Aging inhibitor

Abstract

The purpose of the present invention is to provide an aging inhibitor using a biologically derived material. The present invention is an aging inhibitor comprising a vitamin D binding protein.

Description

Technical Field

[0001] The present invention relates to aging inhibitors. Background Art

[0002] As one of the reasons for the aging of cells, the shortening of telomeres can be cited.Telomeres are repeated sequences of 6 bases of TTAGGG present at the ends of chromosomes, which play a role in maintaining the stability of chromosomes and protecting genetic information.Telomeres shorten with cell division, and the shortening of telomeres causes the aging of cells.Inflammatory cytokines are secreted by aged cells, causing the function of internal organs and tissues to decrease, so it is believed that the shortening of telomeres is the cause of aging diseases on the individual level.It is believed that if telomeres are extended, aging inhibition on the individual level can be achieved, but it is unknown that telomeres can be extended by materials of biological origin.

[0003] On the other hand, vitamin D binding protein (VDBP) that binds to vitamin D is a protein contained in plasma and milk, and is also called vitamin D binding protein. It is known that vitamin D binding protein has the ability to activate macrophages (Patent Document 1).

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: International Publication No. WO2013 / 038997 Summary of the invention

[0007] Technical problem solved by the invention

[0008] The problem underlying the present invention is to provide an ageing inhibitor using materials of biological origin.

[0009] Technical means of solving problems

[0010] The present inventors have discovered that vitamin D binding protein has an aging-suppressing effect, thereby completing the present invention.

[0011] That is, the present invention relates to an aging inhibitor comprising a vitamin D binding protein.

[0012] The vitamin D binding protein is preferably treated with an enzyme, and the enzyme is one or more selected from the group consisting of β-galactosidase, sialidase, and N-acetylgalactosaminidase.

[0013] The vitamin D binding protein is preferably derived from milk or serum.

[0014] The milk is preferably colostrum or normal milk.

[0015] The aging inhibitors are preferably used to extend telomeres.

[0016] The present invention also relates to a method for producing an aging inhibitor, comprising the step of contacting a vitamin D binding protein with one or more enzymes selected from the group consisting of β-galactosidase, sialidase, and N-acetylgalactosaminidase.

[0017] Effects of the Invention

[0018] The aging inhibitor of the present invention contains vitamin D binding protein as a biologically derived material, and therefore has less risk of side effects. The aging inhibitor of the present invention exerts an effect of lengthening telomeres of cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] [ Figure 1 ] is the relative value of telomere length of all subjects.

[0020] [ Figure 2 ] is the relative value of telomere length of each gender of the subjects. DETAILED DESCRIPTION

[0021] <Vitamin D binding protein>

[0022] The aging inhibitor of the present invention comprises vitamin D binding protein. Vitamin D binding protein is a protein present in mammalian body fluids such as plasma and milk, and is also called Gc protein. The vitamin D binding protein used in the present invention is not particularly limited as long as it is derived from mammals. For example, vitamin D binding protein derived from cows, goats, mice, and humans can be cited, and vitamin D binding protein derived from cows is preferred.

[0023] Specific examples of vitamin D binding proteins include polypeptides that have a sequence identity of 85% or more to the amino acid sequence represented by SEQ ID NO.1 in the sequence listing, or polypeptides that comprise an amino acid sequence in which one or more amino acids are deleted, inserted, substituted and / or added in the amino acid sequence represented by SEQ ID NO.1 in the sequence listing.

[0024] The sequence identity with the amino acid sequence represented by SEQ ID NO. 1 is preferably 90% or more, more preferably 95% or more, further preferably 98% or more, and particularly preferably 99% or more.

[0025] In the amino acid sequence represented by SEQ ID NO.1, the number of deleted, inserted, substituted and / or added amino acids is preferably 68 or less, more preferably 45 or less, further preferably 22 or less, further preferably 9 or less, and particularly preferably 4, 3 or 2 or less.

[0026] As for the vitamin D binding protein, the vitamin D binding protein contained in the body fluids such as milk or serum of the above-mentioned mammals can be used. Milk, serum or processed products thereof containing the vitamin D binding protein can be used as an aging inhibitor, and the purified vitamin D binding protein can also be used as an aging inhibitor.

