A ＊ In 02:01 transgenic mice, induction of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was confirmed. Furthermore, similar to the results of Test Example 3, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was greater when the compound synthesized in Example 1 was added to cocktail vaccine a than when the compound synthesized in Reference Example 12 was added to cocktail vaccine a.

[0291] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of induced CTLs, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo. Furthermore, it was shown that the effect of the compound synthesized in Example 1 in increasing CTLs is higher than that of the compound synthesized in Reference Example 12, which does not contain a PEG structure.

[0292] Test Example 5 Enhancement of in vivo tumor growth inhibitory effect by vaccine HLA-A ＊24:02 3-methylcholanthrene suspended in corn oil was administered intradermally to the ventral side of transgenic mice, and HLA-A was detected in the tumors that developed at the administration site. ＊ A cell line (herein also referred to as MCA-A24 / Kb-WT1 tumor cells) in which the WT1 antigen peptide (SEQ ID NO: 2) was stably expressed in these cells was suspended in Hank's balanced salt solution and subjected to HLA-A ＊ The cells were then intradermally transplanted into the ventral skin of 24:02 transgenic mice (5 × 10 per mouse). ５ (100 mice). Three groups were administered: a vehicle group (group a), a vaccine group (group b), and a vaccine containing the compound synthesized in Example 1 (group c). Six mice were used for each group. Seven days before and seven days after tumor cell transplantation, mice in group a received an emulsified composition containing water for injection mixed with an equal volume of Montanide ISA 51 VG, which was then administered intradermally at the base of the tail (0.1 mL per mouse). Mice in group b received an emulsified composition containing the compound represented by formula 4 and the peptide represented by sequence number 3 mixed with an equal volume of Montanide ISA 51 VG, which was then administered intradermally at the base of the tail (300 μg of the compound represented by formula 4 per mouse and 225 μg of the peptide represented by sequence number 3 per mouse). To the mice in group c, a composition containing the compound represented by formula 4, the peptide represented by SEQ ID NO: 3, and the compound synthesized in Example 1 was mixed with an equal amount of Montanide ISA 51 VG to form an emulsion, which was then administered intradermally into the tail base (300 μg of the compound represented by formula 4 per mouse, 225 μg of the peptide represented by SEQ ID NO: 3 per mouse, and 100 ng of the compound synthesized in Example 1 per mouse per administration). 27 days after tumor inoculation, tumor diameters were measured and tumor volumes were calculated.

[0293] The mean tumor volumes of six mice from each group 27 days after tumor inoculation are shown in Figure 2. The vaccine (group b) significantly suppressed tumor cell growth compared to the vehicle (group a) (nonparametric Dunnett's multiple test, *: p<0.05). Furthermore, tumor cell growth was more significantly suppressed by adding the compound synthesized in Example 1 to the vaccine (group c, **: p<0.01).

[0294] These results demonstrate that the addition of the compound of the present invention to a vaccine enhances preventive tumor growth suppression.

[0295] The peptides shown in Table 7 were obtained as trifluoroacetate salts using the corresponding starting materials according to the method described in Reference Example 1. These compounds are not compounds of the present invention and are therefore used as Reference Examples.

[0296] According to the method described in WO 2007 / 063903, the compound shown in Table 8 was obtained as a trifluoroacetate salt (in the formula, the bond between C and C represents a disulfide bond). This compound is not a compound of the present invention, and therefore is used as a reference example.

[0297]

[0298] Test Example 6 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice. Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by mixing the peptide represented by SEQ ID NO: 6 synthesized in Reference Example 6 with Montanide ISA 51 VG (hereinafter referred to as "vaccine c"), and adding the compound synthesized in Example 1 to prepare a vaccine. ＊ The peptide GLYDGMEHL represented by SEQ ID NO: 6 was administered to 02:01 transgenic mice, and its adjuvant activity was evaluated by an antigen-specific CTL induction test. ＊ 02:01-restricted MAGE-A10 protein-derived antigen peptide.

[0299] Whether administration of vaccine c induces CTLs against the antigen peptide (SEQ ID NO: 6) was determined by measuring whether IFNγ was produced when splenocytes derived from the above mice were restimulated with the peptide of interest. Furthermore, whether the compound synthesized in Example 1 exerts adjuvant activity in vivo was determined by comparing the number of CTLs induced by administration of vaccine c with the number of CTLs induced by administration of a vaccine prepared by adding the compound synthesized in Example 1 to vaccine c, and determining whether the number increased.

[0300] Specifically, the peptide represented by SEQ ID NO: 6 was dissolved in DMSO and then mixed with water for injection to a diluted concentration of 0.1 mg / mL. This peptide dilution was mixed with an equal volume of Montanide ISA 51 VG to form an emulsion, and vaccine c was prepared. This vaccine was administered intradermally at the base of the tail of mice so that the peptide represented by SEQ ID NO: 6 was 10 μg / mouse. Alternatively, a vaccine was prepared by adding the compound synthesized in Example 1 to the peptide dilution used in preparing vaccine c. This vaccine was administered intradermally at the base of the tail of mice so that the peptide represented by SEQ ID NO: 6 was 10 μg / mouse and the compound synthesized in Example 1 was 0.4 nmol / mouse. Administration was performed twice, with a one-week interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 6) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0301] The results are shown in Figure 3. In Figure 3, the vertical axis represents the average number of cells that produced IFNγ in response to stimulation in the inoculated cells of three mice in each group, and the horizontal axis represents the vaccine administered to the mice. ＊1 shows the results of culturing splenocytes derived from 02:01 transgenic mice in the presence and absence of the peptide represented by SEQ ID NO: 6. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 6 was found to be greater when the compound synthesized in Example 1 was added to vaccine c than when no compound was added.

