Anti-aging composition comprising bacillus subtilis flagellin variant

Abstract

The present invention relates to an anti-aging composition comprising a flagellin variant and, more specifically, to: an anti-aging composition comprising a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1; a composition for extending lifespan; a composition for preventing inoculating against hair loss; and a composition for preventing or inoculating against a decrease in muscle strength or exercise ability.

Description

Anti-aging composition comprising a Bacillus subtilis flagellin variant

[0001] This application claims the benefit of Korean Patent Application No. 10-2024-0009025, filed on January 19, 2024, and Korean Patent Application No. 10-2025-0007144, filed on January 17, 2025, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to an anti-aging composition comprising a flagellin mutant, and more particularly, to an anti-aging composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1, a composition for extending life, a composition for preventing or preventing hair loss, and a composition for preventing or preventing a decrease in muscle strength or exercise capacity.

[0003]

[0004] Flagellin is a unit protein of filament, one of the components of bacterial flagella. Filaments are formed by regular combinations of flagellins and function by extending from the cell surface to enable bacterial movement. Flagellin acts as a virulence factor in pathogenic bacteria, facilitating their invasion and invasion into host cells. Flagellin is found exclusively in bacteria and is one of the most abundant proteins in flagellated bacteria, making it a key target of host immune surveillance. When bacteria invade the host, flagellin is detected by TLR5 and NAIP5 / NLRC4 in the host, activating the innate immune system that contributes to the immediate elimination of the pathogen. It is known in the art that mammalian TLR5 (Toll-like receptor 5) recognizes flagellin from both Gram-negative and Gram-positive bacteria and activates NF-κB.

[0005] Flagellin has attracted attention as a potential candidate for vaccine carrier protein development or vaccine adjuvant development because it serves as the first line of defense against flagellated pathogenic bacteria. Fusion proteins of antigens and flagellin have proven effective as experimental vaccines against various infectious diseases, and flagellin-mediated TLR5 activation has also been reported to protect hematopoietic cells and gastrointestinal tissues from radiation damage, as well as to influence the survival and growth of cancer cells. However, flagellin itself, when used as a biological therapeutic, poses the problem of side effects due to its ability to act as an antigen for the body's immune response. Therefore, the present inventors have previously developed and disclosed a flagellin with reduced immunogenicity (see Patent Publication No. 10-2024-0022417).

[0006] Meanwhile, TLR5 is an innate immune receptor located on the cell surface and consists of an extracellular leucine-rich repeat (LRR) domain, a transmembrane domain, and an intracellular domain. TLR5 recognizes flagellin as a pathogen-associated molecular pattern using its extracellular domain, activating the MyD88-dependent signaling pathway and NF-κB-mediated inflammatory cytokine production.

[0007] TLR expression is known to be decreased in immune cells of elderly humans and mice, and the age-related decline in TLR1 / 4 contributes to the decreased antibody response to influenza vaccine in elderly patients, and TLR2 signaling is also reduced in aged alveolar macrophages, making them ineffective against bacterial infection. In contrast, the expression and signaling of TLR5 were found to be relatively well preserved in aged mice compared to other types of TLRs, and accordingly, flagellin, a TLR5 ligand and enhancer of antigen-specific immune responses, was confirmed to induce similar levels of inflammatory cytokines in macrophages from both young and aged mice.

[0008] Based on this background, the present inventors sought to determine whether the Bacillus subtilis flagellin mutant has any effect on aging by activating TLR5.

[0009]

[0010] Accordingly, the inventors of the present invention completed the present invention by confirming that administering a Bacillus subtilis flagellin mutant with reduced immunogenicity of wild-type Bacillus subtilis flagellin into the nasal cavity of aged mice had a positive effect on the health and various physiological aspects of aged mice, thereby extending their lifespan.

[0011]

[0012] Accordingly, an object of the present invention is to provide an anti-aging composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0013]

[0014] Another object of the present invention is to provide a life extension composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0015]

[0016] Another object of the present invention is to provide a composition for preventing, improving or treating hair loss, comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0017]

[0018] Another object of the present invention is to provide a composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity, comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0019]

[0020] Another object of the present invention is to provide a use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing an anti-aging composition.

[0021]

[0022] Another object of the present invention is to provide an anti-aging method comprising administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0023]

[0024] Another object of the present invention is to provide a use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for extending lifespan.

[0025]

[0026] Another object of the present invention is to provide a method for extending lifespan, which comprises administering to an individual in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0027]

[0028] Another object of the present invention is to provide a use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, improving or treating hair loss.

[0029]

[0030] Another object of the present invention is to provide a method for preventing, improving or treating hair loss, which comprises administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0031]

[0032] Another object of the present invention is to provide a use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity.

[0033]

[0034] Another object of the present invention is to provide a method for preventing, preventing or improving a decrease in muscle strength or exercise capacity, which comprises administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0035]

[0036] In order to achieve the above purpose, the present invention provides an anti-aging composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0037]

[0038] In order to achieve another object of the present invention, the present invention provides a composition for extending lifespan comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0039]

[0040] In order to achieve another object of the present invention, the present invention provides a composition for preventing, improving or treating hair loss, comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0041]

[0042] In order to achieve another object of the present invention, the present invention provides a composition for preventing, preventing or improving a decrease in muscle strength or exercise ability, comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0043]

[0044] In order to achieve another object of the present invention, the present invention provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing an anti-aging composition.

