Composition for promoting interferon production

Abstract

To provide an orally administered composition which maintains, improves and strengthens immune function by enhancing IFN production.SOLUTION: A lactoferrin decomposition product is contained in an interferon (IFN) production enhancing composition as an active ingredient. It is preferable that the lactoferrin decomposition product derives from the lactoferrin of the cow, and it is more preferred that a peptide containing the amino acid sequence shown in the sequence number 2 is contained, and it is further preferred that a peptide containing the amino acid sequence shown in the sequence number 1 is contained. The composition can be ingested in the form of beverages and medicines.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a composition for promoting the production of interferon (IFN) containing lactoferrin degradation products.

Background Art

[0002] Interferon is a type of cytokine and is mainly classified into three types: type I, type II, and type III. Interferon has a strong antiviral effect, and type I interferon is represented by interferon-α and interferon-β. Type I interferon induces the expression of a wide range of genes that act to suppress virus replication in virus-infected cells, and activates a wide range of immune cells to eliminate viruses and virus-infected cells, thereby directly or indirectly activating the body's defense mechanism against viruses. The action of interferon is very strong, and its recombinant is also used for the treatment of viral diseases. Therefore, inducing the production of interferon in vivo can be an effective means to enhance resistance to pathogens such as viruses, and is also considered to be available for medical use. In vivo, type I interferon is produced from white blood cells such as macrophages, neutrophils, and dendritic cells. In particular, plasmacytoid dendritic cells (pDC), which are a type of dendritic cell, are the main type I interferon-producing cells in white blood cells. In addition, it is known that it is also produced in intestinal epithelial cells and the like. Lactoferrin (LF) is a glycoprotein that is abundant in breast milk, especially colostrum, and was originally thought to be involved in the biological defense of newborns. In recent years, it has been applied to foods and the like, and it has been reported that lactoferrin exhibits a wide range of physiological activities including biological defense and infection defense effects not only in newborns but also in children and adults. As mechanisms related to the infection defense effect of lactoferrin, bactericidal action against bacteria and inhibition of cell entry by attachment to virus particles are known.

[0003] ​​​​

[0004] Lactoferrin is hydrolyzed by digestive enzymes such as pepsin. Some orally ingested lactoferrin is broken down into peptides in the stomach, and it is believed that both undigested lactoferrin and its peptide breakdown products reach the small intestine. The peptide breakdown products of lactoferrin include peptides with various functions, one of which is lactoferricin (LFcin), an antimicrobial peptide located at the N-terminus of lactoferrin. Bovine lactoferricin (LFcinB) is a characteristic peptide with strong basicity, consisting of 26 amino acid residues in the amino acid region from the 17th to the 42nd amino acid from the N-terminus of bovine lactoferrin, i.e., the amino acid sequence shown in SEQ ID NO: 1 (Non-Patent Literature 1). This peptide has two cysteine ​​molecules forming a disulfide bond intramolecularly, and its molecular weight is 3195 kDa, which is about 1 / 25th of the molecular weight of bovine lactoferrin, which is approximately 80,000 kDa. The physiological activity of the peptide consisting of the amino acid sequence shown in SEQ ID NO: 1 remains unclear.

[0005] Traditionally, much research has focused on the antiviral and other physiological activities of interferon, and there has been much study on components that promote interferon production in the body; lactoferrin is one such component. In mice orally administered lactoferrin, interferon-α / β production is enhanced in the intestinal epithelium and Peyer's patches of the small intestine where leukocytes such as dendritic cells accumulate (Non-patent Literature 2). This suggests that lactoferrin indirectly participates in virus elimination by suppressing viral replication in infected cells and activating immune cells. Therefore, it has been proposed to incorporate lactoferrin as an active ingredient in interferon-α production enhancers into foods, beverages, and pharmaceuticals. (Patent Document 1)

