FPR2 agonist containing outer membrane vesicles derived from lactic acid bacteria

Abstract

The present invention provides FPR2 agonists. Specifically, an outer membrane vesicle derived from at least one type of lactic acid bacteria selected from the group consisting of the genus Lactobacillus, the genus Lactobacillus casei, the genus Lactobacillus plantarum, the genus Lactobacillus conjuncticus, the genus Lactococcus, and the subsp. Thermophilus of Streptococcus salivarius is compounded as an active ingredient of the FPR2 agonist, and a method for preparing the FPR2 agonist containing the outer membrane vesicle derived from the outer membrane vesicle derived from at least one type of lactic acid bacteria derived from the outer membrane vesicle derived from the at least one type of lactic acid bacteria.

Description

Technical Field

[0001] The present invention relates to an invention characterized in that outer membrane vesicles derived from lactic acid bacteria are used to stimulate FPR2. Background Art

[0002] Outer membrane vesicles are substances covered by lipid bilayer membranes released by various cells. Outer membrane vesicles can pass through the blood-brain barrier (non-patent literature 5 and 6). Outer membrane vesicles are also stable after being taken up by host cells (non-patent literature 7). Outer membrane vesicles with these characteristics are expected to be applied to drug delivery systems (non-patent literature 6 and 7).

[0003] Bacteria also release outer membrane vesicles. Outer membrane vesicles derived from lactic acid bacteria have physiological activities such as inhibiting the secretion of inflammatory mediators (IL-6, etc.), inducing differentiation into M2 macrophages, inhibiting inflammatory responses caused by macrophages, and enhancing the host's immune response to vancomycin-resistant enterococci (Patent Documents 1 to 6 and Non-Patent Documents 1 to 2).

[0004] In addition, it has been reported that outer membrane vesicles derived from Lactobacillus plantarum can induce IL-10 (Non-Patent Document 14), and outer membrane vesicles derived from Lactobacillus paracasei (old classification name: Lactobacillus paracasei) can reduce intestinal inflammatory response (Non-Patent Document 15).

[0005] Formyl Peptide Receptor (FPR) is a known receptor for the chemokine formylmethionyl peptide.

[0006] There are three types of FPR families in humans (FPR1, FPR2, and FPR3). There are various ligands for FPR, and the selectivity for each FPR varies depending on the ligand (Non-Patent Document 3).

[0007] Various substances that act as agonists or antagonists of FPR2 are known (Non-Patent Document 3).

[0008] FPR2 controls both the innate immunity performed by phagocytes and the acquired immunity performed by B cells and the like (Patent Document 7).

[0009] FPR2 plays an important role in host defense and inflammatory response, and is considered to be a target for drugs for treating inflammatory diseases and the like (Non-Patent Document 4).

[0010] It is known that FPR2 agonists are useful as therapeutic agents for diseases associated with FPR2 (inflammatory diseases, etc.) (Patent Document 7).

[0011] FPR2 (FPRL-1) is also a functional receptor for amyloid β42 (Aβ42), which is important in the pathophysiology of neurotoxicity associated with Alzheimer's disease (Non-Patent Document 8). It is said that the peptide humanin (HN) that binds to FPR2 can exert a neuroprotective effect by competitively inhibiting FPR2's access to Aβ42 (Non-Patent Document 8). The effect of humanin (HN) on FPR2 is stronger than that on FPR1 (Non-Patent Document 9).

[0012] FPR2 is expressed in colonic epithelial cells and mediates N-formyl peptide-dependent epithelial cell proliferation and regeneration. In addition, FPR2 plays an important role in maintaining homeostasis in the colon, mediating inflammation, and repairing the epithelium (Non-Patent Document 10).

[0013] The ligand of FPR2 (FPRL1) has a therapeutic effect on ulcerative colitis, which suggests that FPR2 is useful in the development of therapeutic drugs for chronic inflammatory bowel disease (Non-Patent Document 11).

[0014] It is also disclosed that agonists of FPR2 are beneficial in preclinical models of chronic inflammatory human diseases, including infections, psoriasis, dermatitis, inflammatory bowel disease, Crohn's disease, ocular inflammation, sepsis, pain, metabolism / diabetes, cancer, COPD, asthma and allergic diseases, cystic fibrosis, acute lung injury and fibrosis, rheumatoid arthritis and other joint diseases, Alzheimer's disease, renal fibrosis and organ transplantation (Patent Document 7 and Non-Patent Documents 12 and 13).

[0015] Prior art literature

[0016] Patent Literature

[0017] Patent Document 1: Japanese Patent Application No. 2021-516054

[0018] Patent Document 2: Japanese Patent Application No. 2021-513336

[0019] Patent Document 3: Japanese Patent Application No. 2021-502970

[0020] Patent Document 4: Japanese Patent Application No. 2021-501136

[0021] Patent Document 5: Japanese Patent Application No. 2018-511583

[0022] Patent Document 6: Japanese Patent Application No. 2013-530216

[0023] Patent Document 7: Japanese Patent Application No. 2021-516240

[0024] Non-patent literature

[0025] Non-patent literature 1: Urakami Foundation Research Report, Vol.24, p.46-52, 2017

[0026] Non-patent literature 2: BMC Microbiology, 17:66, 2017

[0027] Non-patent document 3: Molecules, 22, 455, 2017

[0028] Non-patent literature 4: NATURE COMMUNICATIONS, 11: 1208, 2020

[0029] Non-patent literature 5: Int. J. Mol. Sci., 21, 4407, 2020

[0030] Non-patent literature 6: Cell Biosci., 11: 142, 2021

[0031] Non-patent literature 7: Chemistry and Biology, Vol.54, No.11, p.812-819, 2016

[0032] Non-patent document 8: Front. Endocrinol., Volume 5, Article 210, December 2014

[0033] Non-patent document 9: Biochemical and Biophysical Research Communications 324 (2004) 255-261

[0034] Non-patent literature 10: J Clin Invest. 2013; 123(4): 1694-1704

[0035] Non-patent literature 11: Experimental & Molecular Medicine (2013) 45, e40

[0036] Non-patent literature 12: Eur. J. Pharmacol., 2015, 5, 49-63

[0037] Non-patent literature 13: Trends in Pharm. Sci., 2015, 36, 737-755

[0038] Non-patent literature 14: Scientific Reports, Vol. 12, Article number: 13330 (2022)

[0039] Non-patent literature 15: Experimental & Molecular Medicine, 52: 423-437 (2020) Summary of the invention

[0040] Problem that the invention aims to solve

[0041] It is generally believed that substances that agonize FPR2 can be used to treat diseases associated with FPR2.

[0042] Outer membrane vesicles have properties that are advantageous for the above-mentioned handling (such as excellent stability).

[0043] However, the outer membrane vesicles that can activate FPR2 are still unknown.

[0044] Therefore, a problem has been raised to provide outer membrane vesicles capable of stimulating FPR2.

[0045] Solutions for solving problems

[0046] As a result of intensive studies on this subject, the present inventors have found that outer membrane vesicles derived from a specific type of lactic acid bacteria can activate FPR2. The present invention has been made based on this finding.

[0047] That is, the present invention relates to the following [1] to

[36] .

[0048] [1] An FPR2 agonist comprising an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of:

[0049] Lactobacillus spp.,

[0050] Lactobacillus spp.,

[0051] Lactiplantibacillus spp.,

[0052] Combined with Lactobacillus spp.,

[0053] Lactococcus, and

[0054] Streptococcus salivarius subsp. thermophilus.

[0055] [2] The FPR2 agonist according to [1] above, wherein the lactic acid bacteria is at least one selected from the group consisting of:

[0056] Lactobacillus delbrueckii subsp.bulgaricus, Lactobacillus helveticus,

[0057] Lactobacillus acidophilus,

[0058] Lactobacillus rhamnosus,

[0059] Lactobacillus paracasei,

[0060] Lactiplantibacillus plantarum,

[0061] Ligilactobacillus salivarius,

[0062] Lactococcus lactis subsp. lactis,

[0063] Streptococcus salivarius subsp.thermophilus,

[0064] Lactobacillus gasseri,

[0065] Lactobacillus amylovorus,

[0066] Lactobacillus crispatus,

[0067] Lactobacillus johnsonii,

[0068] Lactobacillus casei, and

[0069] Lactiplantibacillus pentosus.

[0070] [3] The FPR2 agonist according to [1] or [2] above, wherein the outer membrane vesicle derived from lactic acid bacteria does not have FPR1 agonist activity.

[0071] [4] A food or beverage for stimulating FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0072] Lactobacillus spp.,

[0073] Lactobacillus spp.,

[0074] Lactiplantibacillus spp.,

[0075] Combined with Lactobacillus spp.,

[0076] Lactococcus, and

[0077] Streptococcus salivarius subsp. thermophilus.

[0078] [5] A pharmaceutical composition for treating a disease associated with FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0079] Lactobacillus spp.,

[0080] Lactobacillus spp.,

[0081] Lactiplantibacillus spp.,

[0082] Combined with Lactobacillus spp.,

[0083] Lactococcus, and

[0084] Streptococcus salivarius subsp. thermophilus.

[0085] [6] A cosmetic composition for stimulating FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0086] Lactobacillus spp.,

[0087] Lactobacillus spp.,

[0088] Lactiplantibacillus spp.,

[0089] Combined with Lactobacillus spp.,

[0090] Lactococcus, and

[0091] Streptococcus salivarius subsp. thermophilus.

[0092] [7] A consumer product composition for stimulating FPR2, comprising outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0093] Lactobacillus spp.,

[0094] Lactobacillus spp.,

[0095] Lactiplantibacillus spp.,

[0096] Combined with Lactobacillus spp.,

[0097] Lactococcus, and

[0098] Streptococcus salivarius subsp. thermophilus.

[0099] [8] The food or beverage described in [4] above, the pharmaceutical composition described in [5] above, the cosmetic composition described in [6] above, or the consumer product composition described in [7] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[0100] [9] Lactobacillus paracasei strain CP3526 with the deposit number NITE BP-03625.

[0101]

[10] A food or beverage for inducing IL-10 by stimulating FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0102] Lactobacillus spp.,

[0103] Lactobacillus spp.,

[0104] Lactiplantibacillus spp.,

[0105] Combined with Lactobacillus spp.,

[0106] Lactococcus, and

[0107] Streptococcus salivarius subsp. thermophilus.

[0108]

[11] A pharmaceutical composition for treating a disease associated with IL-10 by inducing IL-10 by stimulating FPR2, comprising as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of:

[0109] Lactobacillus spp.,

[0110] Lactobacillus spp.,

[0111] Lactiplantibacillus spp.,

[0112] Combined with Lactobacillus spp.,

[0113] Lactococcus, and

[0114] Streptococcus salivarius subsp. thermophilus.

[0115]

[12] A cosmetic composition for inducing IL-10 by stimulating FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0116] Lactobacillus spp.,

[0117] Lactobacillus spp.,

[0118] Lactiplantibacillus spp.,

[0119] Combined with Lactobacillus spp.,

[0120] Lactococcus, and

[0121] Streptococcus salivarius subsp. thermophilus.

[0122]

[13] A consumer product composition for inducing IL-10 by stimulating FPR2, comprising outer membrane vesicles derived from at least one lactic acid bacteria selected from the group consisting of:

[0123] Lactobacillus spp.,

[0124] Lactobacillus spp.,

[0125] Lactiplantibacillus spp.,

[0126] Combined with Lactobacillus spp.,

[0127] Lactococcus, and

[0128] Streptococcus salivarius subsp. thermophilus.

[0129]

[14] The food or beverage described in

[10] above, the pharmaceutical composition described in

[11] above, the cosmetic composition described in

[12] above, or the consumer product composition described in

[13] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[0130]

[15] A food or beverage for protecting the skin from ultraviolet damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0131] Lactobacillus spp.,

[0132] Lactobacillus spp.,

[0133] Lactiplantibacillus spp.,

[0134] Combined with Lactobacillus spp.,

[0135] Lactococcus, and

[0136] Streptococcus salivarius subsp. thermophilus.

[0137]

[16] A pharmaceutical composition for protecting the skin from ultraviolet damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0138] Lactobacillus spp.,

[0139] Lactobacillus spp.,

[0140] Lactiplantibacillus spp.,

[0141] Combined with Lactobacillus spp.,

[0142] Lactococcus, and

[0143] Streptococcus salivarius subsp. thermophilus.