[0027] Serum is not particularly limited as long as it is prepared from blood collected from mammals by conventional methods. When purifying vitamin D binding protein from serum, as a purification method, for example, albumin is removed from serum, vitamin D binding protein is adsorbed and eluted using a vitamin D binding column, and then dialyzed.

[0028] The milk may be colostrum secreted only during a given number of days after delivery. In particular, bovine colostrum refers to milk secreted by a cow from the 10th day after delivery. In addition, the milk may be normal milk. As a milk component containing vitamin D binding protein, for example, whey (also called de-cheese component, Whey) can be used, which is obtained by removing the coagulation component (casein, fat, etc.) of cheese when cheese is made from milk, that is, de-cheese component treatment.

[0029] <Gene recombination>

[0030] As vitamin D binding protein, vitamin D binding protein expressed by gene recombination can be used. As a method for expressing vitamin D binding protein by gene recombination, a method for culturing a host cell into which a gene encoding vitamin D binding protein is introduced, and a method for using a cell-free expression system containing a gene encoding vitamin D binding protein can be cited. The host cell is not particularly limited, and microorganisms such as Streptomyces, Rhodococcus, Escherichia, Bacillus, Pseudomonas, Brevibacterium, Streptococcus, Lactobacillus, Saccharomyces, Kluyveromyces, and cells of higher eukaryotic organisms can be cited. As cells of higher eukaryotic organisms, cells derived from humans, hamsters, chickens, and insects can be cited, and as specific cell strains, CHO cells, HeLa cells, HEK293 cells, sf9 cells, DT40 cells, etc. can be cited. The method of cell culture can be suspension culture or adherent culture, preferably suspension culture. In addition, serum-free medium is preferably used to avoid the mixing of impurities.

[0031] <Enzyme treatment>

[0032] The vitamin D binding protein may be treated with one or more enzymes selected from the group consisting of β-galactosidase, sialidase, and N-acetylgalactosaminidase.

[0033] Examples of β-galactosidase include β-galactosidase derived from Escherichia coli and β-galactosidase derived from bovine liver. Examples of commercially available products include Catalog No. 072-04141 of Wako Pure Chemical Industries, Ltd. and G1875 of SIGMA-ALDRICH.

[0034] The amount of β-galactosidase used and the treatment conditions are not particularly limited. When an O-type sugar chain containing three sugars of sialic acid and galactose bound to GalNAc (N-acetylgalactosamine) is added to the threonine (Thr) at position 418 or 420 of the vitamin D binding protein used, the conditions are preferably such that the galactose is dissociated. The temperature for β-galactosidase treatment is more preferably 35 to 39°C. The enzyme treatment time is more preferably 60 to 200 minutes. For example, when using Catalog No. 072-04141 of Wako Pure Chemical Industries, Ltd. as β-galactosidase, it is preferred to use 65 mU or more of the enzyme for 100 μl of milk containing the vitamin D binding protein and react for more than 3 hours. The activity of β-galactosidase was defined as the amount of enzyme that generates 1 μmol of o-nitrophenol in 1 minute at pH 7.3 and 37°C using o-nitrophenol-β-D-galactopyranoside as a substrate, and 1 unit (U).

[0035] Examples of sialidase include sialidase derived from Clostridium perfringenes, sialidase derived from Streptococcus 6646K, sialidase derived from Vibrio cholerae, sialidase derived from Arthrobacter ureafaciens, etc. Examples of commercially available products include SIGMA Prod. Nos. N2876, N2133, N2904, N3001, and N5631 manufactured by SIGMA-ALDRICH, Code No. 120052 manufactured by Seikagaku BioBusiness, and Catalog Nos. P0720L and P0720S manufactured by BioLabs.

[0036] The amount of sialidase used and the treatment conditions are not particularly limited. When an O-type sugar chain containing 3 sugars of sialic acid and galactose is added to the threonine (Thr) at position 418 or position 420 of the vitamin D binding protein used, it is preferably a condition that dissociates the sialic acid. The temperature of the sialidase treatment is more preferably 35 to 39 ° C. The enzyme treatment time is more preferably 60 to 200 minutes. For example, when the product number (N2876) of SIGMA-ALDRICH is used as a sialidase, it is preferred to use 65mU or more enzymes for 100μl of milk containing vitamin D binding protein and react for more than 3 hours. With regard to the activity of sialidase, bovine submandibular gland-derived mucin is used, and the activity of releasing 1.0 micromole of N-acetylneuraminic acid per minute at pH 5.0 and 37 ° C is taken as 1unit (U).

