Zoster vaccine containing TLR9 agonist

Abstract

The present invention provides immunogenic compositions, kits, and methods suitable for stimulating an immune response to varicella-zoster virus (VZV) in individuals requiring such stimulation. [Solution] An immunogenic composition is provided comprising varicella-zoster virus (VZV) glycoprotein E antigen and a Toll-like receptor 9 (TLR9) agonist, such as an oligonucleotide containing a non-methylated cytidine-phospho-guanosine (CpG) motif.

Description

[Technical Field]

[0001] Cross-references to related applications This application is in relation to U.S. Provisional Application No. 62 / 987,243, filed on March 9, 2020. They claim priority and interest in such disclosure, and the entire disclosure is incorporated by reference.

[0002] Submission of the sequence list as an ASCII text file. The following submission in ASCII text file is fully explained by reference. Included in the detailed document: Sequence listing in computer-readable format (CRF) (filename: 377 882007140SEQLIST.TXT, Recorded: March 9, 2021, Size: 1 0KB).

[0003] This disclosure relates to the surface (VZV) glycoprotein E antigen of varicella-zoster virus, and Toll-like antigen. Receptor 9 (TLR9) agonists, such as unmethylated cytidine-phospho-guanosine (C) This relates to immunogenic compositions containing oligonucleotides containing a pG motif. The substance is suitable for stimulating an immune response to VZV in individuals that require it. ru. [Background technology]

[0004] Shingles (herpes zoster) is a disease affecting the body. Or a burning, tingling, and itching sensation on one side of the face, followed by pain. It is a viral disease characterized by rashes and blisters. (Centers for Disease Control and Prevention (C)) According to DC, approximately 10-13% of elderly people with shingles experience long-term neuralgia as a complication. They experience postherpetic neuralgia. In addition, shingles rashes near the eyes or ears can lead to blindness. It may cause drooping or paralysis of the facial muscles (Ramsay Hunt syndrome). group).

[0005] Shingles is caused by infection with the varicella-zoster virus (VZV), resulting in blisters that resemble chickenpox. It is caused. Shingles occurs in individuals who have chickenpox as a result of a primary infection with VZV. It is caused by the reactivation of latent VZV. (Referring to the 1995 varicella vaccine in the US diet) Before the FDA's approval, most children in the United States had contracted chickenpox, Therefore, there is currently a risk of developing shingles as an adult.

[0006] Two vaccines are available for the prevention of shingles in individuals aged 50 and over. ZOSTAVAX (registered trademark) is a trademark of Merck & Co., Inc. (Whitewood). This is a live attenuated virus vaccine sold by Ouse Station (NJ). SHINGRIX (registered trademark) is a licensed product of GlaxoSmithKline (Resear Recombinant adjuvants sold by Triangle Park, NC It is a subunit vaccine. Although direct comparative clinical trials were not conducted, the actual effects of vaccination The Advisory Committee on the Treatment of Herpes Zoster and Postherpetic Neuralgia in 2017 Based on the effectiveness evaluation, SHINGRIX is superior to ZOSTAVAX(registered trademark). They reluctantly expressed their recommendation for the preferential use of the registered trademark (Dooling et al., MMWR, 67 (103-108, 2018). SHINGRIX (registered trademark) is an adjuvant for AS01B In addition, it contains recombinant VZV glycoprotein E (gE) as an antigen. AS01B is Combined in a liposomal formulation, 3-O-desacyl- derived from Salmonella Minnesota 4'-monophosphoryl lipid A (MPL), and QS-21, a saponin purified from the extract of Quillaja saponaria Molina. The narrow approval vote (8 to 7) of SHINGRIX (R) was related to concerns about reactogenicity. The phase III study of SHINGRI X (R) reported that 16.5% of vaccine recipients had grade 3 adverse events compared to 3.1% of placebo recipients, and 10.8% of vaccine recipients had grade 3 systemic events (muscle pain, fatigue, headache, tremor, fever and gastrointestinal symptoms) compared to 2.4% of placebo recipients (La l et al., N Eng J Med, 372:2087-2096, 2015; and Cunningham et al., N Eng J Med, 375:1019-1032, 2016). The relatively high level of serious adverse reactions after SHINGRIX (R) vaccination was recognized to potentially negatively impact the compliance level of the second dose in patients (Bharucha et al., Human Vaccines & Immunotherapeutics , 13:1789-1797, 2017), which was a significant concern considering the need for patients to receive both doses to be protected against herpes zoster.

[0007] The reactogenicity of protein subunit vaccines such as SHINGRIX (R) can be affected by various factors, among which the most important is the nature of the adjuvant included to improve immunogenicity. Different adjuvants combined with The AS adjuvants were compared by immunization of healthy human subjects. A The S01B adjuvant induces higher levels of local and systemic reactogenicity than the comparator adjuvants (AS01E, AS03A, AS04 and alum) as found (Leroux-Roels et al., Clin Immunol, 169:16-27, 2016). This suggests that AS 01B contributes significantly to the feared reactogenicity profile of SHINGRIX. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0008] Therefore, there is a need for an improved varicella zoster vaccine that is inferior to SHINGRIX® with respect to VZV gE immunogenicity but has an excellent safety profile. MEANS FOR SOLVING THE PROBLEMS

[0009] The present disclosure relates to an immunogenic composition comprising a varicella zoster virus surface (VZV) glycoprotein E antigen and a toll-like receptor 9 (TLR9) agonist, such as an oligonucleotide containing an unmethylated cytidine-phospho-guanosine (CpG) motif. The immunogenic composition is suitable for stimulating an immune response against VZV in an individual who needs it. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] [Figure 1A] Figure 1A shows the levels of anti-VZV gE reactive IgG in sera of recipients of the first dose (prime) of the varicella zoster vaccine and control recipients (CTL and LAV). ​​​​​​​​ [Figure 1B] Figure 1B shows the levels of anti-VZV gE-reactive IgG in the serum of recipients and control recipients (CTLs and LAVs) after the first (prime) and second (boost) doses of the herpes zoster vaccine. [Figure 2A] Figures 2A-2C show IL-2 and / or IFN-γ induced by gE antigen secreted by CD4+ T cells in recipients of the first (prime) and second (boost) doses of the herpes zoster vaccine, as well as control recipients (CTLs and LAVs). [Figure 2B] Same as above [Figure 2C] Same as above [Figure 3A] Figures 3A-3B show IL-2 or IFN-γ induced by gE antigen secreted by CD154+ and CD4+ T cells in recipients of the first (prime) and second (boost) doses of the herpes zoster vaccine, as well as control recipients (CTLs and LAVs). [Figure 3B] Same as above [Modes for carrying out the invention]

[0011] General Techniques and Definitions The implementation of this disclosure shall, unless otherwise indicated, be within the scope of the skills in the art, and shall be in the field of molecular biology. Using conventional techniques in chemistry (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology. Yes, they are.

