LIQUID IMMUNITY INDUCTION-PROMOTING COMPOSITION AND VACCINE PHARMACEUTICAL COMPOSITION THAT INCLUDE THROMBOSIS TREATMENT DRUG

Abstract

The present invention aims to provide a humoral immunity induction-promoting composition and a vaccine pharmaceutical composition which can universally be used for inducing humoral immunity to various antigens and exert a high antibody production inducing effect. The present invention relates to a vaccine pharmaceutical composition for inducing humoral immunity containing an antigen and a humoral immunity induction promoter whose active ingredient is a thrombosis treatment drug.

Claims

1. A humoral immunity induction-promoting composition, comprising: a humoral immunity induction promoter whose active ingredient is a thrombosis treatment drug.

2. The humoral immunity induction-promoting composition according to claim 1, wherein the thrombosis treatment drug is a thrombogenesis-suppressing compound.

3. The humoral immunity induction-promoting composition according to claim 2, wherein the thrombogenesis-suppressing compound is at least one of an anticoagulant or an antiplatelet, the anticoagulant being at least one selected from the group consisting of heparin, dalteparin, fondaparinux, antissasin, TAP (tick anticoagulant peptide), rivaroxaban, apixaban, edoxaban, betrixaban, eribaxaban, YM-150, LY-517717, TAK-442, ximelagatran, dabigatran, argatroban, hirudin, nafamostat, camostat, gabexate, warfarin, and coumarin, the antiplatelet being at least one selected from the group consisting of abciximab, eptifibatide, and tirofiban.

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

5. The vaccine pharmaceutical composition according to claim 4, which is administered to a body surface.

6. The vaccine pharmaceutical composition according to claim 4, which is administered by intradermal injection, subcutaneous injection, or intramuscular injection.

7. A method for promoting humoral immunity induction in a subject in need thereof, comprising: administering to the subject an effective amount of a humoral immunity induction promoter comprising a thrombosis treatment drug.

8. The method for promoting humoral immunity induction according to claim 7, wherein the thrombosis treatment drug is a thrombogenesis-suppressing compound.

9. The method for promoting humoral immunity induction according to claim 8, wherein the thrombogenesis-suppressing compound is at least one of an anticoagulant or an antiplatelet, and the anticoagulant is at least one selected from the group consisting of heparin, dalteparin, fondaparinux, antissasin, TAP (tick anticoagulant peptide), rivaroxaban, apixaban, edoxaban, betrixaban, eribaxaban, YM-150, LY-517717, TAK-442, ximelagatran, dabigatran, argatroban, hirudin, nafamostat, camostat, gabexate, warfarin, and coumarin, and the antiplatelet is at least one selected from the group consisting of abciximab, eptifibatide, and tirofiban.

10. A method for promoting humoral immunity induction in a subject in need thereof, comprising: administering to the subject an effective amount of a vaccine pharmaceutical composition for inducing humoral immunity, which comprises an antigen; and a humoral immunity induction promoter comprising a thrombosis treatment drug.

11. The method for promoting humoral immunity induction according to claim 10, wherein the vaccine is administered to a body surface.

12. The method for promoting humoral immunity induction according to claim 10, wherein the vaccine is administered by intradermal injection, subcutaneous injection, or intramuscular injection.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0120] FIG. 1 shows an antigen (OVA)-specific IgG antibody titer in mouse serum after transnasal administration of solutions for transmucosal administration obtained in Examples 1 to 7 and Comparative Example 1.

[0121] FIG. 2 shows an antigen (OVA)-specific IgG antibody titer in mouse serum after sublingual administration of solutions for transmucosal administration obtained in Examples 8 to 14 and Comparative Example 2.

[0122] FIG. 3 shows an antigen (OVA)-specific IgG antibody titer in mouse serum after sublingual administration of solid formulations for sublingual administration obtained in Examples 15 to 28 and Comparative Examples 3 and 4.

[0123] FIG. 4 shows an antigen (OVA)-specific IgG antibody titer in mouse serum after subcutaneous administration of solutions for subcutaneous administration obtained in Examples 29 to 33 and Comparative Example 5.

[0124] FIG. 5 shows an antigen (OVA)-specific IgG antibody titer in mouse serum after transdermal administration of creams for transdermal administration obtained in Examples 34 to 38 and Comparative Example 6.

DESCRIPTION OF EMBODIMENTS

[0125] The present invention will be specifically described in further detail below. The present invention, however, is not limited to these examples.

Examples 1 to 14, Comparative Examples 1 and 2

(Preparation of Solution for Transmucosal Administration)

[0126] A solution for transmucosal administration (transnasal administration or sublingual administration) was prepared according to the formulation shown in Tables 1 and 2 below. Specifically, an antigen (ovalbumin (OVA)) and a humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug were blended in the amounts shown in Tables 1 and 2. For transnasal administration, saline was added thereto suth 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).

