US-12622962-B2 - Pharmaceutical composition and use thereof

Abstract

The present invention belongs to the field of biopharmaceutics and in particular relates to a pharmaceutical composition. The pharmaceutical composition comprises a hepatitis B surface antigen, a hepatitis B core antigen, and an immunostimulatory composition, wherein the immunostimuilatory composition comprises a saponin and a CpG oligodeoxynucleotide, or consists of an adjuvant comprising a saponin and a CpG oligodeoxynucleotide.

Inventors

• Jianqiang Li
• Xiaoxiao CHEN
• Jingfeng Huang
• Xaiodong Wang
• Yue Chen
• Jun Ge
• Jiaojiao SUN
• Tong Zhou
• Sulin Ren
• Changyao Tan
• Yue Gu
• Hongying HUANG
• Shiwei Wang

Assignees

• GRAND THERAVAC LIFE SCIENCES (NANJING) CO., LTD.

Dates

Publication Date

20260512

Application Date

20201211

Priority Date

20191213

Claims (18)

1. 1 . A pharmaceutical composition comprising: i) a hepatitis B surface antigen, an active fragment of the antigen, a variant of the antigen, or a mixture of at least two of them; ii) a hepatitis B core antigen, an active fragment of the antigen, a variant of the antigen, or a mixture of at least two of them; and iii) an immunostimulatory composition comprising a saponin and a CpG oligodeoxynucleotide, or consisting of an adjuvant comprising a saponin and a CpG oligodeoxynucleotide, wherein the sequence of the CpG oligodeoxynucleotide is any one selected from: CpG T1: TCG TTC GTT CGT TCG TTC GTT (SEQ ID NO: 6); CpG T2: TCG TTC GTT CGT TCG TTC GTT CGT T (SEQ ID NO: 7); and CpG T3: TCG TCG TCG TCG TCG TCG TCG (SEQ ID NO: 8), and the saponin is QS-21.
2. 2 . The pharmaceutical composition according to claim 1 , wherein the CpG oligodeoxynucleotide comprises a phosphorothioate linkage.
3. 3 . The pharmaceutical composition according to claim 1 , wherein the CpG oligodeoxynucleotide is a thio-oligodeoxynucleotide.
4. 4 . The pharmaceutical composition according to claim 1 , wherein the CpG oligodeoxynucleotide is a perthio-oligodeoxynucleotide.
5. 5 . The pharmaceutical composition according to claim 1 , wherein the weight ratio of the CpG oligodeoxynucleotide to the saponin is 1˜40:0.1˜2.
6. 6 . The pharmaceutical composition according to claim 1 , wherein the weight ratio of the CpG oligodeoxynucleotide to the saponin is 2:1.
7. 7 . The pharmaceutical composition according to claim 1 , wherein the Hepatitis B surface antigen comprises or consists of the sequence as shown by SEQ ID NO: 1, and the Hepatitis B core antigen comprises or consists of the sequence as shown by SEQ ID NO: 2.
8. 8 . The pharmaceutical composition according to claim 1 , wherein the active fragment of the hepatitis B core antigen comprises or consists of consecutive amino acids from position 1 to position X of SEQ ID NO: 2, wherein X is an integer between 149 and 183.
9. 9 . The pharmaceutical composition according to claim 8 , wherein the active fragment of the hepatitis B core antigen comprises or consists of consecutive amino acids from position 1 to position X of SEQ ID NO: 2, wherein X is an integer between 152 and 183.
10. 10 . The pharmaceutical composition according to claim 1 , wherein the weight ratio among Components i), ii) and iii) in the pharmaceutical composition is 4:2:1.1˜42.
11. 11 . The pharmaceutical composition according to claim 1 , wherein the weight ratio among Components i), ii) and iii) in the pharmaceutical composition is 4:2:3.
12. 12 . The pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition further comprises: iv) a pharmaceutically acceptable carrier.
13. 13 . A hepatitis B prophylactic or B therapeutic vaccine, comprising the pharmaceutical composition of claim 1 .
14. 14 . A method for preventing and/or treating a hepatitis B virus infection and/or a hepatitis B virus mediated disease, comprising administering to a subject in need thereof a prophylactically and/or therapeutically effective amount of the pharmaceutical composition according to claim 1 .
15. 15 . The method according to claim 14 , wherein the hepatitis B virus infection and/or the hepatitis B virus mediated disease is selected from the group consisting of cirrhosis and liver cancer.
16. 16 . A method for generating a humoral immune response and/or a cellular immune response against hepatitis B virus in a subject, comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition of claim 1 .
17. 17 . A method for switching the subtype of hepatitis B core antibody in a subject, comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition of claim 1 .
18. 18 . A method for breaking through the immune tolerance of hepatitis B virus in a subject, comprising administering to a subject in need thereof an effective amount of the pharmaceutical composition of claim 1 .