[0006] More than 60% of orally ingested lactoferrin passes into the small intestine undigested, and less than 40% is digested in the stomach (Non-Patent Literature 3). The ability to induce interferon-α / β production is impaired by undigested lactoferrin. If ctoferrin is more potent, it is thought that its effect would be enhanced by processing it to prevent digestion in the stomach before ingestion. Conversely, if the ability to induce interferon-α / β production is eliminated in the stomach... If the lactoferrin hydrolysate is more potent, then it is thought that its effectiveness would be further enhanced by completely breaking it down beforehand with pepsin, a digestive enzyme in the stomach, before ingestion. In a study administering unprocessed lactoferrin and liposomal-encapsulated lactoferrin to humans, it was reported that liposomal-encapsulated lactoferrin had a higher capacity to produce interferon-α (Non-Patent Literature 4). Since liposomes exhibit resistance to digestive enzymes, it is hypothesized that the increased interferon-α production was due to the lactoferrin not being digested in the stomach (Non-Patent Literature 5). However, the phospholipids that make up liposomes have a high affinity for cell membranes, and it is possible that the liposomes increased the uptake of lactoferrin into target cells, thereby increasing interferon-α production. In that case, digestion in the stomach may not be relevant, but this possibility is not mentioned in these literatures, and there is absolutely no consideration of the possibility that digested lactoferrin breakdown products may be more effective than undigested lactoferrin. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Japanese Patent Publication No. 2006-083089 [Patent Document 2] Japanese Patent Publication No. 2012-235768 [Non-patent literature]

[0008] [Non-Patent Document 1] Wakabayashi, Hiroyuki, et al. Current Pharmaceutical Design 9.16 (2003): 1277-1287. [Non-Patent Document 2] Kuhara, Tetsuya, et al. Nutrition and Cancer 38.2 (2000): 192-199. [Non-Patent Document 3] Troost, Freddy J., et al. The Journal of nutrition 131.8 (2001): 2101-2104. [Non-Patent Document 4] Ishikado, Atsushi, et al. Biofactors 21.1-4 (2004): 69-72. [Non-Patent Document 5] http: / / www.ec-lactoferrin.org / lactoferrin / PDF / 6-4.pdf [Overview of the Initiative] [Problems that the invention aims to solve]

[0009] The relationship between lactoferrin hydrolysates and IFN production induction is unknown. If lactoferrin hydrolysates are involved in IFN production induction, the hydrolysates obtained by breaking down lactoferrin are a mixture of countless peptides, so it is thought that peptides that contribute to production induction and peptides that do not contribute are present. In addition, lactoferrin pepsin hydrolysates (mixtures) have a bitter taste and affect the flavor of food (Patent Document 2). If lactoferrin hydrolysates are involved in IFN production induction, identifying the peptide that contributes to IFN production promotion activity from among the various peptides contained in lactoferrin hydrolysates and using it alone would allow the desired effect to be achieved with a smaller amount. There are also processing advantages such as minimizing the impact on flavor. In light of these circumstances, it is worthwhile to investigate the IFN production-inducing ability of lactoferrin degradation products. Therefore, the object of the present invention is to provide an orally ingestible composition that can maintain, improve, or enhance immune function by promoting IFN production. [Means for solving the problem]

[0010] As a result of intensive studies to solve the above problems, the present inventors have found that when mononuclear cells collected from human peripheral blood are cultured in the presence of lactoferrin degradation products, IFN production is promoted more than in the presence of lactoferrin.Furthermore, the inventors have found that a specific peptide contained in the lactoferrin degradation products contributes to the action of promoting IFN production, and have thus completed the present invention.

[0011] That is, the present invention is a composition for promoting IFN production, which contains lactoferrin degradation products. Here, the lactoferrin degradation products are preferably derived from bovine lactoferrin, and more preferably contain a peptide having the amino acid sequence shown in SEQ ID NO: 2. Even more preferably, the peptide having the amino acid sequence shown in SEQ ID NO: 2 is a peptide consisting of the amino acid sequence shown in SEQ ID NO: X. In the present invention, the IFN is preferably IFN-α. The composition of the present invention can preferably be in the form of a food or drink.Such a food or drink is preferably ingested to maintain, improve, or enhance the immune function. The composition of the present invention can preferably be in the form of a pharmaceutical.Such a pharmaceutical is preferably administered for diseases and / or conditions that can be prevented, treated, or improved by promoting IFN production.Such diseases and / or conditions include one or more selected from the group consisting of multiple sclerosis, psoriasis, melanoma, demyelinating polyradiculoneuritis, cancer cell growth and / or metastasis, and diseases and / or complications caused by virus growth.