[0302] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0303] Test Example 7 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice. Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine (hereinafter referred to as "vaccine d") was prepared by mixing the peptide represented by SEQ ID NO: 5 synthesized in Reference Example 5 with Montanide ISA 51 VG, and the compound synthesized in Example 1 was added to the vaccine. ＊ The adjuvant activity was evaluated by an antigen-specific CTL induction test. ＊ 02:01-restricted Proteinase-3 protein-derived antigen peptide.

[0304] Whether administration of vaccine d induces CTLs against the antigen peptide (SEQ ID NO: 5) was determined by measuring whether IFNγ was produced when splenocytes derived from the above mice were restimulated with the peptide of interest. Furthermore, whether the compound synthesized in Example 1 exerts adjuvant activity in vivo was determined by comparing the number of CTLs induced by administration of vaccine d with the number of CTLs induced by administration of a vaccine prepared by adding the compound synthesized in Example 1 to vaccine d, and determining whether the number increased.

[0305] Specifically, the peptide represented by SEQ ID NO: 5 was dissolved in DMSO and then mixed with water for injection to a concentration of 2 mg / mL after dilution. This peptide dilution was mixed with an equal volume of Montanide ISA 51 VG to form an emulsion, and vaccine d was prepared. This vaccine was administered intradermally at the base of the tail of mice so that the peptide represented by SEQ ID NO: 5 was 100 μg / mouse. Alternatively, a vaccine was prepared by adding the compound synthesized in Example 1 to the peptide dilution used in preparing vaccine d, and this vaccine was administered intradermally at the base of the tail of mice so that the peptide represented by SEQ ID NO: 5 was 100 μg / mouse and the compound synthesized in Example 1 was 330 ng / mouse. After one week, the mice were inoculated with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 5) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0306] The results are shown in Figure 4. In Figure 4, the vertical axis represents the average number of cells that produced IFNγ in response to stimulation in the inoculated cells of three mice in each group, and the horizontal axis represents the vaccine administered to the mice. ＊ 1 shows the results of culturing splenocytes derived from 02:01 transgenic mice in the presence and absence of the peptide represented by SEQ ID NO: 5. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 5 was found to be greater when the compound synthesized in Example 1 was added to vaccine d than when no compound was added.

[0307] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0308] Test Example 8 HLA-A ＊Evaluation of in vivo adjuvant activity using 24:02 transgenic mice. Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, the compound synthesized in Example 1 was added to a vaccine (hereinafter referred to as "vaccine e") prepared by mixing the peptide represented by SEQ ID NO: 18 synthesized in Reference Example 20 with Montanide ISA 51 VG. ＊ The peptide TYAGCLSQIF represented by SEQ ID NO: 18 was administered to 24:02 transgenic mice, and its adjuvant activity was evaluated by an antigen-specific CTL induction test. ＊ 24:02-restricted Or7c1 protein-derived antigenic peptide.

[0309] Whether administration of vaccine e induces CTLs against the antigen peptide (SEQ ID NO: 18) was determined by measuring whether IFNγ was produced when splenocytes derived from the above mice were restimulated with the peptide of interest. Furthermore, whether the compound synthesized in Example 1 exerts adjuvant activity in vivo was determined by comparing the number of CTLs induced by administration of vaccine e with the number of CTLs induced by administration of a vaccine prepared by adding the compound synthesized in Example 1 to vaccine e, and determining whether the number increased.

[0310] Specifically, the peptide represented by SEQ ID NO: 18 was dissolved in DMSO and then mixed with water for injection to a concentration of 3 mg / mL after dilution. This peptide dilution was mixed with an equal volume of Montanide ISA 51 VG to form an emulsion, and vaccine e was prepared. This vaccine was administered intradermally at the base of the tail of mice so that the peptide represented by SEQ ID NO: 18 was 300 μg / mouse. Alternatively, a vaccine was prepared by adding the compound synthesized in Example 1 to the peptide dilution used in preparing vaccine e. This vaccine was administered intradermally at the base of the tail of mice so that the peptide represented by SEQ ID NO: 18 was 300 μg / mouse and the compound synthesized in Example 1 was 0.4 nmol / mouse. Administration was performed twice, with a one-week interval. One week after the final administration, the mice were immunized with CO ２After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 3, the splenocytes to which the peptide (SEQ ID NO: 18) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0311] The results are shown in Figure 5. In Figure 5, the vertical axis represents the average number of cells that produced IFNγ in response to stimulation in the inoculated cells of three mice in each group, and the horizontal axis represents the vaccine administered to the mice. ＊ 1 shows the results of culturing splenocytes derived from 24:02 transgenic mice in the presence and absence of the peptide represented by SEQ ID NO: 18. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 18 was found to be greater when the compound synthesized in Example 1 was added to vaccine e than when no compound was added.