[0045]

[0046] In order to achieve another object of the present invention, the present invention provides an anti-aging method comprising administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0047]

[0048] In order to achieve another object of the present invention, the present invention provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for extending lifespan.

[0049]

[0050] In order to achieve another object of the present invention, the present invention provides a method for extending life, which comprises administering to an individual in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0051]

[0052] In order to achieve another object of the present invention, the present invention provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, improving or treating hair loss.

[0053]

[0054] In order to achieve another object of the present invention, the present invention provides a method for preventing, improving or treating hair loss, which comprises administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0055]

[0056] In order to achieve another object of the present invention, the present invention provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity.

[0057]

[0058] In order to achieve another object of the present invention, the present invention provides a method for preventing, prophylaxis or improving a decrease in muscle strength or exercise capacity, which comprises administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0059]

[0060] The present invention is described in detail below.

[0061]

[0062] The single (three) letters of amino acids used herein refer to the following amino acids according to standard abbreviation conventions in the field of biochemistry: A (Ala): alanine; C (Cys): cysteine; D (Asp): aspartic acid; E (Glu): glutamic acid; F (Phe): phenylalanine; G (Gly): glycine; H (His): histidine; I (IIe): isoleucine; K (Lys): lysine; L (Leu): leucine; M (Met): methionine; N (Asn): asparagine; O (Pyr): pyrrolysine; P (Pro): proline; Q (Gln): glutamine; R (Arg): arginine; S (Ser): serine; T (Thr): threonine; U (Sec): selenocysteine; V (Val): valine; W (Trp): tryptophan; Y (Tyr): tyrosine.

[0063]

[0064] The present invention provides an anti-aging composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0065]

[0066] In the present invention, the flagellin is a structural protein that assembles into a filament among bacterial flagella as described above, and the Bacillus subtilis flagellin refers to flagellin derived from Bacillus subtilis. The Bacillus subtilis flagellin is known to be able to induce an immune response in an infected host when a flagellar bacterium is infected. More specifically, Toll-like receptor 5 (TLR5) present on the surface of the human cell membrane interacts with the flagellin to induce intracellular signal transduction, thereby increasing the expression of the transcription factor NF-κB, which not only induces innate immune signal activation but also regulates an acquired immune response.

[0067] In the present invention, the wild-type flagellin derived from Bacillus subtilis has an amino acid sequence according to the following sequence number 1.

[0068]

[0069] [Sequence number 1]

[0070] MRINHNIAALNTLNRLGSNNGAAQKNMEKLSSGLRINRAGDDAAGLAISEKMRGQIRGLEMASKNSQDGISLIQTAEGALTETHAILQRMRELTVQAGNTGTQQAEDLGAIKDEMDALIEEIDGISNRTEFNGKKLL DGTNSTDGFTFQIGANAGQQLNVKIDSMSSTALGVNALDVTDFAATAFDDQLKSIDTAINTVSTQRAKLGAVQNRLEHTINNLGASGENLTAAESRIRDVDMAKEMSEFTKNNILSQASQAMLAQANQQPQNVLQLLR

[0071]

[0072] In one aspect of the present invention, the Bacillus subtilis flagellin variant may comprise an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0073] The amino acid sequence having the above sequence identity may be any modification of the amino acid sequence of SEQ ID NO: 1, and may include one or more substitutions, deletions, additions, and / or insertions. Such variants may occur naturally or may be synthetically produced using any number of techniques well known in the art, for example, by modifying or altering one or more of the amino acid sequences of the present invention and evaluating their biological activity as described herein.

[0074] In one embodiment of the present invention, the amino acid sequence having 80% or more sequence identity with SEQ ID NO: 1 comprises conservative substitutions. A 'conservative substitution' is a substitution in which an amino acid is replaced with another amino acid having similar properties, such that a person skilled in the art would predict that the secondary structure and hydropathic nature (hydrophobicity or hydrophilicity) of the peptide are substantially unchanged. Generally, the following amino acid groups exhibit conservative changes: (1) ala, pro, gly, glu, asp, gln, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala, phe; (4) lys, arg, his; and (5) phe, tyr, trp, his.

[0075] The above modifications may be performed within the wild-type Bacillus subtilis flagellin structure according to SEQ ID NO: 1, and may yield functional molecules encoding protein variants or derivatives having desired (desirable) characteristics. When it is desired to change the amino acid sequence of a peptide to produce an equivalent or improved variant of Bacillus subtilis flagellin according to SEQ ID NO: 1, one of ordinary skill in the art can change one or more codons based on protein codon information known in the art.

[0076] Additionally, variants having at least 80% sequence identity with the Bacillus subtilis flagellin according to SEQ ID NO: 1 may include non-conservative changes in amino acids. For example, the variants may be altered by deletion or addition of amino acids that have minimal impact on the secondary structure and hydropathic properties of the Bacillus subtilis flagellin according to SEQ ID NO: 1.