[0127] As the humoral immunity induction promoter whose active ingredient is a thrombosis treatment drug, rivaroxaban (ChemScene, LLC), apixaban (ChemScene, LLC), argatroban (LKT Laboratories), nafamostat mesylate (Wako Pure Chemical Industries, Ltd.), camostat mesylate (Wako Pure Chemical Industries, Ltd.), gabexate mesylate (Wako Pure Chemical Industries, Ltd.), and tirofiban (Sigma-Aldrich) were used.

Examples 15 to 28, Comparative Examples 3 and 4

(Preparation of Solid Formulation for Sublingual Administration)

[0128] A solid formulation (freeze-dry formulation or film) for sublingual administration was prepared according to the formulation shown in Table 3 below. Specifically, an antigen (ovalbumin (OVA)), a humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug, and hydroxypropylcellulose (HPC-SSL, Nippon Soda Co., Ltd.) as abase were blended in the amounts shown in Table 3. 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 humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug was the same as that used for preparing the solution for transmucosal administration.

<Evaluation 1>

[0129] 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)

[0130] 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 FIG. 1, 2, or 3.

(1) Mouse Immunity Test of Solution for Transmucosal Administration or Solid Formulation for Sublingual Administration

[0131] 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 1 to 7 and Comparative Example 1 (Table 1)) or sublingual administration (30 μL, Examples to 14 and Comparative Example 2 (Table 2)). Similarly, the solid formulation for sublingual administration (Examples 15 to 28 and Comparative Examples 3 and 4 (Table 3)) 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))

[0132] 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.

[0133] 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.

[0134] 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 hours at room temperature.

[0135] 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.

[0136] 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.

[0137] Thereafter, 100 μL of a IM 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-00001 TABLE 1 Antigen Immunity induction promoter Immunological IgG antibody Administration Dosage Amount Amount evaluation titer No. route form Name [ug] Name [ug] mouse [Log2 titer] Comp. Transnasal Solution OVA 1 — — BALB/c 4.3 Ex. 1 Ex. 1 Transnasal Solution OVA 1 Rivaroxaban 20 BALB/c 10.5 Ex. 2 Transnasal Solution OVA 1 Apixaban 50 BALB/c 8.1 Ex. 3 Transnasal Solution OVA 1 Argatroban 20 BALB/c 13.3 Ex. 4 Transnasal Solution OVA 1 Nafamostat mesylate 20 BALB/c 12.3 Ex. 5 Transnasal Solution OVA 1 Camostat mesylate 20 BALB/c 10.8 Ex. 6 Transnasal Solution OVA 1 Gabexate mesylate 20 BALB/c 8.5 Ex. 7 Transnasal Solution OVA 1 Tirofiban 20 BALB/c 12.4

TABLE-US-00002 TABLE 2 Antigen Immunity induction promoter Immunological IgG antibody Administration Dosage Amount Amount evaluation titer No. route form Name [ug] Name [ug] mouse [Log2 titer] Comp. Sublingual Solution OVA 1 — — BALB/c 4.1 Ex. 2 Ex. 8 Sublingual Solution OVA 1 Rivaroxaban 100 BALB/c 9.5 Ex. 9 Sublingual Solution OVA 1 Apixaban 100 BALB/c 7.5 Ex. 10 Sublingual Solution OVA 1 Argatroban 100 BALB/c 11.4 Ex. 11 Sublingual Solution OVA 1 Nafamostat mesylate 50 BALB/c 10.9 Ex. 12 Sublingual Solution OVA 1 Camostat mesylate 100 BALB/c 10.3 Ex. 13 Sublingual Solution OVA 1 Gabexate mesylate 100 BALB/c 8.0 Ex. 14 Sublingual Solution OVA 1 Tirofiban 100 BALB/c 11.4

TABLE-US-00003 TABLE 3 Formulation [Parts by weight] Ex. Comp. Ex. Component 15 16 17 18 19 20 21 22 23 24 25 26 27 28 3 4 Antigen OVA 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Immunity Name Rivaroxaban 10 — — — — — — 10 — — — — — — — — induction Apixaban — 10 — — — — — — 10 — — — — — promoter Argatroban — — 10 — — — — — — 10 — — — — Nafamostat — — — 5 — — — — — — 5 — — — — — mesylate Camostat — — — — 10 — — — — — — 10 — — — — mesylate Gabexate — — — — — 10 — — — — — — 10 — — — mesylate Tirofiban — — — — — — 10 — — — — — — 10 — — Base HPC-SSL 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 250 Saline 739.9 739.9 739.9 744.9 739.9 739.9 739.9 739.9 739.9 739.9 744.9 739.9 739.9 739.9 749.9 749.9 Dispensing amount 25 25 25 25 25 25 25 25 25 25 25 25 25 25 25 25 [mg/mouse] IgG antibody titer 9.1 8.0 11.3 10.9 10.5 7.8 11.4 9.1 7.9 11.5 10.5 10.6 7.9 11.1 4.3 4.1 [Log2 titer] Dosage form Solid (freeze dry) Film Solid Film (freeze dry) Administration route Sublingual administration

Examples 29 to 33, Comparative Example 5

(Preparation of Solution for Subcutaneous Administration)

[0138] A formulation for subcutaneous administration was prepared according to the formulation shown in Table 4 below.