[0012] When used herein and in the appended claims, the singular "a" is used. "n" and "the" refer to multiple objects unless otherwise indicated. For example, "a n (one type) excipients include one or more excipients.

[0013] The term "comprising" as used in this specification is open This is an end statement indicating that such an embodiment may include additional elements. In other words, the phrase "consisting of" is a closed word, and Embodiments like the one described above indicate that the product is free of additional elements (except for trace amounts of impurities). The phrase "consisting essentially of" It is partially closed, and such embodiments significantly alter the fundamental features of such embodiments. This indicates that it may include additional elements that remain unchanged.

[0014] As used herein in reference to values, the term "about" means 90% to 11% of that value. Including 0% (for example, approximately 3000 μg of CpG 1018 is equivalent to 2700 μg to 330 μg) (Refers to 0 μg of CpG 1018).

[0015] In this specification, "polynucleotide" and "oligonucleic acid" are used interchangeably. The term "rheotide" refers to single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), Single-stranded RNA (ssRNA) and double-stranded RNA (dsRNA), modified oligonucleotides This includes oligonucleotides and oligonucleotides, or combinations thereof. Oligonucleotides are It may be a linear or cyclic structure, or the oligonucleotide may be linear and cyclic. Oligonucleotides can contain both phospho segments. Oligonucleotides are generally phospho A polymer of nucleosides linked through diester linkages, but alternative linkages, for example Phosphothioate esters may also be used in oligonucleotides. Creosides are purines (adenine (A) or guanine (G)) that are bound to sugars, or (These derivatives), or pyrimidines (thymine (T), cytosine (C), or uracil) (U), or derivatives thereof, consists of bases. The four types of nucleoside units in DNA (ma (or bases) are deoxyadenosine, deoxyguanosine, thymidine and deoxycithin It is called din. A nucleotide is a phosphate ester of a nucleoside.

[0016] The terms "CpG," "CpG motif," and "cytosine-phosphate-guanosine" When used herein, unmethylated cytidine-phospho-guanosine dinucleotide This refers to rheotide, which, when present in oligonucleotides, can be used in vitro, in vivo. and / or contribute to an ex vivo measurable immune response. Examples of measurable immune responses and While not limited to these, antigen-specific antibody production, cytokine secretion, and NK cell secretion are also possible mechanisms of action. Activation of lymphocyte populations such as cells, CD4+ T lymphocytes, CD8+ T lymphocytes, and B lymphocytes. Other examples include expansion and proliferation. Preferably, the CpG oligonucleotide is Th1 type. Prioritize activating the response.

[0017] An "effective amount" or "sufficient amount" of a substance is defined as having beneficial or desired results, including clinical outcomes. It is a sufficient amount to do so, and for example, the "effective amount" depends on the situation in which it is applied. It exists. In situations where an immunogenic composition is administered, the effective amount is to stimulate the immune response. Serum containing sufficient antigen and TLR9 agonist (preferably, an antibody against the antigen) (Protection level).

[0018] The terms "individual" and "subject" refer to mammals. "Mammals" are limited to... It is not something that will be done, but humans, non-human primates (e.g., monkeys), livestock, sporting animals, etc. Rodents (e.g., mice and rats), and pets (e.g., dogs and cats) It can be listed.

[0019] As used herein in reference to immunogenic compositions, the term “dose” is any Sometimes the immune system is ingested by the target (administered to the target, or received by the target). This refers to the measurement portion of the epidemiogenic composition.

[0020] As used herein, the terms “isolated” and “purified” refer to natural Extracted from at least one component related to (for example, from its original environment) It refers to the material (that was extracted). The term "isolated" is used when referring to recombinant proteins. When used, the protein is extracted from the culture medium of the host cell that produced the protein. It refers to quality.

[0021] The "stimulus" of the response or parameter is otherwise the same except for the parameter of interest. When compared to the same condition, or alternatively compared to another condition, the response or parameter - This includes the induction and / or enhancement of (e.g., TLR agonists) (Increased TLR signaling in the presence of a TLR agonist compared to its absence). For example Therefore, "stimulation" of the immune response means an increase in the response. Depending on the parameter being measured, The increase is 5 to 500 times or more, or 5, 10, 50, or 100 to 500 times. It could be double, 1,000 times, 5,000 times, or 10,000 times.

[0022] As used herein, the term "immunization" refers to the interaction of mammals with an antigen. A process that increases the elephant's response, and therefore resists or overcomes infection. It refers to the process of improving one's ability to resist and / or disease.

[0023] As used herein, the term “vaccination” refers to the application of an injection of a vaccine to the body of a mammal. This refers to the introduction of vaccines.

[0024] An "adjuvant," when added to a composition containing an antigen, reacts to the mammalian receptor during exposure. This refers to substances that enhance or strengthen the immune response to an antigen in a patient.

[0025] Detailed explanation This disclosure relates to varicella-zoster virus (VZV) glycoprotein E antigen (gE) and Toll Tolerant receptor 9 (TLR9) agonists, such as unmethylated cytidine phosphoguanosine. This relates to an immunogenic composition containing an oligonucleotide containing a (CpG) motif. VZV gE antigen consists of the signal peptide, transmembrane domain and cytoplasmic domain of the full-length VZV gE antigen. It may also be a cleaved recombinant protein lacking the main protein. The immunogenic composition requires it. It is suitable for stimulating the immune response to the varicella-zoster virus in individuals.

[0026] I. Immunogenic Compositions and Kits This disclosure relates to immunogens for stimulating an immune response to varicella-zoster virus (VZV). Sexual composition comprising VZV glycoprotein E (gE) antigen and Toll-like receptor 9 (TLR 9) Contains an agonist, and the TLR9 agonist is unmethylated cytidine-phosphoguanoside 8-35 containing the cytosine (CpG or cytosine-phosphate-guanosine) motif It is an oligonucleotide of nucleotide length, and the gE antigen and oligonucleotide are nucleotides In mammals, for example, in amounts effective to stimulate an immune response to gE antigen in humans, This relates to immunogenic compositions present in immunogenic compositions. In some embodiments, The immunogenic composition further includes an aluminum salt adjuvant to which the VZV gE antigen is adsorbed. include.