[0077]

[0078] When comparing amino acid sequences of proteins, if the amino acid sequences in the two sequences are identical when the two sequences are aligned with maximum correspondence, as described below, then the two sequences are said to be "identical." Comparison between two sequences is typically performed by comparing the alignments over a comparison window to identify and compare local regions of sequence similarity. As used herein, the term "comparison window" means a segment of at least about 20 consecutive positions, typically 30 to 75, 40 to 50 consecutive positions, in which the sequences can be compared to a reference sequence of the same number of consecutive positions after the two sequences are optimally aligned.

[0079] Optimal alignment of sequences for comparison can be performed using, for example, the MegAlign program in the Lasergene suite of bioinformatics software (DNASTAR, Inc., Madison, Wis.) using default parameters. This program includes several alignment schemes described in the following references: Dayhoff, MO (1978) A model of evolutionary change in proteins-Matrices for detecting distant relationships. In Dayhoff, MO (ed.) Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, Washington DC Vol. 5, Suppl. 3, pp. 345-358; Hein J. (1990) Unified Approach to Alignment and Phylogenes pp. 626-645 Methods in Enzymology vol. 183, Academic Press, Inc., San Diego, Calif.; Higgins, DG and Sharp, PM (1989) CABIOS 5:151-153; Myers, E.W. and Muller W. (1988) CABIOS 4:11-17; Robinson, E. D. (1971) Comb. Theor 11:105; Santou, N. Nes, M. (1987) Mol. Biol. Evol. 4:406-425; Sneath, PHA and Sokal, RR (1973) Numerical Taxonomy—the Principles and Practice of Numerical Taxonomy, Freeman Press, San Francisco, Calif.; Wilbur, W.J. and Lipman, D. J. (1983) Proc. Nat'l Acad., Sci. USA 80:726-730.

[0080] Alternatively, optimal alignment of sequences for comparison may be performed by the partial identity algorithm of Smith and Waterman (1981) Add. APL. Math 2:482, by the identity alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443, by the similarity search method of Pearson and Lipman (1988) Proc. Natl. Acad. Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.), or by inspection.

[0081] Examples of suitable algorithms for determining percent sequence identity and sequence similarity include the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977) Nucl. Acids Res. 25:3389-3402 and Altschul et al. (1990) J. Mol. Biol. 215:403-410, respectively. BLAST and BLAST 2.0 can be used to determine percent sequence identity for the polynucleotides and polypeptides of the present invention, for example, in conjunction with the parameters described herein. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. For amino acid sequences, a scoring matrix can be used to calculate a cumulative score.

[0082] In one exemplary approach, the 'percentage of sequence identity' is determined by comparing two optimally aligned sequences over a comparison window of at least 20 positions, wherein the portion of the polypeptide sequence within the comparison window may contain additions or deletions (i.e., gaps) of no more than 20 percent, typically 5 to 15 percent, or 10 to 12 percent, compared to the reference sequence (which does not include additions or deletions). The percentage of sequence identity is obtained by determining the number of positions at which identical amino acid residues occur in both sequences to obtain the number of matching positions, dividing the number of matching positions by the total number of positions in the reference sequence (i.e., the window size), and multiplying this result by 100.

[0083] The term "percent (%) sequence identity" as used herein is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a reference protein, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percent amino acid identity can be achieved by various methods within the skill in the art, for example, using publicly available computer software programs, such as those described in Current Protocols in Molecular Biology (Ausubel et al., eds., 1987), and using BLAST, BLAST-2, ALIGN, or MegAlign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm necessary to achieve maximum alignment over the full length of the sequences being compared.

[0084] For purposes herein, the percent amino acid sequence identity of a given amino acid sequence A to a given amino acid sequence B is calculated as follows: 100 times the fraction X / Y, where X is the number of amino acid residue scores that are identically matched by a sequence alignment program in the program's alignment of A and B, and Y is the total number of amino acid residues in B. It will be appreciated that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the percent amino acid sequence identity of A to B will not be the same as the percent amino acid sequence identity of B to A.

[0085]

[0086] As a more specific example, according to one aspect of the present invention, the Bacillus subtilis flagellin mutant may mean one in which amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 are deleted from the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO: 1. In this case, the Bacillus subtilis mutant may preferably have an amino acid sequence according to SEQ ID NO: 2, but is not limited thereto.

[0087]

[0088] According to another aspect of the present invention, the Bacillus subtilis flagellin mutant may mean a mutant in which Streptococcus pneumoniae surface protein A (PspA) is bound to the C-terminus of the Bacillus subtilis mutant lacking amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183.

[0089] The above PspA is a surface-exposed molecule found in most strains of pneumococci, and is known as an important toxic molecule of Streptococcus pneumoniae that protects bacteria from the bactericidal action of lactoferricins by binding to lactoferrin, and is a protein that contributes to the toxicity of pneumococci through various mechanisms. In the present invention, the PspA bound to the Bacillus subtilis variant may preferably have an amino acid sequence according to SEQ ID NO: 3, but is not limited thereto.

[0090]

[0091] According to another aspect of the present invention, the Bacillus subtilis flagellin mutant may mean that, in addition to the mutation described above, that is, deletion of amino acids M1 to S32, T100, D144, G145, F146, A181, A182, and T183, amino acids Q266 to R275 are further deleted. In this case, the Bacillus subtilis mutant may have an amino acid sequence according to SEQ ID NO: 4, but is not limited thereto.