[0037] As N-acetylgalactosaminidase, endo-α-N-acetylgalactosaminidase and exo-α-N-acetylgalactosaminidase can be mentioned. As endo-α-N-acetylgalactosaminidase, for example, endo-α-N-acetylgalactosaminidase derived from Escherichia coli, endo-α-N-acetylgalactosaminidase derived from Bifidobacterium longum, endo-α-N-acetylgalactosaminidase derived from Streptococcus pneumoniae, endo-α-N-acetylgalactosaminidase derived from bovine liver, etc. can be mentioned. As commercially available products, for example, catalog #324716 of MERCK company can be mentioned.

[0038] The amount of N-acetylgalactosamidase used and the treatment conditions are not particularly limited. When GalNAc (N-acetylgalactosamine) is added to the threonine (Thr) at position 418 or 420 of the vitamin D binding protein, the conditions for removing the N-acetylgalactosamine are preferred. The amount of endo-α-N-acetylgalactosamidase used is preferably 1 to 500 mU relative to 1 μg of vitamin D binding protein, and more preferably 100 to 200 mU. The temperature of the enzyme treatment is preferably 35 to 39° C. The enzyme treatment time is preferably 60 to 200 minutes.

[0039] The activity of N-acetylgalactosaminidase was defined as the activity of dissociating 1.0 μmol of p-nitrophenol from p-nitrophenyl-2-acetamide-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside within 1 minute at 37°C and pH 5.0, and was defined as 1 unit (U).

[0040] Furthermore, when sialic acid is bound to N-acetylgalactosamine, the cleavage effect of N-acetylgalactosamine by endo-α-N-acetylgalactosaminidase is reduced, so sialic acid is preferably cleaved in advance. Sialic acid cleavage can be performed by treating the vitamin D binding protein with sialidase.

[0041] Removal of N-acetylgalactosamine from vitamin D binding protein by N-acetylgalactosaminidase treatment can be confirmed by whether there is binding to lectin. Examples of lectins include lectins derived from Wisteria floribunda (multiflora wisteria) and lectins derived from Helix pomatia (Roman snail). When vitamin D binding protein does not bind to lectin, it can be determined that N-acetylgalactosamine has been removed.

[0042] The enzyme treatment of the vitamin D binding protein may be performed in the presence of a buffer solution, such as physiological saline or phosphate buffered saline (PBS).

[0043] The enzyme may be inactivated by heat treatment after the enzyme treatment. The heat treatment conditions are not particularly limited as long as the enzyme can be inactivated, for example, heating at a temperature of about 60° C. for about 10 minutes. In addition, the enzyme may be removed from the vitamin D binding protein by combining a vitamin D binding column, a gel filtration column, ultrafiltration, etc. after the enzyme treatment.

[0044] The enzyme treatment can be carried out using an enzyme fixed to a solid phase (immobilized enzyme). Methods for fixing an enzyme to a solid phase are well known to those skilled in the art, and for example, a method for fixing an enzyme to agarose beads using a coupling agent such as cyanogen bromide can be cited. The enzyme reaction can be carried out by applying the vitamin D binding protein to a solid phase on which the enzyme has been immobilized. By using an immobilized enzyme, after the enzyme treatment, it can be recovered without inactivating the enzyme by heat treatment, and inclusions (proteins such as enzymes inactivated by heat treatment, etc.) can be removed.

[0045] When milk or serum containing vitamin D binding protein is subjected to enzyme treatment, it is preferred to perform a pretreatment before the enzyme treatment. As a pretreatment, in addition to concentration to reduce the amount of water, there can be cited a treatment to remove the coagulated components (casein, fat, etc.) of cheese when cheese is made from milk (cheese component removal treatment). The milk obtained by concentration is called "concentrated milk". The milk obtained by removing the cheese component treatment is also called cheese component-removed milk, whey or Whey. The vitamin D binding protein obtained by the above method can be further freeze-dried to be made into a solid or powdery state.