[0027] A. Toll-like receptor 9 (TLR9) agonists Toll-like receptors (TLRs) are found in and on dendritic cells and other innate immune cells. It is one of the most important receptors for stimulating a response to the presence of invading pathogens. Yes. Humans can recognize different parts of viruses or bacteria that are structurally similar. It possesses several types of TLRs. By activating specific TLRs, it enhances adaptive responses. It is possible to stimulate and control specific types of innate immune responses that can be used for this purpose. ru.

[0028] TLR9 (CD289) is an unmethylated cytidine-phosphorus found in microbial DNA. -Recognizes the guanosine (CpG) motif, which is a synthetic CpG-containing oligodeoxynucleotide. This can be mimicked using leotide (CpG-ODN). CpG-ODN is an antibody. It is known to enhance production and stimulate the T helper 1 (Th1) cell response. (Coffman et al., Immunity, 33:492-503, 2010). Based on structure and biological function. CpG-ODN has three common classes: CpG-A, CpG-B, and CpG- It is divided into C (Campbell, Methods Mol Biol, 1494:15-27, 2017). B cell activation The degree of B cell activation is weak CpG-A ODN, good CpG-C ODN, and The class of oligonucleotides TLR9 varies depending on the strength of the CpG-B ODN. The agonist is preferably a good B cell activator (CpG-C ODN), or More preferably, it is a strong B cell activator (CpG-B ODN).

[0029] The optimal oligonucleotide TLR9 agonist is often a purine with the general formula: 5'-purine. -Purin-CG-Pyrimidine-Pyrimidine-3' or 5'-Purin-Purin-CG- Contains a palindromic sequence following pyrimidine-pyrimidine-CG-3' (USA 6,589 (No. 940). TLR9 agonism is also a certain non-palintilized CpG enriched phosphorothiocyanate. Observed by nucleotides, but affected by changes in the nucleotide sequence. In some cases, this can occur. In addition, TLR9 agonism involves cytosine within CpG dinucleotides. It is inactivated by methylation. Therefore, in some embodiments, TLR9 The agonist is an oligonucleotide 8-35 nucleotide long containing the sequence 5'-AACGTTCG-3'. It is a gonucleotide. In some embodiments, the oligonucleotide is 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 Nukureochi Longer than the dot length, the oligonucleotides are 35, 34, 33, 32, 31, 30, 29, The length is less than 28, 27, 26, 25, or 24 nucleotides. In some embodiments, In this case, the TLR9 agonist is sequence 5'-AACGTTCGAG-3' (sequence number 3) It is an oligonucleotide with a length of 10 to 35 nucleotides, which is included in some embodiments. And the oligonucleotides are 10, 11, 12, 13, 14, 15, 16, 17, 18 Oligonucleotides are longer than 19 or 20 nucleotides, and are 35, 34, or 3 Less than 3, 32, 31, 30, 29, 28, 27, 26, 25, or 24 nucleotides in length That is the case.

[0030] Dynavax Technologies Corporation (Emeryv Researchers at ille (CA) have found that they substantially enhance the immune response to co-administered antigens across species. A 22-nucleotide-long phosphorothioate-linked oligodeoxy containing a specific sequence that can be used. The nucleotide CpG 1018 was identified (Campbell, Methods Mol Biol, 1494:15-27). , 2017). CpG 1018(5'-TGACTGTGAA CGTTCGAGAT G A-3' (as indicated by Sequence ID No. 1) demonstrates in vitro and in vivo immunostimulatory activity. Selected after screening a broad panel of oligonucleotides for CpG. 1018 is active Cp in mice, rabbits, dogs, baboons, cynomolgus monkeys, and humans. It is GB ODN. Therefore, in some preferred embodiments, the TLR9 ago NIST is an oligonucleotide containing the sequence of Sequence ID No. 1.

[0031] CpG 1018, an exemplary oligonucleotide TLR9 agonist, is CpG- Although this disclosure is ODN, it is not limited to complete DNA molecules. That is, several actual In terms of application form, TLR9 agonists have CpG and palindromic sequences that are deoxyribonucleic acid. DNA / RNA chimeric molecules in which one or more nucleic acids outside these regions are ribonucleic acids. It is a child. In some embodiments, the CpG oligonucleotide is linear. In other embodiments, the CpG oligonucleotide is cyclic or hairpin It contains CpG oligonucleotides. CpG oligonucleotides can be single-stranded or double-stranded.

[0032] In some embodiments, the CpG oligonucleotide may contain modifications. The modifications are not limited to those involving 3'OH or 5'OH groups, or nucleation. Modifications include modifications of rheotide bases, modifications of sugar components, and modifications of phosphate groups. The modified base is related to its natural complement through Watson-Crick base pairing. As long as the same specificity is maintained (for example, the palindromic part remains self-complementary), It may be included in the palindromic sequence of CpG oligonucleotides. The CpG oligonucleotide contains a non-canonical base. In some embodiments, C The pG oligonucleotide contains a modified nucleoside. In some embodiments, the modified The nucleosides are 2'-deoxy-7-deazaguanosine and 2'-deoxy-6-thio Guanosine, Arabinoguanosine, 2'-deoxy-2'-substituted-arabinoguanosine, Selected from the group consisting of 2'-O-substituted arabinoguanosine. Several implementation forms In this state, the TLR9 agonist has the sequence 5'-TCG1AACG1TTCG1-3'( The oligonucleotide containing (SEQ ID NO: 2), where G1 is 2'-deoxy-7- It is deazaguanosine. In some embodiments, the oligonucleotide is sequence 5 Includes '-TCG1AACG1TTCG1-X-G1CTTG1CAAG1CT-5', Here, G1 is 2'-deoxy-7-deazaguanosine and X is glycerol. It exists (5'-sequence number 2-3'-X-3'-sequence number 2-5').

[0033] CpG oligonucleotides may contain modifications to the phosphate group. For example, phospho In addition to diester linkage, phosphate modification is not limited to methyl phosphate. Fonate, phosphorothioate, phosphoramidate (crosslinked or uncrosslinked), phosphotri Examples include esters and phosphorodithioates, which may be used in any combination. In addition, other non-phosphoric acid linkages may be used. In some embodiments, oligonucleotides The rheotide contains only the phosphorothioate skeleton. In some embodiments, oligo The nucleotides consist only of the phosphodiester skeleton. In some embodiments, the original A nucleotide is a combination of phosphate links in a phosphate backbone, for example, a phosphodiester and Includes combinations of phosphorothioate linkages. Oligonucleotides having a phosphorothioate skeleton. Rheotides may be more immunogenic than those with a phosphodiester skeleton, and to the host It appears to be more resistant to degradation after injection (Braun et al., J Immunol, 141:208). 4-2089, 1988; and Latimer et al., Mol Immunol, 32:1057-1064, 1995). C of this disclosure pG oligonucleotides contain at least one, two, or three internucleotide phosphonucleotides. Contains rothioate ester linkage. In some embodiments, a plurality of CpG oligonucleotides When a cleotide molecule is present in a pharmaceutical composition containing at least one excipient, phospho Both stereoisomers of the rothioate ester linkage are found in multiple CpG oligonucleotide molecules. It is present in some embodiments between the nucleotides of CpG oligonucleotides. All of the linkages are phosphorothioate linkages, or CpG linkages in the same manner as described above. Ligonucleotides have a phosphorothioate skeleton.