[0092]

[0093] Meanwhile, in the present invention, the term "[amino acid single letter][number]" means the [amino acid] identified at the position of the amino acid sequence indicated by the [number] based on the wild-type protein. For example, in the present invention, "M1" means methionine (M, Met), which is the 1st amino acid of the wild-type Bacillus subtilis flagellin represented by SEQ ID NO: 1, and "T183" means threonine (T, Thr), which is the 183rd amino acid of the wild-type Bacillus subtilis flagellin.

[0094]

[0095] In one embodiment of the present invention, when the Bacillus subtilis flagellin mutant was administered to aged mice at two-week intervals for 15 weeks, the survival period was extended in the experimental substance administration group compared to the control group, rapid weight changes due to aging of the animals were alleviated, the condition of the fur was maintained or even improved, and muscle strength or exercise ability was also maintained, confirming that the Bacillus subtilis flagellin mutant has an anti-aging effect.

[0096]

[0097] In the present invention, the aging is a concept encompassing a degenerative change phenomenon that generally occurs due to a decline in the structure and function of the body with age, and the changes due to aging are very diverse, such as a decrease in the weight and body weight of each tissue due to a decrease in the number of actual cells, hair loss or hair loss, a decrease in muscle mass or exercise ability, a change in connective tissue, a change in body composition, a decrease in the elasticity of blood vessels or skin, a decline in the function of each organ, a decrease in the ability to fight diseases including the immune system, a decrease in the function of sensory organs, and a decrease in memory, learning ability, and comparative ability.

[0098] In the present invention, the term "anti-aging" refers to any action that suppresses, inhibits, or delays the aforementioned aging symptoms by administering the composition of the present invention. Specifically, it refers to any action that at least reduces the severity of the aforementioned aging-related parameters, such as symptoms, and includes actions in which aging symptoms are improved, alleviated, or beneficially altered by administering the composition of the present invention. Furthermore, the term may be used interchangeably with "aging inhibition" or "aging delay."

[0099]

[0100] More specifically, for the purposes of the present invention, the aging may include, but is not limited to, one or more aging selected from the group consisting of hair aging, muscle aging, skin aging, vision aging, hearing aging, digestive organ aging, immune aging, and urinary organ aging.

[0101] The above "hair aging" preferably refers to hair loss due to aging, that is, hair falling out from the skin or hair becoming thin and thin due to aging. As one ages, hair loss symptoms may occur due to various causes such as aging of skin blood vessels and hormonal imbalance.

[0102] The above "muscle aging" is a term used interchangeably with "muscle aging", and encompasses muscle wasting or muscle reduction that occurs with aging, such as a decline in muscle function (muscle strength, muscle endurance, muscle power, etc.) or a decrease and / or decline in exercise capacity that accompanies this, or muscle atrophy, etc. The above muscle atrophy refers to a decrease in muscle mass due to a decrease or reduction of muscle cells. In humans, muscle aging can cause a gradual decline in muscle density and function after the age of 30, and falls and fractures can easily occur. The causes of muscle aging can include a decrease in growth hormones and androgens, a decrease in the body's ability to synthesize protein, and a decrease in the ability to absorb protein or calories related to maintaining muscle density.

[0103] The above "skin aging" refers to symptoms such as loss of elasticity, loss of shine, formation of wrinkles, weakened regenerative power, or severe dryness of the skin, and can be caused by the passage of time or external environment.

[0104] The above "aging of vision" refers to a phenomenon of vision decline that accompanies aging due to various causes, such as a decrease in the elasticity of the lens and a decrease in accommodative power with age, a decrease in the elasticity of ciliary muscle fibers, or a hardening of the cornea, and may include the phenomenon commonly referred to as presbyopia and diseases such as dry eye, macular degeneration, hyperopia, myopia, and cataracts caused by it.

[0105] The above "auditory aging" refers to the gradual loss of hearing that accompanies aging, which may be caused by a decrease in the number and function of nerve cells connected to the inner ear, middle ear, and brain, and may be accompanied by symptoms such as tinnitus or hearing loss.

[0106] The above "digestive tract aging" refers to changes in the oral cavity, esophagus, and gastrointestinal tract due to aging, and may be accompanied by symptoms such as decreased secretion of gastric acid and pancreatic juice, decreased motility of the gastrointestinal tract, decreased speed and efficiency of digestion, or a significant decrease in the absorption rate of ingested nutrients such as fat, which may result in symptoms such as indigestion and diarrhea.

[0107] The above "immunosenescence" refers to changes in immune function due to aging, and an example is a phenomenon in which the number of neutrophils increases and the number of lymphocytes decreases. Lymphocytes are a type of white blood cell that is created when hematopoietic stem cells differentiate and mature into lymphoid hematopoietic stem cells through the hematopoietic process, and are involved in specific immune responses. Therefore, a decrease in lymphocytes can be a factor causing a decline in immunity. However, the relationship between immunosenescence and immune decline is not limited to this. The above immunosenescence can cause diseases such as autoimmune diseases, pneumonia, influenza, tetanus, infective endocarditis, and cancer to develop, or the worsening of the symptoms of the above diseases can be accelerated, but is not limited thereto.