[0139] Specifically, an antigen (ovalbumin (OVA)) and a humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug were blended in the amounts shown in Table 4. 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 2>

[0140] 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)

[0141] 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. 4.

(1) Mouse Immunity Test of Formulation for Subcutaneous Administration

[0142] 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

[0143] The antigen (OVA)-specific IgG antibody titer in the mouse serum was determined by ELISA by the same procedure as in

<Evaluation 1>.

[0144]

TABLE-US-00004 TABLE 4 Antigen Immunity induction promoter Immunological IgG antibody Administration Dosage Amount Amount evaluation titer No. route form Name [ug] Name [ug] mouse [Log2 titer] Comp. Subcutaneous Solution OVA 0.05 — — BALB/c 5.4 Ex. 5 Ex. 29 Subcutaneous Solution OVA 0.05 Rivaroxaban 200 BALB/c 8.3 Ex. 30 Subcutaneous Solution OVA 0.05 Argatroban 200 BALB/c 10.4 Ex. 31 Subcutaneous Solution OVA 0.05 Nafamostat mesylate 200 BALB/c 9.5 Ex. 32 Subcutaneous Solution OVA 0.05 Camostat mesylate 200 BALB/c 9.9 Ex. 33 Subcutaneous Solution OVA 0.05 Tirofiban 200 BALB/c 9.1

Examples 34 to 38, Comparative Example 6

(Preparation of Cream for Transdermal Administration)

[0145] A cream for transdermal administration was prepared according to the formulation shown in Table 5 below. Specifically, an antigen (ovalbumin (OVA)) and a humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug were blended in the amounts shown in Table 5 and a base (base cream) was added thereto to achieve a total weight of 100 parts 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 shown in Table 6.

[0146] The humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug 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.

[0147] 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 3>

[0148] 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)

[0149] 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. 5.

(1) Mouse Immunity Test of Cream for Transdermal Administration

[0150] 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

[0151] The antigen (OVA)-specific IgG antibody titer in the mouse serum was determined by ELISA by the same procedure as in

<Evaluation 1>.

[0152]

TABLE-US-00005 TABLE 5 Antigen Immunity induction promoter Amount Amount Immunological IgG antibody Administration Dosage [Parts by [Parts by evaluation titer No. route form Name weight] Name weight] mouse [Log2 titer] Comp. Transdermal Cream OVA 5 — — C57BL6 5.4 Ex. 6 Ex. 34 Transdermal Cream OVA 5 Rivaroxaban 5 C57BL6 9.0 Ex. 35 Transdermal Cream OVA 5 Argatroban 5 C57BL6 12.0 Ex. 36 Transdermal Cream OVA 5 Nafamostat mesylate 5 C57BL6 10.8 Ex. 37 Transdermal Cream OVA 5 Camostat mesylate 5 C57BL6 11.1 Ex. 38 Transdermal Cream OVA 5 Tirofiban 5 C57BL6 11.5

TABLE-US-00006 TABLE 6 Amount Additve [Parts by 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

Examples 39 to 198, Comparative Examples 7 to 46

[0153] A solution for transmucosal administration (transnasal administration or sublingual administration) was prepared according to the formulation shown in Tables 7 to 11 below. Specifically, an antigen and a humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug were blended in the amounts shown in Tables 7 to 11. 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).

[0154] 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).

[0155] As the humoral immunity induction promoter whose active ingredient was a thrombosis treatment drug, argatroban (LKT Laboratories), nafamostat mesylate (Wako Pure Chemical Industries, Ltd.), camostat mesylate (Wako Pure Chemical Industries, Ltd.), and tirofiban (Sigma-Aldrich) were used.

<Evaluation 4>

[0156] 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)

[0157] 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

[0158] Mouse serum was taken by the same procedure as in <Evaluation 1>, an evaluation of solution for transmucosal administration or sublingual administration.

(2) ELISA

[0159] The antigen (OVA)-specific IgG antibody titer in the mouse serum was determined by ELISA by the same procedure as in <Evaluation 1>, an evaluation of solution for transmucosal administration or sublingual administration.