[0034] Typically, the unit dose of an immunogenic composition, which is 0.5 ml, is approximately 375 μg to approximately 60 00 μg of CpG oligonucleotide, preferably about 750 μg to about 3000 μg of C It may contain pG oligonucleotides. In some embodiments, 0.5 ml The immunogenic composition is available in doses of approximately 250 μg, 500 μg, 750 μg, 1000 μg, and This is more than 1250 μg of CpG oligonucleotides, approximately 6000 μg, 5000 μg, Contains 4000 μg, 3000 μg, or less than 2000 μg of CpG oligonucleotides. In some embodiments, a 0.5 ml dose of the immunogenic composition contains approximately 375 μg. 750 μg, 1500 μg, 3000 μg, or 6000 μg CpG oligonucleotides Contains thiosulfate. In some embodiments, a 0.5 ml dose of the immunogenic composition contains about 75 Contains 0 μg of CpG oligonucleotide. In some embodiments, for 0.5 ml The immunogenic composition contains approximately 1500 μg of CpG oligonucleotide. In this embodiment, a 0.5 ml dose of immunogenic composition contains approximately 3000 μg of CpG oligonucleotides. Contains nucleotides.

[0035] The CpG oligonucleotides described herein, unless otherwise indicated, This is a pharmaceutically acceptable form of salt. Exemplary basic salts include ammonium salts and sodium salts. Alkali metal salts such as sodium, lithium, and potassium salts, calcium, and magnesium Alkaline earth metal salts such as um salts, zinc salts, N-Me-D-glucamine, N-[1-( 2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride Organic bases such as choline, tromethamine, dicyclohexylamine, and t-butylamine (For example, salts with organic amines, as well as salts with amino acids such as arginine and lysine.) Examples include: In some embodiments, the CpG oligonucleotide is ammonia It is in the form of a salt of ammonium, sodium, lithium, or potassium. In one preferred embodiment... In this context, CpG oligonucleotides exist in the form of sodium salts.

[0036] B. Varicella-zoster virus (VZV) glycoprotein E (gE) antigen The VZV gE antigen of the immunogenic composition of this disclosure comprises gE or a fragment thereof. Preferred In the embodiment, the gE antigen is recognized by a VZV-reactive antibody, and / or g The peptide fragment of E is recognized by VZV-reactive T cells. In some embodiments... In this embodiment, the gE antigen is a recombinant protein, but in other embodiments, the gE antigen is V It is purified from ZV virion. In some preferred embodiments, the gE antigen is It is an isolated antigen. In some embodiments, the VZV gE antigen is full-length VZV cleaved recombinant gE antigen lacking signal peptide, transmembrane domain, and cytoplasmic domain It is a protein. The typical amino acid sequence of gE is GenBank number AQT341. It is described as 20.1. In some embodiments, the gE antigen is GenBa The amino acid sequence from residues 39-585 of nk number AQT34120.1, or a small portion thereof. At the very least, the amino acid composition is 90%, 95%, 96%, 97%, 98%, or 99% identical. Includes columns.

[0037] In some embodiments, the amino acid sequence of gE is described as SEQ ID NO: 4. ru. MGTVNKPVVG VLMGFGIITG TLRITNPVRA SVLRYDDFHI DEDKLDTNSV YEPYYHSDHA ESSWVNRGES SRKAYDHNSP YIWPRNDYDG FLENAHEHHG VYNQGRGIDS GERLMQPTQM SAQEDLGDDT GIHVIPTLNG DDRHKIVNVD QRQYGDVFKG DLNPKPQGQR LIEVSVEENH PFTLRAPIQR IYGVRYTETW SFLPSLTCTG DAAPAIQHIC LKHTTCFQDV VVDVDCAENT KEDQLAEISY RFQGKKEADQ PWIVVNTSTL FDELELDPPE IEPGVLKVLR TEKQYLGVYI WNMRGGSDGTS TYATFLVTWK GDEKTRNPTP AVTPQPRGAE FHMWNYHSHV FSVGDTFSLA MHLQYKIHEA PFDLLLEWLY VPIDPTCQPM RLYSTCLYHP NAPQCLSHMN SGCTFTSPHL AQRVASTVYQ NCEHADNYTA YCLGISHMEP SFGLILHDGG TTLKFVDTPE SLSGLYVFVV YFNGHVEAVA YTVVSTVDHF VNAIEERGFP PTAGQPPATT KPKEITPVNP GTSPLIRYAA WTGGLA In some embodiments, the gE antigen is the amino acid sequence of SEQ ID NO: 4, or SEQ ID NO: Ami is at least 90%, 95%, 96%, 97%, 98%, or 99% identical to 4. Contains an acid sequence.

[0038] In some embodiments, the amino acid sequence of gE is described as SEQ ID NO: 5. ru. MGTVNKPVVG VLMGFGIITG TLRITNPVRA SVLRYDDFHT DEDKLDTNSV YEPYYHSDHA ESSWVNRGES SRKAYDHNSP YIWPRNDYDG FLENAHEHHG VYNQGRGIDS GERLMQPTQM SAQEDLGDDT GIHVIPTLNG DDRHKIVNVD QRQYGDVFKG DLNPKPQGQR LIEVSVEENH PFTLRAPIQR IYGVRYTETW SFLPSLTCTG DAAPAIQHIC LKHTTCFQDV VVDVDCAENT KEDQLAEISY RFQGKKEADQ PWIVVNTSTL FDELELDPPE IEPGVLKVLR TEKQYLGVYI WNMRGGSDGTS TYATFLVTWK GDEKTRNPTP AVTPQPRGAE FHMWNYHSHV FSVGDTFSLA MHLQYKIHEA PFDLLLEWLY VPIDPTCQPM RLYSTCLYHP NAPQCLSHMN SGCTFTSPHL AQRVASTVYQ NCEHADNYTA YCLGISHMEP SFGLILHDGG TTLKFVDTPE SLSGLYVFVV YFNGHVEAVA YTVVSTVDHF VNAIEERGFP PTAGQPPATT KPKEITPVNP GTSPLLR In some embodiments, the gE antigen is the amino acid sequence of SEQ ID NO: 5, or SEQ ID NO: 5. Ami is at least 90%, 95%, 96%, 97%, 98%, or 99% identical to 5. Contains an acid sequence.