[0312] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0313] Test Example 9 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice. Regarding the in vivo adjuvant activity of the compounds synthesized in Examples 8, 9, and 10 and the compound synthesized in Reference Example 12, a vaccine was prepared by adding the compound synthesized in Examples 8, 9, or 10 or the compound synthesized in Reference Example 12 to cocktail vaccine b, which was prepared by mixing the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 with Montanide ISA51 VG. ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0314] Specifically, the compound represented by formula 4 and the peptide represented by SEQ ID NO:3 were dissolved in DMSO and then mixed with water for injection to achieve a diluted concentration of 3 mg / mL for the compound represented by formula 4 and 2.25 mg / mL for the peptide represented by SEQ ID NO:3. This peptide dilution was mixed with an equal volume of Montanide ISA51 VG to form an emulsion, and cocktail vaccine b was administered intradermally at the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO:3. Alternatively, a vaccine prepared by adding the compound synthesized in Example 8, 9, or 10 or the compound synthesized in Reference Example 12 to the peptide dilution solution used in preparing cocktail vaccine b was administered intradermally at the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4, 225 μg / mouse of the peptide represented by SEQ ID NO:3, and 0.4 nmol / mouse of the compound synthesized in Example 8, 9, or 10 or the compound synthesized in Reference Example 12. The administration was carried out twice at a one-week interval. One week after the final administration, the mice were ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0315] The results are shown in Figures 6 and 7. As a result of this test, the number of peptide-reactive CTLs represented by SEQ ID NO: 1 or SEQ ID NO: 4 was observed to be greater when the compound synthesized in Example 8, 9, or 10 or the compound synthesized in Reference Example 12 was added to cocktail vaccine b, compared to when no compound was added. Furthermore, more peptide-reactive CTLs were observed when the compound synthesized in Example 8, 9, or 10 was added to cocktail vaccine b, compared to when the compound synthesized in Reference Example 12 was added to cocktail vaccine b.

[0316] This demonstrates that the addition of the compounds synthesized in Examples 8, 9, or 10 to vaccines increases the number of induced CTLs, strongly suggesting that the compounds synthesized in Examples 8, 9, or 10 have adjuvant activity in vivo. Furthermore, it was shown that the CTL-increasing effect of the compounds synthesized in Examples 8, 9, or 10 was higher than that of the compound synthesized in Reference Example 12, which does not contain a PEG structure.

[0317] Test Example 10 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice. The in vivo adjuvant activity of the compound synthesized in Example 11 was evaluated by adding the compound synthesized in Example 11 to cocktail vaccine b similar to that in Test Example 9 to prepare a vaccine. ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0318] Specifically, the compound represented by formula 4 and the peptide represented by SEQ ID NO:3 were dissolved in DMSO and then mixed with water for injection to achieve a diluted concentration of 3 mg / mL for the compound represented by formula 4 and 2.25 mg / mL for the peptide represented by SEQ ID NO:3. This peptide dilution was mixed with an equal volume of Montanide ISA51 VG to form an emulsion, and cocktail vaccine b was administered intradermally at the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO:3. Alternatively, a vaccine prepared by adding the compound synthesized in Example 11 to the peptide dilution solution used in preparing cocktail vaccine b was administered intradermally at the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4, 225 μg / mouse of the peptide represented by SEQ ID NO:3, and 0.4 nmol / mouse of the compound synthesized in Example 11. Administration was performed twice, with a one-week interval. One week after the final administration, the mice were immunized with CO ２After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0319] The results are shown in Figures 8 and 9. As a result of this test, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was found to be greater when the compound synthesized in Example 11 was added to cocktail vaccine b than when no compound was added.

[0320] This demonstrates that the addition of the compound synthesized in Example 11 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 11 has adjuvant activity in vivo.

[0321] Test Example 11 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a cocktail vaccine (hereinafter referred to as "cocktail vaccine f") in which the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 were used as emulsified composition 1. The vaccine was then tested against HLA-A ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0322] Specifically, the above-mentioned emulsion composition 1 was prepared as follows. 95.8% (w / w) soybean oil, 3% (w / w) PEG-30 dipolyhydroxystearate, and 1.2% (w / w) polysorbate 80 were mixed to form an oil phase mixture. The preparation amounts were increased or decreased as necessary. The compound represented by formula 4 was mixed with a pH 2.5 buffer solution (10 mM tartaric acid, 10% trehalose) to a concentration of 3 mg / mL and the peptide represented by SEQ ID NO: 3 to a concentration of 2.25 mg / mL. This peptide dilution was mixed with an equal amount of the oil phase mixture to form an emulsion, thereby preparing cocktail vaccine f. This was administered intradermally at the base of the tail of mice at a concentration of 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO: 3. Alternatively, the compound synthesized in Example 1 was added to the peptide dilution solution used to prepare cocktail vaccine f, and the vaccine was then administered intradermally to the tail base of mice so that the compound represented by formula 4 was 300 μg / mouse, the peptide represented by SEQ ID NO: 3 was 225 μg / mouse, and the compound synthesized in Example 1 was 0.04 nmol / mouse or 0.4 nmol / mouse. Administration was performed twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0323] The results are shown in Figures 10 and 11. As a result of this test, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was found to be greater when the compound synthesized in Example 1 was added to cocktail vaccine f than when no compound was added.