[0160] 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 a humoral immunity induction promoter whose active ingredient is a thrombosis treatment drug (Examples 1 to 14) provides a higher antigen-specific IgG antibody titer than the administration of a solution for transmucosal administration free from a humoral immunity induction promoter whose active ingredient is a thrombosis treatment drug (Comparative Examples 1 and 2).

[0161] Accordingly, also when antigens such as those shown in Tables 7 to 11 below are used, the use of a humoral immunity induction promoter whose active ingredient is a thrombosis treatment drug leads to a high antigen-specific IgG antibody titer.

TABLE-US-00007 TABLE 7 Antigen Immunity induction promoter Amount Amount Dosage Administration Amount Name [μg] Name [μg] form route [μL] Comp. A/California/07/2009 [H1N1] 1.0 — — Solution Transnasal 10 Ex. 7 Ex. 39 A/California/07/2009 [H1N1] 1.0 Argatroban 20 Solution Transnasal 10 Ex. 40 A/California/07/2009 [H1N1] 1.0 Nafamostat mesylate 20 Solution Transnasal 10 Ex. 41 A/California/07/2009 [H1N1] 1.0 Camostat mesylate 20 Solution Transnasal 10 Ex. 42 A/California/07/2009 [H1N1] 1.0 Tirofiban 20 Solution Transnasal 10 Ex. 43 A/California/07/2009 [H1N1] 1.0 — — Solution Sublingual 30 Comp. A/California/07/2009 [H1N1] 1.0 Argatroban 100 Solution Sublingual 30 Ex. 8 Ex. 44 A/California/07/2009 [H1N1] 1.0 Nafamostat mesylate 100 Solution Sublingual 30 Ex. 45 A/California/07/2009 [H1N1] 1.0 Camostat mesylate 100 Solution Sublingual 30 Ex. 46 A/California/07/2009 [H1N1] 1.0 Tirofiban 100 Solution Sublingual 30 Comp. A/Victoria361/2011 [H3N2] 1.0 — — Solution Transnasal 10 Ex. 9 Ex. 47 A/Victoria361/2011 [H3N2] 1.0 Argatroban 20 Solution Transnasal 10 Ex. 48 A/Victoria361/2011 [H3N2] 1.0 Nafamostat mesylate 20 Solution Transnasal 10 Ex. 49 A/Victoria361/2011 [H3N2] 1.0 Camostat mesylate 20 Solution Transnasal 10 Ex. 50 A/Victoria361/2011 [H3N2] 1.0 Tirofiban 20 Solution Transnasal 10 Comp. A/Victoria361/2011 [H3N2] 1.0 — — Solution Sublingual 30 Ex. 10 Ex. 51 A/Victoria361/2011 [H3N2] 1.0 Argatroban 100 Solution Sublingual 30 Ex. 52 A/Victoria361/2011 [H3N2] 1.0 Nafamostat mesylate 100 Solution Sublingual 30 Ex. 53 A/Victoria361/2011 [H3N2] 1.0 Camostat mesylate 100 Solution Sublingual 30 Ex. 54 A/Victoria361/2011 [H3N2] 1.0 Tirofiban 100 Solution Sublingual 30 Comp. B/Wisconsin/1/2010 1.0 — — Solution Transnasal 10 Ex. 11 Ex. 55 B/Wisconsin/1/2010 1.0 Argatroban 20 Solution Transnasal 10 Ex. 56 B/Wisconsin/1/2010 1.0 Nafamostat mesylate 20 Solution Transnasal 10 Ex. 57 B/Wisconsin/1/2010 1.0 Camostat mesylate 20 Solution Transnasal 10 Ex. 58 B/Wisconsin/1/2010 1.0 Tirofiban 20 Solution Transnasal 10 Comp. B/Wisconsin/1/2010 1.0 — — Solution Sublingual 30 Ex. 12 Ex. 59 B/Wisconsin/1/2010 1.0 Argatroban 100 Solution Sublingual 30 Ex. 60 B/Wisconsin/1/2010 1.0 Nafamostat mesylate 100 Solution Sublingual 30 Ex. 61 B/Wisconsin/1/2010 1.0 Camostat mesylate 100 Solution Sublingual 30 Ex. 62 B/Wisconsin/1/2010 1.0 Tirofiban 100 Solution Sublingual 30 Comp. B/Brisbane/60/2008 1.0 — — Solution Transnasal 10 Ex. 13 Ex. 63 B/Brisbane/60/2008 1.0 Argatroban 20 Solution Transnasal 10 Ex. 64 B/Brisbane/60/2008 1.0 Nafamostat mesylate 20 Solution Transnasal 10 Ex. 65 B/Brisbane/60/2008 1.0 Camostat mesylate 20 Solution Transnasal 10 Ex. 66 B/Brisbane/60/2008 1.0 Tirofiban 20 Solution Transnasal 10 Comp. B/Brisbane/60/2008 1.0 — — Solution Sublingual 30 Ex. 14 Ex. 67 B/Brisbane/60/2008 1.0 Argatroban 100 Solution Sublingual 30 Ex. 68 B/Brisbane/60/2008 1.0 Nafamostat mesylate 100 Solution Sublingual 30 Ex. 69 B/Brisbane/60/2008 1.0 Camostat mesylate 100 Solution Sublingual 30 Ex. 70 B/Brisbane/60/2008 1.0 Tirofiban 100 Solution Sublingual 30