Effects of the Invention

[0012] According to the present invention, the production of IFN can be efficiently promoted, thereby maintaining, improving, or enhancing the immune function.

Modes for Carrying Out the Invention

[0013] Note: There seems to be a mistake in the original text where "SEQ ID NO: X" should be "SEQ ID NO: 1" in the translation of item ID=7. Also, the etc. tags are left unchanged as per the instruction.Next, the present invention will be described in detail. However, the present invention is not limited to the following embodiments and can be freely changed within the scope of the present invention.

[0014] The composition of the present invention contains lactoferrin degradation product as an active ingredient.

[0015] Lactoferrin is an iron-binding glycoprotein contained in the milk, tears, saliva, blood, etc. of mammals such as sheep, goats, pigs, mice, water buffalo, camels, yaks, horses, donkeys, llamas, cows or humans. The lactoferrin in the present invention may be derived from any mammal and is not particularly limited. However, from the viewpoints of content and availability, lactoferrin derived from milk of, for example, cows, humans, etc. is preferable, and that derived from cow milk is particularly preferable. The milk may be any of colostrum, transitional milk, normal milk, or terminal milk. In addition, the lactoferrin in the present invention may be lactoferrin separated from skim milk, whey, etc., which are processed products of the above-mentioned milk, by a conventional method (for example, ion chromatography, etc.), recombinant lactoferrin produced from microorganisms, animal cells, transgenic animals, etc. by genetic engineering, synthetic lactoferrin, or a mixture thereof. Lactoferrin may also be non-glycosylated or glycosylated. As such lactoferrin, commercially available lactoferrin (for example, manufactured by Morinaga Milk Industry Co., Ltd., etc.) produced on an industrial scale can be used for obtaining the degradation product.

[0016] In this specification, when referring to "derived from bovine lactoferrin" or the like, "derived" means originally possessed by the animal and does not mean the source of collection. For example, lactoferrin or a peptide which is a degradation product thereof encoded by a gene of bovine lactoferrin is introduced into a host organism such as Escherichia coli, and the lactoferrin or peptide produced by expressing the gene is "derived" from cows.

[0017] Herein, an example of a method for preparing lactoferrin (separation and purification of lactoferrin from raw materials such as milk) is shown below, but it is not limited to this method. A bovine-derived milk raw material is passed through a cation exchange column, the eluate is collected, and this eluate is repeatedly passed through the column as needed. Deionized water is passed through this column, followed by saline solution to obtain an eluate of basic proteins adsorbed on the cation exchange column. The proteins are collected from this eluate, washed as needed, dissolved in deionized water, and this solution is filtered through an ultrafiltration membrane. Further desalting and freeze-drying are performed to obtain powdered lactoferrin.

[0018] More specifically, first, an ion exchanger is packed into a column, hydrochloric acid is passed through it, and the ion exchanger is washed with water to equilibrate it. Next, skim milk cooled to 4°C and pH 6.9 is passed through the column, the permeate is collected, and the process is repeated. Then, deionized water is passed through the column, followed by saline solution to obtain an eluate of basic proteins adsorbed onto the ion exchanger. The proteins recovered from this eluate are washed, dissolved with deionized water, and the resulting solution is desalted using an ultrafiltration membrane module and freeze-dried to obtain powdered bovine lactoferrin. In this way, bovine lactoferrin with a purity of 95% by mass or higher is obtained.

[0019] In lactoferrin, genetic mutations such as substitutions, deletions, insertions, additions, or inversions of one or more bases at one or more positions naturally exist depending on the species, genus, individual, etc., and mutations may also occur in the amino acids of the protein encoded by genes with such mutations. The lactoferrin used in this invention may contain such mutations, as long as they do not impair the effects of the present invention.

