IP Library › Granted Patent US 12,589,148
Granted Patent B2
US 12,589,148 · App. 17/280,605 · Granted Mar 31, 2026

Immunity-inducing agent comprising antigen peptide-adjuvant nucleotide conjugate and pharmaceutical composition comprising same

Inventors: Shinichi Mochizuki (Fukuoka, JP); Makoto Koizumi (Tokyo, JP); Koji Morita (Tokyo, JP)
Assignees: The University of Kitakyushu; Daiichi Sankyo Company, Limited
A61K39/39A61K47/548A61K47/549A61K2039/55561
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,589,148
App. No.
17/280,605
Granted
Mar 31, 2026
Kind
B2
Abstract

The present invention provides an immunity-inducing agent comprising, as an active component, a polynucleotide/peptide conjugate in which a single-chain polynucleotide or polynucleotide derivative comprising a CpG motif, and an antigenic peptide are bound via a spacer, wherein the spacer is covalently bound at one end thereof to the polynucleotide or polynucleotide derivative and covalently bound at the other end thereof to the antigenic peptide, as well as a pharmaceutical composition comprising said immunity-inducing agent.

Claims (70)

1 . An immunity-inducing agent comprising, as an active component, a conjugate comprising (i) a single-chain polynucleotide or a single-chain polynucleotide derivative comprising a CpG motif, and (ii) an antigenic peptide bound thereto via a spacer, wherein the spacer is covalently bound at one end thereof to the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif and covalently bound at the other end thereof to the antigenic peptide,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is not complexed with β-1,3-glucan,

wherein the antigenic peptide and the spacer are bound together via a disulfide bond produced by a reaction between a thiol group of a cysteine residue at the N-terminus of the antigenic peptide and a thiol group of the spacer, and

wherein the spacer comprises a repeating unit represented by the formula:

wherein

X represents an oxygen atom or a sulfur atom, wherein each X may be the same or different,

R represents any of (CH 2 ) p O, (CH 2 ) q NH, and (CH 2 CH 2 O) m , wherein m, p and q each independently represent a natural number of not more than 10, and

n represents a natural number of not more than 10.

2 . The immunity-inducing agent according to claim 1 , wherein the antigenic peptide has an amino acid length of not less than 5 but not more than 30.

3 . The immunity-inducing agent according to claim 1 , wherein the antigenic peptide has an amino acid length of not less than 8 but not more than 11.

4 . The immunity-inducing agent according to claim 1 , wherein the single-chain polynucleotide or the single-chain polynucleotide derivative is a polydeoxyribonucleotide (DNA) or a DNA derivative comprising two or more CpG motifs.

5 . The immunity-inducing agent according to claim 1 , wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif has a nucleotide length of not less than 15 but not more than 40.

6 . The immunity-inducing agent according to claim 1 , wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif has a nucleotide length of not less than 20 but not more than 30.

7 . The immunity-inducing agent according to claim 1 , wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is a polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds.

8 . The immunity-inducing agent according to claim 7 , wherein, in the polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds, not less than 50% of the phosphodiester bonds are substituted with phosphorothioate bonds.

9 . The immunity-inducing agent according to claim 7 , wherein, in the polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds, not less than 90% of the phosphodiester bonds are substituted with phosphorothioate bonds.

10 . The immunity-inducing agent according to claim 1 , wherein the spacer comprises a repeating unit represented by the formula:

wherein

X represents an oxygen atom or a sulfur atom, wherein each X may be the same or different,

R represents (CH 2 ) q NH, wherein each q independently represents a natural number of not more than 10, and

n represents a natural number of not more than 10.

11 . The immunity-inducing agent according to claim 1 , wherein the spacer has a structure represented by any of the formulas:

12 . The immunity-inducing agent according to claim 1 , further comprising a substance having immunostimulatory activity as an adjuvant.

13 . An immunity-inducing agent according to claim 1 ,

wherein the antigenic peptide has an amino acid length of not less than 5 but not more than 30,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is a polydeoxyribonucleotide (DNA) or a DNA derivative comprising two or more CpG motifs,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is a polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds, and

wherein the antigenic peptide and the spacer are bound together via a disulfide bond produced by a reaction between a thiol group of a cysteine residue at the N-terminus of the antigenic peptide and a thiol group of the spacer.

