VACCINE PHARMACEUTICAL COMPOSITION FOR CELL-MEDIATED IMMUNITY CONTAINING BISPHOSPHONATES

Abstract

The present invention aims to provide a vaccine pharmaceutical composition universally usable for induction of cellular immunity against various antigens and exerting a high cellular immunity inducing effect. The present invention relates to a vaccine pharmaceutical composition for inducing cellular immunity, containing: an antigen; and a first cellular immunity induction promoter that is a bisphosphonate.

Claims

1. A vaccine pharmaceutical composition for inducing cellular immunity, comprising: an antigen; and a first cellular immunity induction promoter that is a bisphosphonate.

2. The vaccine pharmaceutical composition according to claim 1, further comprising a second cellular immunity induction promoter that is a helper peptide.

3. The vaccine pharmaceutical composition according to claim 1, wherein the first cellular immunity induction promoter that is a bisphosphonate is at least one selected from the group consisting of etidronate, clodronate, tiludronate, pamidronate, alendronate, ibandronate, neridronate, zoledronate, risedronate, and minodronate.

4. The vaccine pharmaceutical composition according to claim 3, further comprising at least one of an antioxidant and an anti-inflammatory drug.

5. The vaccine pharmaceutical composition according to claim 4, wherein the vaccine pharmaceutical composition comprises at least one antioxidant selected from the group consisting of sodium nitrite, ascorbic acid, sodium hydrogen sulfite, cysteine hydrochloride, citric acid hydrate, dibutylhydroxytoluene, soybean lecithin, tocopherol, sodium pyrosulfite, dibutylhydroxyanisole, 1,3-butylene glycol, benzotriazole, propyl gallate, and 2-mercaptobenzimidazole.

6. The vaccine pharmaceutical composition according to claim 4, wherein the vaccine pharmaceutical composition comprises at least one anti-inflammatory drug selected from the group consisting of polyphenols, alkaloids, and phospholipase A2 inhibitors.

7. The vaccine pharmaceutical composition according to claim 4, wherein the vaccine pharmaceutical composition comprises at least one anti-inflammatory drug that is a cyclooxygenase inhibitor.

8. The vaccine pharmaceutical composition according to claim 7, wherein the cyclooxygenase inhibitor is at least one selected from the group consisting of cyclooxygenase non-selective inhibitors, cyclooxygenase-1 selective inhibitors, and cyclooxygenase-2 selective inhibitors.

9. The vaccine pharmaceutical composition according to claim 1, structured and arranged to be administered to a body surface.

10. A method for inducing cellular immunity comprising administering a vaccine pharmaceutical composition, comprising: an antigen; and a first cellular immunity induction promoter that is a bisphosphonate, wherein the administration induces a cellular immune response.

11. The method for inducing cellular immunity according to claim 10, wherein the vaccine pharmaceutical composition further comprises a second cellular immunity induction promoter that is a helper peptide.

12. The method for inducing cellular immunity according to claim 10, wherein the first cellular immunity induction promoter that is a bisphosphonate is at least one selected from the group consisting of etidronate, clodronate, tiludronate, pamidronate, alendronate, ibandronate, neridronate, zoledronate, risedronate, and minodronate.

13. The method for inducing cellular immunity according to claim 10, wherein the vaccine pharmaceutical composition comprises at least one of an antioxidant and an anti-inflammatory drug.

14. The method for inducing cellular immunity according to claim 13, wherein the vaccine pharmaceutical composition comprises at least one antioxidant selected from the group consisting of sodium nitrite, ascorbic acid, sodium hydrogen sulfite, cysteine hydrochloride, citric acid hydrate, dibutylhydroxytoluene, soybean lecithin, tocopherol, sodium pyrosulfite, dibutylhydroxyanisole, 1,3-butylene glycol, benzotriazole, propyl gallate, and 2-mercaptobenzimidazole.

15. The method for inducing cellular immunity according to claim 13, wherein the vaccine pharmaceutical composition comprises at least one anti-inflammatory drug selected from the group consisting of polyphenols, alkaloids, and phospholipase A2 inhibitors.

16. The method for inducing cellular immunity according to claim 13, wherein the vaccine pharmaceutical composition comprises at least one anti-inflammatory drug that is a cyclooxygenase inhibitor.

17. The method for inducing cellular immunity according to claim 16, wherein the cyclooxygenase inhibitor is at least one selected from the group consisting of cyclooxygenase non-selective inhibitors, cyclooxygenase-1 selective inhibitors, and cyclooxygenase-2 selective inhibitors.

18. The method for inducing cellular immunity according to claim 10, comprising administering the vaccine pharmaceutical composition to a body surface.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0169] FIG. 1 is a graph showing the cellular immunity inducing effect upon transdermal administration of creams for transdermal administration obtained in Examples 1 to 10 and Comparative Example 2.

