Adjuvant and composition thereof

Abstract

Provided are: a use of a CBR2 agonist as an adjuvant; and an adjuvant composition.　This antibody induction enhancer contains, as an active ingredient, at least one selected from among (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol.

Description

Adjuvants and compositions thereof

[0001] The present invention relates to the use of cannabinoid receptor type 2 agonists as adjuvants and compositions thereof.

[0002] Cannabis, derived from the cannabis plant and commonly known as marijuana, has long been used to treat pain, anxiety, and loss of appetite. This effect is due to the active compounds contained in cannabis called cannabinoids. In the 1960s, the main active component, Δ ９ Tetrahydrocannabinol (THC) and cannabidiol, which do not exhibit psychotic symptoms such as euphoria, were discovered. In the 1990s, two types of G protein-coupled receptors were identified as the biological target molecules of THC and named cannabinoid receptor type 1 (CBR1) and type 2 (CBR2). Furthermore, anandamide, an endogenous ligand for these receptors, was discovered in 1992, and 2-arachidonoylglycerol in 1995. Cannabinoid receptors are now believed to play important physiological roles.

[0003] These cannabinoid receptors are widely distributed in the central nervous system, circulatory system, digestive system, liver, muscles, bones, and the spleen / immune system, with CBR1 being known to be primarily expressed in the central nervous system, including the brain, and CBR2 being primarily expressed in the spleen / immune system. Therefore, it has been reported that CBR1 suppresses the release of neurotransmitters upon stimulation by agonists, eliciting various pharmacological activities such as analgesia, orthostatic hypotension, and euphoria, while CBR2 is involved in the differentiation and regulation of inflammatory cells and immune cells. Because cannabinoid receptors thus possess a variety of physiological activities, they are targets for pharmaceutical development.

[0004] As CBR1 agonists, the following have been approved in Europe and the United States from the 1990s to the early 2000s: dronabinol, a capsule containing the natural agonist THC dissolved in sesame oil, which is used as an appetite stimulant for AIDS patients and as an antiemetic for cancer chemotherapy; nabilone, a synthetic derivative of THC, which is used as an antiemetic for cancer chemotherapy; and nabiximols, an oral spray containing THC as its main ingredient for the treatment of spasticity and pain in multiple sclerosis and cancer pain. However, these pharmaceuticals have the risk of central nervous system side effects such as orthostatic hypotension and euphoria, as well as dependency, and are therefore subject to regulations requiring restrictions on their use and strict drug management.

[0005] On the other hand, CBR2 is mainly expressed in the peripheral immune system, and CBR2 selective agonists are expected to be safe drugs without the risk of centrally mediated side effects like CBR1, and have been developed as therapeutic agents for systemic sclerosis, atopic dermatitis, osteoarthritis pain, post-herpetic pain, etc. However, although clear analgesic effects have been demonstrated in non-clinical trials such as inflammatory pain models, and a discrepancy with centrally mediated side effects has also been confirmed, due to differences in efficacy due to species differences, only limited CBR2 agonists such as GW842166X, S-777469, and JBT-101 have been clinically tested.

[0006] CBR2 has been reported to be involved in vaccine-induced immunity. Dotsey et al. reported that administering the CBR2 antagonist AM630 or the inverse agonist JTE907 immediately before vaccination enhanced specific antibody responses to vaccine antigens (Non-Patent Document 1). Furthermore, such activity was not observed with the CBR2 agonist AM1241, and the antibody response was reported to be almost the same as that of the control group. Thus, CBR2 is known to be involved in opposing pharmacological activities, such as promoting and suppressing inflammation, and it is believed that inhibiting CBR2 activity is necessary to enhance the efficacy of vaccines.

[0007] In general, vaccines may require adjuvants to improve immunogenicity. In particular, inactivated vaccines that have lost their infectivity and subunit vaccines such as recombinant proteins have low immunogenicity, and adjuvants may be used in combination to enhance the immune response to a level that can protect against infection. Vaccines distributed in Japan also use aluminum gel, monophosphoryl lipid (MPL), and AS01B, a mixed adjuvant combining MPL and QS-21 with liposomes, as adjuvants.