TABLE-US-00008 TABLE 8 Antigen Immunity induction promoter Amount Amount Dosage Administration Amount Name [μg] Name [μg] form route [μL] Comp. Pneumococcal capsular 20 — — Solution Transnasal 10 Ex. 15 polysaccharide Pneumovax NP Ex. 71 Pneumococcal capsular 20 Argatroban 20 Solution Transnasal 10 polysaccharide Pneumovax NP Ex. 72 Pneumococcal capsular 20 Nafamostat mesylate 20 Solution Transnasal 10 polysaccharide Pneumovax NP Ex. 73 Pneumococcal capsular 20 Camostat mesylate 20 Solution Transnasal 10 polysaccharide Pneumovax NP Ex. 74 Pneumococcal capsular 20 Tirofiban 20 Solution Transnasal 10 polysaccharide Pneumovax NP Comp. Pneumococcal capsular 20 — — Solution Sublingual 30 Ex. 16 polysaccharide Pneumovax NP Ex. 75 Pneumococcal capsular 20 Argatroban 100 Solution Sublingual 30 polysaccharide Pneumovax NP Ex. 76 Pneumococcal capsular 20 Nafamostat mesylate 100 Solution Sublingual 30 polysaccharide Pneumovax NP Ex. 77 Pneumococcal capsular 20 Camostat mesylate 100 Solution Sublingual 30 polysaccharide Pneumovax NP Ex. 78 Pneumococcal capsular 20 Tirofiban 100 Solution Sublingual 30 polysaccharide Pneumovax NP Comp. HPV16 recombinant protein 10 — — Solution Transnasal 10 Ex. 17 Ex. 79 HPV16 recombinant protein 10 Argatroban 20 Solution Transnasal 10 Ex. 80 HPV16 recombinant protein 10 Nafamostat mesylate 20 Solution Transnasal 10 Ex. 81 HPV16 recombinant protein 10 Camostat mesylate 20 Solution Transnasal 10 Ex. 82 HPV16 recombinant protein 10 Tirofiban 20 Solution Transnasal 10 Comp. HPV16 recombinant protein 10 — — Solution Sublingual 30 Ex. 18 Ex. 83 HPV16 recombinant protein 10 Argatroban 100 Solution Sublingual 30 Ex. 84 HPV16 recombinant protein 10 Nafamostat mesylate 100 Solution Sublingual 30 Ex. 85 HPV16 recombinant protein 10 Camostat mesylate 100 Solution Sublingual 30 Ex. 86 HPV16 recombinant protein 10 Tirofiban 100 Solution Sublingual 30 Comp. Live attenuated rotavirus 10 — — Solution Transnasal 10 Ex. 19 (RIX4414 strain) Ex. 87 Live attenuated rotavirus 10 Argatroban 20 Solution Transnasal 10 (RIX4414 strain) Ex. 88 Live attenuated rotavirus 10 Nafamostat mesylate 20 Solution Transnasal 10 (RIX4414 strain) Ex. 89 Live attenuated rotavirus 10 Camostat mesylate 20 Solution Transnasal 10 (RIX4414 strain) Ex. 90 Live attenuated rotavirus 10 Tirofiban 20 Solution Transnasal 10 (RIX4414 strain) Comp. Live attenuated rotavirus 10 — — Solution Sublingual 30 Ex. 20 (RIX4414 strain) Ex. 91 Live attenuated rotavirus 10 Argatroban 100 Solution Sublingual 30 (RIX4414 strain) Ex. 92 Live attenuated rotavirus 10 Nafamostat mesylate 100 Solution Sublingual 30 (RIX4414 strain) Ex. 93 Live attenuated rotavirus 10 Camostat mesylate 100 Solution Sublingual 30 (RIX4414 strain) Ex. 94 Live attenuated rotavirus 10 Tirofiban 100 Solution Sublingual 30 (RIX4414 strain) Comp. Inactivated poliovirus Vaccine 100 μL — — Solution Transnasal 10 Ex. 21 (type 1, type 2, and type 3) equivalent Ex. 95 Inactivated poliovirus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 (type 1, type 2, and type 3) equivalent Ex. 96 Inactivated poliovirus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 (type 1, type 2, and type 3) equivalent Ex. 97 Inactivated poliovirus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 (type 1, type 2, and type 3) equivalent Ex. 98 Inactivated poliovirus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 (type 1, type 2, and type 3) equivalent Comp. Inactivated poliovirus Vaccine 100 μL — — Solution Sublingual 30 Ex. 22 (type 1, type 2, and type 3) equivalent Ex. 99 Inactivated poliovirus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 (type 1, type 2, and type 3) equivalent Ex. 100 Inactivated poliovirus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 (type 1, type 2, and type 3) equivalent Ex. 101 Inactivated poliovirus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 (type 1, type 2, and type 3) equivalent Ex. 102 Inactivated poliovirus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 (type 1, type 2, and type 3) equivalent