14 . An immunity-inducing agent according to claim 1 ,

wherein the antigenic peptide has an amino acid length of not less than 8 but not more than 11,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is a polydeoxyribonucleotide (DNA) or a DNA derivative comprising two or more CpG motifs,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif has a nucleotide length of not less than 20 but not more than 30,

wherein, in the polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds, not less than 90% of the phosphodiester bonds are substituted with phosphorothioate bonds, and

the antigenic peptide and the spacer are bound together via a disulfide bond produced by a reaction between a thiol group of a cysteine residue at the N-terminus of the antigenic peptide and a thiol group of the spacer.

15 . An immunity-inducing agent according to claim 1 ,

wherein the antigenic peptide has an amino acid length of not less than 8 but not more than 11,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is a polydeoxyribonucleotide (DNA) or a DNA derivative comprising two or more CpG motifs,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif has a nucleotide length of not less than 20 but not more than 30,

wherein, in the polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds, not less than 90% of the phosphodiester bonds are substituted with phosphorothioate bonds,

wherein the antigenic peptide and the spacer are bound together via a disulfide bond produced by a reaction between a thiol group of a cysteine residue at the N-terminus of the antigenic peptide and a thiol group of the spacer, and

wherein the spacer comprises a repeating unit represented by the formula:

wherein

X represents an oxygen atom or a sulfur atom, wherein each X may be the same or different,

R represents any of (CH 2 ) p O, (CH 2 ) q NH, and (CH 2 CH 2 O) m , wherein m, p and q each independently represent a natural number of not more than 10, and

n represents a natural number of not more than 10.

16 . An immunity-inducing agent according to claim 1 ,

wherein the antigenic peptide has an amino acid length of not less than 8 but not more than 11,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is a polydeoxyribonucleotide (DNA) or a DNA derivative comprising two or more CpG motifs,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif has a nucleotide length of not less than 20 but not more than 30,

wherein, in the polynucleotide derivative in which phosphodiester bonds are at least partially substituted with phosphorothioate bonds, not less than 90% of the phosphodiester bonds are substituted with phosphorothioate bonds,

wherein the antigenic peptide and the spacer are bound together via a disulfide bond produced by a reaction between a thiol group of a cysteine residue at the N-terminus of the antigenic peptide and a thiol group of the spacer, and

wherein the spacer has a structure represented by any of the formulas:

17 . A pharmaceutical composition comprising a conjugate comprising (i) a single-chain polynucleotide or a single-chain polynucleotide derivative comprising a CpG motif, and (ii) an antigenic peptide bound thereto via a spacer, wherein the spacer is covalently bound at one end thereof to the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif and covalently bound at the other end thereof to the antigenic peptide,

wherein the single-chain polynucleotide or the single-chain polynucleotide derivative comprising a CpG motif is not complexed with β-1,3-glucan,

wherein the antigenic peptide and the spacer are bound together via a disulfide bond produced by a reaction between a thiol group of a cysteine residue at the N-terminus of the antigenic peptide and a thiol group of the spacer, and

wherein the spacer comprises a repeating unit represented by the formula:

wherein

X represents an oxygen atom or a sulfur atom, wherein each X may be the same or different,

R represents any of (CH 2 ) p O, (CH 2 ) q NH, and (CH 2 CH 2 O) m , wherein m, p and q each independently represent a natural number of not more than 10, and

n represents a natural number of not more than 10.

18 . A pharmaceutical composition comprising the immunity-inducing agent according to claim 13 .

19 . A pharmaceutical composition comprising the immunity-inducing agent according to claim 14 .

20 . A pharmaceutical composition comprising the immunity-inducing agent according to claim 15 .

21 . A pharmaceutical composition comprising the immunity-inducing agent according to claim 16 .

22 . A method for treating a tumor, comprising administering an effective amount of the immunity-inducing agent according to claim 1 to a subject in need thereof.