[0039] Typically, the unit dose of an immunogenic composition is 0.5 ml, ranging from approximately 10 μg to approximately 100 μg. μg of gE antigen, preferably about 25 μg to about 75 μg of gE antigen, preferably about 40 It may contain μg to approximately 60 μg of gE antigen, or approximately 50 μg of gE antigen.

[0040] C. Additional Components The immunogenic compositions of this disclosure include one or more additional components, for example, one or more. It may contain several excipients, another adjuvant, and / or additional antigens.

[0041] 1. Excipients Examples of pharmaceutically acceptable excipients in this disclosure include solvents, bulking agents, buffering agents, and osmotic agents. Examples include pressure regulators and preservatives (Pramanick et al., Pharma Times, 45:65-77, 20 13) In some embodiments, the immunogenic composition includes a solvent, a bulking agent, a buffering agent, and It may also contain excipients that function as one or more osmotic pressure regulators (for example, food Sodium chloride in saline solution can function as both an aqueous medium and an osmotic regulator. (ru).

[0042] In some embodiments, the immunogenic composition comprises an aqueous medium as a solvent. Examples of media include sterile water, saline solution, phosphate-buffered saline, and Ringer's solution. In some embodiments, the composition is isotonic.

[0043] The immunogenic composition may contain a buffering agent. The buffering agent is used to control the pH during processing. It inhibits the degradation of the active agent during storage and, optionally, during reconstitution. Examples include salts containing acetate, citrate, phosphate, or sulfate. Suitable buffering agents include, for example, arginine, glycine, histidine, and lysine. The amino acids are listed below. The buffering agent further contains hydrochloric acid or sodium hydroxide. It is also possible. In some embodiments, the buffering agent maintains the pH of the composition within the range of 6 to 9. It maintains. In some embodiments, the pH is higher (lower limit) than 6, 7, or 8. In some embodiments, the pH is less than 9, 8, or 7 (upper limit). That is, pH is in the range of approximately 6 to 9, with the lower limit being less than the upper limit.

[0044] The immunogenic composition may contain an osmotic regulator. Suitable osmotic regulators include: For example, dextrose, glycerol, sodium chloride, glycerin, and mannitol One example is [the rule].

[0045] The immunogenic composition may contain a volume extender. The volume extender is frozen before administration of the pharmaceutical composition. It is particularly useful when drying. In some embodiments, the bulking agent is frozen or This helps stabilize the active ingredient and prevent its degradation during spray drying and / or storage. It is a protective agent. Suitable fillers include sugars (monosaccharides, disaccharides, and polysaccharides), such as sucrose. Lactose, trehalose, mannitol, sorbitol, glucose, and raffia It is North.

[0046] The immunogenic composition may contain a preservative. Suitable preservatives include, for example, acid-fast preservatives. Examples include chemical agents and antimicrobial agents. However, in preferred embodiments, immunogenic compounds are used. The product is prepared under sterile conditions and is contained in single-use containers; therefore, the inclusion of preservatives is not necessary. It is not considered to be so.

[0047] 2. Additional adjuvants Adjuvants are known in the art and are not limited to those mentioned above. Van (aluminum salt), oil-in-water emulsion, water-in-oil emulsion, liposomes, and fine particles, such as poly(lactide-co-glycoside) fine particles, are examples. Shah et al., Methods Mol Biol, 1494:1-14, 2017). In some embodiments, immunity The epidemiogenic composition further comprises an aluminum salt adjuvant on which gE antigen is adsorbed. In several embodiments, the aluminum salt adjuvant is amorphous hydroxyphosphate Aluminum sulfate, aluminum hydroxide, aluminum phosphate and aluminum sulfate It comprises one or more of the group consisting of mucurium. In some embodiments, aluminum Aluminum salt adjuvants are either aluminum hydroxide or aluminum phosphate, or both. It includes. In some embodiments, the aluminum salt adjuvant is aluminum hydroxide. It consists of um. In some embodiments, the unit dose of the immunogenic composition (e.g., about 0 0.5 ml contains approximately 0.25 to 0.50 mg of Al 3+ Preferably, about 0.30 to about 0 0.40mg Al 3+ Includes.

[0048] In other embodiments, the immunogenic composition further comprises additional adjuvants. Suitable adjuvants are not limited to, but include, underwater squalene emulsion. (For example, MF59 or AS03), TLR3 agonist (for example, Poly-IC or Poly-ICLC), TLR4 agonist (e.g., saponin, e.g., AS01 or A) Quil A or QS-21 in S02, TLR5 agonists (bacterial fragile Jerin), as well as TLR7 and / or TLR8 agonists (imidazoquinoline inducers). Examples of conductors include imiquimods and resikimods (Coffman et al., Immunity). (33:492-503, 2010). For veterinary use and for antibody production in non-human animals. To that end, we use the Freund adjuvant (both complete and imperfect) schism-promoting components. It is possible.

[0049] D. Kit This disclosure relates to i) VZV gE antigen and Toll-like receptor 9 (TLR9) agonists, For example, immunogenic compositions containing CpG oligonucleotides; and ii) immunogenic compositions Administering this to mammalian subjects, such as humans who require it, against gE antigens The kit also includes a set of instructions for use to stimulate an immune response. i) a first composition containing VZV gE antigen, ii) a TLR9 agonist, for example, Cp iii) A second composition containing a G oligonucleotide, and mixing the first composition with the second composition. Instructions for use for preparing immunogenic compositions, and optionally iv) immunogenic compositions The product is administered to mammals, such as humans who require it, against gE antigens. The kit provides an additional set of instructions for use to stimulate an immune response. In that embodiment, the CpG oligonucleotide has the sequence 5'-AACGTTCG-3 'Includes. In some embodiments, the CpG oligonucleotide has sequence 5'-AA Contains CGTTCGAG-3' (SEQ ID NO: 3). In some preferred embodiments, CpG oligonucleotides are 5'-TGACTGTGAA CGTTCGAGAT G Contains the sequence A-3' (sequence number 1).

[0050] The kit may include appropriately packaged immunogenic compositions. For example, an immunogenic group If the product is a freeze-dried powder, the powder is injected through an elastic stopper into the liquid (e.g., sterile water). Vials with elastic stoppers are usually designed to allow for easy resuspension with saline solution, etc. It is used. In some embodiments, the kit is a device for administration (e.g., Includes syringes and needles. Instructions for use regarding the use of immunogenic compositions are generally intended for use by... This includes information on dosage, schedule, and route of administration for the intended use. In some embodiments, the immunogenic composition stimulates an immune response to VZV. It is a food item.

