IMMUNE-INDUCTION-PROMOTING COMPOSITION INCLUDING NUCLEAR RECEPTOR LIGAND, AND VACCINE PHARMACEUTICAL COMPOSITION

Abstract

The present invention aims to provide a composition for promoting immunity induction and a vaccine pharmaceutical composition which are universally usable for inducing immunity to various antigens and capable of exerting a high immunity inducing effect. The present invention relates to a composition for promoting immunity induction containing at least one nuclear receptor ligand and also relates to a vaccine pharmaceutical composition for inducing immunity containing an antigen and a composition for promoting immunity induction that is at least one nuclear receptor ligand.

Claims

1. A composition for promoting immunity induction, comprising: a nuclear receptor ligand.

2. The composition for promoting immunity induction according to claim 1, wherein the nuclear receptor ligand is at least one selected from the group consisting of a retinoid receptor agonist, a retinoid X receptor agonist, a thyroid hormone receptor agonist, and an estrogen receptor modulator.

3. The composition for promoting immunity induction according to claim 1, wherein the nuclear receptor ligand is at least one selected from the group consisting of a retinoid receptor agonist, a thyroid hormone receptor agonist, and an estrogen receptor modulator and is for inducing humoral immunity.

4. The composition for promoting immunity induction according to claim 1, wherein the nuclear receptor ligand is at least one of a retinoid receptor agonist or a retinoid X receptor agonist and is for inducing cellular immunity.

5. The composition for promoting immunity induction according to claim 1, further comprising a helper peptide.

6. A vaccine pharmaceutical composition, comprising: an antigen for inducing immunity; and the composition for promoting immunity induction according to claim 1.

7. The vaccine pharmaceutical composition according to claim 6, which is administered to a body surface.

8. The vaccine pharmaceutical composition according to claim 6, which is administered by intradermal injection, subcutaneous injection, or intramuscular injection.

9. A method for promoting immunity induction in a subject in need thereof, comprising: administering to the subject an effective amount of a nuclear receptor ligand.

10. The method for promoting immunity induction according to claim 9, wherein the nuclear receptor ligand is at least one selected from the group consisting of a retinoid receptor agonist, a retinoid X receptor agonist, a thyroid hormone receptor agonist, and an estrogen receptor modulator.

11. The method for promoting immunity induction according to claim 9, wherein the nuclear receptor ligand is at least one selected from the group consisting of a retinoid receptor agonist, a thyroid hormone receptor agonist, and an estrogen receptor modulator and is for inducing humoral immunity.

12. The method for promoting immunity induction according to claim 9, wherein the nuclear receptor ligand is at least one of a retinoid receptor agonist or a retinoid X receptor agonist and is for inducing cellular immunity.

13. The method for promoting immunity induction according to claim 9, further comprising administering a helper peptide.

14. A method for promoting immunity induction in a subject in need thereof, comprising: administering to the subject an effective amount of a pharmaceutical composition comprising: an antigen for inducing immunity; and a nuclear receptor ligand.

15. The method according to claim 14, further comprising administering a helper peptide.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0164] FIG. 1 is a graph showing evaluation results of the number of IFN-γ-producing cell spots in mouse spleen cells after transdermal administration of creams for transdermal administration obtained in Examples 1 to 8 and Comparative Examples 1 and 2.

[0165] FIG. 2 is a graph showing evaluation results of the number of IFN-γ-producing cell spots in mouse spleen cells after transdermal administration of tapes for transdermal administration obtained in Examples 9 to 20 and Comparative Examples 3 to 8.

[0166] FIG. 3 is a graph showing an OVA-specific IgG antibody titer in mouse serum after transnasal administration of solutions for transnasal administration obtained in Examples 21 to 28 and Comparative Example 9.

[0167] FIG. 4 is a graph showing an OVA-specific IgG antibody titer in mouse serum after sublingual administration of solutions for sublingual administration obtained in Examples 29 to 36 and Comparative Example 10.

[0168] FIG. 5 is a graph showing an OVA-specific IgG antibody titer in mouse serum after sublingual administration of solid formulations for sublingual administration obtained in Examples 37 to 52 and Comparative Examples 11 and 12.

[0169] FIG. 6 is a graph showing an OVA-specific IgG antibody titer in mouse serum after subcutaneous administration of solutions for subcutaneous administration obtained in Examples 53 to 56 and Comparative Example 13.

[0170] FIG. 7 is a graph showing an OVA-specific IgG antibody titer in mouse serum after transdermal administration of creams obtained in Examples 57 to 60 and Comparative Example 14.

DESCRIPTION OF EMBODIMENTS

[0171] The present invention will be specifically described in further detail below. The present invention, however, is not limited to these examples.

Examples 1 to 8, Comparative Examples 1 and 2

(Preparation of Cream for Transdermal Administration)

[0172] A cream for transdermal administration was prepared according to the formulation shown in Table 1 below. Specifically, 5% by weight of an antigen peptide, 3% by weight of a nuclear receptor ligand, and if necessary 1 part by weight of a helper peptide and 15% by weight of dimethylsulfoxide (DMSO) (listed below) were blended in the amounts shown in Table 1. A base (base cream) was then added to achieve a total amount of 100% by weight, followed by mixing to provide a cream for transdermal administration. The base cream was prepared by blending and mixing materials according to the formulation in Table 14. White Vaseline, sorbitan monostearate, isostearic acid, benzyl alcohol, stearyl alcohol, polysorbate 60, concentrated glycerin, dimethylsulfoxide (DMSO) were purchased from Wako Pure Chemical Industries, Ltd. Cetanol was purchased from Tokyo Chemical Industry Co., Ltd.

[0173] A composite base was prepared by bonding a PET film/PET nonwoven fabric laminate (area: 0.7 cm.sup.2) to the center portion of an adhesive tape for attachment such that the PET film-side faced the tape. The cream for transdermal administration in an amount of 4 mg was applied to the nonwoven fabric portion of the composite base to provide a sample for an immunity test.

(Nuclear Receptor Ligand)

[0174] Tretinoin (all-trans-retinoic acid, Wako Pure Chemical Industries, Ltd.) [0175] Isotretinoin (13-cis-retinoic acid, Sigma-Aldrich) [0176] Alitretinoin (9-cis-retinoic acid, Wako Pure Chemical Industries, Ltd.) [0177] Bexarotene (Sigma-Aldrich)

(Antigen Peptide)

[0178] OVAp (OVA peptide, peptide of 8 amino acids having the sequence Ser Ile Ile Asn Phe Glu Lys Leu (SEQ ID No: 13))

(Helper Peptide)

[0179] PADRE

<Evaluation 1>

[0180] Each of the creams for transdermal administration obtained in the examples and comparative examples were subjected to the following evaluations.