23 . A method for treating a tumor, comprising administering an effective amount of the pharmaceutical composition of according to claim 17 to a subject in need thereof.

24 . A method for treating a tumor, comprising administering an effective amount of the pharmaceutical composition of according to claim 18 to a subject in need thereof.

25 . A method for treating a tumor, comprising administering an effective amount of the pharmaceutical composition of according to claim 19 to a subject in need thereof.

26 . A method for treating a tumor, comprising administering an effective amount of the pharmaceutical composition of according to claim 20 to a subject in need thereof.

27 . A method for treating a tumor, comprising administering an effective amount of the pharmaceutical composition of according to claim 21 to a subject in need thereof.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 5, 2021
From: MOCHIZUKI, SHINICHI; KOIZUMI, MAKOTO; MORITA, KOJI
To: THE UNIVERSITY OF KITAKYUSHU; DAIICHI SANKYO COMPANY, LIMITED
Reel/Frame 055829/0079 →
Priority Claims (1)
JP 2018-186093 · Sep 28, 2018 · national
Continuity (1)
Related Publication 20220031839A1 · Feb 3, 2022
References Cited (75)
US 10195270B2 · Sakurai · 2019 [cited by examiner]
US 11793874B2 · Mochizuki et al. · 2023 [cited by applicant]
US 12220460B2 · Mochizuki et al. · 2025 [cited by applicant]
US 20060084149A1 · Kimura et al. · 2006 [cited by applicant]
US 20080146488A1 · Wettstein · 2008 [cited by applicant]
US 20140051637A1 · Suzumura · 2014 [cited by applicant]
US 20150191730A1 · Levy et al. · 2015 [cited by applicant]
US 20160000906A1 · Diamond · 2016 [cited by applicant]
US 20160186178A1 · Radovic-Moreno · 2016 [cited by examiner]
US 20160208260A1 · Ishii · 2016 [cited by examiner]
US 20170007695A1 · Sakurai · 2017 [cited by examiner]
US 20170035864A1 · Theriault · 2017 [cited by applicant]
US 20210106678A1 · Mochizuki et al. · 2021 [cited by applicant]
US 20220031839A1 · Mochizuki · 2022 [cited by applicant]
CN 101563104A · 2009 [cited by applicant]
CN 107281476A · 2017 [cited by applicant]
EP 1142591A1 · 2001 [cited by applicant]
EP 3858383A1 · 2021 [cited by applicant]
JP 2007070307A · 2007 [cited by applicant]
JP 200850907 · 2008 [cited by applicant]
JP 2008509072 · 2008 [cited by applicant]
JP 2010174107A · 2010 [cited by applicant]
JP 2017500313A · 2017 [cited by applicant]
TW 201639583A · 2016 [cited by applicant]
WO 0134207A1 · 2001 [cited by applicant]
WO 02072152A1 · 2002 [cited by applicant]
WO 2012147805 · 2012 [cited by applicant]
WO 2015089114A1 · 2015 [cited by applicant]
WO WO2015118789A1 · 2015 [cited by examiner]
WO 2016152767A1 · 2016 [cited by applicant]
WO 2017217531A1 · 2017 [cited by applicant]
WO 2020067400 · 2020 [cited by applicant]
Kramer et al., 2017, Intracellular Cleavable CpG Oligodeoxynucleotide-Antigen Conjugate Enhances Anti-tumor Immunity, Molecular Therapy, 25(1): 62-70. [cited by examiner]
Hayashi et al., 2005, Resistance to influenza A virus infection by antigen-conjugated CpG oligonucleotides, a novel antigen-specific immunomodulator, Biochemical and Biophysical Research Communications, 329: 230-236. [cited by examiner]
Kapadia, C.H., et al. “Extending antigen release from particulate vaccines results in enhanced antitumor immune response,” Journal of Controlled Release 2018, vol. 269, pp. 393-404. [cited by applicant]