[0324] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0325] Test Example 12 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a cocktail vaccine (hereinafter referred to as "cocktail vaccine g") in which the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by sequence number 3 synthesized in Reference Example 3 were used as emulsified composition 2. The vaccine was then tested against HLA-A ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0326] Specifically, the above-mentioned emulsion composition 2 was prepared as follows. 95.8% (w / w) of medium-chain fatty acid triglyceride (Miglyol 812), 3% (w / w) of PEG-30 dipolyhydroxystearate, and 1.2% (w / w) of polysorbate 80 were mixed to form an oil phase mixture. The preparation amounts were increased or decreased as necessary. The compound represented by formula 4 was mixed with a pH 2.5 buffer solution (10 mM tartaric acid, 10% trehalose) to a concentration of 3 mg / mL and the peptide represented by SEQ ID NO: 3 to a concentration of 2.25 mg / mL. This diluted peptide solution was mixed with an equal amount of the oil phase mixture to form an emulsion, yielding cocktail vaccine g. This was administered intradermally at the base of the tail of mice at a concentration of 300 μg of the compound represented by formula 4 and 225 μg of the peptide represented by SEQ ID NO: 3 per mouse. Alternatively, the compound synthesized in Example 1 was added to the peptide dilution solution used to prepare cocktail vaccine g, and the vaccine was then administered intradermally to the tail base of mice so that the compound represented by formula 4 was 300 μg / mouse, the peptide represented by SEQ ID NO: 3 was 225 μg / mouse, and the compound synthesized in Example 1 was 0.04 nmol / mouse or 0.4 nmol / mouse. Administration was performed twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0327] The results are shown in Figures 12 and 13. As a result of this test, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was found to be greater when the compound synthesized in Example 1 was added to the cocktail vaccine g than when no compound was added.

[0328] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0329] Test Example 13 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a cocktail vaccine (hereinafter referred to as "cocktail vaccine h") containing the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by sequence number 3 synthesized in Reference Example 3 as emulsion composition 3, and this was used to prepare a vaccine. ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0330] Specifically, the above-mentioned emulsion composition 3 was prepared as follows. 95.8% (w / w) isopropyl myristate, 3% (w / w) PEG-30 dipolyhydroxystearate, and 1.2% (w / w) polysorbate 80 were mixed to form an oil phase mixture. The preparation amounts were increased or decreased as necessary. The compound represented by formula 4 was mixed with a pH 2.5 buffer solution (10 mM tartaric acid, 10% trehalose) to a concentration of 3 mg / mL and the peptide represented by SEQ ID NO: 3 to a concentration of 2.25 mg / mL. This peptide dilution was mixed with an equal amount of the oil phase mixture to form an emulsion, thereby preparing cocktail vaccine h. This was administered intradermally at the base of the tail of mice at a concentration of 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO: 3. Alternatively, the compound synthesized in Example 1 was added to the peptide dilution solution used to prepare cocktail vaccine h, and the vaccine was then administered intradermally to the tail base of mice so that the compound represented by formula 4 was 300 μg / mouse, the peptide represented by SEQ ID NO: 3 was 225 μg / mouse, and the compound synthesized in Example 1 was 0.04 nmol / mouse. Administration was performed twice at a one-week interval. One week after the final administration, the mice were then immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0331] The results are shown in Figure 14. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 1 was found to be greater when the compound synthesized in Example 1 was added to cocktail vaccine h than when no compound was added.

[0332] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0333] The results of measuring the spleen weights of mice in Test Examples 11 to 13 are shown in Figure 26. As a result of this test, it was found that the addition of Example 1 did not cause a significant increase in spleen weight, suggesting that the compound synthesized in Example 1 exhibits adjuvant activity in vivo without causing splenomegaly.

[0334] Test Example 14 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a cocktail vaccine (hereinafter referred to as "cocktail vaccine i") prepared by dissolving the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by sequence number 3 synthesized in Reference Example 3 in an oily suspension. ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0335] Specifically, the oil suspension was prepared by dissolving sucrose fatty acid ester (Ryoto Sugar Ester L-195) in cyclohexane to a concentration of 12.5 mg / mL, adding the compound represented by formula 4 at 0.75 mg / mL and the peptide represented by SEQ ID NO: 3 at 0.5625 mg / mL to form an oil phase mixture. To 2 mL of the oil phase mixture, 1 mL of pH 2.5 buffer (10 mM tartaric acid, 10% trehalose) was added, and the mixture was dispersed using a homogenizer and lyophilized. 1 mL of isopropyl myristate was added to the lyophilized product to obtain Cocktail Vaccine i, an oil suspension. The preparation volume was adjusted as needed. Cocktail Vaccine i was administered intradermally to the base of the tail of mice at a concentration of 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO: 3. Alternatively, the compound synthesized in Example 1 was added to the pH 2.5 buffer solution used to prepare cocktail vaccine i, and the vaccine was administered intradermally to the base of the tail of mice so that the compound represented by formula 4 was 300 μg / mouse, the peptide represented by SEQ ID NO: 3 was 225 μg / mouse, and the compound synthesized in Example 1 was 0.04 nmol / mouse or 0.4 nmol / mouse. Administration was performed twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0336] The results are shown in Figures 15 and 16. As a result of this test, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was found to be greater when the compound synthesized in Example 1 was added to cocktail vaccine i than when no compound was added.