TABLE-US-00009 TABLE 9 Antigen Immunity induction promoter Amount Amount Dosage Administration Amount Name [μg] Name [μg] form route [μL] Comp. Inactivated hepatitis A virus Vaccine 100 μL — — Solution Transnasal 10 Ex. 23 equivalent Ex. 103 Inactivated hepatitis A virus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 equivalent Ex. 104 Inactivated hepatitis A virus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 equivalent Ex. 105 Inactivated hepatitis A virus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 equivalent Ex. 106 Inactivated hepatitis A virus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 equivalent Comp. Inactivated hepatitis A virus Vaccine 100 μL — — Solution Sublingual 30 Ex. 24 equivalent Ex. 107 Inactivated hepatitis A virus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 equivalent Ex. 108 Inactivated hepatitis A virus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 equivalent Ex. 109 Inactivated hepatitis A virus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 equivalent Ex. 110 Inactivated hepatitis A virus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 equivalent Comp. Inactivated Japanese encephalitis Vaccine 100 μL — — Solution Transnasal 10 Ex. 25 virus equivalent Ex. 111 Inactivated Japanese encephalitis Vaccine 100 μL Argatroban 20 Solution Transnasal 10 virus equivalent Ex. 112 Inactivated Japanese encephalitis Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 virus equivalent Ex. 113 Inactivated Japanese encephalitis Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 virus equivalent Ex. 114 Inactivated Japanese encephalitis Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 virus equivalent Comp. Inactivated Japanese encephalitis Vaccine 100 μL — — Solution Sublingual 30 Ex. 26 virus equivalent Ex. 115 Inactivated Japanese encephalitis Vaccine 100 μL Argatroban 100 Solution Sublingual 30 virus equivalent Ex. 116 Inactivated Japanese encephalitis Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 virus equivalent Ex. 117 Inactivated Japanese encephalitis Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 virus equivalent Ex. 118 Inactivated Japanese encephalitis Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 virus equivalent Comp. Live attenuated mumps virus Vaccine 100 μL — — Solution Transnasal 10 Ex. 27 equivalent Ex. 119 Live attenuated mumps virus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 equivalent Ex. 120 Live attenuated mumps virus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 equivalent Ex. 121 Live attenuated mumps virus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 equivalent Ex. 122 Live attenuated mumps virus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 equivalent Comp. Live attenuated mumps virus Vaccine 100 μL — — Solution Sublingual 30 Ex. 28 equivalent Ex. 123 Live attenuated mumps virus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 equivalent Ex. 124 Live attenuated mumps virus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 equivalent Ex. 125 Live attenuated mumps virus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 equivalent Ex. 126 Live attenuated mumps virus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 equivalent Comp. Live attenuated measles virus Vaccine 100 μL — — Solution Transnasal 10 Ex. 29 equivalent Ex. 127 Live attenuated measles virus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 equivalent Ex. 128 Live attenuated measles virus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 equivalent Ex. 129 Live attenuated measles virus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 equivalent Ex. 130 Live attenuated measles virus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 equivalent Comp. Live attenuated measles virus Vaccine 100 μL — — Solution Sublingual 30 Ex. 30 equivalent Ex. 131 Live attenuated measles virus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 equivalent Ex. 132 Live attenuated measles virus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 equivalent Ex. 133 Live attenuated measles virus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 equivalent Ex. 134 Live attenuated measles virus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 equivalent