Office Action mailed Aug. 23, 2022, issued in corresponding Japanese Application No. 2019-509625, filed Mar. 20, 2018, 7 pages. [cited by applicant]
Extended European Search Report mailed Aug. 24, 2022, issued in corresponding European Application No. 19865489.9, filed Sep. 27, 2019, 2018, 14 pages. [cited by applicant]
Maurer, T. et al., “CpG-DNA aided cross-presentation of soluble antigens by dendritic cells”, European Journal of Immunology 32(8): 2356-2364, Aug. 2, 2022. [cited by applicant]
Shirota, H. et al., “Regulation of Murine Airway Eosinophilia and Th2 Cells by Antigen-Conjugated CpG Oligodeoxynucleotides as a Novel Antigen-Specific Immunomodulator”, The Journal of Immunology, 164:5575-5582, Jan. 1,… [cited by applicant]
Wagner, H., “Bacterial CpG DNA Activates Immune Cells to Signal Infectious Danger”, Adv. Immunol., 73, 329-368, 1999. [cited by applicant]
Krieg, A., “CpG Motifs in Bacterial DNA and Their Immune Effects”, Annu. Rev. Immunol., 20, 709-760, 2002. [cited by applicant]
Yamamoto, S., et al., “The discovery of immunostimulatory DNA sequence”, Springer Semin Immunopathol, 22, 11-19, 2000. [cited by applicant]
Taniguchi, M., et al., Standard Immunology, 2nd Edition, 333, 2002. [cited by applicant]
Sakurai, K., et al., “Molecular Recognition of Adenine, Cytosine, and Uracil in a Single-Stranded RNA by a Natural Polysaccharide: Schizophyllan”, J. Am. Chem. Soc., 122, 4520-4521, 2000. [cited by applicant]
Sakurai, K., et al., “Polysaccharide-Polynucleotide Complexes. 2. Complementary Polynucleotide Mimic Behavior of the Natural Polysaccharide Schizophyllan in the Macromolecular Complex with Single-Stranded RNA and DNA”, … [cited by applicant]
Mochizuki, S., et al., “Dectin-1 targeting delivery of TNF-α antisense ODNs complexed with β-1,3-glucan protects mice from LPS-induced hepatitis”, J. Control. Release, 151, 155-161, 2001. [cited by applicant]
Miyoshi, K., et al., “Polysaccharide-Polynucleotide Complexes. Part 32. Structural Analysis of the Curdlan/Poly (cytidylic acid) Complex with Semiempirical Molecular Orbital Calculations”, Biomacromolecules, 6, 1540-154… [cited by applicant]
Mizu, M., et al., “A Polysaccharide Carrier for Immunostimulatory CpG DNAs to Enhance Cytokine Secretion”, J. Am. Chem. Soc., 126, 8372-8373, 2004. [cited by applicant]
Mizu, M., et al., “Protection of polynucleotides against nuclease-mediated hydrolysis by complexation with schizophyllan”, Biomaterials, 25, 15, 3109-3116, 2004. [cited by applicant]
Shimada, N., et al., “Synthesis and in Vitro Characterization of Antigen-Conjugated Polysaccharide as a CpG DNA Carrier”, Bioconjugate Chem., 17, 1136-1140, 2006. [cited by applicant]
Khan, S., et al., “Distinct Uptake Mechanisms but Similar Intracellular Processing of Two Different Toll-like Receptor Ligand-Peptide Conjugates in Dendritic Cells”, J Biol Chem., 282(29), 21145-21159, Jul. 20, 2007. [cited by applicant]
Kramer, K., et al., “Intracellular Cleavable CpG Oligodeoxynucleotide-Antigen Conjugate Enhances Anti-tumor Immunity”, Mol. Ther., 25(1), 62-70, Jan. 4, 2017. [cited by applicant]
Shirota, H., et al., “TLR-9 Agonist Immunostimulatory Sequence Adjuvants Linked to Cancer Antigens”, Methods in Molecular Biology, vol. 1139, 337-344, 2014. [cited by applicant]
Kramer, K., et al., “Comparative Study of 5′- and 3′-Linked CpG-Antigen Conjugates for the Induction of Cellular Immune Responses”, ACS Omega, vol. 2, 227-235, 2017. [cited by applicant]