[0051] II. How to Use This disclosure relates to a method for stimulating an immune response to VZV, wherein the VZV glycoprotein E(gE) antigens and Toll-like receptor 9 (TLR9) agonists, such as CpG antigens. An immunogenic composition containing gnucleotides is used to respond to the immune response to gE antigen in mammals. The present invention relates to a method involving administration to a mammalian subject to stimulate a response. Preferred implementation In this state, the immunogenic composition is administered by intramuscular injection in a volume of approximately 0.5 mL, optionally. For example, it should be administered in a unit dose. In some embodiments, it is administered by intramuscular injection. This is for the deltoid muscle of the upper arm in humans who require it.

[0052] Stimulating the immune response means increasing the immune response, which is a de novo immune response. To induce an epidemic response (e.g., as a result of an initial vaccination regimen), or existing To enhance the immune response (for example, as a result of a booster vaccination regimen) This can occur. In some embodiments, stimulating an immune response is not limited to However, it does not involve stimulating cytokine production, stimulating B lymphocyte proliferation, and antibodies. Stimulating production, stimulating interferon pathway-related gene expression, chemotaxis By stimulating the expression of plasma-related genes and the maturation of plasmacytoid dendritic cells, The group includes one or more of the following. In some preferred embodiments, the immune response Stimulation involves increasing the antigen-specific antibody response in the target.

[0053] List of embodiments 1. Immunogenic compositions for stimulating an immune response to varicella-zoster virus (VZV) The VZV glycoprotein E (gE) antigen and Toll-like receptor 9 (TLR9) It contains a TLR9 agonist, and the TLR9 agonist is unmethylated cytidine-phospho-guanosine (Cp G) An oligonucleotide of 10-35 nucleotides in length containing the motif, gE antigen Oligonucleotides stimulate the immune response to gE antigens in mammals. An immunogenic composition present in an effective amount for that purpose. 2. The oligonucleotide contains the sequence 5'-AACGTTCGAG-3' (SEQ ID NO: 3) The composition described in Embodiment 1. 3. The oligonucleotide is 5'-TGACTGTGAA CGTTCGAGAT GA The composition according to Embodiment 1, comprising the sequence -3'(SEQ ID NO: 1). 4. The oligonucleotide contains a modified nucleoside, and optionally the modified nucleoside is , 2'-deoxy-7-deazaguanosine, 2'-deoxy-6-thioguanosine, ara Binoguanosine, 2'-deoxy-2'-substituted arabinoguanosine, and 2'-O-substituted One of the embodiments 1 to 3, selected from the group consisting of conversion-arabinoguanosine. The composition of the material. 5. Oligonucleotides have the sequence 5'-TCG1AACG1TTCG1-3' (Sequence ID) 2) including, where G1 is 2'-deoxy-7-deazaguanosine, optional And the oligonucleotide has the sequence 5'-TCG1AACG1TTCG1-X-G1CTT It contains G1CAAG1CT-5', where G1 is 2'-deoxy-7-deazaguano It is syn, and X is glycerol (5'-SEQ ID NO: 2-3'-X-3 2-5'), the composition according to Embodiment 4. 6. The oligonucleotide contains at least one phosphorothioate linkage, optionally In Embodiments 1 to 5, all nucleotide linkages are phosphorothioate linkages. The composition described in any one of the following. 7. Embodiments 1-6 in which the oligonucleotide is a single-stranded oligodeoxynucleotide A composition as described in any one of the following. An immunogenic composition in an 8.0.5 ml dose contains approximately 375 μg to 6000 μg of oligonucleotides. Contains either oxytocin or approximately 750 μg to 3000 μg of oligonucleotides, optional. , 0.5 ml doses of immunogenic composition contain approximately 375 μg, approximately 750 μg, and approximately 1500 μg. Embodiments 1 to 7 contain approximately 3000 μg or approximately 6000 μg of oligonucleotide. The composition described in any one of the following. 9. The VZV gE antigen is the signal peptide of the full-length VZV gE antigen, and the transmembrane domain It is a cleaved recombinant protein lacking a cytoplasmic domain, and can be optionally used as an anti-VZV gE agent. Hara, (i) Amino acid sequence of residues 39-585 of GenBank number AQT34120.1, Or at least 90%, 95%, 96%, 97%, 98%, or 99% identical to it. A certain amino acid sequence; or (ii) The amino acid sequence of SEQ ID NO: 4, or at least 90% or 95% of SEQ ID NO: 4, Amino acid sequences that are 96%, 97%, 98%, or 99% identical; or (iii) The amino acid sequence of SEQ ID NO: 5, or at least 90% to 95% of SEQ ID NO: 5 , amino acid sequences that are 96%, 97%, 98%, or 99% identical A composition according to any one of embodiments 1 to 8, comprising: A 10.0.5 ml dose of immunogenic composition contains approximately 25 μg to approximately 75 μg of gE antigen. Optionally, the immunogenic composition contains approximately 40 μg to 60 μg of gE antigen or approximately 50 μg A composition according to any one of Embodiments 1 to 9, comprising the gE antigen. 11. Any one of Embodiments 1 to 10 further comprises an aluminum salt adjuvant. The composition of the material. 12. The aluminum salt adjuvant is amorphous aluminum sulfate hydroxyphosphate. It consists of salt, aluminum hydroxide, aluminum phosphate, and potassium aluminum sulfate. The composition according to Embodiment 11, comprising one or more of the group. 13. The aluminum salt adjuvant comprises aluminum hydroxide, as described in Embodiment 11. The composition of. A 14.0.5 ml dose of immunogenic composition contains approximately 0.25 to 0.50 mg of Al 3+ Includes In addition, optionally, a 0.5 ml dose of immunogenic composition containing approximately 0.30 to 0.40 mg of A l 3+A composition according to any one of embodiments 11 to 13, including the composition described above. 15. The composition according to any one of Embodiments 1 to 14, wherein the mammalian subject is a human subject. thing. 16.i) The immunogenic composition described in any one of Embodiments 1 to 15, and ii) Administer the composition to stimulate an immune response to gE antigen in mammalian subjects. Instructions for use A kit that includes this. 17.i) A first composition comprising the glycoprotein E(gE) antigen of the varicella-zoster virus, ii) A second composition comprising a TLR9 agonist, and iii) Mixing the first composition with the second composition to prepare an immunogenic composition Instructions for use A kit that includes this. 18. iv) Immunogenic composition administered to mammalian subjects to improve immune response to gE antigen Further set of user instructions to stimulate the answer The kit according to Embodiment 17, further comprising: 19. Embodiment 16 further includes a syringe and needle for intramuscular injection of an immunogenic composition. Kits listed in any one of the ~18 items. 20. Stimulate the immune response to varicella-zoster virus (VZV) in mammals. A method for doing so, wherein the immunogenic composition described in any one of Embodiments 1 to 15 is used To stimulate the immune response to gE antigen in mammals, administer to mammals. A method that includes doing so. 21. The method according to Embodiment 20, wherein the immunogenic composition is administered by intramuscular injection. [Examples]

