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EP-4741506-A1 - CPG OLIGODEOXYNUCLEOTIDE CAPABLE OF REGULATING IMMUNE FUNCTION OF BODY AND USE THEREOF

EP4741506A1EP 4741506 A1EP4741506 A1EP 4741506A1EP-4741506-A1

Abstract

The present disclosure discloses a plurality of CpG-containing oligodeoxynucleotides (ODNs) with immunomodulatory properties, and use thereof. The oligodeoxynucleotides provided in present disclosure differ in sequence and structure from the design concept of conventional Type A CpG ODNs, reflecting their own complex drug-forming principles and overcoming the problems of conventional Type A CpG in drug development. The oligodeoxynucleotides exhibit excellent immunomodulatory activity both in vitro and in vivo , and they can stimulate the production of type I, II, and III interferons, upregulate Th1-type immune responses, and inhibit Th2-type and Th17-type inflammatory responses. Therefore, the oligodeoxynucleotides can: prevent and treat allergic diseases caused by Th2-type immune responses, including allergic rhinitis, atopic dermatitis, asthma, chronic obstructive pulmonary disease, eosinophilia, and Th17-related diseases; can prevent and treat infections and transmission of respiratory or non-respiratory pathogens, including viruses, bacteria, fungi, and parasites; can improve the immune levels of the elderly population and individuals with immunodeficiency; can transform "cold tumors" into "hot tumors" and improve the effectiveness of cancer immunotherapy; can prevent and treat central nervous system diseases related to immune dysfunction, such as Alzheimer's disease; and can be applied to other diseases related to immune dysregulation.

Inventors

  • LIU, LIMING
  • PAN, QI
  • FU, YOURONG
  • MAO, HUI

Assignees

  • Nanjing Jsiama Biopharmaceuticals Ltd.

Dates

Publication Date
20260513
Application Date
20240719

Claims (20)