[0020] The degradation products obtained by degrading lactoferrin, usually through hydrolysis, are used as lactoferrin degradation products in this invention. Examples of hydrolysis treatments include the method described in Japanese Patent Publication No. 2012-235768. Specifically, the pH of the lactoferrin solution is adjusted to 2-4, preferably 2.5-3.5, and particularly preferably 3, using an acid such as hydrochloric acid, citric acid, or acetic acid before the enzymatic reaction treatment. After adding a desired amount of proteolytic enzyme to a pH-adjusted lactoferrin solution, the enzyme reaction temperature is maintained at 35-55°C, preferably 40-50°C, more preferably 42-48°C, and the lactoferrin is hydrolyzed while stirring for 6-24 hours, preferably 12-18 hours. Next, for example, the reaction solution is heated to 80°C and maintained at that temperature for 10 minutes to inactivate the enzyme by heating. Furthermore, preferably, an alkaline solution such as sodium hydroxide solution is added to adjust the pH to 5-7, for example, 6. The reaction solution (lactoferrin hydrolysate) after pH adjustment may be used as a solution, but it is preferable to powderize it by freeze-drying or the like. Furthermore, the lactoferrin hydrolysate can also be used after being fractionated by chromatography or ultrafiltration.

[0021] The lactoferrin degradation products obtained in this manner are usually mixtures of various peptides. Among these, it is preferable that they contain a peptide containing the amino acid sequence shown in SEQ ID NO: 2. The amino acid sequence shown in SEQ ID NO: 2 is the active site sequence of lactoferricin and is known to have functional properties such as antibacterial activity, and is considered to be a site that contributes to the IFN production promoting effect according to the present invention (Non-Patent Literature 1). In this invention, the "peptide containing" the amino acid sequence shown in SEQ ID NO: 2 means a peptide in which one or more peptide residues of any amino acid are added to the N-terminus and / or C-terminus of the amino acid sequence, more specifically 1 to 10 residues, preferably 1 to 5 residues, and even more preferably Alternatively, a peptide with 1 to 3 residues added, and a sequence that promotes INF production, is preferable. As a peptide containing the amino acid sequence shown in SEQ ID NO: 2, bovine lactoferricin, i.e., a peptide consisting of the amino acid sequence shown in SEQ ID NO: 1, is particularly preferred. Note that bovine lactoferricin may also refer to a peptide consisting of a 25-residue amino acid sequence in which the C-terminal alanine is missing from the amino acid sequence shown in SEQ ID NO: 1, and this is also preferred as a peptide containing the amino acid sequence shown in SEQ ID NO: 2. As shown in the examples described later, bovine lactoferricin can promote IFN production in mononuclear cells recovered from human peripheral blood more effectively than lactoferrin degradation products (mixtures) containing it.

[0022] In the composition of the present invention, the content of lactoferrin hydrolysate relative to the total composition is preferably 0.001% by mass or more and less than 100% by mass, more preferably 0.005 to 75% by mass, and even more preferably 0.01 to 50% by mass. These amounts may be calculated based on the amount of lactoferrin that is digested in the body after ingestion of the composition of the present invention and converted into lactoferrin hydrolysates. In other words, it is not prevented that the composition of the present invention may contain lactoferrin.

[0023] In the present invention, from the viewpoint of achieving the desired effect, it is desirable that the lactoferrin hydrolysate contains a peptide containing the amino acid sequence shown in SEQ ID NO: 2 in an amount of preferably 0.01% by weight or more, more preferably 0.1% by weight or more, and even more preferably 0.5% by weight or more of the total lactoferrin hydrolysate. There is no particular upper limit, but it is usually 10% by weight or less, and more preferably 3% by weight or less. Furthermore, from the viewpoint of achieving its effects, it is desirable that the lactoferrin hydrolysate contains a peptide consisting of the amino acid sequence shown in SEQ ID NO: 1 in an amount of preferably 0.01% by weight or more, more preferably 0.1% by weight or more, and even more preferably 0.5% by weight or more of the total lactoferrin hydrolysate. There is no particular upper limit, but it is usually 30% by weight or less, and more preferably 10% by weight or less.