(Evaluation of Cellular Immunity Inducing Effect)

[0181] A mouse immunity test using an animal model for immunological evaluation was performed with the cream for transdermal administration by the following procedure. Thereafter, the level of induction of antigen-specific cellular immunity was evaluated by the ELISPOT assay. The results are shown in FIG. 1.

(1) Animal Model for Immunological Evaluation

[0182] The “animal model for immunological evaluation” herein means an animal model for evaluating immunity induction properties of a vaccine pharmaceutical composition (here, the cream for transdermal administration). Specifically, the term means an animal model for evaluating the level of induction of cellular immunity of the cream for transdermal administration.

[0183] The animal model for immunological evaluation used was an animal in which the induction of cellular immunity by the antigen in the cream for transdermal administration can be evaluated, in view of the compatibility of the antigen in the cream for transdermal administration with MHC class 1 molecules of the animal.

[0184] Specifically, for vaccine pharmaceutical compositions containing HLA-A*24 type MHC-restricted class I peptide as the antigen, evaluation was performed using BALB/c mice. For vaccine pharmaceutical compositions containing HLA-A*02 type MHC-restricted peptide as the antigen, evaluation was performed using genetically engineered mice in which the induction of cellular immunity by HLA-A*02 type MHC-restricted peptide could be evaluated. For vaccine pharmaceutical compositions containing other HLA-type MHC-restricted peptides, evaluation was performed using animals in which the induction of cellular immunity by the HLA-type MHC-restricted peptides could be evaluated.

(2) Mouse Immunity Test of Cream for Transdermal Administration

[0185] The back of the mouse shown in Table 1 below was shaved. After a rearing period for recovery from damage from the shaving, 4 mg of the cream for transdermal administration was administered to the skin of the back of the mouse for 24 hours and removed. The mouse was reared for six days. Six days after the administration, the spleen was taken out, and a spleen cell suspension was prepared. Spleen cells (1×10.sup.6 cells/well) and the antigen peptide (100 μM) together with culture medium were put into wells of an ELISPOT plate on which anti-mouse IFN-γ antibody was immobilized. The spleen cells were co-cultured with the antigen at 37° C. and 5% CO.sub.2 for 20 hours. The number of IFN-γ-producing cell spots was evaluated by the ELISPOT assay. The number of IFN-γ-producing spots is shown as “Immunity result” in Table 1.

TABLE-US-00001 TABLE 1 Antigen Nuclear receptor ligand Helper peptide Amount Amount Amount [Parts [Parts [Parts Immunological Administration Dosage by by by evaluation Immunity No. route form Name weight] Name weight] Name weight] mouse result Comp. Ex. 1 Transdermal Cream OVAp 5 — — PADRE — C57BL/6 6.0 Comp. Ex. 2 Transdermal Cream OVAp 5 — — PADRE 1 C57BL/6 9.6 Ex. 1 Transdermal Cream OVAp 5 Tretinoin 3 PADRE — C57BL/6 90.4 Ex. 2 Transdermal Cream OVAp 5 Isotretinoin 3 PADRE — C57BL/6 85.6 Ex. 3 Transdermal Cream OVAp 5 Alitretinoin 3 PADRE — C57BL/6 97.5 Ex. 4 Transdermal Cream OVAp 5 Bexarotene 3 PADRE — C57BL/6 55.4 Ex. 5 Transdermal Cream OVAp 5 Tretinoin 3 PADRE 1 C57BL/6 264.9 Ex. 6 Transdermal Cream OVAp 5 Isotretinoin 3 PADRE 1 C57BL/6 252.0 Ex. 7 Transdermal Cream OVAp 5 Alitretinoin 3 PADRE 1 C57BL/6 248.3 Ex. 8 Transdermal Cream OVAp 5 Bexarotene 3 PADRE I C57BL/6 99.8

Examples 9 to 20, Comparative Examples 3 to 8

(Preparation of Tape for Transdermal Administration)

[0186] A tape for transdermal administration was prepared according to the formulation shown in Table 2 below. Specifically, an antigen peptide, a nuclear receptor ligand, and a helper peptide (listed below) were mixed in amounts shown in Table 2. An adhesive base and an organic solvent (ethyl acetate) shown in Table 2 were then added so that the total amount of the components and the adhesive base after drying the organic solvent was 100% by weight. This was followed by mixing to provide an adhesive solution. The resulting adhesive solution was spread onto a release liner such that the thickness after drying was about 80 μm. The organic solvent was removed by drying, whereby an adhesive layer was formed. The release liner was a polyethylene terephthalate (PET) liner (thickness: 75 μm) release-treated with silicone. A support was attached to the adhesive layer to provide a tape for transdermal administration. The support was a polyethylene terephthalate (PET) film (thickness: 25 μm).

[0187] The tape for transdermal administration was cut into a piece with an area of 0.7 cm.sup.2. The piece was used as a sample for immunity experiment. The release liner was removed before administration.

(Nuclear Receptor Ligand)

[0188] Tretinoin (all-trans-retinoic acid, Wako Pure Chemical Industries, Ltd.)

(Antigen Peptide)

[0189] HER2/neu_E75 (HER2/neu_E75 peptide, cancer antigen peptide) IPEP87 (IPEP87 peptide, infectious pathogen-derived antigen) MAGE-A3_A02 (MAGE3_A02 peptide, cancer antigen peptide)

(Helper Peptide)

[0190] PADRE

(Adhesive Base)

[0191] Acrylic base (acrylic adhesive solution prepared by solution polymerization of 75 parts of 2-ethylhexyl acrylate, 22 parts of N-vinyl-2-pyrrolidone, 3 parts of acrylic acid, and 0.2 parts of azobisisobutyronitrile in ethyl acetate at 60° C. in an innate gas atmosphere) [0192] PIB (PIB adhesive solution prepared by dissolving 24 parts of polyisobutylene (Oppanol B200, BASF), 36 parts by polyisobutylene (Oppanol B12, BASF), 40 parts of alicyclic petroleum resin (Alkon P-100, Arakawa Chemical Industries, Ltd.) in toluene)

<Evaluation 2>

[0193] Each of the tapes for transdermal administration obtained in the examples and the comparative examples were subjected to the following evaluation.

(Evaluation of Cellular Immunity Inducing Effect)

[0194] The level of induction of antigen-specific cellular immunity was evaluated in the same manner as for the cream for transdermal administration. The results are shown in FIG. 2.