[0108] The above "urinary tract aging" includes symptoms that occur due to changes in intra-abdominal pressure, pelvic, urethral, ​​and bladder muscle strength, and urethral mucosal thickening with age, and may be accompanied by diseases such as overactive bladder, urinary incontinence, benign prostatic hyperplasia, lower urinary tract symptoms, glomerulonephritis, and chronic renal failure.

[0109]

[0110] At least one aging-related symptom or condition in a subject can be reduced or delayed by at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%, as compared to a comparable subject not administered the composition of the present invention.

[0111] Alternatively, the lifespan of a subject administered the composition of the present invention may be increased or prolonged by at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% compared to the lifespan of a comparable subject not administered the composition.

[0112]

[0113] Accordingly, the present invention provides a life extension composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0114] In addition, the present invention provides a composition for preventing, improving or treating hair loss, comprising a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0115] In addition, the present invention provides a composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity, comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0116]

[0117] According to one aspect of the present invention, the Bacillus subtilis flagellin mutant may mean one in which amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 are deleted from the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO: 1 as described above, and preferably one having an amino acid sequence according to SEQ ID NO: 2; or one in which Streptococcus pneumoniae surface protein A (PspA) is bound to the C-terminus of the Bacillus subtilis mutant in which amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 are deleted; Alternatively, it may mean that in addition to the deletion of amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 in the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO: 1, amino acids Q266 to R275 are further deleted, and preferably, it may have an amino acid sequence according to SEQ ID NO: 4, but is not limited thereto.

[0118]

[0119] The term "treatment" as used herein refers to temporarily or permanently reversing, alleviating, or inhibiting the progression of the disease or condition to which the term applies, or one or more symptoms of the disease or condition. Furthermore, the terms "prevention" or "prevention" as used herein refer to delaying the onset of the relevant symptoms, and the term "improvement" as used herein refers to any action that improves a parameter associated with an abnormal condition, such as the severity of symptoms.

[0120]

[0121] Therefore, in the composition for preventing, improving, or treating hair loss, preventing hair loss means preventing the occurrence of the aforementioned hair loss symptoms due to aging. In addition, improving hair loss means improving the severity of hair loss symptoms and slowing down the rate of hair loss, and treating hair loss means temporarily or permanently reversing or alleviating it, or inhibiting its progression, thereby reducing the area of ​​hair loss or increasing hair density.

[0122]

[0123] In addition, in the composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity, preventing or preventing a decrease in muscle strength or exercise capacity means suppressing the decrease in muscle and / or the decrease in exercise capacity resulting from the aging described above, or preventing the decrease in muscle or the decrease in exercise capacity from occurring, and improving muscle strength or exercise capacity means improving muscle and / or muscle function, thereby improving muscle strength or exercise capacity.

[0124]

[0125] Meanwhile, the composition according to the present invention may be a pharmaceutical composition comprising a Bacillus subtilis flagellin variant, or a food composition.

[0126]

[0127] In the present invention, the content of the composition is not particularly limited depending on the purpose or aspect of use, and may be, for example, 0.01 to 99 wt%, preferably 0.5 to 50 wt%, and more preferably 1 to 30 wt%, based on the total weight of the composition. In addition, the pharmaceutical composition according to the present invention may further include additives such as pharmaceutically acceptable carriers, excipients, or diluents in addition to the Bacillus subtilis flagellin variant. The pharmaceutical composition of the present invention may include 0.1 to 99.9 wt% of the Bacillus subtilis flagellin variant prepared by the method of the present invention, and may include 99.9% to 0.1 wt% of the carrier.

[0128]

[0129] The pharmaceutical composition of the present invention can be formulated in various ways according to the route of administration by a method known in the art together with a pharmaceutically acceptable carrier in addition to the Bacillus subtilis flagellin variant. "Pharmaceutically acceptable" refers to a non-toxic substance that is physiologically tolerable and does not inhibit the action of the active ingredient when administered to humans and does not typically cause allergic reactions such as gastrointestinal disorders or dizziness or similar reactions. The carrier includes all kinds of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads, and microsomes.

[0130] Administration can be oral or parenteral. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal administration.

[0131] When the pharmaceutical composition of the present invention is administered orally, the pharmaceutical composition of the present invention may be formulated in the form of powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, etc., together with a suitable carrier for oral administration according to a method known in the art. Examples of suitable carriers may include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, and maltitol; starches including corn starch, wheat starch, rice starch, and potato starch; cellulosics including cellulose, methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; fillers such as gelatin and polyvinylpyrrolidone; and, if desired, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant. Furthermore, the pharmaceutical composition may additionally include an anticoagulant, a lubricant, a humectant, a fragrance, an emulsifier, and a preservative.

[0132] When administered parenterally, the pharmaceutical composition of the present invention may be formulated in the form of injections, transdermal administration preparations, and nasal inhalants together with a suitable parenteral carrier according to methods known in the art. The injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. To protect the above injection from microbial contamination, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal, may be additionally included. In addition, the above injection may, in most cases, additionally include an isotonic agent, such as sugar or sodium chloride.