[0008] Dotsey E, Ushach I, Pone E, Nakajima R, Jasinskas A, Argueta DA, Dillon A, DiPatrizio N, Davies H, Zlotnik A, Crompton PD, Felgner PL. Transient Cannabinoid Receptor 2 Blockade during Immunization Heightens Intensity and Breadth of Antigen-specific Antibody Responses in Young and Aged mice. Sci Rep.2017 Feb 17;7:42584.

[0009] The present invention relates to the use of cannabinoid receptor type 2 (CBR2) agonists as adjuvants and provides adjuvant compositions.

[0010] The present inventors have conducted extensive research into substances that enhance vaccine efficacy and have found that a specific CBR2 agonist has an immunostimulatory effect that enhances antibody induction, and that its combined use with a vaccine can enhance the strength of vaccine antigen-specific immunity. This is completely unexpected, given the previous report that inhibiting CBR2 activity exerts an adjuvant-like effect on vaccines (see Non-Patent Document 1).

[0011] That is, the present invention relates to the following 1) to 8): 1) An antibody inducibility enhancer containing one or more active ingredients selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol. 2) An adjuvant composition containing one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol, and a pharmaceutically acceptable carrier. 3) A vaccine composition comprising one or more selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol, a pharmaceutically acceptable carrier, and a vaccine antigen. 4) The vaccine composition of 3), wherein the vaccine antigen is an influenza virus. 5) Use of one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol for the production of an antibody inducibility enhancer.6) Use of one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol for producing an adjuvant. 7) One or more selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol, for enhancing antibody inducibility. 8) A vaccine therapy comprising administering to an animal one or more selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol together with a vaccine antigen.

[0012] According to the present invention, it is possible to enhance the efficacy of vaccines using highly safe CBR2 agonists, and this can make a significant contribution to the pharmaceutical industry by creating high-quality prophylactic drugs.

[0013] IgG antibody titer against the A / Victoria / 1 / 2020 strain (A / H1N1) after a single administration on Day 21. IgG antibody titer against the B / Phuket / 3073 / 2013 strain (B / Yamagata lineage) after a single administration on Day 21. Antibody titers of each IgG subclass against the A / Victoria / 1 / 2020 strain (A / H1N1) after a single administration on Day 21. Antibody titers of IgG subclasses against the B / Phuket / 3073 / 2013 strain (B / Yamagata lineage) after a single administration on Day 21. Evaluation of NF-κB activity by SEAP reporter assay. IgG antibody titer against the A / Victoria / 1 / 2020 strain (A / H1N1) in evaluation of the adjuvant activity of HU-308.

[0014] Preferred embodiments of the present invention will be described in detail below, but the present invention is not limited to the following embodiments.

[0015] In the present invention, (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone is also referred to as GW-405833, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene is also referred to as β-caryophyllene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol is also referred to as HU-308, all of which are compounds known as CBR2 agonists. A CBR2 agonist refers to a substance that binds to CBR2, transmits the signal into cells, and exerts physiological activity via CBR2 activation, and other compounds that have been found include EHP-101, GW-842166X, and LY2828360. 2-Arachidonoylglycerol (2-AG) is also known as an endogenous cannabinoid that binds to CBR2.

[0016] GW-405833, β-caryophyllene, and HU-308 (hereinafter referred to as "compounds of the present invention") have the chemical structures shown in Table 1 below, and can be produced by known chemical synthesis methods or purchased as commercially available products.

[0017]

[0018] The compounds of the present invention can be used either alone or in combination of two or more in the antibody inducibility enhancer, adjuvant composition or vaccine composition of the present invention.