TABLE-US-00002 TABLE 2 Antigen Nuclear receptor ligand Helper peptide Adminis- Amount Amount Amount tration Dosage [% by [% by [% by Immunological Immunity result No. route form Base Name weight] Name weight] Name weight] evaluation mouse Average [cells/well] Comp. Transdermal Tape Acrylic HER2/ 10 — — PADRE 1 Genetically altered  7.5 2 × 10.sup.6 Ex. 3 neu_E75 Ex. 9 Transdermal Tape Acrylic HER2/ 10 Tretinoin 1 — 1 Genetically altered  72.3 neu_E75 Ex. 10 Transdermal Tape Acrylic HER2/ 10 Tretinoin 1 PADRE 1 Genetically altered 130.0 neu_E75 Comp. Transdermal Tape PIB HER2/ 10 — — PADRE 1 Genetically altered  7.5 2 × 10.sup.6 Ex. 4 neu_E75 Ex. 11 Transdermal Tape PIB HER2/ 10 Tretinoin 1 — 1 Genetically altered  74.5 neu_E75 Ex. 12 Transdermal Tape PIB HER2/ 10 Tretinoin 1 PADRE 1 Genetically altered 127.5 neu_E75 Comp. Transdermal Tape Acrylic IPEP87 10 — — PADRE 1 Genetically altered  5.3 2 × 10.sup.6 Ex. 5 Ex. 13 Transdermal Tape Acrylic IPEP87 10 Tretinoin 1 — 1 Genetically altered  76.0 Ex 14 Transdermal Tape Arylic IPEP87 10 Tretinoin 1 PADRE 1 Genetically altered 155.3 Comp. Transdermal Tape PIB IPEP87 10 — — PADRE 1 Genetically altered  3.8 2 × 10.sup.6 Ex. 6 Ex. 15 Transdermal Tape PIB IPEP87 10 Tretinoin 1 — 1 Genetically altered  92.3 Ex. 16 Transdermal Tape PIB IPEP87 10 Tretinoin 1 PADRE 1 Genetically altered 173.3 Comp. Transdermal Tape Acrylic MAGE- 10 — — PADRE 1 Genetically altered  5.8 2 × 10.sup.6 Ex. 7 A3_A02 Ex. 17 Transdermal Tape Acrylic MAGE- 10 Tretinoin 1 — 1 Genetically altered  90.5 A3_A02 Ex. 18 Transdermal Tape Acrylic MAGE- 10 Tretinoin 1 PADRE 1 Genetically altered 175.5 A3_A02 Comp. Transdermal Tape PIB MAGE- 10 — — PADRE 1 Genetically altered  4.3 2 × 10.sup.6 Ex. 8 A3_A02 Ex. 19 Transdermal Tape PIB MAGE- 10 Tretinoin 1 — 1 Genetically altered  96.0 A3_A02 Ex. 20 Transdermal Tape PIB MAGE- 10 Tretinoin 1 PADRE 1 Genetically altered 186.3 A3_A02

Examples 21 to 36, Comparative Examples 9 to 10

(Preparation of Solution for Transmucosal Administration)

[0195] A solution for transmucosal administration (transnasal administration or sublingual administration) was prepared according to the formulation shown in Tables 3 and 4 below. Specifically, an antigen (ovalbumin (OVA)) and an immunity induction promoter that was a nuclear receptor ligand were blended in the amounts shown in Tables 3 and 4. For transnasal administration, saline was added thereto such that the amount of the resulting mixture was 10 μL. For sublingual administration, saline was added such that the amount of the resulting mixture was 30 μL. This was followed by mixing to provide a solution for transmucosal administration (transnasal administration or sublingual administration).

(Nuclear Receptor Ligand)

[0196] Tretinoin (all-trans retinoic acid, Wako Pure Chemical Industries, Ltd.) [0197] Isotretinoin (13-cis-retinoic acid, Sigma-Aldrich) [0198] Alitretinoin (9-cis-retinoic acid, Wako Pure Chemical Industries, Ltd.) [0199] Levothyroxine sodium hydrate (Sigma-Aldrich) [0200] Liothyronine (Sigma-Aldrich) [0201] Clomiphene citrate (Sigma-Aldrich) [0202] Raloxifene hydrochloride (LKT Laboratories) [0203] Tamoxifen citrate (Wako Pure Chemical Industries, Ltd.)

Examples 37 to 52, Comparative Examples 11 and 12

(Preparation of Solid Formulation for Sublingual Administration)

[0204] A solid formulation (freeze dry formulation or film) for sublingual administration was prepared according to the formulation shown in Table 5 below. Specifically, an antigen (ovalbumin (OVA)), an immunity induction promoter that was a nuclear receptor ligand, and hydroxypropylcellulose (HPC-SSL, Nippon Soda Co., Ltd.) as a base were blended. Saline was added thereto, followed by mixing to provide a formulation solution. Thereafter, 25 mg of the formulation solution was dispensed, and the dispensed solution was freeze-dried to provide a freeze dry formulation or dried under reduced pressure to provide a film. The immunity induction promoter that was a nuclear receptor ligand was the same as that used for preparing the solution for transmucosal administration.

<Evaluation 3>

[0205] Each of the solutions for transmucosal administration and solid formulations for sublingual administration obtained in the examples and the comparative examples was subjected to the following evaluation.

(Evaluation of Humoral Immunity Inducing Effect)

[0206] A mouse immunity test using an animal model for immunological evaluation was performed with the solution for transmucosal administration or the solid formulation for sublingual administration by the following procedure. Thereafter, the systemic immune response was evaluated by determining the antigen (OVA)-specific IgG antibody titer in mouse serum. The results are shown in FIGS. 3 to 5.

(1) Mouse immunity test of solution for transmucosal administration or solid formulation for sublingual administration

[0207] A mouse (BALB/c mouse, female, 7 weeks old) was provided in advance. After the mouse was anesthetized, the solution for transmucosal administration was administered to the mouse by transnasal administration (10 μL, Examples 21 to 28 and Comparative Example 9) or sublingual administration (30 μL, Examples 29 to 36 and Comparative Example 10). Similarly, the solid formulation for sublingual administration (Examples 37 to 52 and Comparative Examples 11 and 12) was administered. One week after the administration, the mouse was anesthetized again, and the administration was performed again in the same manner. One week after the second administration, the mouse serum was taken.

(2) ELISA

(Method for Determining Antigen-Specific IgG Antibody Titer in Mouse Serum (ELISA))

[0208] To each well of a 96-well plate for ELISA was added 100 μL of an OVA-containing solution (100 μg/mL) diluted with carbonate buffer, followed by standing overnight.