[0337] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0338] Test Example 15 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a hydrogel formulation of the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 (hereinafter referred to as "cocktail vaccine j"). This vaccine was then used to treat HLA-A ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0339] Specifically, the hydrogel formulation was prepared by mixing 1.5 mg / mL of the compound represented by formula 4, 1.125 mg / mL of the peptide represented by SEQ ID NO: 3, and 200 mg / mL of polyoxyethylene (196) polyoxypropylene (67) glycol with pH 2.5 buffer (10 mM tartaric acid) and cooling on ice to prepare cocktail vaccine j, a hydrogel formulation. The preparation amounts were adjusted as needed. Cocktail vaccine j was administered intradermally to the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4 and 225 μg / mouse of the peptide represented by SEQ ID NO: 3. Alternatively, a vaccine prepared by adding the compound synthesized in Example 1 to the pH 2.5 buffer used to prepare cocktail vaccine j was administered intradermally to the base of the tail of mice at 300 μg / mouse of the compound represented by formula 4, 225 μg / mouse of the peptide represented by SEQ ID NO: 3, and 0.4 nmol / mouse of the compound synthesized in Example 1. Administration was performed twice weekly. One week after the final administration, the mice were ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0340] The results are shown in Figures 17 and 18. As a result of this test, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was found to be greater when the compound synthesized in Example 1 was added to cocktail vaccine j than when no compound was added.

[0341] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0342] The results of measuring the spleen weight of mice in Test Examples 14 and 15 are shown in Figure 27. As a result of this test, it was found that the addition of Example 1 did not cause a significant increase in spleen weight, suggesting that the compound synthesized in Example 1 exhibits adjuvant activity in vivo without causing splenomegaly.

[0343] Test Example 16 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a vaccine (hereinafter referred to as "cocktail vaccine k") in which the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 were used as liposome formulation 1. The vaccine was then tested against HLA-A ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0344] Specifically, liposome preparation 1 was prepared by dissolving 47.1 mg of hydrogenated soybean phosphatidylcholine and 15.47 mg of cholesterol in t-butyl alcohol, lyophilizing the solution, and then adding 2 mL of an aqueous solution containing 2.5 mL of a pH 2.5 buffer solution (10 mM tartaric acid, 10% trehalose) to a concentration of 2.5 mg / mL of the compound represented by formula 4 and 1.875 mg / mL of the peptide represented by SEQ ID NO:3. The solution was then passed through a 0.1 μm polycarbonate membrane using an extruder (Mini-Extruder, Avanti Polar Lipids) heated to approximately 65°C to obtain cocktail vaccine k, which is liposome preparation 1. The preparation amount was adjusted as needed. Cocktail vaccine k was administered intradermally to the base of the tail of mice at a concentration of 500 μg of the compound represented by formula 4 and 375 μg of the peptide represented by SEQ ID NO:3 per mouse. Alternatively, the compound synthesized in Example 1 was added to the pH 2.5 buffer solution used to prepare cocktail vaccine k, and the vaccine was administered intradermally to the base of the tail of mice so that the compound represented by formula 4 was 500 μg / mouse, the peptide represented by SEQ ID NO: 3 was 375 μg / mouse, and the compound synthesized in Example 1 was 0.4 nmol / mouse. The administration was carried out twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0345] The results are shown in Figure 19. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 1 was found to be greater when the compound synthesized in Example 1 was added to cocktail vaccine k than when no compound was added.

[0346] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0347] Test Example 17 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a vaccine (hereinafter referred to as "cocktail vaccine 1") in which the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 were used as liposome formulation 2, and this was used as HLA-A ＊ The vaccine was administered to 02:01 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0348] Specifically, liposome preparation 2 was prepared by dissolving 42.66 mg of sphingomyelin and 15.47 mg of cholesterol in t-butyl alcohol and then lyophilizing the solution. To the lyophilized product, 2 mL of an aqueous solution containing 2.5 mg of the compound represented by formula 4 and 1.875 mg of the peptide represented by SEQ ID NO:3 was added. The solution was then passed through a 0.1 μm polycarbonate membrane using an extruder (Mini-Extruder, Avanti Polar Lipids) heated to approximately 65°C to obtain cocktail vaccine 1, which was liposome preparation 2. The preparation volume was adjusted as needed. Cocktail vaccine 1 was administered intradermally to the base of the tail of mice at 500 μg of the compound represented by formula 4 and 375 μg of the peptide represented by SEQ ID NO:3 per mouse. Alternatively, the compound synthesized in Example 1 was added to the pH 2.5 buffer solution used to prepare cocktail vaccine 1, and the vaccine was administered intradermally to the base of the tail of mice so that the compound represented by formula 4 was 500 μg / mouse, the peptide represented by SEQ ID NO: 3 was 375 μg / mouse, and the compound synthesized in Example 1 was 0.4 nmol / mouse. Administration was performed twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 4) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C. in 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0349] The results are shown in Figures 20 and 21. As a result of this test, the number of CTLs reactive to the peptides represented by SEQ ID NO: 1 or SEQ ID NO: 4 was found to be greater when the compound synthesized in Example 1 was added to cocktail vaccine 1 than when no compound was added.