[0046] <Aging Suppression>

[0047] The aging inhibitor of the present invention can inhibit the aging of an individual by administering it to the individual. Aging inhibition can be evaluated by measuring telomere length. Telomere length can be measured by, for example, extracting chromosomal DNA from cells, reacting with a labeled telomere probe and detecting the luminescence intensity of the label. When the telomere length in the subject group administered with the aging inhibitor is extended compared to the subject group not administered with the aging inhibitor, it can be judged that aging is inhibited. The telomere length in the subject group administered with the aging inhibitor is preferably 1.02 times or more of the telomere length in the subject group not administered with the aging inhibitor, more preferably 1.05 times or more, further preferably 1.1 times or more, and further preferably 1.2 times or more. By administering the aging inhibitor, telomere length can be extended regardless of tissue or cell type. Representative cells include leukocytes, skin cells, mucosal cells, etc. Leukocytes can be collected from whole blood, and mucosal cells can be collected from the oral mucosa and nasal cavity.

[0048] <Macrophage Activation Ability>

[0049] The vitamin D binding protein used in the present invention may have macrophage activation ability. The vitamin D binding protein having macrophage activation ability is also called GcMAF (Gc protein-derived macrophage activating factor). As the vitamin D binding protein having macrophage activation ability, vitamin D binding protein in which the threonine (Thr) at position 418 or 420 is not bound to GalNAc (N-acetylgalactosamine) and vitamin D binding protein in which the threonine (Thr) at position 418 or 420 is only bound to GalNAc can be cited.

[0050] In addition, the vitamin D binding protein used in the present invention may be a form in which an O-type sugar chain containing a trisaccharide of sialic acid and galactose bonded to GalNAc (N-acetylgalactosamine) is bonded to threonine (Thr) at position 418 or 420. This form is considered to have no macrophage activation ability.

[0051] The macrophage activation ability can be evaluated by measuring the phagocytic ability of macrophages. The determination of the phagocytic ability of macrophages is carried out by adding vitamin D binding protein to the culture fluid of macrophages, making the macrophages engulf test substances such as fluorescent latex beads and zymosan, and calculating the proportion of macrophages engulfed and the number of particles of the engulfed test substance. When the proportion of macrophages engulfed and the number of particles of the engulfed test substance are more than the comparison control, it can be judged that it has macrophage activation ability. As macrophage cell strains, for example, RAW264.7, NR8383, J774.1, etc. can be used.

[0052] The aging inhibitor of the present invention may be a substance that does not increase NO production by macrophages. The NO production capacity can be evaluated by adding the aging inhibitor to macrophages and measuring the NO production after standing for about 24 hours. The NO production can be measured by, for example, the Griess method described in the Examples.

[0053] The aging inhibitor of the present invention may be a substance that inhibits NO production by macrophages. The ability to inhibit NO production can be evaluated by adding an aging inhibitor and a NO production stimulator such as lipopolysaccharide (LPS) to macrophages and measuring the amount of NO produced after standing for about 24 hours. Inhibiting NO production by macrophages means having an anti-inflammatory effect.

[0054] The aging inhibitor of the present invention may be a substance that does not increase the production of TNF-α based on macrophages. The ability to produce TNF-α can be evaluated by adding an aging inhibitor, a NO production stimulator such as lipopolysaccharide (LPS), and IFN-γ to macrophages, and measuring the amount of TNF-α produced after standing for about 24 hours. The amount of TNF-α produced can be measured, for example, by the ELISA method described in the examples. Not increasing the production of TNF-α based on macrophages means having an anti-inflammatory effect.

[0055] The aging inhibitor of the present invention may have the ability to differentiate into M2 macrophages. The ability to differentiate into M2 macrophages can be evaluated by adding the aging inhibitor to the macrophages and measuring the expression level of the marker gene in the macrophages after standing for about 24 hours. As a marker gene, for example, ARG-1 can be used. The expression level of the marker gene can be evaluated by observation based on a fluorescence microscope of fixed cells, such as described in the Examples. Generally speaking, as differentiation types of macrophages, inflammatory M1 type and anti-inflammatory M2 type are known.

[0056] <Food composition>

[0057] The aging inhibitor can be combined with various additives commonly used in food, such as auxiliary agents, sweeteners, spices, seasonings, preservatives, sterilizing agents, antioxidants, etc., to prepare a food composition as needed.