[0209] The wells were washed three times with preliminarily prepared wash (Tween 20-containing PBS), and to each well was added 200 μL of a blocking solution prepared by diluting a blocking agent (Block Ace, Sumitomo Dainippon Pharma Co., Ltd.) in purified water to 4 g/100 mL. This was followed by standing for 2 hours at room temperature. The wells were then washed three times with wash.

[0210] The serum taken from the mouse was centrifuged at 4° C. and 3,000 g for 10 minutes, and the supernatant was recovered. The supernatant was diluted in two-fold increments using a solution prepared by diluting a blocking agent in a phosphate buffer (Nacalai Tesque, Inc.) to 0.4 g/100 mL. The diluted solution was added to wells (50 μL for each well), followed by standing for 2 hours at room temperature.

[0211] The wells were then washed three times with wash. An HRP-labeled anti-mouse IgG antibody (Goat-anti mouse IgG Fc HRP, BETHYL) was diluted 10,000-fold using a solution prepared by diluting a blocking agent in a phosphate buffer (Nacalai Tesque, Inc.) to 0.4 g/100 mL. To each well was added 100 μL of the resulting solution, followed by standing for 1 hour at room temperature.

[0212] The wells were then washed three times with wash, and 100 μL of a TMB solution (ELISA POD TMB kit, Nacalai Tesque, Inc.) was added to each well, followed by standing for 30 minutes at dark place.

[0213] Thereafter, 100 μL of a 1M sulfuric acid solution was added to each well, and the 96-well plate was subjected to measurement of absorbance at 450 nm with a microplate reader (Spectra Max M2.sup.e, molecular device). The IgG antibody titer in the mouse serum was determined as Log 2 titer based on the absorbance at the incremental dilution.

TABLE-US-00003 TABLE 3 Antigen Nuclear receptor ligand Immunological IgG antibody Administration Dosage Amount Amount Pharmacological evaluation titer No. route form Name [ug] Name [ug] effect mouse [Log2 titer] Comp. Ex. 9 Transnasal Solution OVA 1 — — — BALB/c  4.6 Ex. 21 Transnasal Solution OVA 1 Tretinoin 50 RAR activation BALB/c  9.4 Ex. 22 Transnasal Solution OVA 1 Isotretinoin 50 RAR activation BALB/c  9.8 Ex. 23 Transnasal Solution OVA 1 Alitretinoin 50 RAR activation BALB/c  9.1 Ex. 24 Transnasal Solution OVA 1 Levothyroxine sodium 20 TR activation BALB/c 12.1 Ex. 25 Transnasal Solution OVA 1 Liothyronine 50 TR activation BALB/c 13.0 Ex. 26 Transnasal Solution OVA 1 Clomiphene citrate 10 ER modulation BALB/c 13.1 Ex. 27 Transnasal Solution OVA 1 Raloxifene hydrochloride 20 ER modulation BALB/c 11.4 Ex. 28 Transnasal Solution OVA 1 Tamoxifen citrate 10 ER modulation BALB/c 12.5

TABLE-US-00004 TABLE 4 Antigen Nuclear receptor ligand Immunological IgG antibody Administration Dosage Amount Amount Pharmacological evaluation titer No. route form Name [ug] Name [ug] effect mouse [Log2 titer] Comp. Ex. 10 Sublingual Solution OVA 1 — — — BALB/c  4.4 Ex. 29 Sublingual Solution OVA 1 Tretinoin 100 RAR activation BALB/c  7.8 Ex. 30 Sublingual Solution OVA 1 Isotretinoin 100 RAR activation BALB/c  8.0 Ex. 31 Sublingual Solution OVA 1 Alitretinoin 100 RAR activation BALB/c  7.9 Ex. 32 Sublingual Solution OVA 1 Levothyroxine sodium 100 TR activation BALB/c  8.9 Ex. 33 Sublingual Solution OVA 1 Liothyronine 100 TR activation BALB/c  9.4 Ex. 34 Sublingual Solution OVA 1 Clomiphene citrate 100 ER modulation BALB/c  9.8 Ex. 35 Sublingual Solution OVA 1 Raloxifene hydrochloride 100 ER modulation BALB/c  9.5 Ex. 36 Sublingual Solution OVA 1 Tamoxifen citrate 100 ER modulation BALB/c 10.5

TABLE-US-00005 TABLE 5 Formulation [Parts by weight] Ex. Component 37 38 39 40 41 42 43 44 45 Antigen OVA 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Immuno- Nuclear Tretinoin 10 — — — — — — — 10 stimulant receptor Isotretinoin — 10 — — — — — — — ligand Alitretinoin — — 10 — — — — — — Levothyroxine — — — 10 — — — — — sodium Liothyronine — — — — 10 — — — — Clomiphene — — — — — 10 — — — citrate Raloxifene — — — — — — 10 — — hydrochloride Tamoxifen — — — — — — — 10 — citrate Base HPC-SSL 250 250 250 250 250 250 250 250 250 Saline solution 739.9 739.9 739.9 739.9 739.9 739.9 739.9 739.9 739.9 Dispensing amount [mg/mouse] 25 25 25 25 25 25 25 25 25 IgGantibody titer [Log2 titer] 7.9 8.0 8.1 8.9 9.6 10.0 9.5 10.8 7.8 Dosage form Solid (freeze dry) Film Administration route Sublingual administration Formulation [Parts by weight] Ex. Comp. Ex. Component 46 47 48 49 50 51 52 11 12 Antigen OVA 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Immuno- Nuclear Tretinoin — — — — — — — — — stimulant receptor Isotretinoin 10 — — — — — — ligand Alitretinoin — 10 — — — — — Levothyroxine — — 10 — — — — — — sodium Liothyronine — — — 10 — — — — — Clomiphene — — — — 10 — — — — citrate Raloxifene — — — — — 10 — — — hydrochloride Tamoxifen — — — — — — 10 — — citrate Base HPC-SSL 250 250 250 250 250 250 250 250 250 Saline solution 739.9 739.9 739.9 739.9 739.9 739.9 739.9 749.9 749.9 Dispensing amount [mg/mouse] 25 25 25 25 25 25 25 25 25 IgGantibody titer [Log2 titer] 7.9 8.1 9.0 9.6 10.0 9.6 10.8 4.3 4.3 Dosage form Film Solid Film (freeze dry) Administration route Sublingual administration

Examples 53 to 56, Comparative Example 13

(Preparation of Solution for Subcutaneous Administration)

[0214] A formulation for subcutaneous administration was prepared according to the formulation shown in Table 6 below.