TABLE-US-00010 TABLE 10 Antigen Immunity induction promoter Amount Amount Dosage Administration Amount Name [μg] Name [μg] form route [μL] Comp. Live attenuated rubella virus Vaccine 100 μL — — Solution Transnasal 10 Ex. 31 equivalent Ex. 135 Live attenuated rubella virus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 equivalent Ex. 136 Live attenuated rubella virus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 equivalent Ex. 137 Live attenuated rubella virus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 equivalent Ex. 138 Live attenuated rubella virus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 equivalent Comp. Live attenuated rubella virus Vaccine 100 μL — — Solution Sublingual 30 Ex. 32 equivalent Ex. 139 Live attenuated rubella virus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 equivalent Ex. 140 Live attenuated rubella virus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 equivalent Ex. 141 Live attenuated rubella virus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 equivalent Ex. 142 Live attenuated rubella virus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 equivalent Comp. Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL — — Solution Transnasal 10 Ex. 33 influenzae type b polysaccharide equivalent Ex. 143 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 influenzae type b polysaccharide equivalent Ex. 144 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 influenzae type b polysaccharide equivalent Ex. 145 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 influenzae type b polysaccharide equivalent Ex. 146 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 influenzae type b polysaccharide equivalent Comp. Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL — — Solution Sublingual 30 Ex. 34 influenzae type b polysaccharide equivalent Ex. 147 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 influenzae type b polysaccharide equivalent Ex. 148 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 influenzae type b polysaccharide equivalent Ex. 149 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 influenzae type b polysaccharide equivalent Ex. 150 Tetanus toxoid-conjugated Haemophilus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 influenzae type b polysaccharide equivalent Comp. Recombinant HBs antigen protein Vaccine 100 μL — — Solution Transnasal 10 Ex. 35 equivalent Ex. 151 Recombinant HBs antigen protein Vaccine 100 μL Argatroban 20 Solution Transnasal 10 equivalent Ex. 152 Recombinant HBs antigen protein Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 equivalent Ex. 153 Recombinant HBs antigen protein Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 equivalent Ex. 154 Recombinant HBs antigen protein Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 equivalent Comp. Recombinant HBs antigen protein Vaccine 100 μL — — Solution Sublingual 30 Ex. 36 equivalent Ex. 155 Recombinant HBs antigen protein Vaccine 100 μL Argatroban 100 Solution Sublingual 30 equivalent Ex. 156 Recombinant HBs antigen protein Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 equivalent Ex. 157 Recombinant HBs antigen protein Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 equivalent Ex. 158 Recombinant HBs antigen protein Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 equivalent Comp. Live attenuated yellow fever virus Vaccine 100 μL — — Solution Transnasal 10 Ex. 37 equivalent Ex. 159 Live attenuated yellow fever virus Vaccine 100 μL Argatroban 20 Solution Transnasal 10 equivalent Ex. 160 Live attenuated yellow fever virus Vaccine 100 μL Nafamostat mesylate 20 Solution Transnasal 10 equivalent Ex. 161 Live attenuated yellow fever virus Vaccine 100 μL Camostat mesylate 20 Solution Transnasal 10 equivalent Ex. 162 Live attenuated yellow fever virus Vaccine 100 μL Tirofiban 20 Solution Transnasal 10 equivalent Comp. Live attenuated yellow fever virus Vaccine 100 μL — — Solution Sublingual 30 Ex. 38 equivalent Ex. 163 Live attenuated yellow fever virus Vaccine 100 μL Argatroban 100 Solution Sublingual 30 equivalent Ex. 164 Live attenuated yellow fever virus Vaccine 100 μL Nafamostat mesylate 100 Solution Sublingual 30 equivalent Ex. 165 Live attenuated yellow fever virus Vaccine 100 μL Camostat mesylate 100 Solution Sublingual 30 equivalent Ex. 166 Live attenuated yellow fever virus Vaccine 100 μL Tirofiban 100 Solution Sublingual 30 equivalent