[0024] The compositions of the present invention can promote the production of IFN, particularly IFN-α. Here, "promotion of IFN production" means enhancing the production of IFN in IFN-producing cells after application of the composition of the present invention compared to before application, or increasing production when the composition of the present invention is applied compared to when it is not applied, or more strongly inducing production. Therefore, the IFN production promoting composition in the present invention may be rephrased as an IFN production enhancing composition or an IFN production induction composition.

[0025] Furthermore, cells or tissues whose IFN production is promoted by stimulation with the IFN production-promoting composition of the present invention include immunotherapeutic cells such as plasmacytoid dendritic cells (pDCs), macrophages, and NK cells. Examples include cells, fibroblasts, epithelial cells, and Peyer's patch tissue (for example, Peyer's patches distributed on the mucosa of the small intestine). The IFN production-promoting composition of the present invention can effectively act on organs that produce these cells and efficiently produce IFN. The amount of IFN produced after applying the composition of the present invention is at least 1.1 times, preferably 1.5 times or more, preferably 2 times or more, preferably 2.5 times or more, preferably 3 times or more, preferably 3.5 times or more, and more preferably 4 times or more, compared to before application. The upper limit is not particularly limited, but may be 8 times or less, 7 times or less, 6 times or less, or 5 times or less. It may also be within a range of non-consistent combinations of the above values, specifically 1.1 to 8 times, 1.5 to 8 times, 2 to 8 times, 2.5 to 8 times, 3 to 8 times, 3.5 to 8 times, or 4 to 8 times. Here, the amount of IFN produced can be confirmed qualitatively or quantitatively by well-known methods, for example, by measuring the expression level of the gene encoding IFN using standard methods.

[0026] Since the composition of the present invention promotes IFN production, it is expected to activate the function of various immune cells and promote IFN production, and therefore is expected to maintain, improve, or strengthen immune function. Maintaining immune function includes maintaining a state in which normal or good immune function is exhibited and preventing a decline in function, and improving immune function includes restoring a declined immune function to a normal state.

[0027] The subjects to whom the composition of the present invention is administered (ingested) are not particularly limited as long as they are animals, but are usually mammals, and humans are preferred. Furthermore, although the subjects are not particularly limited, healthy people are preferred. Here, a healthy person means a person who is not suffering from disease or illness.

[0028] Another aspect of the present invention is the use of lactoferrin hydrolysates in the production of compositions for promoting IFN production. Another aspect of the present invention is the use of lactoferrin degradation products in promoting IFN production. Another aspect of the present invention is lactoferrin degradation products used to promote IFN production. Another aspect of the present invention is a method for promoting IFN production, which involves administering lactoferrin hydrolysates to a target.

[0029] Furthermore, "administering lactoferrin hydrolysates to a target" may be synonymous with "allowing lactoferrin hydrolysates to be ingested by a target." Ingestion may be voluntary (ad libido) or compulsory (forced ingestion). In other words, the administration process may specifically involve, for example, supplying lactoferrin hydrolysates to a target by incorporating them into food, beverages, or animal feed, thereby allowing the target to ingest lactoferrin hydrolysates ad libido.

[0030] The timing and duration of administration of the composition of the present invention are not particularly limited and can be appropriately selected depending on the condition of the recipient.

[0031] The composition of the present invention may be in the form of food or beverages or pharmaceuticals, or it may be included as an additive in food or beverages or pharmaceuticals. The composition of the present invention may be administered orally or parenterally, but is usually administered orally. Parenteral administration methods include transdermal, intravenous, rectal, and inhalation.

[0032] The content of the composition of the present invention when administered orally may be the total content of the composition as described above, or it may be diluted as appropriate. For example, the content of lactoferrin hydrolysate relative to the total composition when administered orally is preferably 0.001 to 100% by mass, more preferably 0.005 to 75% by mass, and even more preferably 0.01 to 50% by mass. These may be within the range of content typically used when distributed as an oral composition.