[0054] Abbreviations: CpG (unmethylated cytidine-phosphoguanosine); CTRL (control); gE (VZV glycoprotein E); LAV (attenuated live virus); mcg (microgram); m cl (microliter); MPL (monophosphoryl lipid A); TLR9 (Thor's Receiver) Symptom 9); and VZV (varicella-zoster virus). [Examples]

[0055] Immunogenicity of CpG adjuvant herpes zoster vaccine in mice This example uses a weakened varicella-zoster virus (VZV) strain (hereinafter referred to as LAV). A preclinical study to evaluate the immunogenicity of herpes zoster vaccine in mice infected with [unspecified pathogen]. Provides clarity.

[0056] Antigen: Recombinant VZV glycoprotein E (gE) has a transmembrane anchor and carboxyl terminus. Genetically modified Chinese hamsters expressing a cleaved version of gE lacking a domain. It is obtained by cell culture of ovarian host cells and is therefore secreted into the supernatant. Cell culture It contains no albumin, antibiotics, or animal-derived proteins, but contains amino acids. Supported by culture medium. gE is purified by chromatography and described Freeze-dried for future use (Haumont et al., Virus Res, 40:199-204, 1996). VZV gE was commercially available for the initial preclinical study.

[0057] gE is either an adjuvant, a pharmaceutically acceptable buffer, or physiological saline. The vaccine was prepared before use by mixing with [a specific substance]. Specifically, gE was mixed before injection. It was adsorbed onto alum (aluminum hydroxide) for 30 minutes, and AS01B and CpG 1 Adjuvant 018 was mixed with gE immediately before injection.

[0058] Experimental design: The adjuvant activity of CpG 1018 was measured at 35 minutes before the start of the immunization schedule. A few days prior (-35 days) a live attenuated VZV vaccine (500 mcl administered via subcutaneous route) was administered. 10 inside 4 C57BL / 6 mice primed with plaque-forming units (LAV) (n=4 Evaluation was performed in groups ~8). The LAV used was from Merck & Co., Inc. VARIVA (Sold by Whitehouse Station, NJ) X (registered trademark) was a live varicella virus vaccine. Mice supported sufficient replication of VZV. Since there is no existing example, this was done to mimic existing VZV infections in humans. Mice were used. On day 0 and day 28, 50 mcl of the vaccine formulation or Immunization was performed using a saline control. The groups, dosages, and sample collections are listed in Table 1-1.

[0059] [Table 1]

[0060] VZV gE contains both B cell and T cell epitopes. Therefore, gE Serum antibodies that respond to this are produced by the first dose of the vaccine (day 21) or the second dose (day 56). After administration of ), use a commercially available EL plate coated with VZV gE antigen. Measurements were taken using ISA. CD3 + CD4 + T cell cytokine response (IL-2 and I) In vitro re-insertion of spleen cell cultures with a duplicated peptide (FN-γ) extending to the length of the gE antigen. After stimulation, measurements were taken by flow cytometry. Three spleen cell subpools were selected per group. Tested (3, 3 or 2 mouse spleens / pool). T cell response, CD4+ Percentage of T cells as, or CD154 + CD4 + evaluated as the percentage of T cells, where CD154 is a molecule transiently upregulated in antigen-stimulated cells, and thus serves as a specific marker for antigen-reactive CD4 + T cells (i.e., cells that respond to gE peptide stimulation ex vivo). Mice were monitored for health parameters including injection site reactions and systemic appearance. All adjuvant groups tested induced high levels of anti-VZV IgG responses after prime and booster vaccinations. On day 21 after the first gE immunization (prime), the anti-VZV IgG response was elevated compared to the background levels observed in non-immunized mice and mice primed with LAV but not further immunized.

[0061] The addition of the adjuvant aluminum hydroxide (alum) alone or CpG 1018 alone increased the anti-VZV IgG response compared to immunization with gE antigen alone. The highest anti-VZV IgG response after the first immunization was observed in mice immunized with CpG-1018 + alum (Figure 1A). The response of the gE + CpG 1018 + alum immunized group was significantly higher than that in mice immunized with gE in combination with CpG-1018 alone or gE + AS01b. The anti-VZV IgG response was observed to be approximately 1 log higher in all gE ± adjuvant groups on day 56 after the second immunization (boost) (Figure 1B), at which point equivalent responses were observed in gE + AS01b and ​​​​​​​​​​​​It was observed in the gE+CpG 1018+alum group and immunized with gE+AS01b. The response in these mice was greater than in mice immunized with gE+CpG 1018. It was highly anticipated (ANOVA with Tukey multiple comparison test, *** P<0.001, ** ** P < 0.0001).

[0062] Ex vivo intracellular cytokine expression response in spleen cells stimulated with gE peptide gE antigen-specific CD4 + Quantitative measurement of the effect of immunotherapy regimens on T cell response Cytokine expression stimulated by gE was modified across all CD4 + As a percentage of T cells and antigen-reactive population (CD154 + CD4 + Measured as a percentage of T cells. IL-2 and IFN-γ expression were observed in the control (CTL), LAV prime only, and gE. CD4 from the alum adjuvant group + In T cells, they are either absent or The levels were very low. In contrast, AS01b or CpG as adjuvants The use of 1018 was observed in IL-2 (Figure 2A) and IFN as measured after ex vivo restimulation. -γ expression (Figure 2B) induced a similar increase in T cells. IL-2 and IFN-γ expression were From the gE+CpG 1018+alum immunization group, restimulated CD4 + T cells tend to be lower CD4+ T cells co-expressing IL-2 and IFN-γ were AS01b adjuvants. Cells from band mice showed the highest results, followed by those from CpG 1018 adjuvant mice and The cells were from CpG 1018+ alum-adjuvant mice (Figure 2C). Response The pattern is from AS01b and CpG 1018 adjuvant mice, gE peptide Antigen reactivity (CD154) with equivalent cytokine expression in restimulated T cells + )C D4 + Similarity was observed within the T cell population (Figures 3A-3B). IFN-γ expression was observed in AS01b adjuvants. Antigens from CpG 1018 adjuvant mice compared to T cells from banded mice. The tendency is higher in specific T cells, and both intracellular IL-2 and IFN-γ expression are elevated. In mice in which CpG 1018 + alum was used as an adjuvant for the gE response, It was lower than expected.