[0215] Specifically, an antigen (ovalbumin (OVA)) and an immunity induction promoter that was a nuclear receptor ligand were blended in the amounts shown in Table 6. Saline was added thereto such that the amount of the resulting mixture was 200 μL, followed by mixing to provide a solution for subcutaneous administration.

<Evaluation 4>

[0216] Each of the formulations for subcutaneous administration obtained in the examples and the comparative examples was subjected to the following evaluation.

(Evaluation of Humoral Immunity Inducing Effect)

[0217] A mouse immunity test using an animal model for immunological evaluation was performed with the formulation for subcutaneous administration by the following procedure. Thereafter, the systemic immune response was evaluated by determining the antigen (OVA)-specific IgG antibody in mouse serum. The results are shown in FIG. 6.

(1) Mouse Immunity Test of Formulation for Subcutaneous Administration

[0218] A mouse (BALB/c mouse, female, 7 weeks old) was provided in advance. After the mouse was anesthetized, 200 μL of the formulation was subcutaneously administered to the skin of the back of the mouse. One week after the administration, the mouse was anesthetized again, and administration was performed again in the same manner. One week after the second administration, the mouse serum was taken.

(2) ELISA

[0219] The antigen (OVA)-specific IgG antibody titer in the mouse serum was determined by ELISA by the same procedure as in <Evaluation 3>.

TABLE-US-00006 TABLE 6 Antigen Nuclear receptor ligand Immunological IgG antibody Administration Dosage Amount Amount Pharmacological evaluation titer No. route form Name [ug] Name [ug] effect mouse [Log2 titer] Comp. Ex. 13 Subcutaneous Solution OVA 0.05 — — — BALB/c  5.1 Ex. 53 Subcutaneous Solution OVA 0.05 Tretinoin 200 RAR activation BALB/c 10.4 Ex. 54 Subcutaneous Solution OVA 0.05 Levothyroxine sodium 200 TR activation BALB/c  9.1 Ex. 55 Subcutaneous Solution OVA 0.05 Liothyronine 200 TR activation BALB/c  9.6 Ex. 56 Subcutaneous Solution OVA 0.05 Raloxifene hydrochloride 200 ER modulation BALB/c 10.5

Examples 57 to 60, Comparative Example 14

(Preparation of Cream for Transdermal Administration)

[0220] A cream for transdermal administration was prepared according to the formulation shown in Table 7 below. Specifically, an antigen (ovalbumin (OVA)) and a nuclear receptor ligand were blended in the amounts shown in Table 7 and a base (base cream) was added thereto to achieve a total weight of 100 parts by weight. This was followed by mixing to provide a cream for transdermal administration. The base cream was prepared by blending and mixing materials according to the formulation shown in Table 14.

[0221] The immunity induction promoter that was a nuclear receptor ligand was the same as that used for preparing the solutions for transnasal or sublingual administration. White Vaseline, sorbitan monostearate, isostearic acid, benzyl alcohol, stearyl alcohol, polysorbate 60, and concentrated glycerin were purchased from Wako Pure Chemical Industries, Ltd. Cetanol was purchased from Tokyo Chemical Industry Co., Ltd.

[0222] A composite base was prepared by bonding a PET film/PET nonwoven fabric laminate (area: 0.7 cm.sup.2) to the center portion of an adhesive tape for attachment such that the PET-film side faced the tape. The cream in an amount of 4 mg was applied to the nonwoven fabric portion of the composite base to provide a sample for a mouse immunity test.

<Evaluation 5>

[0223] Each of the creams for transdermal administration obtained in the examples and the comparative examples was subjected to the following evaluation.

(Evaluation of Humoral Immunity Inducing Effect)

[0224] A mouse immunity test using an animal model for immunological evaluation was performed with the cream for transdermal administration by the following procedure. Thereafter, the systemic immune response was evaluated by determining the antigen (OVA)-specific IgG antibody in mouse serum. The results are shown in FIG. 7.

(1) Mouse Immunity Test of Cream for Transdermal Administration

[0225] The right back of a mouse (C57BL6 NCr mouse, female, 7 weeks old) was shaved in advance. After a rearing period for recovery from the skin damage caused by the shaving, 4 mg of the cream for transdermal administration was administered to the skin of the right back of the mouse, and the left back was shaved at the same time. Twenty-four hours later, the cream for transdermal administration on the right back was removed. One week after the administration, the cream for transdermal administration was administered to the skin of the left back of the mouse in the same manner and removed 24 hours later. One week after the second administration, the mouse serum was taken.

(2) ELISA

[0226] The antigen (OVA)-specific IgG antibody titer in the mouse serum was determined by ELISA by the same procedure as in <Evaluation 3>.

TABLE-US-00007 TABLE 7 Antigen Nuclear receptor ligand IgG Amount Amount Immunological antibody Administration Dosage [Parts by [Parts by Pharmacological evaluation titer No. route form Name weight] Name weight] effect mouse [Log2 titer] Comp. Ex. 14 Transdermal Cream OVA 5 — — — C57BL6  5.1 Ex. 57 Transdermal Cream OVA 5 Tretinoin 5 RAR activation C57BL6  9.0 Ex. 58 Transdermal Cream OVA 5 Levothyroxine sodium 5 TR activation C57BL6 11.3 Ex. 59 Transdermal Cream OVA 5 Liothyronine 5 TR activation C57BL6 11.5 Ex. 60 Transdermal Cream OVA 5 Raloxifene hydrochloride 5 ER modulation C57BL6 10.9

Examples 61 to 180, Comparative Examples 15 to 54

[0227] A solution for transmucosal administration (transnasal administration or sublingual administration) was prepared according to the formulation shown in Tables 8 to 12 below. Specifically, an antigen and a nuclear receptor ligand were blended in the amounts shown in Tables 8 to 12. For transnasal administration, saline was added thereto so that the amount of the resulting mixture was 10 μL. For sublingual administration, saline was added so that the amount of the resulting mixture was 30 μL. This was followed by mixing to provide a solution for transmucosal administration (transnasal administration or sublingual administration).