DESCRIPTION OF EMBODIMENTS

[0170] The present invention is described specifically in detail in the following with reference to, but not limited to, examples.

Examples 1 to 64, Comparative Examples 1 to 51

(Preparation of Creams for Transdermal Administration)

[0171] Creams for transdermal administration having a composition as shown in Tables 1 to 4 and 6 were prepared. Specifically, an antigen peptide, a first cellular immunity induction promoter that is a bisphosphonate, and if needed, a second cellular immunity induction promoter that is a helper peptide, an antioxidant, and an anti-inflammatory drug (including COX inhibitor) mentioned below, and 15% by weight of dimethyl sulfoxide (DMSO) were mixed in amounts specified in Tables 1 to 4 and 6. To the resulting mixture, a base (base cream) was added to obtain a total weight of 100% by weight, and mixed to give a cream for transdermal administration. The base cream used was prepared by mixing materials shown in Table 5 in amounts as specified. White Vaseline, sorbitan monostearate, isostearic acid, benzyl alcohol, stearyl alcohol, polysorbate 60, concentrated glycerin, and dimethyl sulfoxide (DMSO) were purchased from Wako Pure Chemical Industries, Ltd. Cetanol was purchased from Tokyo Chemical Industry Co., Ltd.

[0172] A PET film/PET nonwoven fabric laminate (area: 0.7 cm.sup.2) was attached to the center of an adhesive tape for fixing in such a manner that the PET film is in contact with the tape, thereby preparing a complex base. To a nonwoven fabric portion of the obtained complex base, 4 mg of the cream for transdermal administration was applied. This was used as an administration sample in an immunity test.

(First Cellular Immunity Induction Promoter that is a Bisphosphonate)

[0173] Etidronate (LKT Labolatories, Inc.)

[0174] Clodronate (LKT Labolatories, Inc.)

[0175] Tiludronate (Sigma-Aldrich Co. LLC)

[0176] Pamidronate (Tokyo Chemical Industry Co., Ltd.)

[0177] Alendronate (Medichem)

[0178] Ibandronate (URQUIMA S.A.)

[0179] Neridronate (Sigma-Aldrich Co. LLC)

[0180] Zoledronate (Konan Chemical Industry co., ltd.)

[0181] Risedronate (Propharma S.A.)

[0182] Minodronate (Ava Chem Scientific)

(Antioxidant)

[0183] Sodium nitrite (Wako Pure Chemical Industries, Ltd.)

[0184] Ascorbic acid (Wako Pure Chemical Industries, Ltd.)

[0185] Sodium hydrogen sulfite (Wako Pure Chemical Industries, Ltd.)

[0186] Cysteine hydrochloride (Kyowa Hakko Bio Co., Ltd.)

[0187] Citric acid hydrate (Komatsuya Corporation)

[0188] BHT (dibutylhydroxytoluene, Wako Pure Chemical Industries, Ltd.)

[0189] Soybean lecithin (Wako Pure Chemical Industries, Ltd.)

[0190] Tocopherol (Kanto Chemical Co., Inc.)

[0191] Sodium pyrosulfite (Wako Pure Chemical Industries, Ltd.)

[0192] BHA (dibutylhydroxyanisole, Kanto Chemical Co., Inc.)

[0193] 1,3-butylene glycol (1,3-butanediol, Wako Pure Chemical Industries, Ltd.)

[0194] Benzotriazole (Wako Pure Chemical Industries, Ltd.)

[0195] Propyl gallate (DSP Gokyo Food & Chemical Co., Ltd.)

[0196] 2-MBI (2-mercaptobenzimidazole, Wako Pure Chemical Industries, Ltd.)

(Anti-Inflammatory Drug)

[0197] Naringenin (polyphenol, LKT Labolatories, Inc.)

[0198] Epicatechin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0199] Apigenin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0200] Chrysin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0201] Myricetin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0202] Rutin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0203] Genistein (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0204] Nobiletin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0205] Curcumin (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0206] Resveratrol (polyphenol, Wako Pure Chemical Industries, Ltd.)

[0207] Coumarin (alkaloid, Wako Pure Chemical Industries, Ltd.)

[0208] Berberine (berberine chloride n-hydrate, alkaloid, Wako Pure Chemical Industries, Ltd.)

[0209] Glycyrrhetinic acid (phospholipase A2 inhibitor, Wako Pure Chemical Industries, Ltd.)

[0210] (COX inhibitor (anti-inflammatory drug))

[0211] Etodolac (Wako Pure Chemical Industries, Ltd.)

[0212] Loxoprofen (loxoprofen Na, Yoshindo Inc.)

[0213] Diclofenac (diclofenac sodium, Wako Pure Chemical Industries, Ltd.)