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application is the U.S. national phase of PCT/CN2020/135570, filed on Dec. 11, 2020, which claims priority to Chinese Application No. 201911279536.0, filed on Dec. 13, 2019, each expressly incorporated herein by reference in its entirety. STATEMENT REGARDING SEQUENCE LISTING The sequence listing associated with this application is provided in text format in lieu of a paper copy and is hereby incorporated by reference into the specification. The name of the text file containing the sequence listing is 3053-P31USPNP_Seq_List_20221221_ST25_Revised.txt. The text file is 11 KB; was created on Dec. 21, 2022, contains no new matter, and is being submitted via Patent Center. TECHNICAL FIELD The present invention belongs to the field of biopharmaceutics. In particular, the present invention relates to a pharmaceutical composition comprising a hepatitis B surface antigen, a hepatitis B core antigen and an immunostimulatory composition, wherein the immunostimulatory composition comprises a saponin and a CpG oligodeoxynucleotide, or consists of an adjuvant comprising a saponin and a CpG oligodeoxynucleotide, and the sequence of the CpG oligodeoxynucleotide has two or more copies of 5′-TTCGTT-3′ (SEQ ID NO: 31) motif or 5′-TCGTCGTCG-3′ (SEQ ID NO: 32) motif. The present invention also relates to use of the pharmaceutical composition for treating a hepatitis B virus infection and/or a hepatitis B virus mediated disease. BACKGROUND ART Hepatitis B virus (HBV) infection is one of serious public health problems worldwide. HBV infection is a significant cause of chronic hepatitis B, cirrhosis and hepatocellular carcinoma (Fatt ovich G. J. Hepatol. 2008; 48: 335-352). Common drugs for the clinical treatment of chronic HBV infection mainly include nucleoside analogues and interferons. Nucleoside analogues cannot completely eliminate cccDNA in hepatocytes, and their long-term use is prone to the emergence of drug-resistant mutants and rebound after drug withdrawal (Kwon H, Lok A S. Nat Rev Gastroenterol Hepatol. 2011; 8: 275-284). Interferons are not suitable for asymptomatic HBV carriers. In chronic HBV patients, the seroconversion incidence of HBeAg is only 33% after half a year of use. Also, the application of interferons is limited by their larger side effects (Tang S X, Yu G L. Lancet 1990; 335 (8684): 302). At present, the widely used hepatitis B protein vaccines achieve the goal of prevention by inducing humoral immunity and producing protective neutralizing antibodies. Many studies have found that the neutralizing antibodies can only eliminate extracellular viral particles, while elimination of intracellularly infected viruses mainly relies on specific cellular immune responses, Th1-type cytokines such as IFN-γ produced by helper T cells, CD4+ T cells, especially virus-specific cytotoxic T lymphocytes (CTL) (ChinR, Lacamini S. Rev Med Viorl. 2003: 13 (4): 255-72). The intensity of cellular immune response directly determines the prognosis of hepatitis B. Therefore, an ideal therapeutic hepatitis B vaccine needs to induce both specific humoral and cellular immunity to break through the immune tolerance of hepatitis B. For example, Chinese patent CN104043120B provides a therapeutic hepatitis B vaccine comprising a hepatitis B surface antigen (HBsAg), a hepatitis B core antigen (HBcAg) and an oligodeoxynucleotide (CpG), which can break through the immune tolerance of hepatitis B and be used for treating viral hepatitis B, especially chronic hepatitis B. The present inventors have intensively studied the prior art and found that adjuvants play an important role in the therapeutic effect of therapeutic hepatitis B vaccine. The adjuvants, oligodeoxynucleotides (CpGs), are commonly used as immunostimulatory compounds, and their chemical nature is an oligodeoxynucleotide containing cytosine-guanine dinucleotide, which have a similar immune response to the natural pattern recognition receptors for CpG, and can bind to Toll-like receptors on cell membrane, effectively triggering mammalian immune response through TLR9 signaling pathway. Saponin adjuvants are a class of glycosides, the aglycons of which are triterpene or spirostane compounds, and belong to plant-derived adjuvants. Among them, quillaja saponin (QS) is the saponins extracted from quillaja, and QS-21 is the most widely reported adjuvant in QS series. However, QS-21 may induce cell hemolysis and has some systemic and local toxic/side effects. Alving et al.’ study (ALVING CR, MATYAS G, BECK Z, et al. Revue Roumaine de Chimie, 2016, 61(8): 631-635) found that ALF liposomes in combination with MPLA and QS-21 as an adjuvant against HIVgp140 protein can effectively increase the antibody titer in serum. Ng et al. (NG H, FERNANDO G J P, DEPELSENAIRE A C I, et al. Scientific Reports, 2016, 6(1): 228-230) used a subcutaneous delivery technique, a nano-patch, to form an adjuvant complex with QS-21. The results showed, compared with tr