[0228] Influenza vaccine antigens used were an influenza vaccine antigen-containing solution H1N1 (A/California/07/2009, The Research Foundation for Microbial Diseases of Osaka University), H3N2 (A/Victoria361/2011, The Research Foundation for Microbial Diseases of Osaka University), Influenza B virus (B/Wisconsin/1/2010, The Research Foundation for Microbial Diseases of Osaka University), Influenza B virus (B/Brisbane/60/2008, The Research Foundation for Microbial Diseases of Osaka University) were used. Also used were a pneumococcal capsular polysaccharide-containing solution (Pneumovax NP, MSD), HPV16 recombinant protein-containing solution (HPV16, PROSPEC), a live attenuated rotavirus-containing solution (RotaTeq Oral Solution, MSD), an inactivated poliovirus-containing solution (IMOVAX POLIO for subcutaneous injection, Sanofi), an inactivated hepatitis A virus-containing solution (Aimmugen, The Chemo-Sero-Therapeutic Research Institute), an inactivated Japanese encephalitis virus-containing solution (Encevac for subcutaneous injection, The Chemo-Sero-Therapeutic Research Institute), a live attenuated mumps virus-containing solution (live mumps vaccine, Kitasato Daiichi Sankyo Vaccine Co., Ltd), a live attenuated measles virus-containing solution (live measles vaccine, Kitasato Daiichi Sankyo Vaccine Co., Ltd), a live attenuated rubella virus-containing solution (dried live attenuated rubella vaccine, Kitasato Daiichi Sankyo Vaccine Co., Ltd), a solution containing tetanus toxoid-conjugated haemophilus influenzae type b polysaccharide (ActHIB, Sanofi), a recombinant HBs antigen protein-containing solution (Bimmugen, The Chemo-Sero-Therapeutic Research Institute), a live attenuated yellow fever virus-containing solution (yellow fever vaccine, Sanofi), a tetanus toxoid-containing solution (tetanus toxoid, Denka Seiken Co., Ltd.), a live attenuated varicella virus-containing solution (dried live attenuated varicella vaccine, The Research Foundation for Microbial Diseases of Osaka University), a live BCG-containing solution (dried BCG vaccine, Japan BCG Laboratory), and an inactivated rabies virus-containing solution (tissue-cultured inactivated rabies vaccine, The Chemo-Sero-Therapeutic Research Institute).

[0229] As the immunity induction promoter that is a nuclear receptor ligand, tretinoin (all-trans-retinoic acid, Wako Pure Chemical Industries, Ltd.), liothyronine (Sigma-Aldrich), and tamoxifen citrate (Wako Pure Chemical Industries, Ltd.) were used.

<Evaluation 6>

[0230] Each of the solutions for transmucosal administration obtained in the examples and the comparative examples was subjected to the following evaluation.

(Evaluation of Humoral Immunity Inducing Effect)

[0231] A mouse immunity test using an animal model for immunological evaluation was performed with the solution for transmucosal administration by the following procedure. Thereafter, the systemic immune response was evaluated by determining the antigen (OVA)-specific IgG antibody in mouse serum.

(1) Mouse Immunity Test of Solution for Transmucosal Administration

[0232] Mouse serum was taken by the same procedure as in <Evaluation 3>, an evaluation of solution for transmucosal administration or sublingual administration.

(2) ELISA

[0233] The antigen (OVA)-specific IgG antibody titer in the mouse serum was determined by ELISA by the same procedure as in <Evaluation 3>, an evaluation of solution for transmucosal administration or sublingual administration.

[0234] The evaluation of the humoral immunity inducing effect shows that the transmucosal administration (transnasal administration or sublingual administration) of a solution for transmucosal administration containing an immunity induction promoter that is a nuclear receptor ligand (Examples 21 to 36) provides a higher antigen-specific IgG antibody titer than the administration of a solution for transmucosal administration free from an immunity induction promoter that is a nuclear receptor ligand (Comparative Examples 9 and 10).

[0235] Accordingly, also when antigens such as those shown in Tables 8 to 12 below are used, the use of an immunity induction promoter that is a nuclear receptor ligand leads to a high antigen-specific IgG antibody titer.

TABLE-US-00008 TABLE 8 Antigen Nuclear receptor ligand Amount Amount Administration Amount Name [μg] Name [μg] Formulation route [μL] Comp. Ex. 15 A/California/07/2009 [H1N1] 1.0 — — Solution Transnasal 10 Ex. 61 A/California/07/2009 [H1N1] 1.0 Tretinoin 50 Solution Transnasal 10 Ex. 62 A/California/07/2009 [H1N1] 1.0 Liothyronine 50 Solution Transnasal 10 Ex. 63 A/California/07/2009 [H1N1] 1.0 Tamoxifen 10 Solution Transnasal 10 Comp. Ex. 16 A/California/07/2009 [H1N1] 1.0 — — Solution Sublingual 30 Ex. 64 A/California/07/2009 [H1N1] 1.0 Tretinoin 100 Solution Sublingual 30 Ex. 65 A/California/07/2009 [H1N1] 1,0 Liothyronine 100 Solution Sublingual 30 Ex. 66 A/California/07/2009 [H1N1] 1.0 Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 17 A/Victoria361/2011 [H3N2] 1.0 — — Solution Transnasal 10 Ex. 67 A/Victoria361/2011 [H3N2] 1.0 Tretinoin 50 Solution Transnasal 10 Ex. 68 A/Victoria361/2011 [H3N2] 1.0 Liothyronine 50 Solution Transnasal 10 Ex. 69 A/Victoria361/2011 [H3N2] 1.0 Tamoxifen 10 Solution Transnasal 10 Comp. Ex. 18 A/Victoria361/2011 [H3N2] 1.0 — — Solution Sublingual 30 Ex. 70 A/Victoria361/2011 [H3N2] 1.0 Tretinoin 100 Solution Sublingual 30 Ex. 71 A/Victoria361/2011 [H3N2] 1.0 Liothyronine 100 Solution Sublingual 30 Ex. 72 A/Victoria361/2011 [H3N2] 1.0 Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 19 B/Wisconsin/1/2010 1.0 — — Solution Transnasal 10 Ex. 73 B/Wisconsin/1/2010 1.0 Tretinoin 50 Solution Transnasal 10 Ex. 74 B/Wisconsin/1/2010 1.0 Liothyronine 50 Solution Transnasal 10 Ex. 75 B/Wisconsin/1/2010 1.0 Tamoxifen 10 Solution Transnasal 10 Comp. Ex. 20 B/Wisconsin/1/2010 1.0 — — Solution Sublingual 30 Ex. 76 B/Wisconsin/1/2010 1.0 Tretinoin 100 Solution Sublingual 30 Ex. 77 B/Wisconsin/1/2010 1.0 Liothyronine 100 Solution Sublingual 30 Ex. 78 B/Wisconsin/1/2010 1.0 Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 21 B/Brisbane/60/2008 1.0 — — Solution Transnasal 10 Ex. 79 B/Brisbane/60/2008 1.0 Tretinoin 50 Solution Transnasal 10 Ex. 80 B/Brisbane/60/2008 1.0 Liothyronine 50 Solution Transnasal 10 Ex. 81 B/Brisbane/60/2008 1.0 Tamoxifen 10 Solution Transnasal 10 Comp. Ex. 22 B/Brisbane/60/2008 1.0 — — Solution Sublingual 30 Ex. 82 B/Brisbane/60/2008 1.0 Tretinoin 100 Solution Sublingual 30 Ex. 83 B/Brisbane/60/2008 1.0 Liothyronine 100 Solution Sublingual 30 Ex. 84 B/Brisbane/60/2008 1.0 Tamoxifen 100 Solution Sublingual 30