Kupihar, Z., et al., “Synthesis and Application of a Novel, Crystalline Phosphoramidite Monomer with Thiol Terminus, Suitable for the Synthesis of DNA Conjugates”, Bioorganic & Medicinal Chemistry, vol. 9, 1241-1247, 20… [cited by applicant]
International Search Report mailed Nov. 19, 2019, issued in corresponding International Application No. PCT/JP2019/038090, filed Sep. 27, 2019. [cited by applicant]
Mochizuki, S., et al., “Immunization with antigenic peptides complexed with β-glucan induces potent cytotoxic T-lymphocyte activity in combination with CpG-ODNs”, Journal of Controlled Release, 220, 495-502, 2015. [cited by applicant]
Tighe, H., et al., “Conjugation of immunostimulatory DNA to the short ragweed allergen Amb a 1 enhances its immunogenicity and reduces its allergenicity”, J. Allergy. Clin. Immunol., 106 (1 Pt. 1), 124-134, Jul. 2000. [cited by applicant]
Mochizuki, S., et al., “Complex Consisting of β-Glucan and Antigenic Peptides with Cleavage Site for Glutathione and Aminopeptidases Induces Potent Cytotoxic T Lymphocytes”, Bioconjugate Chem., 28, 2246-2253, Jul. 24, 2… [cited by applicant]
International Search Report mailed May 1, 2018, issued in corresponding International Application No. PCT/JP2018/011201, filed Mar. 20, 2018, 2 pages. [cited by applicant]
Motohiko Suzuki, et al., International Immunopharmacology, vol. 7, Issue 1, Jan. 2007, pp. 46-54. [cited by applicant]
Mochizuki, S., et al., “Dectin-1 targeting delivery of TNF-α antisense ODNs complexed with β-1,3-glucan protects mice from LPS-induced hepatitis”, J. Control. Release, 151, 155-161, 2011. [cited by applicant]
Irie, et al., Bioconjugate Chem. 2020, 31, 2585-2595. [cited by applicant]
Rapin, Nicolas et al., Immunogenetics, 2008, vol. 60, No. 12, pp. 759-765. [cited by applicant]
Search Report mailed Apr. 20, 2021, issued in related International Application No. PCT/JP2021/012787, filed Mar. 26, 2021, 3 pages. [cited by applicant]
Aurisicchio, L. et al., “A novel minigene scaffold for therapeutic cancer vaccines,” Oncolmmunology 3, e27529-1-e27529-13; Jan. 2014. [cited by applicant]
Extended European Search Report mailed Jul. 12, 2024, issued in Application No. EP 21776683.1, filed Mar. 26, 2021, 10 pages. [cited by applicant]
Daftarian et al., “Novel conjugates of epitope fusion peptides with CpG-ODN display enhanced immunogenicity and HIV recognition,” Vaccine (2005), vol. 23, No. 26, pp. 3453-3468. [cited by applicant]
First Chinese Office Action mailed on Feb. 19, 2025, issued in Chinese App No. 202180024931.3; 18 pages. [cited by applicant]
Office Action mailed Jun. 5, 2025, issued in U.S. Appl. No. 17/907,531, filed Sep. 27, 2022, 53 pages. [cited by applicant]
Tung, et al., “Preparation and Applications of Peptide-Oligonucleotide Conjugates;” Bioconjugate Chemistry 11 (5): 605-618 (2000). [cited by applicant]
Final Office Action mailed Oct. 23, 2025, issued in U.S. Appl. No. 17/907,531, filed Sep. 27, 2022, 33 pages. [cited by applicant]
Bolcato, V., et al., “Healthcare-acquired Sars-Cov2 infection: A viable legal category?”, International Journal of Risk & Safety in Medicine 34: 129-134, 2023. [cited by applicant]
Palshof, F.K., et al. “Non-preventable cases of breast, prostate, lung and colorectal cancer in 2050 in an elimination scenario of modifiable risk factors,” Nature 14:8577, 2024. [cited by applicant]
Israelsen, A., et al., “Preventing Allergies in Infants: What Foods to Introduce and When,” Utah State University Extension, pp. 1-6, 2020. [cited by applicant]