[0350] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0351] Test Example 18 HLA-A ＊ Evaluation of in vivo adjuvant activity using 02:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine was prepared by adding the compound synthesized in Example 1 to a vaccine (hereinafter referred to as "cocktail vaccine m") in which the compound represented by formula 4 synthesized in Reference Example 8 and the peptide represented by SEQ ID NO: 3 synthesized in Reference Example 3 were used as liposome formulation 3, and this was used as a vaccine for HLA-A ＊ The vaccine was administered to 02:01 transgenic mice and evaluated by an antigen-specific CTL induction test.

[0352] Specifically, liposome preparation 3 was prepared by dissolving 36.7 mg of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 7.58 mg of 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine sodium salt, and 15.47 mg of cholesterol in a t-butyl alcohol / water mixture, followed by lyophilization. To the lyophilized product, 2 mL of an aqueous solution containing the compound represented by formula 4 at 2.5 mg / mL and the peptide represented by SEQ ID NO: 3 at 1.875 mg / mL, mixed with a pH 2.5 buffer solution (10 mM tartaric acid, 10% trehalose), was added. The mixture was then passed through a 0.1 μm polycarbonate membrane using an extruder (Mini-Extruder, Avanti Polar Lipids) heated to approximately 65°C to obtain cocktail vaccine m, which is liposome preparation 3. The amounts prepared were adjusted as needed. Cocktail vaccine m was administered intradermally to the base of the tail of mice so that the compound represented by formula number 4 was 500 μg / mouse and the peptide represented by sequence number 3 was 375 μg / mouse. Alternatively, a vaccine prepared by adding the compound synthesized in Example 1 to the pH 2.5 buffer used to prepare cocktail vaccine m was administered intradermally to the base of the tail of mice so that the compound represented by formula number 4 was 500 μg / mouse, the peptide represented by sequence number 3 was 375 μg / mouse, and the compound synthesized in Example 1 was 0.4 nmol / mouse. Administration was performed twice at a one-week interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0353] The results are shown in Figure 22. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 1 was found to be greater when the compound synthesized in Example 1 was added to the cocktail vaccine m than when no compound was added.

[0354] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0355] The results of measuring the spleen weights of mice in Test Examples 16 to 18 are shown in Figure 28. As a result of this test, it was found that the addition of Example 1 did not cause a significant increase in spleen weight, suggesting that the compound synthesized in Example 1 exhibits adjuvant activity in vivo without causing splenomegaly.

[0356] Test Example 19 HLA-A ＊ 02:01 / HLA-DRB1 ＊ Evaluation of in vivo adjuvant activity using 01:01 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a cocktail vaccine (hereinafter referred to as "cocktail vaccine n") was prepared by mixing the peptide represented by SEQ ID NO: 1 synthesized in Reference Example 1 and the peptide represented by SEQ ID NO: 17 synthesized in Reference Example 20 with a pre-emulsified composition, and the compound synthesized in Example 1 was added to the cocktail vaccine to prepare a vaccine. ＊ 02:01 / HLA-DRB1 ＊ The peptide was administered to 01:01 transgenic mice (C57BL / 6CrHLA-A2.1DR1) and evaluated by antigen-specific CTL and antigen-specific helper T cell induction tests. The peptide represented by SEQ ID NO: 17 is a helper peptide derived from the WT1 protein.

[0357] Specifically, the peptide represented by SEQ ID NO: 1 and the peptide represented by SEQ ID NO: 17 were dissolved in DMSO and then mixed with water for injection to give a diluted concentration of 2 mg / mL for the peptide represented by SEQ ID NO: 1 and 4 mg / mL for the peptide represented by SEQ ID NO: 17. This peptide dilution was mixed with an equal amount of a pre-emulsified composition prepared in the same manner as in Test Example 4 to form an emulsion, and the resulting cocktail vaccine n was administered intradermally at the base of the tail of mice at 200 μg / mouse of the peptide represented by SEQ ID NO: 1 and 400 μg / mouse of the peptide represented by SEQ ID NO: 17. Alternatively, the compound synthesized in Example 1 was added to the peptide dilution solution used in preparing cocktail vaccine n, and the resulting vaccine was administered intradermally at the base of the tail of mice at 200 μg / mouse of the peptide represented by SEQ ID NO: 1, 400 μg / mouse of the peptide represented by SEQ ID NO: 17, and 0.04 nmol / mouse or 0.4 nmol / mouse of the compound synthesized in Example 1. Administration was performed twice at weekly intervals. One week after the final administration, the mice were subjected to CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 4, the splenocytes to which the peptide (SEQ ID NO: 1 or SEQ ID NO: 17) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0358] The results are shown in Figures 23 and 24. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 1 was observed to be greater when the compound synthesized in Example 1 was added to cocktail vaccine n than when no compound was added. Furthermore, the number of helper T cells reactive to the peptide represented by SEQ ID NO: 17 was observed to be greater when the compound synthesized in Example 1 was added to cocktail vaccine n than when no compound was added.