[0063] In summary, all adjuvants tested showed high efficacy in C57BL / 6 mice. Inducing Bell's anti-VZV IgG response, followed by gE + CpG 1018 + alum after initial immunization. In immunized mice, the highest levels were observed in gE+AS01b immunized mice after booster immunization. Although there was an antibody response, the levels induced with CpG 1018 + alum were comparable. CD4 in response to gE immunization. + T cell response after ex vivo gE peptide restorative stimulation Evaluate and identify all CDs in the antigen-reactive population by CD154 expression. 4 + When measured as a percentage of cells, CpG 1018 and AS01b The highest response was observed in the adjuvant group. This data is from the combination with alum. CpG 1018, either as an adjuvant or as an independent adjuvant, can be used to improve immunity. In mice, the mice induced antibody and T cell responses equivalent to those of AS01b. This was demonstrated. Since the T cell response is thought to be the main cause of the rapid increase in shingles, The data supports the use of CpG 1018 as an adjuvant in gE-based vaccines. It is maintained. CpG 1018 alone or in combination with alum is A Compared to S01b, this is intended to obtain a more favorable reactance profile.

[0064] The foregoing disclosure is described in some detail by examples and embodiments for the purpose of clarity and understanding. Although this has been done, it will be obvious to those skilled in the art that certain changes and modifications may be made. Therefore, the examples do not limit the scope of this disclosure as detailed by the appended claims. It should not be interpreted as meaning that it does so.

Claims

1. An immunogenic composition for stimulating an immune response to varicella-zoster virus (VZV) There are VZV glycoprotein E (gE) antigen and Toll-like receptor 9 (TLR9) agonists. It contains a TLR9 agonist, which is unmethylated cytidine-phosphoguanosine (CpG ) is an oligonucleotide 10 to 35 nucleotides long containing the motif, and the γE antigen and Oligonucleotides stimulate the immune response to geE antigens in mammals. An immunogenic composition present in an effective amount for the purpose of immunogenicity.

2. The oligonucleotide contains the sequence 5'-AACGTTCGAG-3' (SEQ ID NO: 3) The immunogenic composition according to claim 1.

3. The oligonucleotide has the sequence 5'-TGACTGTGAA CGTTCGAGAT G The immunogenic composition according to claim 1, comprising the sequence A-3' (SEQ ID NO: 1).

4. The oligonucleotide contains a modified nucleoside, and optionally, the modified nucleoside is 2'-deoxy-7-deazaguanosine, 2'-deoxy-6-thioguanosine, Arabic Noguanosine, 2'-deoxy-2' substituted-arabinoguanosine, and 2'-O-substituted - An immunogenic composition according to claim 1, selected from the group consisting of arabinoguanosine.

5. The oligonucleotide is sequence 5'-TCG １ AACG １ TTCG １ -3' (Sequence ID 2) ) including, here, G １ It is 2'-deoxy-7-deazaguanosine, and is available by choice. , the oligonucleotide is sequence 5'-TCG １ AACG １ TTCG １ -X-G １ CTTG １ CAAG １ including CT-5', where G １ is 2'-deoxy-7-deazaguanosine X is glycerol (5'-SEQ ID NO: 2-3'-X-3'-SEQ ID NO: 2) -5'), the immunogenic composition according to claim 4.

6. The oligonucleotide contains at least one phosphorothioate linkage, or The immunogenicity according to claim 3, wherein all nucleotide linkages are phosphorothioate linkages. composition.

7. The oligonucleotide is a single-stranded oligodeoxynucleotide, according to claim 6. Immunogenic composition.

8. A 0.5 ml dose of immunogenic composition contains approximately 375 μg to 6000 μg of oligonucleotides. Contains thiosulfate or approximately 750 μg to approximately 3000 μg of oligonucleotide, or 0. The immunogenic composition in 5 ml doses is approximately 375 μg, approximately 750 μg, approximately 1500 μg, and approximately 30 The immunogen according to claim 6, comprising 00 μg or about 6000 μg of oligonucleotide. sexual composition.

9. The VZV gE antigen, the signal peptide of the full-length VZV gE antigen, the transmembrane domain and The immunogenic composition according to claim 8, which is a cleaved recombinant protein lacking a cytoplasmic domain. thing.

10. A 0.5 ml dose of the immunogenic composition contains approximately 25 μg to approximately 75 μg of GE antigen, or Alternatively, the immunogenic composition contains approximately 40 μg to 60 μg of gE antigen or approximately 50 μg of gE antigen The immunogenic composition according to claim 9, comprising the original.

11. The immunogenic composition according to claim 10, further comprising an aluminum salt adjuvant.

12. The aluminum salt adjuvant is amorphous aluminum sulfate hydroxyphosphate. A group consisting of aluminum hydroxide, aluminum phosphate, and potassium aluminum sulfate. The immunogenic composition according to claim 11, comprising one or more types.

13. The immunoassay according to claim 11, wherein the aluminum salt adjuvant comprises aluminum hydroxide. genic composition.

14. A 0.5 ml dose of the immunogenic composition contains approximately 0.25 to 0.50 mg of Al ３＋ Does it include , or an immunogenic composition in a 0.5 ml dose containing approximately 0.30 to 0.40 mg of Al ３＋ of The immunogenic composition according to claim 11, including the immunogenic composition described in claim 11.

15. The immunogenic group according to any one of claims 1 to 14, wherein the mammalian target is a human target. Finished product.

16. i) The immunogenic composition according to claim 15, and ii) Administer an immunogenic composition to induce an immune response to GE antigen in mammalian subjects. Instructions for use to stimulate A kit that includes this.

17. i) A first composition comprising the glycoprotein E (gE) antigen of the varicella-zoster virus, ii) A second composition comprising a TLR9 agonist, and iii) Mixing the first composition with the second composition to prepare an immunogenic composition Instructions for use A kit that includes this.

18. iv) Administer an immunogenic composition to induce an immune response to GE antigen in mammalian subjects. Further set of instructions for use to stimulate The kit according to claim 17, further comprising:

19. Claim 18 further comprises a syringe and needle for intramuscular injection of an immunogenic composition. The kit.

20. To stimulate the immune response to varicella-zoster virus (VZV) in mammals A method for which the immunogenic composition according to any one of claims 1 to 15 is used in mammals To stimulate an immune response to geE antigens in mammalian subjects, administer to them. A method that includes this.

21. The method according to claim 20, wherein the immunogenic composition is administered by intramuscular injection.

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