[0019] As shown in the Examples below, when mice were subcutaneously injected with a vaccine composition prepared by adding a compound of the present invention to an influenza virus split vaccine distributed in Japan, the titer of antigen-specific IgG induced was significantly higher than that of a group administered only with the vaccine. Furthermore, a comparison of the geometric mean titers (GMT) of IgG subclasses also showed that the addition of these CBR2 agonists significantly increased antibody titers of IgG2a and IgG2b in addition to IgG1. That is, the compounds of the present invention serve as antibody induction enhancers that enhance the strength of antigen-specific immunity in vaccine therapy involving the administration of an immunogen (antigen), and these compounds can be used as adjuvants. Furthermore, a composition containing the compounds of the present invention and a pharmaceutically acceptable carrier can be used as an adjuvant composition. Furthermore, the compounds of the present invention can be used to produce antibody induction enhancers or adjuvant compositions. Here, "enhancing antibody induction" refers to the effect of enhancing the strength of antigen-specific immunity, specifically, the effect of enhancing antigen-specific IgG antibody titers.

[0020] Such an antibody inducibility enhancer can be a pharmaceutical material or formulation for humans or animals that enhances the antibody inducibility of an antigen. The adjuvant composition can be a pharmaceutical formulation for humans or animals that enhances the antibody inducibility of an antigen, i.e., has adjuvant activity. Such pharmaceutical formulations can be prepared by appropriately combining the compound of the present invention with a pharmaceutically acceptable carrier, such as an excipient, binder, filler, disintegrant, lubricant, dispersant, buffer, flavoring, diluent, etc.

[0021] The antibody induction enhancer and adjuvant composition of the present invention can be administered in combination with an antigen, and may be administered simultaneously with, before, or after the administration of the antigen. The dose of the adjuvant composition of the present invention can be appropriately determined depending on the subject, administration method, administration form, and type of antigen.

[0022] The antibody induction enhancer and adjuvant composition of the present invention can be combined with an antigen to form a vaccine composition for administration to animals, including humans, and can be further formulated into an appropriate formulation by appropriately adding the above-mentioned pharmaceutically acceptable carrier. In the vaccine composition of the present invention, the compound of the present invention may be chemically bound to an antigen or other component, or may exist in a free molecular state.

[0023] Antigens include natural products purified from pathogens, or proteins or peptides artificially produced by techniques such as genetic recombination. Specific examples include virions (complete virus particles), incomplete virus particles, viral structural proteins, viral nonstructural proteins, whole pathogen cells, pathogen-derived proteins, glycoproteins, capsular polysaccharides, protective antigens, and neutralizing epitopes. These antigens include both infectious and non-infectious antigens (inactivated antigens). Inactivated antigens include, but are not limited to, antigens inactivated by physical (X-ray irradiation, UV irradiation, heat, ultrasound) or chemical (formalin, beta-propiolactone, binary ethyleneimine, mercury, alcohol, chlorine) procedures. From the perspective of safety, pathogen-derived antigens are preferably inactivated antigens derived from the viruses or pathogens listed above.

[0024] Examples of viruses include measles virus, rubella virus, mumps virus, poliovirus, rotavirus, norovirus, adenovirus, enterovirus, herpes virus, varicella virus, severe acute respiratory syndrome (SARS) virus, novel coronavirus (SARS-CoV-2), human immunodeficiency virus (HIV), human T-cell leukemia virus (HTLV-1), human papillomavirus, Japanese encephalitis virus, West Nile virus, yellow fever virus, dengue virus, Zika virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, respiratory syncytial virus, and influenza virus, with influenza virus being preferred.

[0025] Examples of pathogenic bacteria include Corynebacterium diphtheriae, Corynebacterium tetani, Bordetella pertussis, Neisseria meningitidis, Hemophilus influenzae type b, Streptococcus pneumoniae, Vibrio cholerae, Mycobacterium tuberculosis, and periodontal disease bacteria.