[0133] Transdermal administration formulations include ointments, creams, lotions, gels, topical solutions, pastes, liniments, and aerosols. As used herein, "transdermal administration" refers to topically administering a pharmaceutical composition to the skin, thereby delivering an effective amount of the active ingredient contained in the pharmaceutical composition into the skin. For example, the pharmaceutical composition of the present invention may be prepared in an injectable formulation and administered by lightly pricking the skin with a 30-gauge thin needle or by directly applying the formulation to the skin. These formulations are generally known in pharmaceutical chemistry.

[0134] For inhalation administration, the flagellin variants may be conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer, for example, using dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or another suitable gas. For pressurized aerosols, the dosage unit may be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in inhalers or insufflators may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0135] Other pharmaceutically acceptable carriers known in the art may be referred to.

[0136]

[0137] Additionally, the pharmaceutical composition according to the present invention may further comprise one or more buffers (e.g., saline or PBS), carbohydrates (e.g., glucose, mannose, sucrose or dextran), antioxidants, bacteriostats, chelating agents (e.g., EDTA or glutathione), adjuvants (e.g., aluminum hydroxide), suspending agents, thickening agents and / or preservatives.

[0138] Additionally, the pharmaceutical composition of the present invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.

[0139] In addition, the pharmaceutical composition of the present invention can be administered in combination with a known substance that has the effect of preventing or treating each of the symptoms listed above.

[0140]

[0141] In the food composition of the present invention, "food" includes all foods in the conventional sense, including all forms such as functional foods, nutritional supplements, health foods, and food additives. The above-mentioned types of food compositions can be manufactured in various forms according to conventional methods known in the art.

[0142] The food composition of the present invention may include a health functional food. The term "health functional food" as used herein refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients having functionality useful to the human body. Here, "functionality" means obtaining a beneficial effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological effects. The health functional food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, it can be manufactured by adding raw materials and ingredients commonly added in the art.

[0143] In addition, the formulation of the health functional food can be manufactured without limitation as long as it is a formulation recognized as a health functional food. The food composition of the present invention can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time as it uses food as a raw material, and has excellent portability, so the health functional food of the present invention can be consumed as a supplement to enhance the anti-aging effect. For example, the health functional food can be consumed in the form of tea, juice, and drink containing a composition including the Bacillus subtilis flagellin variant according to the present invention, or can be consumed in the form of granules, capsules, and powder.

[0144] In addition, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in mixtures. The ratio of these additives is not particularly important, but is generally selected from the range of 0.01 to 0.3 parts by weight per 100 parts by weight of the food composition of the present invention, but is not limited thereto.

[0145] In addition, the food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. The carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, a natural sweetener such as thaumatin or stevia extract, or a synthetic sweetener such as saccharin or aspartame may be used. The proportion of the natural carbohydrate may generally be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention, but is not limited thereto.

[0146]

[0147] The present invention also provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing an anti-aging composition.

[0148] In addition, the present invention provides an anti-aging method comprising administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0149] The present invention also provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for extending lifespan.

[0150] In addition, the present invention provides a method for extending lifespan, comprising administering to a subject in need thereof an effective amount of a composition comprising, as an active ingredient, a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

[0151] In addition, the present invention provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, improving or treating hair loss.

[0152] In addition, the present invention provides a method for preventing, improving or treating hair loss, which comprises administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity with the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0153] The present invention also provides the use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity.

[0154] In addition, the present invention provides a method for preventing, preventing or improving a decrease in muscle strength or exercise capacity, comprising administering to a subject in need thereof an effective amount of a composition comprising a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 as an active ingredient.

[0155]

[0156] The 'effective amount' of the present invention refers to an amount that, when administered to an individual, exhibits the effects of anti-aging, life extension, hair loss prevention, hair loss improvement, hair loss treatment, prevention, prevention or improvement of a decrease in muscle strength or exercise ability, and the 'individual' may be an animal, preferably a mammal, particularly an animal including a human, and may also be a cell, tissue, organ, etc. derived from an animal. The individual may be a patient in need of the above effect.

[0157]

[0158] A composition comprising a Bacillus subtilis flagellin variant according to the present invention, when administered to an aged animal, extends the survival period of the animal and suppresses weight changes, hair loss, and decline in muscle strength and exercise ability due to aging, and thus can be usefully used in the development of an anti-aging composition.

[0159]

[0160] Figure 1 is a schematic diagram of flagellin mutant BM03 and BM03_PspA proteins according to the present invention.

[0161] Figure 2 illustrates a schematic diagram of an experiment to confirm the anti-aging activity of BM03 and BM03_PspA.

[0162] Figures 3a and 3b show the results of evaluating the viability of mice in the BM03 or BM03_PspA administration group.

[0163] Figure 4 shows a schematic diagram of an experiment to confirm the anti-aging activity of BM03 and BM12.

[0164] Figures 5a to 5d show the body weight changes of mice administered BM03 or BM12 by week. Figures 5a and 5c show the body weight values ​​of mice by week, and Figures 5b and 5d show the amount of body weight change, respectively. Meanwhile, Figures 5a and 5b show error bars, while Figures 5c and 5d do not show error bars.