[0058] The food composition can be made into so-called health food, health drink, functional food, nutritional functional food, health supplement food, nutritional supplement food (supplement), special purpose food, specific health food, etc. As the form of the food composition, powder, granule, tablet (including sublingual tablet, etc.), enteric capsule, gastric capsule, oral capsule, pill, internal liquid (including suspension, emulsion, syrup, paste, cream, etc.) and the like can be mentioned.

[0059] The dosage of the food composition varies according to the age, sex, body weight, symptoms, administration method, etc. of the ingestor. As a typical example, for example, the total amount of protein contained in the food composition is preferably 0.005 mg to 1.0 mg, more preferably 0.01 mg to 0.5 mg, and further preferably 0.01 mg to 0.1 mg per kg of body weight per administration. In addition, the dosage of vitamin D-binding protein is preferably 0.0001 mg to 1.0 mg, more preferably 0.001 mg to 0.5 mg per kg of body weight per administration.

[0060] In the case where the food composition is administered with the dosage per 1 time, the administration interval and the number of administrations are preferably once a week to 3 times a day, more preferably 2 times a week to 2 times a day, and further preferably 1 to 2 times a day. The administration is preferably continuous for more than 1 month, more preferably continuous for more than 2 months, further preferably continuous for more than 3 months, further preferably continuous for more than 4 months, and particularly preferably continuous for more than 6 months.

[0061] <Pharmaceutical Composition>

[0062] The aging inhibitor can be combined with pharmaceutically acceptable additives to prepare a pharmaceutical composition as needed. Examples of pharmaceutically acceptable additives include diluents, stabilizers, preservatives, buffers, and the like.

[0063] The form of the pharmaceutical composition is not particularly limited, and examples thereof include an injectable composition, an oral composition, an eye drop composition, an infusion composition, a nasal drop composition, an ear drop composition, a suppository, and an enteral nutritional composition. Examples of oral compositions include powders, granules, tablets (including sublingual tablets, etc.), capsules, pills, enteric solvents, internal liquids (including suspensions, emulsions, syrups, etc.), inhalants, etc. Examples of injections include intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, and intraperitoneal injections.

[0064] The dosage of the pharmaceutical composition varies depending on the patient's age, sex, body weight, symptoms, administration method, etc. As a typical example, for example, the total amount of protein contained in the pharmaceutical composition is preferably 0.005 mg to 1.0 mg per kg of body weight per administration, more preferably 0.01 mg to 0.5 mg, and further preferably 0.01 mg to 0.1 mg. In addition, the dosage of vitamin D binding protein is preferably 0.0001 mg to 1.0 mg per kg of body weight per administration, and more preferably 0.001 mg to 0.5 mg.

[0065] When the pharmaceutical composition is administered in the dosage per administration, the administration interval and the number of administrations are preferably once a week to three times a day, more preferably twice a week to twice a day, and further preferably once or twice a day. The administration is preferably continuous for more than 1 month, more preferably continuous for more than 2 months, further preferably continuous for more than 3 months, further preferably continuous for more than 4 months, and particularly preferably continuous for more than 6 months.

[0066] <Adaptive symptoms>

[0067] Vitamin D binding protein has the ability to activate macrophages, so the pharmaceutical composition of the present invention can not only have an aging inhibitory effect, but also have a disease therapeutic effect based on macrophage activation. Examples of diseases include cancer, infectious diseases, autoimmune diseases, autism, inflammatory diseases, brain / neural degenerative diseases, skin diseases, and heart diseases.

[0068] Cancer includes any of carcinoma, sarcoma, and other malignant tumors, for example, skin cancer, bronchial cancer, lung cancer, non-small cell lung cancer, breast cancer, ovarian cancer, tongue cancer, pharyngeal cancer, esophageal cancer, gastric cancer, small intestine cancer, large intestine cancer, rectal cancer, colon cancer, liver cancer, pancreatic cancer, kidney cancer, renal cell carcinoma, bladder cancer, prostate cancer, uterine cancer, cervical cancer, Wilms' tumor, malignant melanoma, meningioma, neuroblastoma, osteosarcoma, Kaposi's polysarcoma, lymphoma, leukemia, etc. It should be noted that cancer includes not only these malignant tumors, but also metastatic tumors thereof.