  1. A CpG oligodeoxynucleotide with immunomodulatory properties, comprising a sequence having the structure of 5'-(G) n -core sequence-(N) q -(G)m-3', wherein the core sequence is a palindromic sequence, N is any one selected from the group consisting of adenine deoxyribonucleotide (A), thymine deoxyribonucleotide (T), and cytosine deoxyribonucleotide (C); G is guanine deoxyribonucleotide; q, n, and m are all integers, in which n ranges from 2 to 11, q ranges from 0 to 10, and m ranges from 2 to 11.
  2. The CpG oligodeoxynucleotide of claim 1, wherein n ranges from 3 to 4, and q ranges from 1 to 5.
  3. The CpG oligodeoxynucleotide of claim 1, wherein the core sequence is at least one selected from the group consisting of CGCGACGCGTCGCG, ACGATCGAGATCGT, AGGATCGATCCT, TTCGATCGATCGAA, CGATCGATCG, GACGATCGTC, and TGCATCGATGCA; wherein, if the core sequence is CGCGACGCGTCGCG, q ranges from 0 to 10, n ranges from 0 to 3, and m ranges from 2 to 11; or if the core sequence is ACGATCGAGATCGT, q ranges from 0 to 10, n ranges from 2 to 11, and m ranges from 2 to 11; or if the core sequence is AGGATCGATCCT, q ranges from 0 to 10, n ranges from 2 to 11, and m ranges from 2 to 11; or if the core sequence is TTCGATCGATCGAA, q ranges from 0 to 10, n ranges from 2 to 11, and m ranges from 2 to 11; or if the core sequence is CGATCGATCG, q ranges from 0 to 10, n ranges from 1 to 11, and m ranges from 2 to 11; or if the core sequence is GACGATCGTC, then q is 0, n ranges from 0 to 3, and m ranges from 2 to 11, or then q ranges from 1 to 10, n ranges from 2 to 11, and m ranges from 2 to 11; or if the core sequence is TGCATCGATGCA, then q is 0, n has a value of 1, 3, 4 or 5, and m ranges from 2 to 11, or then q ranges from 1 to 10, n ranges from 2 to 11, and m ranges from 2 to 11.
  4. The CpG oligodeoxynucleotide of claim 1, comprising at least one selected from the group consisting of SEQ ID NO.1 - SEQ ID NO.82.
  5. The CpG oligodeoxynucleotide of any one of claims 1-4, wherein the CpG oligodeoxynucleotide comprises at least one selected from the group consisting of: thio-modification, fluorination-modification, methoxy-modification, locked nucleic acid modification, or nanoparticle modification.
  6. The CpG oligodeoxynucleotide of claim 5, wherein the thio-modification occurs at the 5' and/or 3' ends of polyguanosine.
  7. The CpG oligodeoxynucleotide of claim 5, wherein nanoparticles used in the nanoparticle modification include at least one selected from the group consisting of PLGA, chitosan, lipid nanoparticles, and liposomes.
  8. A CpG oligodeoxynucleotide solution formulation, comprising the CpG oligodeoxynucleotide of any one of claims 1 to 7 and excipients, wherein the excipients comprise at least one selected from the group consisting of: pH buffer pair, glycine, trehalose, mannitol, sucrose, arginine, lysine, histidine, glycerol, propylene glycol, Pluronic F127, Pluronic F68, Tween 20, Tween 80, benzalkonium chloride, disodium edetate, sodium citrate, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, methyl betacyclodextrin, and polyethylene glycol.
  9. The formulation of claim 8, wherein metal ions in the CpG oligodeoxynucleotide formulation include sodium, potassium, magnesium, calcium, zinc, and iron, with a total concentration ranging from 0.1 to 90 mM.
  10. The formulation of claim 8, wherein the pH of the CpG oligodeoxynucleotide formulation ranges from 7 to 9.
  11. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of drugs for regulating respiratory immune responses.
  12. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of drugs for prevention and treatment of respiratory diseases, wherein the respiratory diseases include at least one selected from the group consisting of respiratory viral infections, respiratory bacterial infections, respiratory fungal infections, respiratory parasitic infections, and respiratory allergic diseases.
  13. The use of claim 12, wherein the respiratory viruses include at least one selected from the group consisting of: COVID-19 virus, influenza virus, RSV virus, and SARS-CoV.
  14. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of drugs for prevention and treatment of non-respiratory tract infections, wherein the non-respiratory tract infections include at least one selected from the group consisting of HIV infection, HBV infection, and HCV infection.
  15. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of drugs for regulating proliferation of immune cells or release of immune cell cytokines.
  16. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of immunoadjuvants.
  17. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of drugs for prevention and treatment of viral infections.
  18. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of drugs for treatment of tumors, wherein the CpG oligodeoxynucleotide is the only anti-tumor active ingredient.
  19. An anti-tumor composition, comprising the CpG oligodeoxynucleotide of any one of claims 1 to 6 and an anti-tumor drug, wherein the anti-tumor drug includes, but is not limited to, PD-1 anti-tumor drugs, PDL-1 anti-tumor drugs and anti-tumor cell therapy, wherein the CpG oligodeoxynucleotide in the composition serves as a component with anti-tumor efficacy or as an immunoadjuvant.
  20. Use of the CpG oligodeoxynucleotide of any one of claims 1 to 7 in preparation of human or animal vaccines.