[0144]

[17] A cosmetic composition for protecting the skin from ultraviolet damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0145] Lactobacillus spp.,

[0146] Lactobacillus spp.,

[0147] Lactiplantibacillus spp.,

[0148] Combined with Lactobacillus spp.,

[0149] Lactococcus, and

[0150] Streptococcus salivarius subsp. thermophilus.

[0151]

[18] A consumer product composition for protecting the skin from ultraviolet damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0152] Lactobacillus spp.,

[0153] Lactobacillus spp.,

[0154] Lactiplantibacillus spp.,

[0155] Combined with Lactobacillus spp.,

[0156] Lactococcus, and

[0157] Streptococcus salivarius subsp. thermophilus.

[0158]

[19] The food or beverage described in

[15] above, the pharmaceutical composition described in

[16] above, the cosmetic composition described in

[17] above, or the consumer product composition described in

[18] above, which protects the skin from ultraviolet damage by stimulating FPR2.

[0159]

[20] The food or beverage described in

[15] above, the pharmaceutical composition described in

[16] above, the cosmetic composition described in

[17] above, or the consumer product composition described in

[18] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[0160]

[21] A food or beverage for protecting nerve cells from neurotoxic damage, comprising outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0161] Lactobacillus spp.,

[0162] Lactobacillus spp.,

[0163] Lactiplantibacillus spp.,

[0164] Combined with Lactobacillus spp.,

[0165] Lactococcus, and

[0166] Streptococcus salivarius subsp. thermophilus.

[0167]

[22] A pharmaceutical composition for protecting nerve cells from neurotoxic damage, comprising outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0168] Lactobacillus spp.,

[0169] Lactobacillus spp.,

[0170] Lactiplantibacillus spp.,

[0171] Combined with Lactobacillus spp.,

[0172] Lactococcus, and

[0173] Streptococcus salivarius subsp. thermophilus.

[0174]

[23] A cosmetic composition for protecting nerve cells from neurotoxic damage, comprising outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0175] Lactobacillus spp.,

[0176] Lactobacillus spp.,

[0177] Lactiplantibacillus spp.,

[0178] Combined with Lactobacillus spp.,

[0179] Lactococcus, and

[0180] Streptococcus salivarius subsp. thermophilus.

[0181]

[24] A consumer product composition for protecting nerve cells from neurotoxic damage, comprising outer membrane vesicles derived from at least one lactic acid bacteria selected from the group consisting of:

[0182] Lactobacillus spp.,

[0183] Lactobacillus spp.,

[0184] Lactiplantibacillus spp.,

[0185] Combined with Lactobacillus spp.,

[0186] Lactococcus, and

[0187] Streptococcus salivarius subsp. thermophilus.

[0188]

[25] The food or beverage described in

[21] , the pharmaceutical composition described in

[22] , the cosmetic composition described in

[23] or the consumer product composition described in

[24] , which protects neurons from neurotoxic damage by stimulating FPR2.

[0189]

[26] The food or beverage described in

[21] above, the pharmaceutical composition described in

[22] above, the cosmetic composition described in

[23] above, or the consumer product composition described in

[24] above, wherein the neurotoxicity is neurotoxicity caused by amyloid β.

[0190]

[27] The food or beverage described in

[21] above, the pharmaceutical composition described in

[22] above, the cosmetic composition described in

[23] above, or the consumer product composition described in

[24] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[0191]

[28] A method for stimulating FPR2 in a subject, comprising administering to the subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of:

[0192] Lactobacillus spp.,

[0193] Lactobacillus spp.,

[0194] Lactiplantibacillus spp.,

[0195] Combined with Lactobacillus spp.,

[0196] Lactococcus, and

[0197] Streptococcus salivarius subsp. thermophilus.

[0198]

[29] A method for treating a disease associated with FPR2 in a subject, comprising the step of administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0199] Lactobacillus spp.,

[0200] Lactobacillus spp.,

[0201] Lactiplantibacillus spp.,

[0202] Combined with Lactobacillus spp.,

[0203] Lactococcus, and

[0204] Streptococcus salivarius subsp. thermophilus.

[0205]

[30] A method for inducing IL-10 in a subject by agonizing FPR2, comprising administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0206] Lactobacillus spp.,

[0207] Lactobacillus spp.,

[0208] Lactiplantibacillus spp.,

[0209] Combined with Lactobacillus spp.,

[0210] Lactococcus, and

[0211] Streptococcus salivarius subsp. thermophilus.

[0212]

[31] A method for protecting a subject's skin from ultraviolet damage, comprising administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0213] Lactobacillus spp.,

[0214] Lactobacillus spp.,

[0215] Lactiplantibacillus spp.,

[0216] Combined with Lactobacillus spp.,

[0217] Lactococcus, and

[0218] Streptococcus salivarius subsp. thermophilus.

[0219]

[32] The method according to

[31] above, wherein the skin is protected from UV damage by stimulating FPR2.

[0220]

[33] A method for protecting neurons from neurotoxic damage in a subject, comprising administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of:

[0221] Lactobacillus spp.,

[0222] Lactobacillus spp.,

[0223] Lactiplantibacillus spp.,

[0224] Combined with Lactobacillus spp.,

[0225] Lactococcus, and

[0226] Streptococcus salivarius subsp. thermophilus.

[0227]

[34] The method according to the above

[33] , which protects neurons from neurotoxic damage by stimulating FPR2.

[0228]

[35] The method according to any one of

[28] to

[34] above, wherein the lactic acid bacteria is at least one selected from the group consisting of:

[0229] Lactobacillus delbrueckii subsp.bulgaricus, Lactobacillus helveticus,

[0230] Lactobacillus acidophilus,

[0231] Lactobacillus rhamnosus,

[0232] Lactobacillus paracasei,

[0233] Lactiplantibacillus plantarum,

[0234] Ligilactobacillus salivarius,

[0235] Lactococcus lactis subsp. lactis,

[0236] Streptococcus salivarius subsp.thermophilus,

[0237] Lactobacillus gasseri,

[0238] Lactobacillus amylovorus,

[0239] Lactobacillus crispatus,

[0240] Lactobacillus johnsonii,

[0241] Lactobacillus casei, and

[0242] Lactiplantibacillus pentosus.

[0243]

[36] The method according to any one of

[28] to

[35] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[0244] Effects of the Invention

[0245] As shown in the examples described below, the outer membrane vesicles derived from specific lactic acid bacteria can stimulate FPR2. Furthermore, as shown in the examples described below, the outer membrane vesicles derived from specific lactic acid bacteria can induce IL-10 by stimulating FPR2, thereby protecting the skin from ultraviolet damage and protecting nerve cells from neurotoxic damage. The present invention using the outer membrane vesicles as an active ingredient is useful for treating diseases related to FPR2, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0246] Figure 1 The FPR2 agonistic activity of outer membrane vesicles derived from 26 bacteria is shown.

[0247] Figure 2 The FPR1 agonistic activity of outer membrane vesicles derived from 26 bacteria is shown.

[0248] Figure 3 The effect of a known FPR2 agonist on IL-10 production by M2 macrophages is shown.

[0249] Figure 4 The IL-10 inducing activity of outer membrane vesicles derived from 17 bacteria is shown.

[0250] Figure 5 Shown is the effect of FPR2 antagonists on the IL-10 inducing capacity of outer membrane vesicles with FPR2 agonistic activity.

[0251] Figure 6 The role of outer membrane vesicles with FPR2 agonistic activity in protecting the skin from UV damage is shown.

[0252] Figure 7 The effect of outer membrane vesicles with FPR2 agonistic activity on protecting neural cells from neurotoxic damage is shown. DETAILED DESCRIPTION

[0253] 〔Lactic acid bacteria〕

[0254] The outer membrane vesicles used in the present invention are derived from the following lactic acid bacteria (1) to (6).

[0255] (1) Lactobacillus

[0256] (2) Lacticaseibacillus

[0257] (3) Lactiplantibacillus

[0258] (4) Ligilactobacillus

[0259] (5) Lactococcus

[0260] (6) Streptococcus salivarius subsp.thermophilus

[0261] The classification of lactic acid bacteria in this specification follows the reclassification published in 2020. Details of the reclassification are described in Int J Syst Evol Microbiol. 2020 Apr; 70(4): 2782-2858, the homepage of the Society of Enteric Bacteria (URL: https: / / bifidus-fund.jp / keyword / kw054.shtml), and the like.

[0262] Known lactic acid strains belonging to any of the above (1) to (6) and secreting outer membrane vesicles can be used in the present invention without particular limitation. Since the outer membrane vesicles exert their effects by administration or ingestion, strains whose safety has been confirmed in animals are preferred.

[0263] Preferred lactic acid bacteria are lactic acid bacteria of the genus (1) Lactobacillus, (2) Lactobacillus, (3) Lactobacillus, (4) Lactobacillus, or (5) Lactococcus.

[0264] More preferred lactic acid bacteria are lactic acid bacteria of the genus (1) Lactobacillus, (2) Lactobacillus casei, (3) Lactobacillus plantarum, or (4) Lactobacillus syngenes.

[0265] More preferably, the lactic acid bacteria are lactic acid bacteria of the genus (1) Lactobacillus, (2) Lactobacillus casei, or (4) Lactobacillus.

[0266] Particularly preferred lactic acid bacteria are lactic acid bacteria of the genus (1) Lactobacillus or (2) Lactobacillus.

[0267] (1) Lactobacillus lactic acid bacteria

[0268] Examples of lactic acid bacteria of the genus Lactobacillus include the following.

[0269] Lactobacillus delbrueckii subsp.bulgaricus

[0270] Lactobacillus helveticus

[0271] Lactobacillus acidophilus

[0272] Lactobacillus delbrueckii

[0273] Lactobacillus crispatus

[0274] Lactobacillus gasseri

[0275] Lactobacillus paragasseri

[0276] Lactobacillus amylovorus

[0277] Lactobacillus johnsonii

[0278] Preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus delbrueckii or Lactobacillus gasseri.

[0279] In one embodiment of the present invention, a preferred lactic acid bacterium of the genus Lactobacillus is Lactobacillus gasseri, Lactobacillus amylovora, Lactobacillus crispatus, or Lactobacillus johnsonii.

[0280] In another embodiment of the present invention, the preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus gasseri, Lactobacillus amylovora, Lactobacillus crispatus or Lactobacillus johnsonii.

[0281] Further preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, or Lactobacillus acidophilus.

[0282] In one embodiment of the present invention, more preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus amylovora, Lactobacillus crispatus, or Lactobacillus johnsonii.

[0283] Still more preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus gasseri or Lactobacillus johnsonii.

[0284] The Lactobacillus delbrueckii subspecies bulgaricus may be any available strain, and may include the model strain Lactobacillus delbrueckii subspecies bulgaricus T.

[0285] The Lactobacillus helveticus may be any available strain, and may include the model strain Lactobacillus helveticus T.

[0286] Lactobacillus acidophilus can include the model strain Lactobacillus acidophilus T as long as it is an available strain. Lactobacillus acidophilus can include other strains, such as JCM1023 strain, CP1613 strain, etc. Lactobacillus acidophilus CP1613 strain is one of the lactic acid bacteria belonging to Lactobacillus acidophilus, and was internationally deposited on January 18, 2013 with the deposit number: NITE BP-1513 in the International Depository Center for Microorganisms of the Independent Administrative Institution for Product Evaluation Technology Foundation (Room 122, 2-5-8, Kazusa Kamashi, Kisarazu City, Chiba Prefecture, Japan, 〒292-0818) based on the international depository institution stipulated in the Budapest Treaty.

[0287] Lactobacillus gasseri can include the model strain Lactobacillus gasseri T as long as it is an obtainable strain. Lactobacillus gasseri can include other strains, such as CP2305 strain, etc. Lactobacillus gasseri CP2305 strain is one of the lactic acid bacteria belonging to Lactobacillus gasseri, and was internationally deposited in the International Depository of the Budapest Treaty - the Patent Organism Depository of the Independent Administrative Institution for Evaluation of Products Technology Foundation (Room 120, 2-5-8, Kazusa Kamashi, Kisarazu City, Chiba Prefecture, Japan, 292-0818) with a deposit number: FERM BP-11331 (deposit date: September 11, 2007).