[0033] The amount of the composition of the present invention to be ingested (administered) is appropriately selected depending on the age (months), sex, condition, and other conditions of the person receiving the intake (administration). The amount of the composition of the present invention to be ingested (administered) is, for example, preferably 0.01 to 1000 mg / kg / day, more preferably 0.1 to 500 mg / kg, as an intake of lactoferrin hydrolysate. A guideline for the amount should be g / day, more preferably in the range of 1 to 300 mg / kg / day. Regardless of the amount or duration of intake (administration), the composition of the present invention can be administered once a day or in multiple divided doses.

[0034] When the composition of the present invention is intended for oral intake, it is preferable to provide it in the form of a food or beverage. As for food and beverages, there are no particular restrictions on their form or properties as long as they do not impair the effects of the present invention and can be taken orally. Except for containing lactoferrin hydrolysate, they can be manufactured using ordinary methods with raw materials commonly used in food and beverages.

[0035] Food and beverages include, regardless of form (liquid, paste, gel, solid, powder, etc.), such as tablets; liquid foods (nutritional foods for tube feeding); wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken batter, breadcrumbs; instant noodles, cup noodles, retort / prepared foods, canned prepared foods, microwaveable foods, instant soups / stews, instant miso soup / clear soup, canned soups, freeze-dried foods, and other instant foods; canned agricultural products, canned fruits, jams, etc. Processed agricultural products such as marmalades, pickles, boiled beans, dried agricultural products, and cereals (grain processed products); processed marine products such as canned seafood, fish ham and sausages, processed seafood products, seafood delicacies, and tsukudani (simmered seafood); processed livestock products such as canned and paste meats, and meat ham and sausages; processed milk, milk beverages, yogurt (fermented milk), lactic acid bacteria beverages, cheese, ice cream, cream, and other dairy products; fats and oils such as butter, margarine, and vegetable oil; soy sauce, miso, and sauces. Basic seasonings such as tomato-based seasonings, mirin (sweet rice wine), and vinegars; compound seasonings and food products such as cooking mixes, curry bases, sauces, dressings, noodle soup bases, spices, and other compound seasonings; frozen foods such as raw frozen foods, semi-prepared frozen foods, and pre-cooked frozen foods; and confectionery such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice crackers, bean sweets, dessert sweets, jellies, and other sweets. Examples include carbonated beverages, natural fruit juices, fruit juice drinks, fruit juice-containing soft drinks, fruit pulp drinks, fruit juice drinks with fruit pulp, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic beverages, and other beverages; baby food, furikake (rice seasoning), nori (rice seasoning for ochazuke), and other commercially available foods; supplements, prepared milk (including powdered milk, liquid milk, etc.), and other nutritional compositions; enteral nutrition foods; and functional foods (foods for specified health uses, foods with nutritional function claims).

[0036] Furthermore, when formulated as a supplement for food and beverage use, it may be enterically coated by an enteric coating or the like. It can be formulated as a solid preparation such as a powder, granules, tablet, or capsule; or as a liquid preparation such as a solution, syrup, suspension, or emulsion. When formulating in this manner, the explanation of the components, carriers, and methods related to the formulation of pharmaceuticals described later may be followed.

[0037] Furthermore, it can also be used as animal feed as a form of food or beverage. Examples of animal feed include pet food, livestock feed, and fish feed. The form of the feed is not particularly limited, and in addition to lactoferrin hydrolysate, it may contain, for example, cereals such as corn, wheat, barley, rye, and milo; vegetable oil cakes such as soybean oil cake, rapeseed oil cake, coconut oil cake, and linseed oil cake; brans such as wheat bran, wheat bran, rice bran, and defatted rice bran; manufacturing residues such as corn gluten meal and corn jam meal; animal feeds such as fish meal, skim milk powder, whey, yellow grease, and taro; yeasts such as Torula yeast and brewer's yeast; mineral feeds such as tricalcium phosphate and calcium carbonate; oils and fats; single amino acids; sugars, etc.