[0026] Dosage forms of the vaccine composition include, for example, liquids, suspensions, freeze-dried preparations, etc. Liquids include those dissolved in purified water or buffer solutions, etc. Suspensions include those suspended in purified water or buffer solutions together with methylcellulose, hydroxymethylcellulose, polyvinylpyrrolidone, gelatin, casein, etc. These preparations can contain commonly used absorption enhancers, surfactants, preservatives, stabilizers, moisture-proofing agents, solubilizers, etc., as needed.

[0027] The vaccine composition of the present invention may contain an adjuvant substance other than the compound of the present invention, as long as it does not impair the immunogenicity and safety of the vaccine. Examples of such adjuvant substances include aluminum salts, squalene emulsion, 3-O-deacylated-4'-monophosphoryl lipid A (MPL), CpG-ODN, and QS-21, which are used in already approved vaccines.

[0028] The amount of antigen contained in the vaccine composition of the present invention is not particularly limited as long as it is an amount sufficient to induce a specific antibody response, and can be appropriately determined taking into consideration the ratio with respect to the adjuvant composition used in combination. For example, when an influenza virus split antigen is used as the antigen, the content may be within the range of 1 to 60 μg HA (HA equivalent), which is the single dose administered, with 9 to 15 μg HA (HA equivalent) being more preferable. The concentration is a value obtained by measuring the concentration of HA protein using a test method established by WHO or national standards, such as a single radial immunodiffusion test.

[0029] Furthermore, the content of the compound of the present invention in the vaccine composition can be adjusted appropriately taking into consideration the antibody titer, but it may be contained within the range of, for example, 150 to 1600 μg, which is the single administration dose, and more preferably 300 to 800 μg.

[0030] The route of administration of the vaccine composition of the present invention is not particularly limited, and may be any of administration by injection (subcutaneous administration, intramuscular administration, intradermal administration, intravenous administration), oral administration, and parenteral administration (e.g., nasal administration, ocular administration, vaginal administration, sublingual administration, transdermal administration), and may be administered, for example, by dropping, spraying, or instilling into the nasal cavity.

[0031] The adjuvant composition or vaccine composition of the present invention can be administered to humans and mammals other than humans, preferably humans, including mice, rats, hamsters, guinea pigs, rabbits, pigs, cows, horses, goats, sheep, dogs, cats, rhesus monkeys, cynomolgus monkeys, orangutans, and chimpanzees.

[0032] The present invention will be specifically explained below with reference to examples, but the present invention is not limited to these examples.

[0033] <Test Compounds> The test compounds used in the following examples are shown below.・GW-405833 Chemical name: (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone Distributor: Tokyo Kasei Kogyo Co., Ltd. ・β-Caryophyllene Chemical name: (1R,1E,9S)-4,11,11-trimethyl-8-methylene-bicyclo[7.2.0]undec-4-ene Distributor: MedChemExpress LLC ・HU-308 Chemical name: 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3. 1.1]hept-2-ene-2-methanol Distributor: abcam ・EHP-101: Chemical name: (1'R,6'R)-3-(Benzylamine)-6-hydroxy-3'-methyl-4-pentyl-6'-(prop-1-en-2-yl)[1,1'-bi(cyclohexane)]-2',3,6-triene-2,5-dione Sold by: MedChemExpress LLC. GW-842166X Chemical name: 2-(2,4-dichloroanilino)-N-(tetrahydropyran-4-ylmethyl)-4-(trifluoromethyl)pyrimidine-5-carboxamide Sold by: Fujifilm Wako Pure Chemical Corporation. LY2828360 Chemical name: 8-(2-chlorophenyl)-2-methyl-6-(4-methylpiperazin-1-yl)-9-tetrahydropyran-4-yl-purine Sold by: MedChemExpress LLC ・2-AG Chemical name: 2-arachidonoyl glycerol Sold by: Tocris Bioscience ・O-1602 (GPR55 agonist) Chemical name: 5-Methyl-4-[(1R,6R)-3-methyl-6-(1-methylethenyl)-2-cyclohexen-1-yl]-1,3-benzenediol Sold by: abcam