[0288] The Lactobacillus amylovora may be any strain as long as it is available, and may include the model strain Lactobacillus amylovora T.

[0289] The Lactobacillus crispatus may be any available strain, and may include the model strain Lactobacillus crispatus T.

[0290] The Lactobacillus johnsonii may be any available strain, and may include the model strain Lactobacillus johnsonii T.

[0291] (2) Lactic acid bacteria of the genus Lactobacillus

[0292] Examples of lactic acid bacteria belonging to the genus Lactobacillus include the following.

[0293] Lacticaseibacillus rhamnosus

[0294] Lacticaseibacillus paracasei

[0295] Lacticaseibacillus casei

[0296] Lacticaseibacillus paracaseisubsp.paracasei

[0297] Preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus rhamnosus, Lactobacillus paracasei, or Lactobacillus casei.

[0298] More preferably, the lactic acid bacteria of the genus Lactobacillus are Lactobacillus paracasei or Lactobacillus casei.

[0299] The Lactobacillus rhamnosus may be any strain as long as it is available, and may include the model strain Lactobacillus rhamnosus T.

[0300] As long as the strain can be obtained, the type strain Lactobacillus paracasei T can be included. Lactobacillus paracasei can include other strains, such as JCM1133 strain, CP3526 strain, etc. Lactobacillus paracasei CP3526 strain is one of the lactic acid bacteria belonging to Lactobacillus paracasei, and was internationally deposited on March 18, 2022 with the deposit number NITE BP-03625 at the International Depository Center for Microorganisms of the Independent Administrative Agency for Product Evaluation Technology Foundation (Room 122, 2-5-8, Kazusa Kamata, Kisarazu City, Chiba Prefecture, Japan, 〒292-0818).

[0301] The Lactobacillus casei may be any available strain, and may include the model strain Lactobacillus casei T.

[0302] In one embodiment of the present invention, the lactic acid bacteria of the genus Lactobacillus may be lactic acid bacteria other than Lactobacillus paracasei.

[0303] (3) Lactobacillus lactic acid bacteria

[0304] Examples of lactic acid bacteria of the genus Lactobacillus include the following.

[0305] Lactiplantibacillus plantarum

[0306] Lactiplantibacillus pentosus

[0307] Lactiplantibacillus paraplantarum

[0308] Preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus plantarum or Lactobacillus pentosaceus.

[0309] A further preferred lactic acid bacterium of the genus Lactobacillus is Lactobacillus plantarum.

[0310] The Lactobacillus plantarum may be any available strain, and may include the model strain Lactobacillus plantarum T.

[0311] The Lactobacillus pentosus may be any strain as long as it is available, and may include the model strain Lactobacillus pentosus T.

[0312] In one embodiment of the present invention, the lactic acid bacteria of the genus Lactobacillus may be lactic acid bacteria other than Lactobacillus plantarum.

[0313] (4) Lactic acid bacteria of the genus Lactobacillus

[0314] Examples of lactic acid bacteria belonging to the genus Lactobacillus include the following.

[0315] Ligilactobacillus salivarius

[0316] Ligilactobacillus acidipiscis

[0317] A preferred lactic acid bacterium of the genus Lactobacillus is Lactobacillus salivarius.

[0318] The Lactobacillus salivarius may be any available strain, and may include the model strain Lactobacillus salivarius T.

[0319] (5) Lactococcus lactic acid bacteria

[0320] Examples of lactic acid bacteria of the genus Lactococcus include the following.

[0321] Lactococcus lactis subsp. lactis

[0322] Lactococcus lactis subsp.cremoris

[0323] Lactococcus lactis

[0324] Preferred lactic acid bacteria of the genus Lactococcus are Lactococcus lactis subsp. lactis or Lactococcus lactis subsp. cremoris.

[0325] A further preferred lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis.

[0326] The Lactococcus lactis subsp. lactis may be any available strain, and may include the model strain Lactococcus lactis subsp. lactis T.

[0327] (6) Streptococcus salivarius subsp.thermophilus

[0328] The Streptococcus salivarius subsp. thermophilus may be any available strain, and may include the model strain Streptococcus salivarius subsp. thermophilus T.

[0329] Mutants or derivatives of the above-mentioned specific strains may also be used as long as they secrete outer membrane vesicles.

[0330] Preferred lactic acid bacteria are at least one selected from the group consisting of:

[0331] Lactobacillus delbrueckii subsp.bulgaricus

[0332] Lactobacillus helveticus

[0333] Lactobacillus acidophilus

[0334] Lactobacillus rhamnosus

[0335] Lactobacillus paracasei

[0336] Lactobacillus plantarum

[0337] Lactobacillus salivae

[0338] Lactococcus lactis subsp. lactis, and

[0339] Streptococcus salivarius thermophilus subsp.

[0340] In one embodiment of the present invention, the preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0341] Lactobacillus gasseri

[0342] Lactobacillus amylovora

[0343] Lactobacillus crispatus

[0344] Lactobacillus johnsonii

[0345] Lactobacillus casei, and

[0346] Lactobacillus pentosaceus.

[0347] In another embodiment of the present invention, the preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0348] Lactobacillus delbrueckii subsp.bulgaricus

[0349] Lactobacillus helveticus

[0350] Lactobacillus acidophilus

[0351] Lactobacillus rhamnosus

[0352] Lactobacillus paracasei

[0353] Lactobacillus plantarum

[0354] Lactobacillus salivae

[0355] Lactococcus lactis subsp. lactis

[0356] Streptococcus salivarius subsp. thermophilus

[0357] Lactobacillus gasseri

[0358] Lactobacillus amylovora

[0359] Lactobacillus crispatus

[0360] Lactobacillus johnsonii

[0361] Lactobacillus casei, and

[0362] Lactobacillus pentosaceus.

[0363] More preferably, the lactic acid bacteria is at least one selected from the group consisting of the following options:

[0364] Lactobacillus delbrueckii subsp.bulgaricus

[0365] Lactobacillus helveticus

[0366] Lactobacillus acidophilus

[0367] Lactobacillus rhamnosus

[0368] Lactobacillus paracasei

[0369] Lactobacillus plantarum

[0370] Saliva combined with Lactobacillus, and

[0371] Lactococcus lactis subsp. lactis.

[0372] In one embodiment of the present invention, more preferably the lactic acid bacteria is at least one selected from the group consisting of the following options:

[0373] Lactobacillus gasseri

[0374] Lactobacillus amylovora

[0375] Lactobacillus crispatus

[0376] Lactobacillus johnsonii

[0377] Lactobacillus casei, and

[0378] Lactobacillus pentosaceus.

[0379] In another embodiment of the present invention, the more preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0380] Lactobacillus delbrueckii subsp.bulgaricus

[0381] Lactobacillus helveticus

[0382] Lactobacillus acidophilus

[0383] Lactobacillus rhamnosus

[0384] Lactobacillus paracasei

[0385] Lactobacillus plantarum

[0386] Lactobacillus salivae

[0387] Lactococcus lactis subsp. lactis

[0388] Lactobacillus gasseri

[0389] Lactobacillus amylovora

[0390] Lactobacillus crispatus

[0391] Lactobacillus johnsonii

[0392] Lactobacillus casei, and

[0393] Lactobacillus pentosaceus.

[0394] Further preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0395] Lactobacillus delbrueckii subsp.bulgaricus

[0396] Lactobacillus helveticus

[0397] Lactobacillus acidophilus

[0398] Lactobacillus rhamnosus

[0399] Lactobacillus paracasei

[0400] Lactobacillus plantarum, and

[0401] Saliva-associated Lactobacillus.

[0402] In one embodiment of the present invention, more preferably the lactic acid bacteria is at least one selected from the group consisting of the following options:

[0403] Lactobacillus gasseri

[0404] Lactobacillus amylovora

[0405] Lactobacillus crispatus

[0406] Lactobacillus johnsonii

[0407] Lactobacillus casei, and

[0408] Lactobacillus pentosaceus.

[0409] In another embodiment of the present invention, the more preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0410] Lactobacillus delbrueckii subsp.bulgaricus

[0411] Lactobacillus helveticus

[0412] Lactobacillus acidophilus

[0413] Lactobacillus rhamnosus

[0414] Lactobacillus paracasei

[0415] Lactobacillus plantarum

[0416] Lactobacillus salivae

[0417] Lactobacillus gasseri

[0418] Lactobacillus amylovora

[0419] Lactobacillus crispatus

[0420] Lactobacillus johnsonii

[0421] Lactobacillus casei, and

[0422] Lactobacillus pentosaceus.

[0423] More preferably, the lactic acid bacteria is at least one selected from the group consisting of the following options:

[0424] Lactobacillus delbrueckii subsp.bulgaricus

[0425] Lactobacillus helveticus

[0426] Lactobacillus acidophilus

[0427] Lactobacillus rhamnosus

[0428] Lactobacillus casei, and

[0429] Saliva-associated Lactobacillus.

[0430] In one embodiment of the present invention, more preferably the lactic acid bacteria is at least one selected from the group consisting of the following options:

[0431] Lactobacillus gasseri

[0432] Lactobacillus amylovora

[0433] Lactobacillus crispatus

[0434] Lactobacillus johnsonii, and

[0435] Lactobacillus casei.

[0436] In another embodiment of the present invention, the lactic acid bacteria is preferably at least one selected from the group consisting of the following options:

[0437] Lactobacillus delbrueckii subsp.bulgaricus

[0438] Lactobacillus helveticus

[0439] Lactobacillus acidophilus

[0440] Lactobacillus rhamnosus

[0441] Lactobacillus paracasei

[0442] Lactobacillus salivae

[0443] Lactobacillus gasseri

[0444] Lactobacillus amylovora

[0445] Lactobacillus crispatus

[0446] Lactobacillus johnsonii, and

[0447] Lactobacillus casei.

[0448] Particularly preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0449] Lactobacillus delbrueckii subsp.bulgaricus

[0450] Lactobacillus helveticus

[0451] Lactobacillus acidophilus

[0452] Lactobacillus rhamnosus, and

[0453] Lactobacillus casei.

[0454] In one embodiment of the present invention, particularly preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0455] Lactobacillus gasseri

[0456] Lactobacillus amylovora

[0457] Lactobacillus crispatus

[0458] Lactobacillus johnsonii, and

[0459] Lactobacillus casei.

[0460] In another embodiment of the present invention, the lactic acid bacteria are preferably at least one selected from the group consisting of the following options:

[0461] Lactobacillus delbrueckii subsp.bulgaricus

[0462] Lactobacillus helveticus

[0463] Lactobacillus acidophilus

[0464] Lactobacillus rhamnosus

[0465] Lactobacillus paracasei

[0466] Lactobacillus gasseri

[0467] Lactobacillus amylovora

[0468] Lactobacillus crispatus

[0469] Lactobacillus johnsonii, and

[0470] Lactobacillus casei.

[0471] Further particularly preferred lactic acid bacteria are at least one selected from the group consisting of the following options:

[0472] Lactobacillus acidophilus

[0473] Lactobacillus casei, and

[0474] Lactobacillus gasseri.

[0475] The most preferred lactic acid bacteria are at least one selected from the group consisting of:

[0476] Lactobacillus acidophilus, and

[0477] Lactobacillus casei.

[0478] The lactic acid bacteria may be used alone or in combination of two or more. Therefore, outer membrane vesicles derived from two or more lactic acid bacteria may be used in the present invention.

[0479] [Outer membrane vesicles from lactic acid bacteria]

[0480] Bacterial-derived outer membrane vesicles are substances secreted by bacteria.

[0481] Outer membrane vesicles derived from lactic acid bacteria are spherical structures composed of the cell membrane (lipid bilayer membrane) of lactic acid bacteria, and their diameter is generally around 20 to 500 nm. The spherical structure sometimes contains nucleic acids (DNA, RNA) or proteins (enzymes, etc.). Outer membrane vesicles derived from the cell membrane of lactic acid bacteria may also contain membrane proteins, peptidoglycan (PG), etc. present in the lipid bilayer membrane of lactic acid bacteria (Journal of the Japanese Society of Lactobacilli, 27(1), 10-16, 2016, and Microbiol Spectr., 7(1)., 2019).