[0359] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of induced CTLs and helper T cells, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0360] Test Example 20 HLA-A ＊ Evaluation of in vivo adjuvant activity using 24:02 transgenic mice Regarding the in vivo adjuvant activity of the compound synthesized in Example 1, a vaccine (hereinafter referred to as "vaccine o") was prepared by mixing the compound represented by formula 5 synthesized in Reference Example 22 with a pre-emulsified composition, and the compound synthesized in Example 1 was added to the vaccine. ＊ The vaccine was administered to 24:02 transgenic mice, and the adjuvant activity was evaluated by an antigen-specific CTL induction test.

[0361] Specifically, the compound represented by formula number 5 was dissolved in DMSO, and then mixed with water for injection so that the concentration of the compound represented by formula number 5 after dilution was 3 mg / mL. This compound dilution was mixed with an equal amount of a pre-emulsified composition prepared in the same manner as in Test Example 4 to emulsify, and vaccine o was prepared. This vaccine was administered intradermally at the base of the tail of mice so that the compound represented by formula number 5 was 300 μg / mouse. Alternatively, the compound synthesized in Example 1 was added to the compound dilution solution used in preparing vaccine o, and this vaccine was administered intradermally at the base of the tail of mice so that the compound represented by formula number 5 was 300 μg / mouse and the compound synthesized in Example 1 was 0.4 nmol / mouse. Administration was performed twice at a weekly interval. One week after the final administration, the mice were immunized with CO ２ After euthanasia by gas, the spleen was removed and splenocytes were prepared. As in Test Example 3, the splenocytes to which the peptide (SEQ ID NO: 2) was added at a final concentration of 10 μg / mL were seeded on an ELISPOT plate and incubated at 37° C., 5% CO ２ After incubation, the supernatant was removed and the number of spots on the colored ELISPOT plate was counted.

[0362] The results are shown in Figure 25. As a result of this test, the number of CTLs reactive to the peptide represented by SEQ ID NO: 2 was found to be greater when the compound synthesized in Example 1 was added to vaccine o than when no compound was added.

[0363] This demonstrates that the addition of the compound synthesized in Example 1 to a vaccine increases the number of CTLs induced, strongly suggesting that the compound synthesized in Example 1 has adjuvant activity in vivo.

[0364] Test Example 21 A human TLR7 reporter gene assay was carried out according to the method of Test Example 1, and the results shown in the table below were obtained.

[0365] The results of Test Example 21 suggest that the example compounds of the present invention act as agonists of human TLR7.

[0366] Test Example 22 Human TLR7 reporter gene assay was carried out according to the method of Test Example 1, and the results in the table below were obtained. ＴＭ The hTLR7 cell line (Invivogen) was used. HEK-Blue ＴＭ The hTLR7 cell line is a stable co-transfected cell line expressing full-length human TLR7 and a secreted SEAP reporter gene under the transcriptional control of the NF-κB response element. TLR7 expression in this cell line was tested by RT-PCR. Stable transfectants were selected using the antibiotics blasticidin and zeocin.

[0367]

Claims

1. Formula (1): [wherein X is methylene, an oxygen atom, a sulfur atom, SO, SO ２ , or NR ５ (R ５ is a hydrogen atom or C １－６ R represents alkyl; １ is C １－６ alkyl, wherein the alkyl is selected from halogen, hydroxy, and C １－６ alkoxy), R ２ and R ３ are each independently a hydrogen atom or C １－６ Alkyl (the alkyl is not limited to halogen, hydroxy, C １－６ alkoxy), R ４ is a hydrogen atom, halogen, hydroxy, C １－６ alkyl (the alkyl may be substituted with 1 to 3 of the same or different halogens), C １－６ represents alkoxy (which may be substituted with 1 to 3 of the same or different halogens) or cyano; L represents a linker; Y １ is -(CH ２ CH ２ O) ｍ -R ６ (R ６ is a hydrogen atom or C １－６ and m represents an integer of 3 to 100.] or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein X is methylene.

3. R １ But C １－３ 3. The compound according to claim 1 or 2, wherein the alkyl is optionally substituted with 1 to 3 of the same or different halogens, or a pharmaceutically acceptable salt thereof.

4. R １ 4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein is methyl.

5. R ４ is a hydrogen atom, hydroxy, C １－３ Alkyl, or C １－３ The compound according to any one of claims 1 to 4, which is alkoxy, or a pharmaceutically acceptable salt thereof.

6. R ４ The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein is a hydrogen atom, hydroxy, or methoxy.

7. R ２ But C １－６ The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein R is alkyl.

8. R ３ is a hydrogen atom, or C １－３ 8. The compound according to any one of claims 1 to 7, wherein the alkyl is alkyl (the alkyl may be substituted with 1 to 3 hydroxyl groups), or a pharmaceutically acceptable salt thereof.

9. L is -O-, -NR Ｙ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR Ｙ -, -NR Ｙ C(O)-, -CH ２ NR Ｙ -, -CH ２ O-, -OC(O)O-, -NR ７ C(O)O-, -OC(O)NR Ｙ -, -NR ７ C(O)NR Ｙ -, -OC(S)NR Ｙ - or -NR ７ C(S)NR Ｙ - (where R ７ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; R Ｙ represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or Y ２ (Y ２ is -(CH ２ CH ２ O) ｎ -R ８ (R ８ represents a hydrogen atom or alkyl having 1 to 6 carbon atoms, and n represents an integer of 3 to 100), or a pharmaceutically acceptable salt thereof.