[0214] Celecoxib (Sigma-Aldrich Co., LLC)

[0215] Valdecoxib (Sigma-Aldrich Co., LLC)

[0216] Piroxicam (Sigma-Aldrich Co., LLC)

[0217] Aspirin (acetylsalicylic acid, Wako Pure Chemical Industries, Ltd.)

[0218] Indomethacin (Wako Pure Chemical Industries, Ltd.)

[0219] Ketoprofen (Wako Pure Chemical Industries, Ltd.)

[0220] Ibuprofen (Wako Pure Chemical Industries, Ltd.)

[0221] Naproxen (Wako Pure Chemical Industries, Ltd.)

(Antigen Peptide)

[0222] OVAp (OVA peptide, 8-amino acid peptide having the sequence Ser Ile Ile Asn Phe Glu Lys Leu (SEQ ID No: 16))

[0223] Survivin-2B (survivin 2B peptide, cancer antigen peptide)

[0224] GPC3 (GPC3 peptide, cancer antigen peptide)

[0225] HER2/neu_A24 (HER2/neu_A24 peptide, cancer antigen peptide)

[0226] MAGE-A3_A24 (MAGE3_A24 peptide, cancer antigen peptide)

[0227] IPEP87 (IPEP87 peptide, infectious pathogen antigen)

[0228] PR1 (PR1 peptide, cancer antigen peptide)

[0229] HER2_A02 (HER2/neu_A02 peptide, cancer antigen peptide)

[0230] MAGE-A3_A02 (MAGE3_A02 peptide, cancer antigen peptide)

[0231] HBVenv (HBVenv peptide, infectious pathogen antigen)

[0232] HER2/neu_E75 (HER2/neu_E75 peptide, cancer antigen peptide)

[0233] MUC1 (MUC1 peptide, cancer antigen peptide)

(Helper Peptide)

[0234] Peptide-25 (Pep25)

[0235] Peptide-25B (Pep25B)

[0236] PADRE

<Evaluation 1>

[0237] The creams for transdermal administration obtained in the examples and comparative examples were evaluated as follows.

(Evaluation of Cellular Immunity Inducing Effect)

[0238] According to the procedure described below, the cream for transdermal administration was used to carry out a mouse immunity test using an animal model for immunological evaluation. Subsequently, the level of induction of antigen-specific cellular immunity was evaluated by ELISPOT assay.

(1) Animal Model for Immunological Evaluation

[0239] The “animal model for immunological evaluation” herein refers to an animal model for evaluating the immunity inducing properties of a vaccine pharmaceutical composition (in the present case, a cream for transdermal administration), and specifically refers to an animal model for evaluating the level of the cellular immunity induced by the cream for transdermal administration.

[0240] In consideration of the compatibility between the antigen in the cream for transdermal administration and MHC class I molecules of the animal, the animal model used for immunological evaluation was an animal with which induction of the cellular immunity by the antigen in the cream for transdermal administration can be evaluated.

[0241] Specifically, in a case where the antigen was a HLA-A*24-type MHC class 1 restriction peptide, the animal used for the evaluation was a BALB/c mouse. In a case where the antigen was a HLA-A*02-type MHC restriction peptide, the animal used was a genetically altered mouse with which induction of the cellular immunity by the HLA-A*02-type MHC restriction peptide can be evaluated. In a case where the antigen was another HLA-type MHC restriction peptide, the animal used was an animal with which induction of the cellular immunity by that HLA-type MHC restriction peptide can be evaluated. In the case of a protein antigen, the animal used was an animal having a MHC compatible with a class 1 epitope corresponding to the cellular immunity intended to be induced among class 1 epitopes contained in the amino acid sequence of the protein antigen.

(2) Mouse Immunity Test of Creams for Transdermal Administration

(Examples 1 to 64, Comparative Examples 1 to 51)

[0242] According to Tables 1 to 4 and 6, a mouse was prepared and its back was shaved. After a certain rearing period for recovery from skin damage caused by the shaving, 4 mg of the cream for transdermal administration was administered to the skin of the back for 24 hours and then removed the cream therefrom. The mouse was reared for six days. Six days after the administration, the spleen was extracted, and a spleen cell suspension was prepared. Spleen cells (5×10.sup.5 cells/well) and an antigen peptide (100 μM) together with a culture fluid were poured into wells of an ELISPOT plate on which an anti-mouse IFN-γ antibody was immobilized, and co-cultured under the culture conditions of 37° C. and 5% CO.sub.2 for 20 hours. The number of IFN-γ-producing cell spots was evaluated by the ELISPOT assay. Tables 1 to 4 and 6 show the number of IFN-γ-producing cell spots as the “immunity result”. FIG. 1 shows the immunity results of Comparative Example 2 (w/o Adjuvant) and Examples 1 to 10.