[0069] Infectious diseases include, for example, viral infectious diseases, bacterial infectious diseases, and the like. Specifically, in addition to the novel coronavirus infectious disease (COVID-19), HIV infectious disease, and AIDS, there are hepatitis B, hepatitis C, herpes, influenza, pneumonia, tuberculosis, and Epstein-Barr virus infectious diseases.

[0070] Autism is a behavioral disorder characterized by difficulty forming social relationships with others and slow language development.

[0071] Inflammatory diseases are diseases caused by inflammation in the defense response of the organism caused by physical stimulation, chemical stimulation, or infection by microorganisms. In addition, the pharmaceutical composition of the present invention is effective against autoinflammatory diseases that naturally produce inflammatory responses due to abnormalities in natural immunity and lead to damage to internal organs.

[0072] Examples of brain / neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, spinocerebellar degeneration, amyotrophic lateral sclerosis, and Lewy body dementia.

[0073] Examples of skin improvement include skin whitening, suppression or improvement of pigmentation, exfoliation or promotion of keratin metabolism, anti-aging, suppression or improvement of wrinkles, moisturizing, regeneration, and treatment or prevention of alopecia.

[0074] Examples of the heart disease include congestive heart failure, arrhythmia, angina pectoris, and myocardial infarction.

[0075] <Compositions for quasi drugs>

[0076] The aging inhibitor can be combined with an adjuvant as needed to prepare a quasi-drug composition. The quasi-drug composition can be prepared in various forms such as a solution, a suspension, a syrup, a granule, a cream, a paste, a jelly, etc., and can be prepared in a desired shape as needed. The quasi-drug composition can be prepared by any conventional method.

[0077] The amount of the Gc protein used in the quasi-drug composition is not particularly limited, and may be the same as that in the case of the pharmaceutical composition, or may be an amount appropriately set with reference to the above contents.

[0078] <Method for producing aging inhibitor>

[0079] The method for producing an aging inhibitor of the present invention is characterized by comprising a step of contacting a vitamin D binding protein with one or more enzymes selected from β-galactosidase, sialidase, and N-acetylgalactosaminidase. β-galactosidase, sialidase, N-acetylgalactosaminidase, and enzyme treatment conditions are as described above with respect to the aging inhibitor.

[0080] In the method for producing an aging inhibitor of the present invention, when sialidase and N-acetylgalactosamidase are used, it is preferred that the vitamin D binding protein is first treated with sialidase and then treated with N-acetylgalactosamidase. This is because when the vitamin D binding protein is bound to sialic acid via N-acetylgalactosamine, the efficiency of cleavage of N-acetylgalactosamine by N-acetylgalactosamidase is reduced. The efficiency of cleavage of N-acetylgalactosamine can be increased by cleaving sialic acid in advance.

[0081] Example

[0082] (1) Production of capsules containing vitamin D binding protein (Production Example 1)

[0083] The crude cheese-free milk (whey taken from Zao Dairy Center) from which the cheese components have been removed was centrifuged at 8000 rpm and 4°C for 1 hour, and the precipitate was noted to recover the supernatant. An equal amount of distilled water (Mill-Q grade. In the following text, the same grade is used unless otherwise specified) was added to the recovered supernatant, and dialyzed using a PENCIL SCALEMODULE UF membrane (manufactured by FUNAKOSHI, AIP-0013D). At this time, dialysis was performed until the filtrate was equal to the amount of distilled water added to obtain cheese-free milk (whey). The protein concentration was determined by absorbance measurement at a wavelength of 570 nm (calibration curve made using BSA (bovine serum albumin, SIGMA, A4503)).

[0084] The de-cheese component milk obtained by the above was packaged in a manner in which its protein content was 6 g, and 4 g / 6000 U of β-galactosidase (made by Oriental Yeast Industry (Co., Ltd.), derived from Escherichia coli) immobilized by formyl resin (made by TOYOPEARL, Formy-650M) was added to the de-cheese component milk, and incubated at 37°C for 1 hour. After incubation, the reaction solution was filtered through a glass filter to separate the formyl resin. The formyl resin was washed with distilled water and stored for repeated use. The filtrate was filter sterilized using LABDISC (made by ADVANTEC TOYO (Co., Ltd.), 50CP020AS). The sterilized filtrate was freeze-dried and powdered to obtain a powdered lactase-treated product. As for the inclusion of vitamin D binding protein in the lactase-treated product, anti-vitamin D binding protein antibodies were used to confirm the protein blotting method described in International Publication No. WO2016 / 194914.