[0165] Figure 6 shows the results of evaluating the survival potential of mice in the BM03 or BM12 administration group.

[0166] Figure 7 shows the standard grade for evaluating the condition of mouse fur.

[0167] Figures 8a to 8c show the results of evaluating the condition of hair in the BM03 or BM12 administration group.

[0168] Figure 9 shows the results of evaluating exercise ability and muscle strength in the BM03 or BM12 administration group.

[0169]

[0170] The present invention is described in detail below.

[0171] However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0172]

[0173] Example 1. Preparation of flagellin mutants and fusion proteins containing the same

[0174] A mutant BM03 (SEQ ID NO: 2) was constructed from wild-type Bacillus subtilis flagellin, lacking M1-S32, T100, D144, G145, F146, A181, A182, and T183. For detailed construction methods, please refer to the inventor's prior invention, Republic of Korea Patent Publication No. 10-2024-0022417.

[0175] Next, a fusion protein BM03_PspA was produced in which PspA (Streptococcus pneumoniae surface protein, SEQ ID NO: 3) of SEQ ID NO: 2 was fused to the C-terminus of BM03. A schematic diagram of the produced BM03 and BM03_PspA flagellin proteins is shown in Fig. 1.

[0176] In addition, a mutant BM12 (SEQ ID NO: 4) was produced in which M1 to S32, T100, D144, G145, F146, A181, A182, T183, and Q266 to R275 were deleted from the wild-type Bacillus subtilis flagellin using the same method as BM03.

[0177] The amino acid sequences of the above-mentioned produced flagellin mutants and fusion proteins are as shown in [Table 1] below.

[0178] Name Amino Acid Sequence SEQ ID NO BM03SGLRINRAGD DAAGLAISEK MRGQIRGLEM ASKNSQDGIS LIQTAEGALT ETHAILQRMR ELTVQAGNGT QQAEDLGAIK DEMDALIEEI DGISNRTEFN GKKLLDGTNS TTFQIGANAG QQLNVKIDSM SSTALGVNAL DVTDFAFDDQ LKSIDTAINT VSTQRAKLGA VQNRLEHTIN NLGASGENLT AAESRIRDVD MAKEMSEFTK NNILSQASQA MLAQANQQPQ NVLQLLR2PspASPVASQSKAE KDYDAAKKDA KNAKKAVEDA QKALDDAKAA QKKYDEDQKK TEEKAALEKA ASEEMDKAVA AVQQAYLAYQ QATDKAAKDA ADKMIDEAKK REEEAKTKFN TVRAMVVPEP EQLAETKKKS EEAKQKAPEL TKKLEEAKAK LEEAEKKATE AKQKVDAEEV APQAKIAELE NQVHRLEQEL KEIDESESED YAKEGFRAPL QSKLDAKKAK LSKLEELSDK IDE3BM12SGLRINRAGD DAAGLAISEK MRGQIRGLEM ASKNSQDGIS LIQTAEGALT ETHAILQRMR ELTVQAGNGT QQAEDLGAIK DEMDALIEEI DGISNRTEFN GKKLLDGTNS TTFQIGANAG QQLNVKIDSM SSTALGVNAL DVTDFAFDDQ LKSIDTAINT VSTQRAKLGA VQNRLEHTIN NLGASGENLT AAESRIRDVD MAKEMSEFTK NNILSQASQA MLAQANQ4

[0179]

[0180] Example 2. Evaluation of anti-aging activity of BM03 and BM03_PspA

[0181] Male and female C57BL / 6J mice aged 760 days were divided into control, BM03, and BM03_PspA groups, respectively, and each experimental substance was administered intranasally at 6.5 ug / head at two-week intervals, and survival was observed. The experimental schematic is shown in Fig. 2.

[0182] As a result, as can be seen in Figures 3a and 3b, it was confirmed that the survival period of mice, regardless of male or female, was extended in the BM03 administration group and the BM03_PspA administration group compared to the control group.

[0183]

[0184] Example 3. Evaluation of anti-aging activity of BM03 and BM12 (1)

[0185] Female C57BL / 6J mice, 840 days old, were divided into control, BM03, and BM12 groups, and each experimental substance was administered intranasally at 5 ug / head at two-week intervals, and body weight was observed weekly. The experimental schematic is shown in Fig. 4.

[0186] As a result, as can be seen in Figures 5a to 5d, the BM03 administration group and the BM12 administration group showed a reduction in the rapid weight changes due to aging of the animals compared to the control group. In addition, as can be seen in Figure 6, the survival period of the animals in the BM03 administration group and the BM12 administration group was confirmed to be extended compared to the control group.

[0187]

[0188] Example 4. Evaluation of anti-aging activity of BM03 and BM12 (2)

[0189] During the above Example 3, the fur condition of each mouse was checked at weeks 0 and 7 (after the 4th administration). The fur condition was scored for each mouse from 1 point for the best to 4 points for the worst based on the fur grades shown in Figure 7, and the results are shown in Figures 8a to 8c.

[0190] As can be seen in Figures 8a to 8c, the hair condition of the BM03 administration group and the BM12 administration group was maintained compared to the control group, and in particular, some individuals in the BM03 administration group were confirmed to have improved.