TABLE-US-00001 TABLE 1 Helper Immuno- Antigen Bisphosphonate peptide logical Dosage Amount Amount Amount evaluation Immunity form Name [wt %] Name [wt %] Name [wt %] mouse result Comparative Cream OVAp 2.5 — — Pep25 — C57BL/6 2.5 Example 1 Comparative Cream OVAp 2.5 — — Pep25 1 C57BL/6 7.0 Example 2 Example 1 Cream OVAp 2.5 Etidronate 2 Pep25 1 C57BL/6 33.0 Example 2 Cream OVAp 2.5 Clodronate 2 Pep25 1 C57BL/6 30.8 Example 3 Cream OVAp 2.5 Tiludronate 2 Pep25 1 C57BL/6 37.5 Example 4 Cream OVAp 2.5 Pamidronate 1 Pep25 1 C57BL/6 78.8 Example 5 Cream OVAp 2.5 Alendronate 1 Pep25 1 C57BL/6 135.5 Example 6 Cream OVAp 2.5 Ibandronate 1 Pep25 1 C57BL/6 124.3 Example 7 Cream OVAp 2.5 Neridronate 1 Pep25 1 C57BL/6 171.3 Example 8 Cream OVAp 2.5 Zoledronate 0.5 Pep25 1 C57BL/6 130.5 Example 9 Cream OVAp 2.5 Risedronate 0.5 Pep25 1 C57BL/6 158.0 Example 10 Cream OVAp 2.5 Minodronate 0.5 Pep25 1 C57BL/6 198.8 Example 11 Cream OVAp 2.5 Alendronate 1 — — C57BL/6 64.2 Example 12 Cream OVAp 2.5 Neridronate 1 — — C57BL/6 82.4 Example 13 Cream OVAp 2.5 Zoledronate 0.5 — — C57BL/6 75.6 Example 14 Cream OVAp 2.5 Risedronate 0.5 — — C57BL/6 90.6 Example 15 Cream OVAp 2.5 Minodronate 0.5 — — C57BL/6 97.8

TABLE-US-00002 TABLE 2 Immuno- Antigen Bisphosphonate Antioxidant Helper peptide logical Dosage Amount Amount Amount Amount evaluation Immunity form Name [wt %] Name [wt %] Name [wt %] Name [wt %] mouse result Comparative Cream OVAp 2.5 — — Sodium nitrite 1 Pep25 1 C57BL/6 6.5 Example 3 Comparative Cream OVAp 2.5 — — Ascorbic acid 1 Pep25 1 C57BL/6 6.5 Example 4 Comparative Cream OVAp 2.5 — — Sodium hydrogen 1 Pep25 1 C57BL/6 7.0 Example 5 sulfite Comparative Cream OVAp 2.5 — — Cysteine hydrochloride 1 Pep25 1 C57BL/6 5.8 Example 6 Comparative Cream OVAp 2.5 — — Citric acid hydrate 1 Pep25 1 C57BL/6 7.3 Example 7 Comparative Cream OVAp 2.5 — — BHT 1 Pep25 1 C57BL/6 6.0 Example 8 Comparative Cream OVAp 2.5 — — Soybean lecithin 1 Pep25 1 C57BL/6 7.8 Example 9 Comparative Cream OVAp 2.5 — — Tocopherol 1 Pep25 1 C57BL/6 6.3 Example 10 Comparative Cream OVAp 2.5 — — Sodium pyrosulfite 1 Pep25 1 C57BL/6 5.3 Example 11 Comparative Cream OVAp 2.5 — — BHA 1 Pep25 1 C57BL/6 6.3 Example 12 Comparative Cream OVAp 2.5 — — 1,3-butylene glycol 1 Pep25 1 C57BL/6 6.0 Example 13 Comparative Cream OVAp 2.5 — — Benzotriazole 1 Pep25 1 C57BL/6 5.5 Example 14 Comparative Cream OVAp 2.5 — — Propyl gallate 1 Pep25 1 C57BL/6 7.0 Example 15 Comparative Cream OVAp 2.5 — — 2-MBI 1 Pep25 1 C57BL/6 15.0 Example 16 Example 16 Cream OVAp 2.5 Zoledronate 0.5 Sodium nitrite 0.5 Pep25 1 C57BL/6 201.3 Example 17 Cream OVAp 2.5 Zoledronate 0.5 Ascorbic acid 0.5 Pep25 1 C57BL/6 175.0 Example 18 Cream OVAp 2.5 Zoledronate 0.5 Sodium hydrogen sulfite 0.5 Pep25 1 C57BL/6 189.8 Example 19 Cream OVAp 2.5 Zoledronate 0.5 Cysteine hydrochloride 0.5 Pep25 1 C57BL/6 166.3 Example 20 Cream OVAp 2.5 Zoledronate 0.5 Citric acid hydrate 0.5 Pep25 1 C57BL/6 163.3 Example 21 Cream OVAp 2.5 Zoledronate 0.5 BHT 0.5 Pep25 1 C57BL/6 201.3 Example 22 Cream OVAp 2.5 Zoledronate 0.5 Soybean lecithin 0.5 Pep25 1 C57BL/6 234.3 Example 23 Cream OVAp 2.5 Zoledronate 0.5 Tocopherol 0.5 Pep25 1 C57BL/6 160.5 Example 24 Cream OVAp 2.5 Zoledronate 0.5 Sodium pyrosulfite 0.5 Pep25 1 C57BL/6 208.3 Example 25 Cream OVAp 2.5 Zoledronate 0.5 BHA 0.5 Pep25 1 C57BL/6 285.3 Example 26 Cream OVAp 2.5 Zoledronate 0.5 1,3-butylene glycol 0.5 Pep25 1 C57BL/6 168.0 Example 27 Cream OVAp 2.5 Zoledronate 0.5 Benzatriazole 0.5 Pep25 1 C57BL/6 161.0 Example 28 Cream OVAp 2.5 Zoledronate 0.5 Propyl gallate 0.5 Pep25 1 C57BL/6 176.3 Example 29 Cream OVAp 2.5 Zoledronate 0.5 2-MBI 0.5 Pep25 1 C57BL/6 229.3