[0034] Example 1 Evaluation of Adjuvant Activity of CBR2 Ligands EHP-101, GW-405833, β-Caryophyllene, GW-842166X, LY2828360, and the endogenous cannabinoid 2-arachidonoylglycerol (2-AG) were dissolved in dimethyl sulfoxide (DMSO) to a concentration of 30 mg / mL. Similarly, LY2828360 was dissolved in DMSO to a concentration of 20 mg / mL. The DMSO solutions of these CBR2 ligands were used as adjuvant solutions. Furthermore, stock solutions of influenza HA vaccines of the A / H1N1 (A / Victoria / 1 / 2020 strain) and B / Yamagata lineage (B / Phuket / 3073 / 2013 strain) were used as antigen solutions. The adjuvant solution, antigen solution, and phosphate buffered saline containing 0.1% polysorbate 80 were mixed to adjust the concentration of each CBR2 ligand to 0.4 mg and the antigen to 1 μg HA of each strain per 0.3 mL. Note that an adjuvant-free test substance was also prepared by further adding phosphate buffered saline containing 0.1% polysorbate 80 instead of the adjuvant solution, and this served as a control.

[0035] BALB / c mice (female, 4 weeks old) were subcutaneously administered 0.3 mL of each influenza vaccine containing CBR2 ligand or the control vaccine in the dorsal region (8 mice per group), and exsanguination was performed 3 weeks after administration. Serum was prepared from the collected blood by centrifugation, and the serum IgG (total IgG) titers that specifically bind to the A / Victoria / 1 / 2020 strain and the B / Phuket / 3073 / 2013 strain were measured. The titers of IgG subclasses specific to each influenza antigen were also measured.

[0036] The IgG titer for the A / Victoria / 1 / 2020 strain (A / H1N1) is shown in Figure 1. Compared to the control group without adjuvant, the IgG titer tended to be higher in the group administered with the vaccine containing EHP-101, GW-405833, and β-caryophyllene, and a significant increase was observed, particularly in the group administered with the vaccine containing GW-405833 (Mann-Whitney U test, p<0.05). Furthermore, as shown in Figure 2, the IgG titer for the B / Phuket / 3073 / 2013 strain (B / Yamagata lineage) tended to be higher in the group administered with the vaccine containing EHP-101, GW-405833, and β-caryophyllene, similar to the A / H1N1 group. Furthermore, for the B / Yamagata lineage, the vaccine containing β-caryophyllene showed significantly higher IgG titers than the control (Mann-Whitney U test, p<0.05). Therefore, adding GW-405833 and β-caryophyllene to the vaccine was thought to enhance vaccine-induced immunity. The results for each IgG subclass specific to influenza antigens are shown in Figure 3A (A / Victoria / 1 / 2020 strain (A / H1N1)) and Figure 3B (B / Phuket / 3073 / 2013 strain (B / Yamagata lineage)). As shown in Figures 3A and 3B, the addition of GW-405833 or β-caryophyllene to the vaccine increased not only the primarily induced IgG1, but also the induction of IgG2a and IgG2b, suggesting that these CBR2 agonists enhance the induction of various subclasses. Since the induction of Th1 is thought to be important in defense against influenza virus infection, increasing the induction of IgG2a, a Th1-restricted subclass, is expected to improve the vaccine's protective effect against infection.

[0037] Example 2 Evaluation of NF-κB activation by CBR2 ligand 1 × 10 ６GW-405833 or β-caryophyllene was added to Raw264.7 Reporter Cell Line (Novus biologicals, NBP2-26261) at a final concentration of 1, 10, 100, or 1000 μM. The cells were incubated in a 5% CO atmosphere. ２ After culturing for 24 hours at 37°C, the culture medium was collected. The collected culture medium was centrifuged at 12,000 x g for 30 seconds, and the alkaline phosphatase activity of the supernatant was measured. The Secreted Alkaline Phosphatase Reporter Gene Assay Kit, Luminometric, SensoLyte (AnaSpec, Inc.) was used for alkaline phosphatase measurement. Furthermore, a 10% FBS-containing DMEM medium was added as a negative control (NC), and Imiquimod (final concentration: 5 μg / mL), an agonist of Toll-like receptor 7, was added as a positive control (PC), and comparative evaluations were performed.