[0085] (2) Administration test of capsules containing vitamin D binding protein (Example)

[0086] Enteric capsules adjusted to 1.0 mg of protein were prepared by conventional methods using the lactase-treated product prepared above. The enteric capsules were orally administered to 167 healthy subjects (118 females and 49 males) twice a day, in the morning and evening, one capsule each time, i.e., two capsules per day.

[0087] It should be noted that the age composition of the subjects is as follows: 60 subjects in the 40s, 52 subjects in the 50s, 28 subjects in the 60s, 24 subjects in the 70s, 2 subjects in the 80s, and 1 subject in the 90s.

[0088] Blood was collected from the subjects at the beginning of the dosing trial and 3 and 6 months after the start of the dosing trial. Chromosomal DNA was extracted from blood cells, and telomere length was measured by real-time qPCR based on the description of the following paper. Then, the average telomere length of all subjects was calculated.

[0089] Journal of Affective Disorders 273 (2020) 453-461 Zeyue Liu et al. “Inverse changes in telomere length between the blood and brain indepressive-like mice”

[0090] The relative values ​​of telomere length at the start of the drug administration test and 3 months and 6 months after the start of the drug administration test are as follows Figure 1 As shown. After 3 months and 6 months from the start of the drug administration test, the relative value of telomere length increased according to the drug administration period. The relative value of telomere length was confirmed to be significantly different at p < 0.01 between the start of drug administration and 3 months after drug administration, the start of drug administration and 6 months after drug administration, and 3 months after drug administration and 6 months after drug administration.

[0091] In addition, the telomere length of each sex of the subjects was Figure 2 As shown. Three months and six months after the start of the dosing test, the relative values ​​of telomere length increased in both men and women according to the dosing period. A significant difference of p<0.01 was confirmed at the beginning of dosing and three months after dosing, and at three months after dosing and six months after dosing for women. In addition, a significant difference of p<0.01 was confirmed at the beginning of dosing and three months after dosing for men. It should be noted that there was no significant difference in telomere length between men and women, confirming that the aging inhibitor of the present invention is effective regardless of gender.

[0092] The present disclosure (1) is an aging inhibitor comprising a vitamin D binding protein.

[0093] The present disclosure (2) is the aging inhibitor according to the present disclosure (1), wherein the vitamin D binding protein is treated with an enzyme, and the enzyme is one or more selected from β-galactosidase, sialidase and N-acetylgalactosaminidase.

[0094] The present disclosure (3) is the aging inhibitor according to the present disclosure (1) or (2), wherein the vitamin D binding protein is derived from milk or serum.

[0095] The present disclosure (4) is the aging inhibitor according to the present disclosure (3), wherein the milk is colostrum or normal milk.

[0096] The present disclosure (5) is an aging inhibitor according to any one of the present disclosures (1) to (4), which is used for extending telomeres.

[0097] The present disclosure (6) is a method for producing an aging inhibitor, comprising the step of contacting a vitamin D binding protein with one or more enzymes selected from the group consisting of β-galactosidase, sialidase, and N-acetylgalactosaminidase.

Claims

1. An aging inhibitor comprising: Vitamin D binding protein.

2. The aging inhibitor according to claim 1, wherein: The vitamin D binding protein is treated with an enzyme, and the enzyme is one or more selected from the group consisting of β-galactosidase, sialidase, and N-acetylgalactosaminidase.

3. The aging inhibitor according to claim 1 or 2, wherein: Vitamin D binding protein is derived from milk or serum.

4. The aging inhibitor according to claim 3, wherein: The milk is colostrum or normal milk. The aging inhibitor according to claim 1 or 2, which is used for extending telomeres.

6. A method for producing an aging inhibitor, comprising: A step of contacting the vitamin D binding protein with one or more enzymes selected from the group consisting of β-galactosidase, sialidase and N-acetylgalactosaminidase.

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