[0482] Outer membrane vesicles derived from bacteria are also called membrane vesicles, extracellular vesicles (EV), extracellular vesicles, microvesicles, membrane vesicles (MV), outer membrane vesicles (OMV), etc.

[0483] The secretion of outer membrane vesicles does not require special culture conditions, and outer membrane vesicles are released into the culture medium by culturing under conditions commonly used in lactic acid bacteria culture.

[0484] The culture medium preferably contains a carbon source, a nitrogen source, inorganic salts, etc. and can efficiently culture lactic acid bacteria. The culture medium may be any of a natural culture medium and a synthetic culture medium, and a known culture medium suitable for the strain used can be appropriately selected.

[0485] Examples of the carbon source include lactose, glucose, sucrose, fructose, galactose, and molasses.

[0486] Examples of the nitrogen source include organic nitrogen-containing substances such as casein hydrolyzate, whey protein hydrolyzate, and soy protein hydrolyzate.

[0487] Examples of the inorganic salts include phosphates, sodium, potassium, and magnesium.

[0488] Examples of a culture medium suitable for lactic acid bacteria include MRS culture medium, GLM culture medium, GAM culture medium, BL culture medium, Briggs Liver Broth, animal milk, skim milk, and milk whey. Preferred are MRS culture medium and GLM culture medium.

[0489] Furthermore, when the outer membrane vesicles of lactic acid bacteria are used for food purposes, a culture medium consisting only of food raw materials and food additives can be used.

[0490] The culture of lactic acid bacteria can be carried out, for example, at a temperature of 20 to 50° C. (preferably 30 to 40° C.) under static or anaerobic conditions. The culture temperature can be adjusted by a thermostatic bath, a jacket heater, a jacket, or the like.

[0491] Anaerobic conditions refer to a low-oxygen environment to the extent that lactic acid bacteria can proliferate. Anaerobic conditions can be set by using an anaerobic chamber, an anaerobic box, or a sealed container or bag containing a deoxidizer.

[0492] Examples of the culture format include static culture, stirring culture, and tank culture.

[0493] The culture time is, for example, 3 to 96 hours, preferably 12 to 32 hours.

[0494] The outer membrane vesicles can be obtained from the lactic acid bacteria culture solution by appropriately combining one or more known separation and purification means according to the quality and size of the outer membrane vesicles.

[0495] Examples of separation and purification means include centrifugation, ultracentrifugation, and filtration.

[0496] For example, the following steps can be used to obtain outer membrane vesicles.

[0497] The lactic acid bacteria culture solution is subjected to centrifugal separation (for example, the centrifugal force is 3,000 to 15,000 g, preferably 8,000 to 10,000 g) to precipitate the bacterial cells. The supernatant after centrifugation is filtered with a filter having a pore size that allows outer membrane vesicles to pass (for example, 220 to 1,000 nm, preferably 450 nm (0.45 μm)). The filtrate containing the outer membrane vesicles is subjected to ultracentrifugation (for example, the centrifugal force is 100,000 to 300,000 g, preferably 150,000 to 300,000 g) to obtain the outer membrane vesicles as a precipitate. The precipitate can be further filtered.

[0498] [FPR2 (formyl peptide receptor 2)]

[0499] FPR2 (Formyl Peptide Receptor 2) is a receptor for peptides having N-formylmethionine (eg, f-Met-Leu-Phe (FMLP)), and is a G protein-coupled receptor.

[0500] FPR2 is one of the FPR family (FPR1, FPR2 and FPR3) present in humans.

[0501] FPR2 is sometimes also called FPRL-1 or FPRL1.

[0502] FPR1 is called formyl peptide receptor, and FPR2 and FPR3 are called formyl peptide-like receptors. Compared with FPR1, FPR2 and FPR3 have high sequence homology (Pharmacol Rev 61: 119-161, 2009).

[0503] [FPR2 agonist]

[0504] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to agonize FPR2 (activate FPR2 by binding to it).

[0505] Therefore, one embodiment of the present invention is an FPR2 agonist containing outer membrane vesicles derived from specific lactic acid bacteria as an active ingredient. In addition, use of outer membrane vesicles derived from specific lactic acid bacteria for the preparation of an FPR2 agonist is also one embodiment of the present invention.

[0506] It is known that low molecular weight compounds, peptides and the like have agonist activity against FPR2, but there is no report that the outer membrane vesicles derived from lactic acid bacteria excite FPR2. The present invention uses the outer membrane vesicles derived from specific lactic acid bacteria that can excite FPR2 as FPR2 agonists. The outer membrane vesicles derived from lactic acid bacteria used as the active ingredient of the FPR2 agonist in the present invention are very stable, and are known to have the property of remaining for a long time even if taken into cells and not being completely decomposed. Therefore, it is believed that it is different from the previous substances with agonist activity against FPR2, and it itself has both the function of exciting FPR2 and the function of being stably delivered to the body. In addition, the outer membrane vesicles derived from lactic acid bacteria can be easily obtained by culturing lactic acid bacteria as described above.

[0507] FPR2 agonists have been shown to be beneficial in astringing chronic inflammation and in preclinical models of chronic inflammatory human diseases including infections, psoriasis, dermatitis, inflammatory bowel disease, Crohn's disease, ocular inflammation, sepsis, pain, metabolism / diabetes, cancer, COPD, asthma and allergic diseases, cystic fibrosis, acute lung injury and fibrosis, rheumatoid arthritis and other joint diseases, Alzheimer's disease, renal fibrosis and organ transplantation (Patent Document 7 and Non-Patent Documents 12 and 13), etc. Therefore, it is believed that the outer membrane vesicles derived from specific lactic acid bacteria of the present invention are useful for the treatment of diseases associated with FPR2.

[0508] FPR2 agonist activity can be measured using a known test system (e.g., a report test described in International Publication No. WO2020 / 026979) that can measure the activation caused by a ligand of a G protein-coupled receptor (GPCR). The report test can be provided as a commissioned service (e.g., a GPCR commissioned test by Tanso Biosciences of Cosmobio).

[0509] The outer membrane vesicles based on the lactic acid bacteria of the present invention preferably selectively stimulate FPR2.

[0510] "Selectively activating FPR2" means having agonistic activity on FPR2 but not on FPR1. FPR1 is involved in the induction of inflammatory response (Non-patent Document 3), so FPR2 agonists that "selectively activate FPR2" can treat diseases associated with FPR2 without inducing inflammatory response.

[0511] [Diseases associated with FPR2]

[0512] FPR2 agonists can be used to treat diseases associated with FPR2.

[0513] “Diseases associated with FPR2” include not only diseases that are the target of drugs but also diseases that are the target of functional foods and drinks and functional cosmetics.

[0514] “Treatment” includes not only actions for the purpose of treatment and prevention (use of FPR2 agonists as drugs or quasi-drugs), but also actions for the purpose of health improvement (use of FPR2 agonists as functional foods and beverages, functional cosmetics).

[0515] The FPR2 agonist can be used for both therapeutic purposes (medical use) and non-therapeutic purposes (non-medical use).

[0516] Examples of "diseases associated with FPR2" include the following diseases.

[0517] Inflammatory diseases, heart disease, chronic respiratory diseases, cancer, sepsis, allergic symptoms, circulatory disorders, neuroinflammation, neurological disorders, pain, prion diseases, amyloid diseases, immune disorders, Behcet's disease, Sweet's disease, systemic lupus erythematosus (SLE), Wegener's granulomatosis, viral infections, diabetes, bacterial infections, physical injuries, physical disorders including radiation exposure, vasoconstriction, allergic reactions, allergic reactions, rhinitis, shock (endotoxic, hemorrhagic, traumatic, visceral ischemic and circulatory shock), rheumatoid arthritis, gout , benign prostatic hypertrophy, myocardial ischemia, myocardial infarction, heart failure, brain injury, lung disease, COPD, COAD, COLD, acute lung injury, acute respiratory distress syndrome, chronic bronchitis, emphysema, asthma (allergic and non-allergic), cystic fibrosis, renal fibrosis, renal damage, glomerular disease, ulcerative colitis, IBD, Crohn's disease, periodontitis, pain, Alzheimer's disease, AIDS, uveitis glaucoma, conjunctivitis, Sjögren's syndrome, atherosclerosis, neurological diseases including multiple sclerosis Inflammation, flare-ups, skin diseases (rosacea, sunburn, psoriasis, flushing, flushing and redness with flushing, erythema with flushing, UV aging, seborrheic dermatitis, acne, allergic dermatitis, vasodilation, hypertrophic rosacea, rash, skin erythema, skin hyperactivity with vasodilation, Lyell syndrome, Stevens-Johnson syndrome, itching and discomfort associated with hemorrhoids, hemorrhoids, erythema multiforme, erythema multiforme, erythema nodosum, eye swelling, urticaria, pruritus, purpura, varicose veins, contact dermatitis, atopic dermatitis Dermatitis, nummular dermatitis, systemic exfoliative dermatitis, stasis dermatitis, neurodermatitis, perioral dermatitis, pseudofolliculitis barbae, annular granuloma, solar keratosis, basal cell carcinoma, squamous cell carcinoma, eczema, skin wound healing, hypertrophic scars, keloids, burns, solar keratosis, melanoma, viral warts, UV-induced aging, photodamage, melanosis, post-inflammatory pigmentation, other hyperpigmentation, hair loss), eye inflammation, enteritis, inflammatory bowel disease, dermatitis, tonsillitis, laryngitis, pharyngitis, sinusitis, esophagitis, gastritis, etc.

[0518] The above-mentioned diseases can be classified, for example, as follows.

[0519] (1) Inflammatory diseases (such as tonsillitis, laryngitis, pharyngitis, eye inflammation, rhinitis, sinusitis, periodontitis, conjunctivitis, neuritis, chronic bronchitis, Behcet's disease, etc.)

[0520] (2) Digestive system diseases (such as esophagitis, gastritis, enteritis, inflammatory bowel disease, ulcerative colitis, etc.)

[0521] (3) Skin diseases (such as dermatitis (including allergic dermatitis, atopic dermatitis, seborrheic dermatitis), acne, rash, sunburn, hot flashes, eczema, ultraviolet aging, itching and discomfort, perioral dermatitis, hyperpigmentation, hair loss, etc.)

[0522] (4) Brain diseases (such as dementia (including alcoholic dementia), migraine, Alzheimer's disease (including early symptoms such as amnesia), prion disease, etc.)

[0523] (5) Other diseases (respiratory diseases (such as asthma, chronic obstructive pulmonary disease (COPD), emphysema, etc.), circulatory system diseases (such as atherosclerosis, heart disease, circulatory disorders, myocardial ischemia, myocardial infarction, heart failure, etc.), joint diseases (such as rheumatoid arthritis, etc.), nervous system diseases (such as neuritis, nerve disorders, pain, etc.), metabolic diseases (such as diabetes, gout, renal dysfunction, etc.), infections (such as bacterial infections, viral infections, sepsis caused by infections, etc.), immune disorders (such as allergic symptoms, AIDS, etc.), others (such as physical injuries, cancer, Crohn's disease, etc.)

[0524] It is known that FPR2 is related to brain neurotoxicity, homeostasis, inflammation, proliferation and regeneration of intestinal epithelial cells (the above-mentioned non-patent documents 8 to 11). In addition, epithelial cells are also present in the skin. In addition, outer membrane vesicles can pass through the blood-brain barrier (non-patent documents 5 and 6). Therefore, FPR2 agonists can be preferably used to treat brain diseases (such as Alzheimer's disease (including early symptoms such as amnesia), dementia (including alcoholic dementia), migraine), digestive system diseases (such as esophagitis, gastritis, enteritis, inflammatory bowel disease, ulcerative colitis), skin diseases (such as dermatitis (including allergic dermatitis, atopic dermatitis, seborrheic dermatitis), acne, rash, sunburn, hot flashes, eczema, aging caused by ultraviolet rays, itching and discomfort, perioral dermatitis, hyperpigmentation, hair loss) by binding to FPR2 and activating FPR2.

[0525] The content of the outer membrane vesicles derived from lactic acid bacteria in the FPR2 agonist can be appropriately set according to the state, symptoms, etc. of the subject to whom the agonist is to be applied.

[0526] The outer membrane vesicle content (number) of each 1 mL of FPR2 agonist is, for example, 2×10 3 ~2×10 14 / mL, preferably 2×10 4 ~2×10 12 / mL, more preferably 2×10 5 ~2×10 10 Pieces / mL.