10. L is -C(O)NR Ｙ -, -CH ２ NR Ｙ -, -C(O)O- or -CH ２ 10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein R is O-.

11. L is -CH ２ NR Ｙ - and R Ｙ is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or Y ２ 10. The compound of claim 9, wherein:

12. Y １ But -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ The compound according to any one of claims 1 to 11, wherein m is alkyl and m is an integer of 3 to 40, or a pharmaceutically acceptable salt thereof.

13. Formula (2): [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl (the alkyl may be substituted with 1 to 3 hydroxy groups); R ４ is a hydrogen atom, hydroxy, or methoxy; L is —CH ２ NR Ｙ - and R Ｙ is a hydrogen atom or C １－６ is alkyl, and Y １ is -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ and m is an integer of 3 to 40.] or a pharmaceutically acceptable salt thereof.

14. Formula (2): [In the formula, R ２ is C １－６ alkyl, R ３ is a hydrogen atom or C １－３ alkyl, which may be substituted with one hydroxyl; R ４ is a hydrogen atom or methoxy, and L is —CH ２ NR Ｙ - and R Ｙ is a hydrogen atom or C １－６ is alkyl, and Y １ is -(CH ２ CH ２ O) ｍ -R ６ and R ６ is a hydrogen atom or C １－６ and m is an integer of 3 to 40.] or a pharmaceutically acceptable salt thereof.

15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, selected from the following group of compounds: 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol, 1-{4-[(2-amino-4-{[(3S)-1-hydroxyhexan-3-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxyphenyl}-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol, 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol, 1-(3-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-4-hydroxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol, 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-N-(20-hydroxy-3,6,9,12,15,18-hexaoxaicosan-1-yl)-3-methoxybenzamide, 2,5,8,11-tetraoxatridecan-13-yl 4-[(2-amino-4-{[(2S)-1-hydroxypentan-2-yl]amino}-6-methylpyrimidin-5-yl)methyl]-3-methoxybenzoate, 5-{[2-methoxy-4-(2,5,8,11,14-pentaoxapentadecan-1-yl)phenyl]methyl}-6-methyl-N ４ -pentylpyrimidine-2,4-diamine, 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29-nonaoxa-2-azahentriacontan-31-ol, 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71-tricosaoxa-2-azatriheptacontan-73-ol, 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92,95,98,101,104,107-pentatriacontaoxa-2-azanonahexan-109-ol, and 12-[(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3,6,9,15,18,21-hexaoxa-12-azatricosane-1,23-diol.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from the following group of compounds: 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14-tetraoxa-2-azahexadecan-16-ol, 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29-nonaoxa-2-azahentriacontan-31-ol, 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71-tricosaoxa-2-azatriheptacontan-73-ol, 1-(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)-2-methyl-5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71,74,77,80,83,86,89,92,95,98,101,104,107-pentatriacontaoxa-2-azanonahexan-109-ol, and 12-[(4-{[2-amino-4-methyl-6-(pentylamino)pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3,6,9,15,18,21-hexaoxa-12-azatricosane-1,23-diol.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

18. The pharmaceutical composition according to claim 17, which is an emulsion formulation, an oily suspension formulation, a hydrogel formulation, or a lipid formulation.

19. The pharmaceutical composition according to claim 18, which is an emulsion formulation.

20. The pharmaceutical composition according to claim 19, wherein the emulsion formulation is a water-in-oil emulsion.

21. The pharmaceutical composition according to claim 20, wherein the emulsion formulation contains (1) ethyl oleate, octyldodecyl myristate, sorbitan monooleate, glyceryl monooleate, polyoxyethylene hydrogenated castor oil 20, glycerin, and sodium dihydrogen phosphate, or (2) Montanide ISA 51VG.

22. The pharmaceutical composition of claim 18, which is a lipid formulation.

23. The pharmaceutical composition according to claim 22, wherein the lipid formulation is a liposome formulation containing phospholipids.

24. The pharmaceutical composition according to claim 22 or 23, wherein the lipid formulation is a liposome formulation containing sterols.

25. The pharmaceutical composition according to claim 24, wherein the sterol is cholesterol.

26. The pharmaceutical composition according to any one of Items 23 to 25, wherein the liposome preparation contains one or more additives selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.

27. The pharmaceutical composition according to any one of claims 17 to 26, further comprising a tumor antigen.

28. The pharmaceutical composition according to claim 27, wherein the tumor antigen is a tumor antigen peptide.

29. A tumor antigen peptide represented by formula (4): [wherein the bond between C and C represents a disulfide bond.] or a pharmaceutically acceptable salt thereof, and a peptide represented by the amino acid sequence of SEQ ID NO: 3: WAPVLDFAPPGASAYGSL or a pharmaceutically acceptable salt thereof.

30. A vaccine adjuvant for a cancer vaccine, comprising the compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

31. A kit comprising: a) a compound represented by formula (1) of claim 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound represented by formula (1) or a pharmaceutically acceptable salt thereof; and b) a tumor antigen or a pharmaceutical composition containing a tumor antigen.

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