TABLE-US-00011 TABLE 11 Antigen Immunity induction promoter Amount Amount Dosage Administration Amount Name [μg] Name [μg] form route [μL] Comp. Ex. 39 Tetanus toxoid Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 167 Tetanus toxoid Vaccine 100 μL equivalent Argatroban  20 Solution Transnasal 10 Ex. 168 Tetanus toxoid Vaccine 100 μL equivalent Nafamostat mesylate  20 Solution Transnasal 10 Ex. 169 Tetanus toxoid Vaccine 100 μL equivalent Camostat mesylate  20 Solution Transnasal 10 Ex. 170 Tetanus toxoid Vaccine 100 μL equivalent Tirofiban  20 Solution Transnasal 10 Comp. Ex. 40 Tetanus toxoid Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 171 Tetanus toxoid Vaccine 100 μL equivalent Argatroban 100 Solution Sublingual 30 Ex. 172 Tetanus toxoid Vaccine 100 μL equivalent Nafamostat mesylate 100 Solution Sublingual 30 Ex. 173 Tetanus toxoid Vaccine 100 μL equivalent Camostat mesylate 100 Solution Sublingual 30 Ex. 174 Tetanus toxoid Vaccine 100 μL equivalent Tirofiban 100 Solution Sublingual 30 Comp. Ex. 41 Live attenuated varicella virus Vaccine 100 μL equivalent — — Solution Transnasal 10 Ex. 175 Live attenuated varicella virus Vaccine 100 μL equivalent Argatroban  20 Solution Transnasal 10 Ex. 176 Live attenuated varicella virus Vaccine 100 μL equivalent Nafamostat mesylate  20 Solution Transnasal 10 Ex. 177 Live attenuated varicella virus Vaccine 100 μL equivalent Camostat mesylate  20 Solution Transnasal 10 Ex. 178 Live attenuated varicella virus Vaccine 100 μL equivalent Tirofiban  20 Solution Transnasal 10 Comp. Ex. 42 Live attenuated varicella virus Vaccine 100 μL equivalent — — Solution Sublingual 30 Ex. 179 Live attenuated varicella virus Vaccine 100 μL equivalent Argatroban 100 Solution Sublingual 30 Ex. 180 Live attenuated varicella virus Vaccine 100 μL equivalent Nafamostat mesylate 100 Solution Sublingual 30 Ex. 181 Live attenuated varicella virus Vaccine 100 μL equivalent Camostat mesylate 100 Solution Sublingual 30 Ex. 182 Live attenuated varicella virus Vaccine 100 μL equivalent Tirofiban 100 Solution Sublingual 30 Comp. Ex. 43 Live BCG Vaccine 30 μL equivalent — — Solution Transnasal 10 Ex. 183 Live BCG Vaccine 30 μL equivalent Argatroban  20 Solution Transnasal 10 Ex. 184 Live BCG Vaccine 30 μL equivalent Nafamostat mesylate  20 Solution Transnasal 10 Ex. 185 Live BCG Vaccine 30 μL equivalent Camostat mesylate  20 Solution Transnasal 10 Ex. 186 Live BCG Vaccine 30 μL equivalent Tirofiban  20 Solution Transnasal 10 Comp. Ex. 44 Live BCG Vaccine 30 μL equivalent — — Solution Sublingual 30 Ex. 187 Live BCG Vaccine 30 μL equivalent Argatroban 100 Solution Sublingual 30 Ex. 188 Live BCG Vaccine 30 μL equivalent Nafamostat mesylate 100 Solution Sublingual 30 Ex. 189 Live BCG Vaccine 30 μL equivalent Camostat mesylate 100 Solution Sublingual 30 Ex. 190 Live BCG Vaccine 30 μL equivalent Tirofiban 100 Solution Sublingual 30 Comp. Ex. 45 Inactivated rabies virus Vaccine 200 μL equivalent — — Solution Transnasal 10 Ex. 191 Inactivated rabies virus Vaccine 200 μL equivalent Argatroban  20 Solution Transnasal 10 Ex. 192 Inactivated rabies virus Vaccine 200 μL equivalent Nafamostat mesylate  20 Solution Transnasal 10 Ex. 193 Inactivated rabies virus Vaccine 200 μL equivalent Camostat mesylate  20 Solution Transnasal 10 Ex. 194 Inactivated rabies virus Vaccine 200 μL equivalent Tirofiban  20 Solution Transnasal 10 Comp. Ex. 46 Inactivated rabies virus Vaccine 200 μL equivalent — — Solution Sublingual 30 Ex. 195 Inactivated rabies virus Vaccine 200 μL equivalent Argatroban 100 Solution Sublingual 30 Ex. 196 Inactivated rabies virus Vaccine 200 μL equivalent Nafamostat mesylate 100 Solution Sublingual 30 Ex. 197 Inactivated rabies virus Vaccine 200 μL equivalent Camostat mesylate 100 Solution Sublingual 30 Ex. 198 Inactivated rabies virus Vaccine 200 μL equivalent Tirofiban 100 Solution Sublingual 30

Examples 199 to 203, Comparative Example 47

[0162] A cream for transdermal administration was prepared according to the formulation shown in Table 12 in the same manner as the cream for transdermal administration of Table 5. 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 the immunization using the minimally invasive administration, humoral immunity specific to the administered antigen can be induced.

TABLE-US-00012 TABLE 12 Antigen Immunity induction promoter Amount Amount Immunological Administration Dosage [Parts by [Parts by evaluation No. route form Name weight] Name weight] mouse Comp. Transdermal Cream OVA 5 — — C57BL6 Ex. 47 (minimally invasive) Ex. 199 Transdermal Cream OVA 5 Rivaroxaban 5 C57BL6 (minimally invasive) Ex. 200 Transdermal Cream OVA 5 Argatroban 5 C57BL6 (minimally invasive) Ex. 201 Transdermal Cream OVA 5 Nafamostat mesylate 5 C57BL6 (minimally invasive) Ex. 202 Transdermal Cream OVA 5 Camostat mesylate 5 C57BL6 (minimally invasive) Ex. 203 Transdermal Cream OVA 5 Tirofiban 5 C57BL6 (minimally invasive)

INDUSTRIAL APPLICABILITY

[0163] The humoral immunity induction-promoting composition and vaccine pharmaceutical composition of the present invention can universally be used for inducing humoral immunity to various antigens, exert a high antibody production inducing effect, and can be suitably used for transdermal administration or transmucosal administration.