[0527] FPR2 agonists can be applied to animals expressing FPR2 without limitation. The application subjects are preferably mammals (humans and non-human mammals (e.g., horses, cattle)), more preferably humans. In addition, the gender and age of the application subjects are not limited.

[0528] [Induction of IL-10]

[0529] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to induce the production and secretion of IL-10 (hereinafter also referred to as "IL-10 induction"). This embodiment can be understood as an IL-10 inducer containing the outer membrane vesicles of the lactic acid bacteria of the present invention as an active ingredient.

[0530] In one embodiment of the present invention, outer membrane vesicles induce IL-10 by stimulating FPR2. This embodiment can be understood as an IL-10 inducer based on stimulating FPR2, containing outer membrane vesicles based on the lactic acid bacteria of the present invention as an active ingredient.

[0531] IL-10 is an anti-inflammatory cytokine produced and secreted by immune cells such as macrophages, helper T cells, and regulatory T cells.

[0532] IL-10 inducers (including those that induce IL-10 by stimulating FPR2. The same applies hereinafter) can be used to treat diseases associated with IL-10.

[0533] “Diseases associated with IL-10” include not only diseases that are the target of drugs but also diseases that are the target of functional foods and beverages and functional cosmetics.

[0534] “Treatment” includes not only actions for the purpose of treatment and prevention (use of IL-10 inducers as drugs or quasi-drugs), but also actions for the purpose of health improvement (use of IL-10 inducers as functional foods and beverages, functional cosmetics).

[0535] The IL-10 inducer can be used for both therapeutic purposes (medical use) and non-therapeutic purposes (non-medical use).

[0536] Examples of "diseases associated with IL-10" include the above-mentioned (1) inflammatory diseases, (2) digestive system diseases, (3) skin diseases, (4) brain diseases, and (5) other diseases.

[0537] The content of outer membrane vesicles derived from lactic acid bacteria in the IL-10 inducer can be appropriately set according to the state, symptoms, etc. of the subject to whom the IL-10 inducer is to be applied.

[0538] The outer membrane vesicle content (number) per 1 mL of IL-10 inducer is, for example, 2×10 3 ~2×1014 / mL, preferably 2×10 4 ~2×10 12 / mL, more preferably 2×10 5 ~2×10 10 Pieces / mL.

[0539] The IL-10 inducer can be applied to animals without limitation. The application object is preferably a mammal (human and non-human mammal (e.g., horse, cattle)), more preferably a human. In addition, the gender and age of the application object are not limited.

[0540] 〔Protect the skin from UV damage〕

[0541] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to protect the skin from ultraviolet damage. This aspect can be understood as a skin protective agent for preventing ultraviolet damage, containing the outer membrane vesicles of the lactic acid bacteria of the present invention as an active ingredient.

[0542] In one embodiment of the present invention, the outer membrane vesicles protect the skin from ultraviolet damage by stimulating FPR2. This embodiment can be understood as a skin protectant that contains the outer membrane vesicles of the lactic acid bacteria of the present invention as an active ingredient and protects the skin from ultraviolet damage by stimulating FPR2.

[0543] Skin protectants against UV damage (including those that protect the skin by stimulating FPR2. The same applies below) can be used to treat UV-related skin diseases.

[0544] “Ultraviolet-related skin diseases” include not only diseases that are the target of drugs but also diseases that are the target of functional foods and beverages and functional cosmetics.

[0545] “Treatment” includes not only actions for the purpose of treatment and prevention (use of skin protectants as drugs or quasi-drugs), but also actions for the purpose of health improvement (use of skin protectants as functional foods and beverages, functional cosmetics).

[0546] The skin protectant may be used for either therapeutic purposes (medical use) or non-therapeutic purposes (non-medical use).

[0547] Examples of "skin diseases associated with ultraviolet rays" include the above-mentioned (3) skin diseases.

[0548] The content of the outer membrane vesicles derived from lactic acid bacteria in the skin protectant can be appropriately set according to the state, symptoms, etc. of the subject to whom the skin protectant is applied.

[0549] The content (number) of outer membrane vesicles per 1 mL of skin protective agent is, for example, 2×10 3 ~2×1014 / mL, preferably 2×10 4 ~2×10 12 / mL, more preferably 2×10 5 ~2×10 10 Pieces / mL.

[0550] The skin protectant can be applied to animals without limitation. The application object is preferably a mammal (human and non-human mammal (e.g., horse, cow)), more preferably a human. In addition, the gender and age of the application object are not limited.

[0551] [Protect nerve cells from neurotoxicity]

[0552] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to protect neurons from neurotoxicity. This aspect can be understood as a neuroprotective agent against neurotoxicity containing outer membrane vesicles derived from the lactic acid bacteria of the present invention as an active ingredient.

[0553] In one embodiment of the present invention, outer membrane vesicles protect neurons from neurotoxic damage by stimulating FPR2. This embodiment can be understood as a neuroprotective agent containing outer membrane vesicles based on the lactic acid bacteria of the present invention as an active ingredient and preventing neurotoxic damage by stimulating FPR2.

[0554] Examples of neuronal cell toxicity include damage caused by endogenous toxins (amyloid β, oxygen free radicals, etc.) and damage caused by exogenous toxins (ethanol, etc.).

[0555] Neuroprotective agents that prevent neuronal cell toxicity (including methods for protecting neurons by stimulating FPR2, the same below) can be used to treat diseases associated with neuronal cell toxicity.

[0556] “Diseases associated with neuronal cell toxicity” include not only diseases that are the target of drugs but also diseases that are the target of functional foods and beverages and functional cosmetics.

[0557] “Treatment” includes not only actions for the purpose of treatment and prevention (use of neuroprotective agents as drugs or quasi-drugs), but also actions for the purpose of health improvement (use of neuroprotective agents as functional foods and beverages, functional cosmetics).

[0558] The neuron cell protective agent can be used for both therapeutic purposes (medical use) and non-therapeutic purposes (non-medical use).

[0559] Examples of "diseases associated with neuronal cell toxicity" include the above-mentioned (4) brain diseases and the like.

[0560] The content of the outer membrane vesicles derived from lactic acid bacteria in the neuron-protective agent can be appropriately set according to the state, symptoms, etc. of the subject to whom the neuron-protective agent is to be applied.

[0561] The content (number) of outer membrane vesicles per 1 mL of the neuron protective agent is, for example, 2×10 3 ~2×10 14 / mL, preferably 2×10 4 ~2×10 12 / mL, more preferably 2×10 5 ~2×10 10 Pieces / mL.

[0562] The neuron cell protective agent can be applied to animals without limitation. The application object is preferably a mammal (human and non-human mammal (such as horse, cow)), more preferably a human. In addition, the gender and age of the application object are not limited.

[0563] 〔Optional ingredients〕

[0564] The FPR2 agonist may contain one or more optional components within a range that does not impair the action of the active ingredient. The optional components may be appropriately selected according to the usage mode of the FPR2 agonist, etc.

[0565] The optional component may be an additive for medicines, cosmetics, foods and beverages, and the like.

[0566] Examples of additives include oils and fats (vegetable oils (soybean oil, corn oil, safflower oil, olive oil, etc.); animal fats and fats (beef tallow, sardine oil, etc.), herbal medicines (royal jelly, ginseng, etc.), amino acids (glutamine, cysteine, leucine, arginine, etc.), polyols (ethylene glycol, polyethylene glycol, propylene glycol, glycerol, sugar alcohols (sorbitol, erythritol, xylitol, maltitol, mannitol, etc.)), natural polymers (gum arabic, agar, water-soluble corn fiber, gelatin, xanthan gum, cresol, etc.), protein, gluten or gluten hydrolysate, lecithin, starch, dextrin, etc.), vitamins (such as vitamin C, B vitamins, etc.), minerals (calcium, magnesium, zinc, iron, etc.), dietary fiber (mannan, pectin, hemicellulose, etc.), surfactants (glycerol fatty acid esters, sorbitan fatty acid esters, etc.), purified water, excipients (glucose, corn starch, lactose, dextrin, etc.); stabilizers; pH adjusters; antioxidants; sweeteners; flavoring ingredients; acidulants; coloring materials, and flavors, etc.

[0567] In addition, the FPR2 agonist may optionally contain functional ingredients other than the active ingredient (outer membrane vesicles derived from lactic acid bacteria) (for example, taurine, glutathione, carnitine, creatine, coenzyme Q, glucuronic acid, glucuronolactone, pepper extract, ginger extract, cocoa extract, guarana extract, garcinia cambogia extract, theanine, γ-aminobutyric acid, capsaicin, capsaicin, various organic acids, flavonoids, polyphenols, catechins, xanthine derivatives, indigestible oligosaccharides such as oligofructose, polyvinyl pyrrolidone, etc.).

[0568] Any component is a known substance and can be easily obtained from the market or can be prepared.

[0569] The optional components may be used alone or in combination of two or more.

[0570] The content of the optional components can be appropriately set depending on the purpose of compounding and the like.

[0571] The above explanation about the optional ingredients can also be applied to the IL-10 inducer, skin protective agent, and nerve cell protective agent according to the present invention.

[0572] [Drug composition]

[0573] The outer membrane vesicles derived from specific lactic acid bacteria of the present invention can be used as medicines or quasi-drugs. Therefore, one embodiment of the present invention is a pharmaceutical composition with the outer membrane vesicles derived from lactic acid bacteria as an active ingredient. Furthermore, the use of the outer membrane vesicles of lactic acid bacteria for preparing a pharmaceutical composition is also one embodiment of the present invention.

[0574] The pharmaceutical composition can be used to treat the above-mentioned "diseases associated with FPR2".

[0575] The pharmaceutical composition can be used as an active ingredient in a method for treating a "disease associated with FPR2" comprising the step of administering an effective amount to a subject for treating the above-mentioned "disease associated with FPR2".

[0576] The dosage form of the pharmaceutical composition is not particularly limited. Examples of dosage forms include oral preparations (tablets, capsules, granules, powders, syrups, dry syrups, liquids, suspensions, inhalants, etc.); enteral preparations (suppositories, etc.); drips; injections; external preparations (aerosols, liquids, emulsions, creams, ointments, gels, lotions, papules, patches, etc.), etc. Among them, oral preparations and external preparations are preferred.

[0577] Liquid preparations such as solutions and suspensions may be dissolved or suspended in water or other suitable media immediately before administration.

[0578] The surfaces of tablets, granules, etc. can be coated by a known method.

[0579] The pharmaceutical composition can be a sustained-release preparation, delayed-release preparation or immediate-release preparation prepared according to known controlled-release technology.

[0580] The pharmaceutical composition may optionally contain pharmaceutically acceptable carriers and additives.

[0581] Examples of pharmaceutically acceptable carriers and additives include water, organic solvents, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymers, sodium alginate, water-soluble dextran, water-soluble dextrin, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerol, propylene glycol, polyethylene glycol, vaseline, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, surfactants, and artificial cell structures (liposomes, etc.).

[0582] The pharmaceutical composition can be used in animals expressing FPR2 without limitation. The application object is preferably a mammal (human and non-human mammal (eg, horse, cow)), more preferably a human. In addition, the gender and age of the application object are not limited.

[0583] The dosage of the pharmaceutical composition can be appropriately set according to the type of target disease, the age and weight of the target, the administration route, the number of administrations, the purpose of administration, and the like.

[0584] When administered orally, the number of outer membrane vesicles per kg of body weight of the subject is, for example, 2×10 6 ~2×10 14 / kg, preferably 2×10 8 ~2×10 13 Pieces / kg.

[0585] The administration interval can be appropriately set depending on the type of the target disease, etc., and may be once a day or divided into several administrations.

[0586] The above description about the pharmaceutical composition can also be applied to the aspects of the present invention regarding the treatment of "diseases associated with IL-10", "skin diseases associated with ultraviolet rays" and "diseases associated with neuronal cell toxicity".

[0587] [Cosmetic composition]

[0588] The outer membrane vesicles derived from the specific lactic acid bacteria of the present invention can be used as an active ingredient of a cosmetic composition. Therefore, one embodiment of the present invention is a cosmetic composition containing the outer membrane vesicles derived from lactic acid bacteria as an active ingredient.