TABLE-US-00003 TABLE 3 Anti-inflammatory Immuno- Antigen Bisphosphonate drug Helper peptide logical Dosage Amount Amount Amount Amount evaluation Immunity form Name [wt %] Name [wt %] Name [wt %] Name [wt %] mouse result Comparative Cream OVAp 2.5 — — Naringenin 5 Pep25 1 C57BL/6 8.5 Example 17 Comparative Cream OVAp 2.5 — — Epicatechin 5 Pep25 I C57BL/6 5.0 Example 18 Comparative Cream OVAp 2.5 — — Apigenin 5 Pep25 C57BL/6 7.8 Example 19 Comparative Cream OVAp 2.5 — — Chrysin 5 Pep25 I C57BL/6 7.0 Example 20 Comparative Cream OVAp 2.5 — — Myricetin 5 Pep25 1 C57BL/6 7.8 Example 21 Comparative Cream OVAp 2.5 — — Rutin 5 Pep25 I C57BL/6 5.8 Example 22 Comparative Cream OVAp 2.5 — — Genistein 5 Pep25 1 C57BL/6 4.5 Example 23 Comparative Cream OVAp 2.5 — — Nobiletin 5 Pep25 1 C57BL/6 4.8 Example 24 Comparative Cream OVAp 2.5 — — Curcumin 5 Pep25 I C57BL/6 10.0 Example 25 Comparative Cream OVAp 2.5 — — Resveratrol 5 Pep25 1 C57BL/6 8.0 Example 26 Comparative Cream OVAp 2.5 — — Coumarin 5 Pep25 1 C57BL/6 8.0 Example 27 Comparative Cream OVAp 2.5 — — Berberine 5 Pep25 1 C57BL/6 11.3 Example 28 Comaprative Cream OVAp 2.5 — — Glycyrrhetinic 5 Pep25 1 C57BL/6 11.3 Example 29 acid Example 30 Cream OVAp 2.5 Zoledronate 0.5 Naringenin 4.5 Pep25 1 C57BL/6 195.5 Example 31 Cream OVAp 2.5 Zoledronate 0.5 Epicatechin 4.5 Pep25 1 C57BL/6 220.8 Example 32 Cream OVAp 2.5 Zoledronate 0.5 Apigenin 4.5 Pep25 1 C57BL/6 160.5 Example 33 Cream OVAp 2.5 Zoledronate 0.5 Chrysin 4.5 Pep25 1 C57BL/6 238.8 Example 34 Cream OVAp 2.5 Zoledronate 0.5 Myricetin 4.5 Pep25 1 C57BL/6 243.0 Example 35 Cream OVAp 2,5 Zoledronate 0.5 Rutin 4.5 Pep25 1 C57BL/6 198.5 Example 36 Cream OVAp 2:5 Zoledronate 0.5 Genistein 4.5 Pep25 1 C57BL/6 162.8 Example 37 Cream OVAp 2,5 Zoledronate 0.5 Nobiletin 4.5 Pep25 1 C57BL/6 185.8 Example 38 Cream OVAp 2.5 Zoledronate 0.5 Curcumin 4.5 Pep25 1 C57BL/6 163.0 Example 39 Cream OVAp 2.5 Zoledronate 0.5 Resveratrol 4.5 Pep25 1 C57BL/6 256.3 Example 40 Cream OVAp 2.5 Zoledronate 0.5 Coumarin 4.5 Pep25 1 C57BL/6 270.3 Example 41 Cream OVAp 2.5 Zoledronate 0.5 Berberine 4.5 Pep25 1 C57BL/6 191.5 Example 42 Cream OVAp 2.5 Zoledronate 0.5 Glycyrrhetinic 4.5 Pep25 1 C57BL/6 279.5 acid