TABLE-US-00009 TABLE 9 Antigen Nuclear receptor ligand Adminis- Amount Amout tration Amount Name [μg] Name [μg] Formulation route [μL] Comp. Pneumococcal capsular polysaccharide 20 — — Solution Transnasal 10 Ex. 23 Pneumovax NP Ex. 86 Pneumococcal capsular polysaccharide 20 Tretinoin  50 Solution Transnasal 10 Pneumovax NP Ex. 86 Pneumococcal capsular polysaccharide 20 Liothyronine  50 Solution Transnasal 10 Pneumovax NP Ex. 87 Pneumococcal capsular polysaccharide 20 Tamoxifen  10 Solution Transnasal 10 Pneumovax NP Comp. Pneumococcal capsular polysaccharide 20 — — Solution Sublingual 30 Ex. 24 Pneumovax NP Ex. 88 Pneumococcal capsular polysaccharide 20 Tretinoin 100 Solution Sublingual 30 Pneumovax NP Ex. 89 Pneumococcal capsular polysaccharide 20 Liothyronine 100 Solution Sublingual 30 Pneumovax NP Ex. 90 Pneumococcal capsular polysaccharide 20 Tamoxifen 100 Solution Sublingual 30 Pneumovax NP Comp. HPV16 recombinant protein 10 — — Solution Transnasal 10 Ex. 25 Ex. 91 HPV16 recombinant protein 10 Tretinoin  50 Solution Transnasal 10 Ex 92 HPV16 recombinant protein 10 Liothyronine  50 Solution Transnasal 10 Ex. 93 HPV16 recombinant protein 10 Tamoxifen  10 Solution Transnasal 10 Comp. HPV16 recombinant protein 10 — — Solution Sublingual 30 Ex. 26 Ex. 94 HPV16 recombinant protein 10 Tretinoin 100 Solution Sublingual 30 Ex. 95 HPV16 recombinant protein 10 Liothyronine 100 Solution Sublingual 30 Ex. 96 HPV16 recombinant protein 10 Tamoxifen 100 Solution Sublingual 30 Comp. Live attenuated rotavirus (RIX4414 strain) 10 — — Solution Transnasal 10 Ex. 27 Ex. 97 Live attenuated rotavirus (RIX4414 strain) 10 Tretinoin  50 Solution Trananasal 10 Ex. 98 Live attenuated rotavirus (RIX4414 strain) 10 Liothyronine  50 Solution Transnasal 10 Ex. 99 Live attenuated rotavirus (RIX4414 strain) 10 Tamoxifen  10 Solution Transnasal 10 Comp. Live attenuated rotavirus (RIX4414 strain) 10 — — Solution Sublingual 30 Ex. 28 Ex. 100 Live attenuated rotavirus (RIX4414 atrain 10 Tretinoin 100 Solution Sublingual 30 Ex. 101 Live attenuated rotavirus (RIX4414 strain) 10 Liothyronine 100 Solution Sublingual 30 Ex. 102 Live attenuated rotavirus (RIX4414 strain) 10 Tamoxifen 100 Solution Sublingual 30 Comp. Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL — — Solution Transnasal 10 Ex. 29 type 3) equivalent Ex. 103 Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL Tretinoin  50 Solution Transnasal 10 type 3) equivalent Ex. 104 Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL Liothyronine  50 Solution Transnasal 10 type 3) equivalent Ex. 105 Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL Tamoxifen  10 Solution Transnasal 10 type 3) equivalent Comp. Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL — — Solution Sublingual 30 Ex. 30 type 3) equivalent Ex. 106 Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL Tretinoin 100 Solution Sublingual 30 type 3) equivalent Ex. 107 Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL Liothyronine 100 Solution Sublingual 30 type 3) equivalent Ex. 108 Inactivated poliovirus (type 1, type 2, and Vaccine 100 μL Tamoxifen 100 Solution Sublingual 30 type 3) equivalent

TABLE-US-00010 TABLE 10 Antigen Nuclear receptor ligand Amount Amount Administration Amount Name [μg] Name [μg] Formulation route [μL] Comp. Ex. 31 Inactivated hepatitis A virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 109 Inactivated hepatitis A virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 110 Inactivated hepatitis A virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 111 Inactivated hepatitis A virus Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 32 Inactivated hepatitis A virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 112 Inactivated hepatitis A virus Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 113 Inactivated hepatitis A virus Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 114 Inactivated hepatitis A virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 33 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 115 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 116 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 117 Inactivated Japanese encephalitis virus Vaccine 100 μL equivaient Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 34 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 118 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 119 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 120 Inactivated Japanese encephalitis virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 35 Live attenuated mumps virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 121 Live attenuated mumps virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 122 Live attenuated mumps virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 123 Live attenuated mumps virus Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 36 Live attenuated mumps virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 124 Live attenuated mumps virus Vaccine 100 μL equivalent Tretinoin 100 Solution Subliagual 30 Ex. 125 Live attenuated mumps virus Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 126 Live attenuated mumps virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 37 Live attenuated measles virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 127 Live attenuated measles virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 128 Live attenuated measles virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 129 Live attenuated measles virus Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 38 Live attenuated measles virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 130 Live attenuated measles virus Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 131 Live attenuated measles virus Vaccine 100 μL equivalent Liothyronixe 100 Solution Sublingual 30 Ex. 132 Live attenuated measles virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30