[0589] The cosmetic composition of the present invention is a cosmetic composition that exerts certain functions (such as anti-inflammatory effects, regeneration and repair effects of epithelial cells, etc.) on the organism by stimulating FPR2 through the outer membrane vesicles derived from the specific lactic acid bacteria of the present invention, that is, a cosmetic composition for stimulating FPR2.

[0590] The cosmetic composition may optionally contain additives permitted for use in cosmetic compositions in addition to the outer membrane vesicles derived from lactic acid bacteria. Examples of additives include antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances and other auxiliary agents, carriers and the like.

[0591] Examples of the product form of the cosmetic composition include lotions, various creams, essences, foundations and other cosmetics, facial cleansers, soaps, shampoos, care solutions, beauty serums, etc. Examples of the dosage form of the cosmetic composition include lotions, creams, essences, soaps, shampoos, care solutions, foams, foundations and the like.

[0592] The content of outer membrane vesicles in the cosmetic composition can be appropriately set in consideration of the form of the cosmetic composition and the required function. For example, the content (number) of outer membrane vesicles per 1 mL of the cosmetic composition is 2×10 3 ~2×10 14 / mL, preferably 2×10 4 ~2×10 12 Pieces / mL.

[0593] The above description about the cosmetic composition can also be applied to the case where the IL-10 inducer, skin protective agent, and nerve cell protective agent according to the present invention are used as a cosmetic composition.

[0594] 〔Food and Beverage〕

[0595] The outer membrane vesicles derived from specific lactic acid bacteria of the present invention can be used in the form of food and beverages (food and beverages). Therefore, one embodiment of the present invention is a food and beverage containing outer membrane vesicles derived from lactic acid bacteria. Food and beverages include all food and beverages that can be combined with outer membrane vesicles derived from lactic acid bacteria.

[0596] As a specific example of food and drink, functional food and drink (tablet candy, tablet, chewable tablet, tablet, powder, powder, capsule, granule and supplement etc. are in various forms) can be listed; Beverage (tea beverage (green tea, oolong tea, black tea etc.), refreshing drink, jelly drink, sports drink, milk drink, carbonated drink, vegetable drink, fruit juice drink, fermented vegetable drink, fermented fruit juice drink, fermented milk drink (yogurt etc.), lactobacillus drink, milk drink (coffee milk, fruit milk etc.), powdered drink, cocoa drink, milk and purified water etc.); Spread (butter, jam, topping, margarine etc.); Mayonnaise; Shortening; Custard cream; Sauce; Bread; Rice; Noodles; Pasta; Miso soup; Tofu; Yogurt; Soup or sauce; Dessert (biscuit, cookie, chocolate, candy, cake, ice cream, chewing gum, table candy etc.), liquid food (enteric nutrient etc. through tube) etc. Among them, preferred functional food and drink.

[0597] Functional foods and beverages refer to foods and beverages in which the outer membrane vesicles derived from specific lactic acid bacteria of the present invention exert certain functions on the organism (such as anti-inflammatory effects, epithelial cell regeneration and repair effects, brain function improvement effects, etc.) by stimulating FPR2, that is, foods and beverages used to stimulate FPR2.

[0598] Functional food and drink products include all of so-called healthy food and drink products. As examples of functional food and drink products, health functional foods (specific health food products (including conditional specific health food products), functional labeled foods, nutritional functional foods), special purpose food and drink products, nutritional supplement food and drink products, health supplement food and drink products, supplements (including various dosage forms such as tablets, coated tablets, sugar-coated tablets, capsules, liquids) and beauty food and drink products (weight loss food and drink products, etc.) etc. can be listed.

[0599] Functional foods and drinks include health foods and drinks to which health claims are applied based on food standards of CODEX (Joint Food Commission of FAO / WHO).

[0600] The food or beverage may optionally contain additives permitted to be incorporated into food or beverages in addition to the outer membrane vesicles derived from lactic acid bacteria.

[0601] Examples of additives include color developers (sodium nitrite, etc.); colorants (gardenia pigment, red 102, etc.); flavors (orange flavor, etc.); sweeteners (stevia, aspartame, etc.); preservatives (sodium acetate, sorbic acid, etc.); emulsifiers (sodium chondroitin sulfate, propylene glycol fatty acid esters, etc.); antioxidants (disodium EDTA, vitamin C, etc.); pH adjusters (citric acid, etc.); chemical seasonings (sodium inosinate, etc.); thickeners (xanthan gum, etc.); leavening agents (calcium carbonate, etc.); defoaming agents (calcium phosphate); binding agents (sodium polyphosphate, etc.); nutritional enhancers (calcium enhancers, vitamin A, etc.); excipients (water-soluble dextrin, etc.), etc.

[0602] Foods and beverages containing outer membrane vesicles derived from lactic acid bacteria can be produced by means available in the food field.

[0603] For example, the outer membrane vesicles prepared in liquid, gel, solid, powder or granular form can be added to food or beverages. Alternatively, the outer membrane vesicles can be directly mixed, dissolved or suspended in food or beverage raw materials.

[0604] Outer membrane vesicles can be coated, covered, infiltrated or insufflated into food and beverages.

[0605] Outer membrane vesicles can be dispersed evenly or unevenly in food and beverage.

[0606] In the case of functional foods and beverages, the outer membrane vesicles may be coated with a capsule, an edible film, an edible coating agent, or the like.

[0607] In the case of functional foods and beverages, appropriate excipients and the like may be added to the outer membrane vesicles and then molded into a tablet or the like.

[0608] Foods and beverages containing outer membrane vesicles can be further processed, and such processed products are also included in the scope of the present invention.

[0609] The content of outer membrane vesicles in food and beverages can be appropriately set in consideration of the form of the food and beverage, the desired taste, etc. In the case of beverages, the content (number) of outer membrane vesicles per 1 mL of the beverage is, for example, 2×10 3 ~2×10 14 / mL, preferably 2×10 4 ~2×10 12 Pieces / mL.

[0610] The intake interval can be appropriately set according to the type of food or drink, and may be once a day or several times a day.

[0611] The above description about foods and drinks can also be applied to the case where the IL-10 inducer, skin protective agent, and nerve cell protective agent according to the present invention are used as foods and drinks.

[0612] 〔feed〕

[0613] The outer membrane vesicles derived from the specific lactic acid bacteria of the present invention can also be used as animal feed.

[0614] Examples of feed include feed for livestock (cattle, pigs, etc.), racehorses, and pets (dogs, cats, etc.).

[0615] Feed is basically the same as food and drink except that the target is animals other than humans, so the above description about food and drink can also be applied to feed.

[0616] The above description about feed can also be applied to the case where the IL-10 inducer, skin protective agent, and nerve cell protective agent according to the present invention are used as feed.

[0617] 〔Consumer product composition〕

[0618] The outer membrane vesicles derived from the specific lactic acid bacteria of the present invention can be used as an active ingredient of a consumer product composition. Therefore, one embodiment of the present invention is a consumer product composition containing the outer membrane vesicles derived from lactic acid bacteria as an active ingredient.

[0619] The consumer product composition of the present invention refers to a consumer product composition that exerts certain functions on the organism (such as anti-inflammatory effects, regeneration and repair of epithelial cells, etc.) by stimulating FPR2 through the outer membrane vesicles derived from specific lactic acid bacteria of the present invention, that is, a consumer product composition for stimulating FPR2.

[0620] Examples of consumer product compositions include deodorants, antibacterial agents, antiperspirants, fine fragrances such as perfumes, compositions for treating hard surfaces such as cloths or floors, and products such as wipes, with deodorants and antibacterial agents being preferred. Examples of dosage forms of consumer product compositions include liquid, solid, semisolid, gel, spray, foam, liquid film, and foam.

[0621] In addition to the outer membrane vesicles derived from lactic acid bacteria, the consumer product composition may optionally contain additives that are allowed to be formulated in the consumer product composition. Examples of additives include surfactants, chelating agents, dispersants, enzymes, enzyme stabilizers, whitening agents, foam suppressants, dyes, fragrances, pigments, fabric softeners, anti-agglomerants, and the like.

[0622] The content of the outer membrane vesicles in the consumer product composition can be appropriately set in consideration of the dosage form. For example, the content (number) of the outer membrane vesicles per 1 mL of the consumer product composition is 2×10 3 ~2×10 14 / mL, preferably 2×10 4 ~2×10 12 Pieces / mL.

[0623] The above description about the consumer product composition can also be applied to the case where the IL-10 inducer, skin protectant and nerve cell protectant based on the present invention are used as a consumer product composition.

[0624] Example

[0625] Next, the effects of the present invention will be specifically described by way of examples, but the present invention is not limited to the examples.

[0626] 〔Test strain〕

[0627] The 26 types of bacteria shown in Table 1 were used.

[0628]

[0629] The bacteria Nos. 1 to 14 and 20 to 26 are lactic acid-producing bacilli.

[0630] The bacteria No. 15 to 16 are lactic acid-producing cocci.

[0631] The bacteria numbered 17 to 19 are Escherichia coli.

[0632] Bacteria Nos. 12 to 14 and 17 to 19 were used as comparative examples.

[0633] [Preparation Example: Preparation of outer membrane vesicles derived from lactic acid bacteria]

[0634] [Preparation of outer membrane vesicles from lactic acid-producing bacilli]

[0635] (1) Lactic acid-producing bacilli were subjected to pre-culture (static culture at 37° C. for 16 hours) using MRS liquid medium (manufactured by BD Japan).

[0636] The pre-culture solution was inoculated into MRS liquid medium (inoculation amount: 5 volume %) and subjected to main culture (static culture at 37°C for 16 hours).

[0637] The main culture solution was subjected to centrifugation (TOMY SEIKO, MX-301, 10,000 g for 5 minutes).

[0638] (2) The supernatant after centrifugation was filtered through a polyvinylidene fluoride (PVDF) membrane filter (pore size: 0.45 μm).

[0639] The filtrate was subjected to ultracentrifugation (Optima XE-90 manufactured by Beckman Coulter, 150,000 g for 2 hours).

[0640] After ultracentrifugation, the supernatant was removed and the obtained precipitate was suspended in phosphate buffered saline (PBS) (2.5 ml) that had been previously filtered through a PVDF membrane filter (pore size: 0.1 μm) to obtain an outer membrane vesicle suspension.

[0641] The outer membrane vesicle suspension and MRS liquid culture medium (manufactured by BD Japan) filtered in advance using a PVDF membrane filter (pore size: 0.1 μm) were mixed at a volume ratio of 1:1.

[0642] The mixed solution was treated with ExoQuick-TC (registered trademark) ULTRA (manufactured by System Biosciences, LLC), and then filtered through a PVDF membrane filter (pore size: 0.45 μm) to obtain a purified outer membrane vesicle suspension.

[0643] (3) Based on the number of outer membrane vesicles determined by the quantitative procedure described below, the purified outer membrane vesicle suspension was appropriately diluted with the buffer attached to ExoQuick-TC (registered trademark) ULTRA (manufactured by System Biosciences, LLC) to prepare an outer membrane vesicle concentration of 6×10 8 This suspension was stored frozen (-20°C) until it was used in the activity test described below.

[0644] 〔Quantification of outer membrane vesicles〕

[0645] The purified outer membrane vesicle suspension (100 μl) was diluted 10-fold with phosphate-buffered saline (PBS) containing 5.56 μM 3,3'-dioctadecyloxocarbocyanine perchlorate (biological membrane labeling fluorescent probe), and then reacted in the dark (37°C for 2 hours) to label the outer membrane vesicles.

[0646] The reaction solution was filtered through a PVDF membrane filter (pore size: 0.45 μm), and the filtrate was diluted 100-fold with PBS.

[0647] The number of outer membrane vesicles in the dilution solution was measured using the “Small particle detection mode” of an ultra-high-sensitivity high-end tabletop flow cytometer (Cell Stream, manufactured by Luminex).

[0648] In the measurement, the output of laser light at wavelengths of 488 nm and 642 nm was set to 100%, and the output of forward scattered light and side scattered light was set to 0%.

[0649] The number of particles detected at an excitation wavelength of 488 nm and a fluorescence wavelength of 528 nm was measured for 180 seconds, and the number of particles detected was regarded as the number of outer membrane vesicles contained in the purified outer membrane vesicle suspension.