TABLE-US-00004 TABLE 4 COX inhibitor (anti- Helper Immuno- Antigen Bisphosphonate inflammatory drug) peptide logical Dosage Amount Amount Amount Amount evaluation Immunity Form Name [wt %] Name [wt %] Name [wt %] Name [wt %] mouse result Comparative Cream OVAp 2.5 — — Etodolac 5 Pep25 1 C57BL/6 26.8 Example 30 Comparative Cream OVAp 2.5 — — Loxoprofen 5 Pep25 1 C57BL/6 26.3 Example 31 Comparative Cream OVAp 2.5 — — Diclofenac 5 Pep25 1 C57BL/6 24.8 Example 32 Comparative Cream OVAp 2.5 — — Celecoxib 5 Pep25 1 C57BL/6 28.5 Example 33 Comparative Cream OVAp 2.5 — — Valdepoxib 5 Pep25 1 C57BL/6 30.5 Example 34 Comparative Cream OVAp 2.5 — — Piroxicam 5 Pep25 1 C57BL/6 14.3 Example 35 Comparative Cream OVAp 2.5 — — Aspirin 5 Pep25 1 C57BL/6 6.5 Example 36 Comparative Cream OVAp 2.5 — — Indomethacin 5 Pep25 1 C57BL/6 15.5 Example 37 Comparative Cream OVAp 2.5 — — Ketoprofen 5 Pep25 1 C57BL/6 18.0 Example 38 Comparative Cream OVAp 2.5 — — Ibuprofen Pep25 1 C57BL/6 11.8 Example 39 Comaprative Cream OVAp 2.5 — — Naproxen 5 Pep25 1 C57BL/6 16.5 Example 40 Example 43 Cream OVAp 2.5 Zoledronate 0.5 Etodolac 4.5 Pep25 1 C57BL/6 195.3 Example 44 Cream OVAp 2.5 Zoledronate 0.5 Loxoprofen 4.5 Pep25 1 C57BL/6 243.3 Example 45 Cream OVAp 2.5 Zoledronate 0.5 Diclofenac 4.5 Pep25 1 C57BL/6 227.3 Example 46 Cream OVAp 2.5 Zoledronate 0.5 Celecoxib 4.5 Pep25 1 C57BL/6 231.0 Example 47 Cream OVAp 2.5 Zoledronate 0.5 Valdecoxib 4.5 Pep25 1 C57BL/6 242.8 Example 48 Cream OVAp 2.5 Zoledronate 0.5 Piroxicam 4.5 Pep25 1 C57BL/6 322.3 Example 49 Cream OVAp 2.5 Zoledronate 0.5 Aspirin 4.5 Pep25 1 C57BL/6 428.3 Example 50 Cream OVAp 2.5 Zoledronate 0.5 Indomethacin 4.5 Pep25 1 C57BL/6 325.0 Example 51 Cream OVAp 2.5 Zoledronate 0.5 Ketoprofen 4.5 Pep25 1 C57BL/6 204.5 Example 52 Cream OVAp 2.5 Zoledronate 0.5 Ibuprofen 4.5 Pep25 1 C57BL/6 222.3 Example 53 Cream OVAp 2.5 Zoledronate 0.5 Naproxen 4.5 Pep25 1 C57BL/6 294.8

TABLE-US-00005 TABLE 5 Amount [parts by Additive weight] Base White Vaseline 60.7 cream 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