Description

The present application claims priority of application No. 202310902755.X filed on July 21, 2023 with China National Intellectual Property Administration, entitled "CPG OLIGODEOXYNUCLEOTIDES WITH IMMUNOMODULATORY PROPERTIES AND THE APPLICATIONS THEREOF", the entire content of which is incorporated herein by reference. TECHNICAL FIELD The present disclosure belongs to the field of oligodeoxynucleotides, specifically relating to a CpG oligodeoxynucleotides with immunomodulatory properties and the applications thereof. BACKGROUND OF THE INVENTION TLR9 is an innate immune receptor in mammals, located in endosomes. Its ligands are oligodeoxynucleotides (ODNs) containing unmethylated Cytosine-Phosphorothioate-Guanine (CpG). Depending on their type and structure, CpG-ODNs, after entering the endosome, can bind to TLR9 and activate the IRF-7 or NF-κB signaling pathways, producing inflammatory factors such as type I interferons or IL-6 to intervene in the immune response. Based on structural characteristics and immune effects, CpG ODNs can be classified into three classes: A, B, and C. Type A CpG enters the early endosomes of plasmacytoid dendritic cells (pDCs), binds to the TLR9 receptor therein, activates the IRF-7 signaling pathway, and induces the production of large amounts of type I interferons (IFN-α/IFN-β) and type III interferon (IFN-λ). Type I interferons, as a general immunomodulatory cytokine, acts on downstream immune cells, including but not limited to NK cells, CD8+ T cells, CD4+ T cells, γ-δ T cells, and macrophages, inducing the production of cytokines such as type II interferon (IFN-γ). Type B CpG is a fully thiolated linear CpG ODN, primarily targeting B lymphocytes. Type B CpG enters late endosomes and binds to TLR9, activating NF-κB and generating a series of immune responses, including the secretion of inflammatory cytokines such as IL-6, IL-1, and TNF-α, and promoting B cell proliferation. Type B CpG can be used as a vaccine adjuvant, significantly increasing antibody levels. Class C CpG is a fully thiolated CpG ODN, containing partial structures of Type A CpG and Type B CpG. It possesses both palindromic sequences capable of forming dimers and non-palindromic linear structures. Class C CpG can enter both early and late endosomes and bind to TLR9. Therefore, Class C CPG possesses the activity of both Type A and Type B CpG-ODNs, activating both the IRF-7 signaling pathway and the NF-κB pathway to produce type I interferons and inflammatory cytokines. While Class C CPG combines the characteristics of both Type A and Type B CpGs, its ability to induce interferons and inflammatory molecules is significantly lower than that of either Type A or Type B CpGs individually. Preclinical animal studies of Type B CpG have demonstrated its successful treatment of allergic rhinitis, asthma, various microbial infections, and anti-tumor effects. Over the past 20 years, more than 100 publications have reported on the preclinical treatment of various diseases with CpGs. CPG-1018, a representative Type B CpG, was successfully approved for marketing as an adjuvant for hepatitis B vaccines in 2017. Current research indicates that when administered via different routes, Type A CpG ODN can induce robust innate immunity (type I/II/III interferons) and Th1 immune responses (activation of DCs, NK and CD8+ T cells, and IL-12 production) by targeting TLR9 in pDCs, and can also modulate immune function, thereby inhibiting Th2 and Th17 immune responses. These immune responses can be used to treat diseases caused by allergic and non-allergic inflammation, combat pathogenic microbial infections, directly or indirectly kill tumor cells, reduce Aβ plaque formation, and enhance vaccine immunogenicity, among other effects. This also laid the foundation for its application in infectious diseases, cancer, immunoadjuvants, allergic rhinitis, atopic dermatitis, asthma, chronic obstructive pulmonary disease, eosinophilic disease, Alzheimer's disease, wound healing, and wound infection prevention and control (Montamat et al., Fronteers in Immunol. 2021. doi: 10.3389/fimmu.2021.590054; Scheiermann et al., Vaccine 2014. 32:6377-6389; Shirota and Klinman., Immunopotentiators in Modern Vaccines ISBN 978-0-12-804019-5, http://dx.doi.org/10.1016/B978-0-12-804019-5.00009-8; Beeh et al., J Allergy Clin Immunol 2013;131:866-74; Hanagata N. et al., Int J Nanomedicine. 2012;7:2181-95; Kayraklioglu et al., Methods Mol Biol. 2021;2197:51-85; Fan et al., Clin Cancer Res. 2012 Oct 15;18(20):5628-38; Senti et al., Clin Exp Allergy. 2009 Apr;39(4):562-70; Scholtzova et al., Acta Neuropathol Commun. 2014 Sep 2;2:101; and Wanke-Jellinek et al., J Immunol. 2016 Jan 15;196(2):767-77). Due to the tendency of Type A CpG to multimerize, its drugability is very poor. As a result, more than 20 years after its discovery, there have been very few clinical trials for Type A CpG, and no drugs have been approved for marketing. Historically, three institutions have a