[0650] [Preparation of outer membrane vesicles from lactic acid-producing cocci]

[0651] Lactococcus was subjected to pre-culture using GLM17 medium (static culture at 30° C. for 16 hours). GLM17 medium was prepared by adding 0.5% by mass of glucose and 0.5% by mass of lactose to M17 liquid medium (manufactured by BD Biosciences).

[0652] The preculture solution was inoculated into GLM17 medium (inoculation amount: 5 volume %) and subjected to main culture (static culture at 30°C for 16 hours).

[0653] The main culture solution was subjected to centrifugation (TOMY SEIKO, MX-301, 10,000 g for 5 minutes).

[0654] The supernatant after centrifugation was subjected to the steps (2) and (3) of the above-mentioned [Preparation of outer membrane vesicles from lactic acid-producing bacilli] to obtain an outer membrane vesicle number concentration of 6×10 8 This suspension was stored frozen (-20°C) until it was used for the activity test.

[0655] 〔Preparation of outer membrane vesicles from Escherichia coli〕

[0656] Escherichia coli was subjected to preculture (shaking culture at 37°C for 16 hours) using LB medium (manufactured by Kanto Chemical).

[0657] The preculture solution was inoculated into LB medium (inoculation amount: 5 volume %) and subjected to main culture (shaking culture at 37°C for 16 hours).

[0658] The main culture solution was subjected to centrifugation (TOMY SEIKO, MX-301, 8,000 g for 5 minutes).

[0659] The supernatant after centrifugation was subjected to the steps (2) and (3) of the above-mentioned [Preparation of outer membrane vesicles from lactic acid-producing bacilli] to obtain an outer membrane vesicle number concentration of 6×10 8 This suspension was stored frozen (-20°C) until it was used for the activity test.

[0660] [Example 1: FPR2 agonist activity]

[0661] The FPR2 agonist activity of outer membrane vesicles was evaluated using the Tanso Biosciences GPCR contract test (Cosmobio). In this test, a report test using the measurement principle described in International Publication No. WO2020 / 026979 was performed.

[0662] As the evaluation sample, an outer membrane vesicle suspension (final concentration of outer membrane vesicles: 2×10 6 / mL).

[0663] The blank sample was prepared according to the following steps. MRS liquid culture medium (manufactured by BD Japan) filtered through a PVDF membrane filter (pore size: 0.1 μm) and phosphate buffered saline (PBS) filtered through a PVDF membrane filter (pore size: 0.1 μm) were mixed at a volume ratio of 1:1. The mixed solution was treated with ExoQuick-TC (registered trademark) ULTRA (manufactured by System Biosciences, LLC) to obtain a blank sample.

[0664] In the positive control sample, the peptide WKYMVM (Trp-Lys-Tyr-Met-Val-Met-NH2) (SEQ ID NO: 1) was used instead of the outer membrane vesicle. This peptide is known to have FPR2 agonist activity (Molecules 2017, 22, 455). The sample containing only the solvent used in the experiment was used as the negative control sample.

[0665] The test was performed three times (the number of sample data (n) = 3).

[0666] The FPR2 activation rate was calculated according to the following formula: The activation rate result was expressed as the average of the values ​​calculated in three experiments.

[0667] FPR2 activation rate (%) = [(reported luminescence amount of the test sample - reported luminescence amount of the negative control sample) / (reported luminescence amount of the positive control sample - reported luminescence amount of the negative control sample)] × 100

[0668] The Student's t test was used for statistical analysis. The test was performed by comparing the excitation rate of the outer membrane vesicles derived from Escherichia coli T (bacterial number 17 in Table 1). The level of p < 0.05 was considered to be significantly different. The results are shown in Tables 2 and Figure 1 .

[0669] Table 2

[0670]

[0671] As shown in Table 2 and Figure 1 As shown, the outer membrane vesicles derived from the lactic acid bacteria (bacterial numbers 1 to 11, 15 and 16 and 20 to 26) based on the present invention have FPR2 agonist activity. On the other hand, the outer membrane vesicles derived from lactic acid bacteria other than the lactic acid bacteria based on the present invention and the outer membrane vesicles derived from Escherichia coli do not have FPR2 agonist activity. This shows that among the outer membrane vesicles derived from bacteria, the outer membrane vesicles derived from the specific species of lactic acid bacteria based on the present invention are useful as FPR2 agonists.

[0672] [Example 2: FPR1 agonistic activity]

[0673] The FPR1 agonist activity of outer membrane vesicles was evaluated by Tanso Biosciences GPCR commissioned test (Cosmobio). In this test, a report test using the measurement principle described in International Publication No. WO2020 / 026979 was performed.

[0674] As the evaluation sample, an outer membrane vesicle suspension (final concentration of outer membrane vesicles: 2×10 6 / mL).

[0675] The blank sample was prepared according to the following steps. MRS liquid culture medium (manufactured by BD Japan) filtered through a PVDF membrane filter (pore size: 0.1 μm) and phosphate buffered saline (PBS) filtered through a PVDF membrane filter (pore size: 0.1 μm) were mixed at a volume ratio of 1:1. The mixed solution was treated with ExoQuick-TC (registered trademark) ULTRA (manufactured by System Biosciences, LLC) to obtain a blank sample.

[0676] For the positive control sample, N-formylmethionyl-leucyl-phenylalanine (fMLP) was used instead of outer membrane vesicles. This peptide is known to have FPR1 agonist activity (literature name: Molecules 2017, 22, 455). The sample containing only the solvent used in the test was used as a negative control sample.

[0677] The test was carried out three times (the number of sample data (n) = 3).

[0678] The FPR1 activation rate was calculated according to the following formula. The activation rate results were expressed as the average of the values ​​calculated in 3 experiments. The results are shown in Tables 3 and Figure 2 .

[0679] FPR1 activation rate (%) = [(reported luminescence amount of the test sample - reported luminescence amount of the negative control sample) / (reported luminescence amount of the positive control sample - reported luminescence amount of the negative control sample)] × 100

[0680] Table 3

[0681]

[0682] As shown in Table 3 and Figure 2 As shown, the outer membrane vesicles derived from the lactic acid bacteria of the present invention (bacteria Nos. 1 to 11, 15 and 16, and 20 to 26) do not have FPR1 agonist activity. This indicates that the outer membrane vesicles selectively agonize FPR2 and are useful as FPR2 selective agonists.

[0683] [Reference Example 1: Effect of a known FPR2 agonist on IL-10 production by M2 macrophages]

[0684] The ability of M2 macrophages differentiated from human monocyte-derived cells (THP-1, JCRB cell bank) to induce the anti-inflammatory cytokine IL-10 was evaluated by the following procedure, which was an improvement of the method described in the literature (BMC Cancer 15: 577 (2015)).

[0685] THP-1 cells were subcultured in a medium (growth medium) prepared by adding 10 vol% bovine serum albumin (hiFBS, Cytiva) and 1 vol% penicillin-streptomycin solution (Fujifilm Wako Pure Chemical Industries, Ltd.) inactivated at 56°C for 30 minutes to RPMI 1640 medium (Nacalai Tesque) and incubated at 1×10 6 Cells were suspended at a concentration of 10 cells / ml in a growth medium supplemented with 150 nM phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich), seeded in 96-well plates at 0.1 ml each, and cultured at 37°C in the presence of 5 vol% CO2 for 24 hours.

[0686] After the culture, the medium was removed, the cells were washed twice with a growth medium, 0.1 ml of the growth medium was added, and the cells were cultured at 37°C for 24 hours in the presence of 5% by volume CO2.

[0687] After the culture, the medium was removed, and 0.1 ml of a culture medium prepared by adding 20 ng / ml human IL-4 (R&D Systems) and 20 ng / ml human IL-13 (Thermo Fisher Scientific) to a proliferation medium was added, and the cells were cultured at 37°C in the presence of 5 vol% CO2 for 24 hours.

[0688] After the culture, the cells were washed twice with a growth medium, 0.1 ml of the growth medium was added, 2 vol% of a test substance (described later) was added, and the cells were cultured at 37°C for 24 hours in the presence of 5% CO2.

[0689] After the culture, the culture supernatant was recovered, and the IL-10 concentration in the supernatant was calculated using Human IL-10 DuoSet ELISA (R&D Systems).

[0690] It is known that the IL-10 induction ability of M2 macrophages is induced by Toll-like receptor (TLR2) agonists (Arthritis Research & Therapy 19: 245 (2017)). Therefore, as test samples, the following three types were used: FSL-1 (InvivoGen) 500ng / ml as a TLR2 agonist (FSL-1 group); a mixture of FSL-1 500ng / ml and WKYMVm (Trp-Lys-Tyr-Met-Val-D-Met-NH2 (SEQ ID NO. 2), Abcam) 5000nM as a FPR2 agonist (FSL-1+WKYMVm group); and, solvent only (negative control group).

[0691] The test sample was added to the cells in the test state at 2% by volume, so that the final concentration of the test sample during the test was 1 / 50 of that during preparation.

[0692] The Student's t-test was used for statistical analysis. The IL-10 concentration in the FSL-1 group was compared with that in the FSL-1 group. A level of p < 0.05 was considered to be significantly different. The number of sample data (n) = 6. The results of IL-10 concentration are expressed as the average of the values ​​measured in 6 tests. The results are shown in Tables 4 and Figure 3 .

[0693] Table 4

[0694]

[0695] As shown in Table 4 and Figure 3 As shown, the co-treatment of FSL-1 and FPR2 agonist induced the secretion of anti-inflammatory cytokine IL-10 at a significantly higher level than the treatment of FSL-1 alone as a TLR2 agonist. This suggests that FPR2 agonistic activity is important in the anti-inflammatory action of immune cells.

[0696] [Example 3: IL-10 inducing activity]

[0697] The outer membrane vesicle concentration was set to 5 × 10 8 A purified outer membrane vesicle suspension derived from lactic acid bacteria (test sample) was prepared according to the procedure described in the above "Preparation Example" except that the above-mentioned procedure had a concentration of 100 cells / mL.

[0698] The IL-10 induction activity of the test sample was evaluated according to the procedure described in the above "Reference Example 1". The blank sample was prepared by the method described in the above "Example 1". FSL-1 (InvivoGen) 500 ng / ml was used as a positive control sample. The sample containing only the solvent used in the test was used as a negative control sample.

[0699] 2% by volume of the test sample was added to the cells in the test state, so that the final concentration of the outer membrane vesicles in the test became 1 / 50 of that in the preparation.

[0700] The IL-10 induction rate was calculated according to the following formula.

[0701] IL-10 induction rate (%) = [(IL-10 concentration when adding test sample - IL-10 concentration when adding negative control sample) / (IL-10 concentration when adding positive control sample - IL-10 concentration when adding negative control sample)] × 100

[0702] The Student's t-test was used for statistical analysis. The results were compared with the induction rate of outer membrane vesicles derived from Lactobacillus reuteri (bacterial number 12 in Table 1). The level of p < 0.05 was considered to be significantly different. The number of data for the sample (n) = 6. The results of the IL-10 induction rate are expressed as the average of the values ​​calculated in 6 experiments. The results are shown in Tables 5 and Figure 4 .

[0703] Table 5

[0704]

[0705] As shown in Table 5 and Figure 4 As shown, the outer membrane vesicles derived from lactic acid bacteria with FPR2 agonist activity (bacterial numbers 1 to 4, 20, 21, 24, 25, 7, 9, 10, 26, 11 and 15 (Example 1)) showed a significantly higher IL-10 induction rate than the outer membrane vesicles of lactic acid bacteria without FPR2 agonist activity (bacterial number 12). This shows that among the outer membrane vesicles derived from bacteria, the outer membrane vesicles derived from the specific species of lactic acid bacteria based on the present invention are useful as IL-10 inducers based on the agonism of FPR2. This shows that the agonist activity of FPR2 is important for the anti-inflammatory effect brought about by the outer membrane vesicles derived from the specific species of lactic acid bacteria based on the present invention.

[0706] [Example 4: Effect of FPR2 antagonists on the IL-10 induction ability of outer membrane vesicles with FPR2 agonist activity]

[0707] The outer membrane vesicle concentration was set to 2.5 × 10 9 A purified outer membrane vesicle suspension derived from Lactobacillus paracasei strain CP3526 (bacterial number 9) was prepared according to the procedure described in the above "Preparation Example" except that the above-mentioned procedure had a concentration of 100 cells / mL.