[0038] The results of alkaline phosphatase measurements are shown in Figure 4. In the groups treated with 1 to 1000 μM GW-405833 or β-caryophyllene, alkaline phosphatase levels equivalent to or lower than those in NC were detected, and it can be seen that these levels were extremely low compared to the Imiquimod-treated PC group. In the Raw264.7 Reporter Cell Line, a gene encoding secretory alkaline phosphatase was inserted downstream of the NF-κB transcription response element, and the fact that the amount of alkaline phosphatase in the culture supernatant was similar to that in NC suggests that these CBR2 ligands do not have the ability to activate NF-κB. Therefore, although the mechanism of the adjuvant activity of GW-405833 and β-Caryophyllene has not been clearly elucidated, it is presumed that they do not cross-react with receptors of the innate immune system, such as TLR, that promote the activation of NF-κB.

[0039] Example 3 Evaluation of the adjuvant activity of CBR2 ligand HU-308 As in Example 1, HU-308, a CBR2 ligand, and O-1602, a ligand for GPR55, which is thought to be a cannabinoid receptor, were each dissolved in DMSO to a concentration of 30 mg / mL. This solution was mixed with a stock solution of A / H1N1 (A / Victoria / 1 / 2020 strain) influenza HA vaccine and phosphate-buffered saline containing 0.1% polysorbate 80 to adjust the concentration of the CBR2 ligand or O-1602 to 25, 100, or 400 μg, and the influenza vaccine to 1 μg HA per 0.3 mL. BALB / c mice (female, 4 weeks old) were subcutaneously administered 0.3 mL of these test substances in the dorsal region (8 mice per group), and whole blood was collected 3 weeks after administration. Serum was prepared from the obtained blood by centrifugation, and the titer of IgG (total IgG) that specifically binds to the A / Victoria / 1 / 2020 strain in the serum was measured.

[0040] The results of IgG titer measurements are shown in Figure 5. Addition of the CBR2 ligand HU-308 resulted in a dose-dependent increase in IgG titer, with a significant increase at 400 μg compared to the control influenza vaccine antigen alone (Mann-Whitney U test, p<0.01). On the other hand, with the GPR55 ligand O-1602, the IgG titer was comparable to the control at any dose of 25 to 400 μg, and no adjuvant activity to enhance vaccine antibody induction was confirmed. Therefore, it was thought that a CBR2 ligand, which is expressed less in the central nervous system and more in peripheral immune tissues, would be a suitable target for adjuvants targeting cannabinoid receptors.

Claims

1. An antibody induction enhancer comprising one or more active ingredients selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol.

2. An adjuvant composition comprising one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol, and a pharma- ceutically acceptable carrier.

3. A vaccine composition comprising one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol, a pharma- ceutically acceptable carrier, and a vaccine antigen.

4. The vaccine composition according to claim 3, wherein the vaccine antigen is an influenza virus.

5. Use of one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol for the production of an antibody induction enhancer.

6. Use of one or more members selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol for producing an adjuvant.

7. One or more selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol for promoting antibody inducing ability.

8. A vaccine therapy comprising administering to an animal one or more selected from (2,3-dichlorophenyl)-[5-methoxy-2-methyl-3-(2-morpholin-4-ylethyl)indol-1-yl]methanone, (1R,4E,9S)-4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene, and 4-[4-(1,1-dimethylheptyl)-2,6-dimethoxyphenyl]-6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-methanol together with a vaccine antigen.