TABLE-US-00006 TABLE 6 Immuno- Antigen Bisphosphonate Helper peptide logical Immunity result Dosage Amount Amount Amount evaluation Average [cells/ form Name [wt %] Name [wt %] Name [wt %] mouse value well] Comparative Cream Survivin-2B 10 — — Pep25B 1 BALB/c 19.3 1 × 10.sup.6 Example 41 Example 54 Cream Survivin-2B 10 Zoledronate 0.5 Pep25B 1 BALB/c 188.8 Comparative Cream GPC3 20 — — Pep25B 1 BALB/c 1.5 3 × 10.sup.6 Example 42 Example 55 Cream GPC3 20 Zoledronate 0.5 Pep25B 1 BALB/c 19.0 Comparative Cream HER2/neu_A24 5 — — Pep25B 1 BALB/c 31.0 1 × 10.sup.6 Example 43 Example 56 Cream HER2/neu_A24 5 Zoledronate 0.5 Pep25B 1 BALB/c 236.8 Comparative Cream MAGE-A3_A24 10 — — Pep25B 1 BALB/c 19.0 1 × 10.sup.6 Example 44 Example 57 Cream MAGE-A3_A24 10 Zoledronate 0.5 Pep25B 1 BALB/c 215.3 Comparative Cream IPEP87 10 — — PADRE 1 Gene-altered 25.8 1 × 10.sup.6 Example 45 Example 58 Cream IPEP87 10 Zoledronate 0.5 PADRE 1 Gene-altered 291.5 Comparative Cream PR1 10 — — PADRE 1 Gene-altered 15.3 1 × 10.sup.6 Example 46 Example 59 Cream PR1 10 Zoledronate 0.5 PADRE 1 Gene-altered 186.3 Comparative Cream HER2_A02 10 — — PADRE 1 Gene-altered 13.8 2 × 10.sup.6 Example 47 Example 60 Cream HER2_A02 10 Zoledronate 0.5 PADRE 1 Gene-altered 186.0 Comparative Cream MAGE-A3_A02 10 — — PADRE 1 Gene-altered 18.0 1 × 10.sup.6 Example 48 Example 61 Cream MAGE-A3_A02 10 Zoledronate 0.5 PADRE 1 Gene-altered 214.5 Comparative Cream HBVenv 20 — — PADRE 1 Gene-altered 9.5 2 × 10.sup.6 Example 49 Example 62 Cream HBVenv 20 Zoledronate 0.5 PADRE 1 Gene-altered 120.8 Comparative Cream HER2/neu_E75 10 — — PADRE 1 Gene-altered 14.5 2 × 10.sup.6 Example 50 Example 63 Cream HER2/neu_E75 10 Zoledronate 0.5 PADRE 1 Gene-altered 188.3 Comparative Cream MUC1 20 — — PADRE 1 Gene-altered 0.8 3 × 10.sup.6 Example 51 Example 64 Cream MUC1 20 Zoledronate 0.5 PADRE 1 Gene-altered 18.3

(3) Mouse Immunity Test of Tapes for Transdermal Administration

Examples 65 to 76, Comparative Examples 52 to 57

(Preparation of Tapes for Transdermal Administration)

[0243] A tape for transdermal administration having a composition shown in Table 7 was prepared. Specifically, an antigen peptide, a first cellular immunity induction promoter that is a bisphosphonate, and if necessary, a second cellular immunity induction promoter that is a helper peptide mentioned above were blended. To the mixture, an adhesive and an organic solvent (ethyl acetate when the adhesive is an acrylic, toluene when the adhesive is PIB) shown in Table 7 were added to obtain the total amount of the components and the adhesive after drying of the organic solvent of 100% by weight, and mixed to prepare an adhesive solution. The obtained adhesive solution was casted on a release liner to the thickness after drying of about 80 μm. The organic solvent was removed by drying, thereby forming an adhesive layer. The release liner used was a polyethylene terephthalate (PET) liner (thickness: 75 μm) subjected to silicon release treatment. The resulting adhesive layer was attached to a support, thereby preparing a tape for transdermal administration. The support used was a polyethylene terephthalate (PET) film (thickness: 25 μm).

[0244] The tape for transdermal administration was cut to give a piece with an area of 0.7 cm.sup.2, and the piece was used as an administration sample in the immunity test. Upon administration, the release liner was removed.

(Adhesive)

[0245] Acrylic adhesive (an acrylic adhesive solution prepared by solution-polymerizing 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 inert gas atmosphere)

[0246] PIB adhesive (PIB adhesive solution prepared by dissolving 24 parts of polyisobutylene (Oppanol B200, Basf SE), 36 parts of polyisobutylene (Oppanol B12, Basf SE), and 40 parts of an alicyclic petroleum resin (ARKON P-100, Arakawa Chemical Industries, LTD.) in toluene)

<Evaluation 2>

[0247] The tapes for transdermal administration obtained in the examples and comparative examples were evaluated as follows.

(Evaluation on Cellular Immunity Inducing Effect)

[0248] The level of inducing the antigen-specific cellular immunity was evaluated in the same manner as in the evaluation of the creams for transdermal administration. Table 7 shows the results as the “Immunity result”.