[0708] The following three types were used as test samples: outer membrane vesicles only (CP3526 group); a mixture of outer membrane vesicles and 50000nM WRW4 (H-Trp-Arg-Trp-Trp-Trp-Trp-Trp-NH2 (sequence number 3), Bachem AG) as an FPR2 antagonist (CP3526+WRW4 group); and, solvent only (negative control group).

[0709] 2% by volume of the test sample was added to the cells in the test state, so that the final concentration of the outer membrane vesicles in the test became 1 / 50 of that in the preparation.

[0710] The IL-10 concentration was evaluated according to the procedure described in the above-mentioned "Reference Example 1".

[0711] Student's t-test was used for statistical processing. The IL-10 concentration in the CP3526 group was compared with that in the CP3526 group. A level of p < 0.05 was considered to be significantly different. The number of sample data (n) = 6. The results of IL-10 concentration are expressed as the average of the values ​​measured in 6 tests. The results are shown in Tables 6 and Figure 5 .

[0712] Table 6

[0713]

[0714] As shown in Table 6 and Figure 5 As shown, the IL-10 concentration when the FPR2 antagonist was added (CP3526+WRW4 group) was significantly reduced compared to the non-addition (CP3526 group). This shows that the outer membrane vesicles derived from the Lactobacillus paracasei CP3526 strain based on the present invention induce IL-10 by stimulating FPR2. This shows that the FPR2 agonist activity is necessary for the anti-inflammatory effect of the outer membrane vesicles derived from the specific species of lactic acid bacteria based on the present invention on immune cells.

[0715] [Example 5: Effect of protecting skin from ultraviolet damage]

[0716] The outer membrane vesicle concentration was set to 1.0 × 10 10 A purified outer membrane vesicle suspension derived from Lactobacillus paracasei strain CP3526 (bacterial number 9) was prepared according to the procedure described in the above "Preparation Example" except that the above-mentioned procedure had a concentration of 100 cells / mL.

[0717] As test samples, the following three types were used: outer membrane vesicles alone (CP3526 group); a mixture of outer membrane vesicles and 50,000 nM WRW4 (Bachem AG), an antagonist of FPR2 (CP3526+WRW4 group); and a solvent alone (negative control group).

[0718] Proliferated normal human adult epidermal keratinocytes (NHEK, KURABO) were cultured at 1.0×10 5 The cells were suspended in Humedia-KG2 (KURABO) at a concentration of 10 cells / ml, 0.1 ml of the suspension was seeded in a 96-well black plate, and cultured at 37°C in the presence of 5 vol% CO2 for 24 hours.

[0719] After incubation, the cells were washed with PBS, 0.02 ml of PBS was added, and irradiation was performed at 75 mJ / cm 2 of UV-B.

[0720] After irradiation, PBS was removed and 0.1 ml of Humedia-KG2 containing 2% by volume of the test sample was added, so that the final concentration of outer membrane vesicles during the test became 1 / 50 of that during the preparation.

[0721] After adding the test sample, the cells were cultured for 24 hours, and the number of viable cells was measured by the WST-8 method.

[0722] The determination using the WST-8 method is as follows. The culture medium is exchanged with Humedia-KG2 containing 10% by volume of the cell counting reagent SF (Nacalai tesque), cultured in a CO2 incubator, and the change in absorbance (450nm) within 60 minutes from 30 minutes to 90 minutes is measured. The value of each change is divided by the average value of the change in absorbance when not irradiated with UV-B measured by 3 tests, and then multiplied by 100, and the resulting value is used as the cell survival rate.

[0723] The Student's t-test was used for statistical analysis. The cell viability was compared with that of the CP3526 group. A level of p < 0.05 was considered to be significantly different. The number of sample data (n) = 3. The results of cell viability are expressed as the average of the values ​​calculated in 3 tests. The results are shown in Tables 7 and Figure 6 .

[0724] Table 7

[0725]

[0726] As shown in Table 7 and Figure 6 As shown, by adding outer membrane vesicles, the survival rate of skin cells after ultraviolet irradiation was significantly improved compared with no addition. Furthermore, by adding FPR2 antagonists, the survival rate was significantly reduced compared with no addition. This shows that the outer membrane vesicles derived from the Lactobacillus paracasei CP3526 strain based on the present invention protect the skin from ultraviolet damage by exciting FPR2. This shows that FPR2 agonist activity is necessary for the protective effect of outer membrane vesicles derived from the specific species of lactic acid bacteria based on the present invention on skin cells.

[0727] [Example 6: Effect of protecting nerve cells from nerve cell toxicity]

[0728] The outer membrane vesicle concentration was set to 1.0 × 10 10 A purified outer membrane vesicle suspension derived from Lactobacillus paracasei strain CP3526 (bacterial number 9) was prepared according to the procedure described in the above "Preparation Example" except that the above-mentioned procedure had a concentration of 100 cells / mL.

[0729] As test samples, two types of samples were used: outer membrane vesicles alone (CP3526 group) and a solvent alone (negative control group).

[0730] 8×10 50 μL of DMEM (1.0 g / l glucose) (containing L-glutamine and pyruvic acid) (liquid) (DMEM (LG), Nacalaitesque) was added with 5 vol% bovine serum albumin (FBS, Themo Fisher Scientific), 10 vol% horse serum (hiHS, Themo Fisher Scientific) inactivated at 56°C for 30 minutes, and 1 vol% penicillin-streptomycin solution (P / S) (Nacalai tesque) (growth induction medium). 4 Proliferated PC-12 cells (ECACC) were suspended at a concentration of 10 cells / ml, 0.1 ml of which was seeded in a 96-well plate (Corning) coated with type IV collagen, and cultured at 37°C in the presence of 5% by volume CO2.

[0731] On the day after seeding and on the 4th day, the medium was replaced with differentiation medium (a medium prepared by adding 0.1 volume % FBS, 0.1 volume % hiHS, 1 volume % P / S and 50 ng / ml Mouse NGF 2.5S Native Protein (NGF, Themo Fisher Scientific) to DMEM (LG)) to induce neural differentiation.

[0732] On the 5th day after seeding, the medium was exchanged with a differentiation medium containing 2 vol % of the test sample, and culture was carried out at 37° C. for 1 hour in the presence of 5 vol % CO 2 .

[0733] Thereafter, the medium was replaced with a differentiation medium containing 60 μM β-amyloid 1-42 (amyloid β, Sigma-Aldrich) (neuronal cell toxicity) and 2 vol % of the test sample, and cultured at 37° C. for 24 hours in the presence of 5 vol % CO 2 .

[0734] After incubation, measure the amount of intracellular ATP. The measurement method is as follows. Add "Cell" ATP Assayreagent Ver.2 (Toyo B-Net) at 0.1 ml / well and stir for 1 minute using a microplate shaker. Incubate at 23°C for 10 minutes, transfer 100 μl to a 96-well black plate, and measure the luminescence. Divide each luminescence by the average luminescence measured in 5 tests without amyloid β treatment, multiply by 100, and use the resulting value as the cell survival rate.

[0735] The Student's t test was used for statistical analysis. The cell viability was compared with that of the CP3526 group. A level of p < 0.05 was considered to be significantly different. The number of sample data (n) = 5. The results of cell viability are expressed as the average of the values ​​calculated in 5 tests. The results are shown in Table 8 and Figure 7 .

[0736] Table 8

[0737]

[0738] As shown in Table 8 and Figure 7 As shown, by adding outer membrane vesicles, the survival rate of neurons treated with amyloid β was significantly improved compared with that without addition. This shows that the outer membrane vesicles derived from the Lactobacillus paracasei CP3526 strain based on the present invention protect neurons from neurotoxic damage. This shows that the outer membrane vesicles derived from the specific species of lactic acid bacteria based on the present invention have a useful effect on the damage of neurons to neurons by neurotoxicity.

[0739] INDUSTRIAL APPLICABILITY The present invention can be used in medicines, foods and drinks, and the like.

Claims

1. An FPR2 agonist comprising an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

2. The FPR2 agonist according to claim 1, wherein The lactic acid bacteria is at least one selected from the group consisting of: Lactobacillus delbrueckii subsp.bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Lactococcus lactis subsp. lactis, Streptococcus salivarius subsp.thermophilus, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lactobacillus casei, and Lactiplantibacillus pentosus.

3. The FPR2 agonist according to claim 1 or 2, wherein Outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

4. A food or beverage for stimulating FPR2, comprising as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

5. A pharmaceutical composition for treating a disease associated with FPR2, comprising as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

6. A cosmetic composition for exciting FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

7. A consumer product composition for exciting FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

8. The food or beverage according to claim 4, the pharmaceutical composition according to claim 5, the cosmetic composition according to claim 6 or the consumer product composition according to claim 7, wherein: Outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

9. Lacticaseibacillus paracasei CP3526 strain with the deposit number NITE BP-03625.

10. A food or beverage for inducing IL-10 by stimulating FPR2, comprising, as an effective ingredient, outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

11. A pharmaceutical composition for treating a disease associated with IL-10 by inducing IL-10 by stimulating FPR2, comprising as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

12. A cosmetic composition for inducing IL-10 by stimulating FPR2, comprising, as an effective ingredient, outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

13. A consumer product composition for inducing IL-10 by agonizing FPR2, comprising outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

14. The food or beverage according to claim 10, the pharmaceutical composition according to claim 11, the cosmetic composition according to claim 12 or the consumer product composition according to claim 13, wherein: Outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

15. A food or beverage for protecting the skin from ultraviolet damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

16. A pharmaceutical composition for protecting the skin from ultraviolet damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

17. A cosmetic composition for protecting the skin from ultraviolet rays, comprising as an effective ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

18. A consumer product composition for protecting the skin from UV damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

19. The food or beverage according to claim 15, the pharmaceutical composition according to claim 16, the cosmetic composition according to claim 17 or the consumer composition according to claim 18, which protects the skin from ultraviolet damage by stimulating FPR2.

20. The food or beverage according to claim 15, the pharmaceutical composition according to claim 16, the cosmetic composition according to claim 17 or the consumer product composition according to claim 18, wherein: Outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

21. A food or beverage for protecting nerve cells from nerve cell toxicity, comprising as an effective ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

22. A pharmaceutical composition for protecting nerve cells from nerve cell toxicity, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

23. A cosmetic composition for protecting nerve cells from nerve cell toxicity, comprising as an effective ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

24. A consumer product composition for protecting nerve cells from neurotoxic damage, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

25. The food or beverage according to claim 21, the pharmaceutical composition according to claim 22, the cosmetic composition according to claim 23, or the consumer composition according to claim 24, which protects neurons from neurotoxic damage by stimulating FPR2.

26. The food or beverage according to claim 21, the pharmaceutical composition according to claim 22, the cosmetic composition according to claim 23 or the consumer product composition according to claim 24, wherein: Neurotoxicity is the neurotoxicity caused by amyloid β.

27. The food or beverage according to claim 21, the pharmaceutical composition according to claim 22, the cosmetic composition according to claim 23 or the consumer product composition according to claim 24, wherein: Outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

28. A method for agonizing FPR2 in a subject, comprising the step of administering to the subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

29. A method for treating a disease associated with FPR2 in a subject, comprising the step of administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

30. A method for inducing IL-10 in a subject by agonizing FPR2, comprising the step of administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

31. A method for protecting a subject's skin from ultraviolet damage, comprising the step of administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

32. The method according to claim 31, wherein: Protects skin from UV damage by stimulating FPR2.

33. A method for protecting neural cells from neural cell toxicity in a subject, comprising the step of administering to the subject outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of: Lactobacillus spp., Lactobacillus spp., Lactiplantibacillus spp., Combined with Lactobacillus spp., Lactococcus, and Streptococcus salivarius subsp. thermophilus.

34. The method of claim 33, which protects neural cells from neural cytotoxicity by agonizing FPR2.

35. The method according to any one of claims 28 to 34, wherein: The lactic acid bacteria is at least one selected from the group consisting of: Lactobacillus delbrueckii subsp.bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Lactococcus lactis subsp. lactis, Streptococcus salivarius subsp.thermophilus, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lactobacillus casei, and Lactiplantibacillus pentosus.

36. The method according to any one of claims 28 to 35, wherein: Outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

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