TABLE-US-00011 TABLE 11 Antigen Nuclear receptor ligand Amount Amount Administration Amount Name [μg] Name [μg] Formulation route [μL] Comp. Ex. 39 Live attenuated rubella virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 133 Live attenuated rubella virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 134 Live attenuated rubella virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 135 Live attenuated rubella virus Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex 40 Live attenuated rubella virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 138 Live attenuated rubella virus Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 137 Live attenuated rubella virus Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 138 Live attenuated rubella virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 41 Tetanus toxoid-conjugated Vaccine 100 μL equivalent — — Solution Transnasal 10 Haemophilus influenzae type b polysaccharide Ex. 139 Tetanus toxoid-conjugated Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Haemophilus influenzae type b polysaccharide Ex. 140 Tetanus toxoid-conjugated Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Haemophilus influenzae type b polysaccharide Ex. 141 Tetanus toxoid-conjugated Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Haemophilus influenzae type b polysacchride Comp. Ex. 42 Tetanus toxoid-conjugated Vaccine 100 μL equivalent — — Solution Sublingual 30 Haemophilus influenzae type b polysaccharide Ex. 142 Tetanus toxoid-conjugated Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Haemophilus influenzae type b polysaccharide Ex. 143 Tetanus toxoid-conjugated Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Haemophilus influenzae type b polysaccharide Ex. 144 Tetanus toxoid-conjugated Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Haemophilus influenzae type b polysaccharide Comp. Ex. 43 Recombinant HBs antigen protein Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 145 Recombinant HBs antigen protein Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 146 Recombinant HBs antigen protein Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 147 Recombinant HBs antigen protein Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 44 Recombinant HBs antigen protein Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 148 Recombinant HBs antigen protein Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 149 Recombinant HBs antigen protein Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 150 Recombinant HBs antigen protein Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 45 Live attenuated yellow fever virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 151 Live attenuated yellow fever virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 152 Live attenuated yellow fever virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 153 Live attenuated yellow fever virus Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 46 Live attenuated yellow fever virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 154 Live attenuated yellow fever virus Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 155 Live attenuated yellow fever virus Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 156 Live attenuated yellow fever virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30

TABLE-US-00012 TABLE 12 Antigen Nuclear receptor ligand Amount Amount Administration Amount Name [μg] Name [μg] Formulation route [μL] Comp. Ex. 47 Tetanus toxoid Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 157 Tetanus toxoid Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 158 Tetanus toxoid Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 159 Tetanus toxoid Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 48 Tetanus toxoid Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 160 Tetanus toxoid Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 161 Tetanus toxoid Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 162 Tetanus toxoid Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 49 Live attenuated varicella virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 163 Live attenuated varicella virus Vaccine 100 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 164 Live attenuated varicella virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 165 Live attenuated varicella virus Vaccine 100 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 50 Live attenuated varicella virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 166 Live attenuated varicella virus Vaccine 100 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 167 Live attenuated varicella virus Vaccine 100 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 168 Live attenuated varicella virus Vaccine 100 μL equivalent Tamoxifen 100 Solution Sublingual 30 Comp. Ex. 51 Live BCG Vaccine 30 μL equivalent — — Solution Transnasal 10 Ex. 169 Live BCG Vaccine 30 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 170 Live BCG Vaccine 30 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 171 Live BCG Vaccine 30 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 52 Live BCG Vaccine 30 μL equivalent — — Solution Sublingual 30 Ex. 172 Live BCG Vaccine 30 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 173 Live BCG Vaccine 30 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 174 Live BCG Vaccine 30 μL equivalent Tainoxifen 100 Solution Sublingual 30 Comp. Ex. 53 Inactivated rabies virus Vaccine 200 μL equivalent — — Solution Transnasal 10 Ex. 175 Inactivated rabies virus Vaccine 200 μL equivalent Tretinoin  50 Solution Transnasal 10 Ex. 176 Inactivated rabies virus Vaccine 100 μL equivalent Liothyronine  50 Solution Transnasal 10 Ex. 177 inactivated rabies virus Vaccine 200 μL equivalent Tamoxifen  10 Solution Transnasal 10 Comp. Ex. 54 Inactivated rabies virus Vaccine 200 μL equivalent — — Solution Sublingual 30 Ex. 178 Inactivated rabies virus Vaccine 200 μL equivalent Tretinoin 100 Solution Sublingual 30 Ex. 179 Inactivated rabies virus Vaccine 200 μL equivalent Liothyronine 100 Solution Sublingual 30 Ex. 180 Inactivated rabies virus Vaccine 200 μL equivalent Tamoxifen 100 Solution Sublingual 30

Examples 181 to 184, Comparative Example 55

[0236] A cream for transdermal administration was prepared according to the formulation shown in Table 13 in the same manner as the cream for transdermal administration of Table 7. The right back of a mouse (C57BL6 NCr mouse, female, 7 weeks old) was shaved, and after the skin was subjected to a corneum removing treatment five times with an OPP tape (EZ Dunplon No. 3301EZ, Nitto Denko Corporation), the cream was administered to the skin (minimally invasive administration), and the left back was shaved at the same time. Twenty-four hours later, the cream for transdermal administration on the right back was removed. One week after the administration, the skin of the left back of the mouse was subjected to a corneum removing treatment in the same manner as above, and the cream for transdermal administration was administered thereto. The cream was removed 24 hours later. One week after the second administration, the mouse serum was taken, and the antigen (OVA)-specific IgG antibody in the serum was determined by ELISA. Also in this immunization using the minimally invasive administration, humoral immunity specific to the administered antigen can be induced.

TABLE-US-00013 TABLE 13 Antigen Nuclear receptor ligand Amount Amount [Parts [Parts Immunological Dosage by by Pharmacological evaluation No. Administration route form Name weight] Name weight] effect mouse Comp. Ex. 55 Transdermal (minimally invasive) Cream OVA 5 — — — C57BL6 Ex. 181 Transdermal (minimally invasive) Cream OVA 5 Tretinoin 5 RAR activation C57BL6 Ex. 182 Transdermal (minimally invasive) Cream OVA 5 Levothyroxine sodium 5 TR activation C57BL6 Ex. 183 Transdermal (minimally invasive) Cream OVA 5 Liothyronine 5 TR activation C57BL6 Ex. 184 Transdermal (minimally invasive) Cream OVA 5 Raloxifene hydrochloride 5 ER modulation C57BL6

TABLE-US-00014 TABLE 14 Amount [Parts by Additve weight] White Vaseline 60.7 Sorbitan monostearate 0.7 Isostearic acid 12 Benzyl alcohol 2.4 Cetanol 2.4 Stearyl alcohol 3.5 Polysorbate 60 3.5 Concentrated glycerin 2.4 Purified water 12.4 Total 100

INDUSTRIAL APPLICABILITY

[0237] The composition for promoting immunity induction and vaccine pharmaceutical composition of the present invention are universally usable for inducing immunity to various antigens and suitable not only for subcutaneous administration but also for transdermal administration or transmucosal administration.