TABLE-US-00007 TABLE 7 Immuno- Antigen Bisphosphonate Helper peptide logical Immunity result Dosage Amount Amount Amount evaluation Average [cells/ form Adhesive Name [wt %] Name [wt %] Name [wt %] mouse value well] Comparative Tape Acrylic HER2/neu_E75 10 — — PADRE 1 Gene-altered 4.3 2 × 10.sup.6 Example 52 Example 65 Tape Acrylic HER2/neu_E75 10 Zoledronate 1 — — Gene-altered 23.5 Example 66 Tape Acrylic HER2/nerr_E75 10 Zoledronate 1 PADRE 1 Gene-altered 49.8 Comparative Tape PIB HER2/neu_E75 10 — — PADRE 1 Gene-altered 5.3 2 × 10.sup.6 Example 53 Example 67 Tape PIB HER2/neu_E75 10 Zoledronate 1 — — Gene-altered 22.8 Example 68 Tape PIB HER2/neu_E75 10 Zoledronate 1 PADRE 1 Gene-altered 52.0 Comparative Tape Acrylic IPEP87 10 — — PADRE 1 Gene-altered 5.8 2 × 10.sup.6 Example 54 Example 69 Tape Acrylic IPEP87 10 Zoledronate 1 — — Gene-altered 23.5 Example 70 Tape Acrylic IPEP87 10 Zoledronate 1 PADRE 1 Gene-altered 53.5 Comparative Tape PIB IPEP87 10 — — PADRE 1 Gene-altered 8.5 2 × 10.sup.6 Example 55 Example 71 Tape PIB IPEP87 10 Zoledronate 1 — — Gene-altered 33.3 Example 72 Tape PIB IPEP87 10 Zoledronate 1 PADRE 1 Gene-altered 69.8 Comparative Tape Acrylic MAGE-A3_A02 10 — — PADRE 1 Gene-altered 10.5 2 × 10.sup.6 Example 56 Example 73 Tape Acrylic MAGE-A3_A02 10 Zoledronate 1 — — Gene--altered 32.8 Example 74 Tape Acrylic MAGE-A3_A02 10 Zoledronate 1 PADRE 1 Gene-altered 68.0 Comparative Tape PIB MAGE-A3_A02 10 — — PADRE 1 Gene-altered 9.5 2 × 10.sup.6 Example 57 Example 75 Tape PIB MAGE-A3_A02 10 Zoledronate 1 — — Gene-altered 36.3 Example 76 Tape PIB MAGE-A3_A02 10 Zoledronate 1 PADRE 1 Gene-altered 73.5

(4) Mouse Immunity Test of Creams for Transdermal Administration (Minimally Invasive Administration)

Examples 77 to 91, Comparative Examples 58 to 59

[0249] Creams for transdermal administration having a composition shown in Table 8 were prepared in the same manner as in the case of the creams for transdermal administration shown in Table 1. A mouse was prepared and its right back was shaved. Corneum exfoliation treatment was performed thereon five times using an OPP tape (EZDunplon No. 3301EZ) produced by Nitto Denko Corporation. The cream was administered to the treated skin (minimally invasive administration). Twenty-four hours later, the cream for transdermal administration was removed, and the mouse was reared for six days. Six days after the administration, the spleen was extracted, and antigen-specific IFN-γ-producing cells were analyzed by the ELISPOT assay.

[0250] Also by an immunization method utilizing minimally invasive administration as shown in Table 8, cellular immunity specific to the administered antigen can be induced.

TABLE-US-00008 TABLE 8 Antigen Bisphosphonate Helper peptide Immunological Dosage Amount Amount Amount evaluation form Name [wt %] Name [wt %] Name [wt %] mouse Comparative Cream OVAp 2.5 — — Pep25 — C57BL/6 Example 58 Comparative Cream OVAp 2.5 — — Pep25 1 C57BL/6 Example 59 Example 77 Cream OVAp 2.5 Etidronate 2 Pep25 1 C57BL/6 Example 78 Cream OVAp 2.5 Clodronate 2 Pep25 1 C57BL/6 Example 79 Cream OVAp 2.5 Tiludronate 2 Pep25 1 C57BL/6 Example 80 Cream OVAp 2.5 Pamidronate 1 Pep25 1 C57BL/6 Example 81 Cream OVAp 2.5 Alendronate 1 Pep25 1 C57BL/6 Example 82 Cream OVAp 2.5 Ibandronate 1 Pep25 1 C57BL/6 Example 83 Cream OVAp 2.5 Neridronate 1 Pep25 1 C57BL/6 Example 84 Cream OVAp 2.5 Zoledronate 0.5 Pep25 1 C57BL/6 Example 85 Cream OVAp 2.5 Risedronate 0.5 Pep25 1 C57BL/6 Example 86 Cream OVAp 2.5 Minodronate 0.5 Pep25 1 C57BL/6 Example 87 Cream OVAp 2.5 Alendronate 1 — — C57BL/6 Example 88 Cream OVAp 2.5 Neridronate 1 — — C57BL/6 Example 89 Cream OVAp 2.5 Zoledronate 0.5 — — C57BL/6 Example 90 Cream OVAp 2.5 Risedronate 0.5 — — C57BL/6 Example 91 Cream OVAp 2.5 Minodronate 0.5 — — C57BL/6

INDUSTRIAL APPLICABILITY

[0251] The vaccine pharmaceutical composition of the present invention is universally usable for induction of cellular immunity against various antigens, exerts a high cellular immunity inducing effect, and is suitably used for transdermal administration or transmucosal administration.