[0191]

[0192] Example 5. Evaluation of anti-aging activity of BM03 and BM12 (3)

[0193] During the above Example 3, the muscle strength and motor coordination ability (hanging test) of each mouse were observed.

[0194] The experiment for the hanging test was conducted according to the following procedures:

[0195] 1) It is conducted in a safe and stress-free environment.

[0196] 2) Let the mouse hang on the wire mesh.

[0197] 3) Measure the time the mouse naturally hangs on the wire mesh.

[0198] After measuring the time that the mice in each experimental group were hanging on the wire mesh according to the above procedure, the results are shown in Figure 9.

[0199] As can be seen in Figure 9, the BM03 and BM12 administration groups showed superior exercise ability and muscle strength compared to the control group.

[0200]

[0201] As described above, the composition comprising the Bacillus subtilis flagellin variant according to the present invention, when administered to an aged animal, has the effect of extending the survival period of the animal, suppressing weight changes, hair loss, and decline in muscle strength and exercise ability due to aging, and thus can be usefully used in the development of an anti-aging composition.

Claims

1. An anti-aging composition comprising a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of sequence number 1.

2. An anti-aging composition according to claim 1, wherein the Bacillus subtilis flagellin mutant has amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 deleted from the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO:

1.

3. An anti-aging composition according to claim 2, wherein the Bacillus subtilis flagellin mutant further lacks amino acids Q266 to R275.

4. An anti-aging composition according to claim 2, wherein the Bacillus subtilis flagellin mutant has Streptococcus pneumoniae surface protein A (PspA) bound to the C-terminus.

5. An anti-aging composition according to claim 2, characterized in that the Bacillus subtilis flagellin mutant has an amino acid sequence according to sequence number 2.

6. An anti-aging composition according to claim 3, characterized in that the Bacillus subtilis flagellin mutant has an amino acid sequence according to SEQ ID NO:

4.

7. An anti-aging composition according to claim 4, characterized in that the PspA has an amino acid sequence according to sequence number 3.

8. An anti-aging pharmaceutical composition comprising an anti-aging composition according to any one of claims 1 to 7.

9. An anti-aging food composition comprising an anti-aging composition according to any one of claims 1 to 7.

10. A composition for extending life span comprising a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of sequence number 1.

11. A composition for extending lifespan in claim 10, wherein the Bacillus subtilis flagellin mutant is characterized in that amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 are deleted from the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO:

1.

12. A composition for extending lifespan in claim 11, wherein the Bacillus subtilis flagellin mutant is further characterized by a deletion of amino acids Q266 to R275.

13. A composition for extending lifespan in claim 11, wherein the Bacillus subtilis flagellin mutant has Streptococcus pneumoniae surface protein (PspA) bound to the C-terminus.

14. A composition for preventing, improving or treating hair loss, comprising a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of sequence number 1.

15. A composition for preventing, improving or treating hair loss in claim 14, wherein the Bacillus subtilis flagellin mutant is characterized in that amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 are deleted from the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO:

1.

16. A composition for preventing, improving or treating hair loss, characterized in that the Bacillus subtilis flagellin mutant in claim 15 further lacks amino acids Q266 to R275.

17. A composition for preventing, improving or treating hair loss, characterized in that the Bacillus subtilis flagellin variant has Streptococcus pneumoniae surface protein (PspA) bound to the C-terminus of the composition in claim 15.

18. A composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity, comprising a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO:

1.

19. A composition for preventing, preventing or improving a decrease in muscle strength or exercise ability, wherein the Bacillus subtilis flagellin mutant is characterized in that amino acids M1 to S32, T100, D144, G145, F146, A181, A182 and T183 are deleted from the wild-type Bacillus subtilis flagellin sequence of SEQ ID NO:

1.

20. A composition for preventing, preventing or improving a decrease in exercise capacity, characterized in that the Bacillus subtilis flagellin mutant in claim 19 further has a deletion of amino acids Q266 to R275.

21. A composition for preventing, preventing or improving a decrease in muscle strength or exercise ability, characterized in that the Bacillus subtilis flagellin variant has Streptococcus pneumoniae surface protein (PspA) bound to the C-terminus of the composition in claim 19.

22. Use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of sequence number 1 for preparing an anti-aging composition.

23. An anti-aging method comprising administering to a subject in need thereof an effective amount of a composition comprising as an active ingredient a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of sequence number 1.

24. Use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of sequence number 1 for preparing a composition for extending lifespan.

25. A method for extending lifespan, comprising administering to a subject in need thereof an effective amount of a composition comprising as an active ingredient a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO:

1.

26. Use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, improving or treating hair loss.

27. A method for preventing, improving or treating hair loss, comprising administering to a subject in need thereof an effective amount of a composition comprising, as an active ingredient, a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO:

1.

28. Use of a Bacillus subtilis flagellin mutant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1 for preparing a composition for preventing, preventing or improving a decrease in muscle strength or exercise capacity.

29. A method for preventing, preventing or improving a decrease in muscle strength or exercise capacity, comprising administering to a subject in need thereof an effective amount of a composition comprising as an active ingredient a Bacillus subtilis flagellin variant having at least 80% identity to the wild-type Bacillus subtilis flagellin of SEQ ID NO: 1.

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