[0038] When the composition of the present invention is in the form of food or beverage (including animal feed), it can be provided and sold as food or beverage with indications of its use, such as maintaining immune function. Furthermore, the lactoferrin hydrolysate described herein can be used for the manufacture of such food or beverage products.

[0039] Such "display" acts include all acts that inform consumers of the aforementioned uses, and any expression that can evoke or infer the aforementioned uses, regardless of the purpose of the display, the content of the display, or the object or medium to which it is displayed, falls under the category of "display" acts in this invention. Furthermore, the "display" will be made using language that allows consumers to directly recognize the above-mentioned uses. This is preferable. Specifically, examples include transferring, delivering, displaying for transfer or delivery, or importing food and beverage products or product packaging on which the above-mentioned use is described; displaying or distributing advertisements, price lists, or transaction documents related to products on which the above-mentioned use is described; or providing information containing such materials on which the above-mentioned use is described by electromagnetic means (such as the Internet).

[0040] On the other hand, the content of the display is preferably a display approved by the government or other administrative body (for example, a display approved based on various systems established by the government and made in accordance with such approval). Furthermore, it is preferable to attach such display content to packaging, containers, catalogs, brochures, point-of-sale (POP) displays and other promotional materials used at sales sites, and other documents.

[0041] Furthermore, "labeling" also includes labels for health foods, functional foods, enteral nutrition foods, foods for special dietary uses, health functional foods, foods for specified health uses, nutrient function foods, foods with functional claims, quasi-drugs, etc. In particular, labels approved by the Consumer Affairs Agency include, for example, labels approved under the systems for foods for specified health uses, foods with nutrient function, or foods with functional claims, or similar systems. Specifically, these include labels for foods for specified health uses, labels for conditionally specified health uses, labels that affect the structure or function of the body, labels that reduce disease risk, and labels that show scientifically based functionality. More specifically, typical examples include labels for foods for specified health uses (especially labels for health uses) and similar labels as defined in the Cabinet Office Ordinance concerning the permission of special use labeling, etc., as stipulated in the Health Promotion Act (Cabinet Office Ordinance No. 57 of August 31, 2009). Such claims include, for example, "increases interferon," "acts on plasmacytoid dendritic cells (pDCs)," "activates immune cells," and "improves the immune function of healthy people." "Helps maintain," "Enhances immune function," "Supports immune function," "Supports the maintenance of immune function in healthy people," "Acts on plasmacytoid dendritic cells (pDCs) and in healthy people Examples of labeling include statements such as "Helps maintain immune function."

[0042] The composition of the present invention can also be used in the form of a pharmaceutical product. Such pharmaceuticals are preferably those administered for diseases involving IFN, specifically diseases and / or conditions that can be prevented, treated, or improved by promoting IFN production. More specifically, prevention and treatment of multiple sclerosis, psoriasis, melanoma, demyelinating polyradiculopathy (Guillan-Barré syndrome), etc.; cancer cells (myeloma, non-Hodgkin lymphoma, malignant lymphoma) It can be a pharmaceutical drug administered to suppress the proliferation and metastasis of tumors (such as mammary glands, breast cancer, hepatocellular carcinoma, osteosarcoma, melanoma, oral papilloma, and acute leukemia); to treat or prevent various diseases and complications caused by the proliferation of viruses (such as herpes simplex virus, cytomegalovirus, and SARS virus); and to reduce the risk of these diseases and complications.

[0043] The route of administration of the drug may be either orally or parenterally, but oral administration is preferred. Parenteral administration methods include transdermal, intravenous, rectal, and inhalation. In terms of pharmaceutical form, the drug can be formulated into any desired dosage form depending on the method of administration. For example, for oral administration, it can be formulated into solid preparations such as powders, granules, tablets, and capsules; or into liquid preparations such as solutions, syrups, suspensions, and emulsions. It can also be made into an enteric-coated preparation by enteric coating, etc. For parenteral administration, it can be formulated into suppositories, ointments, injections, etc. In formulation, in addition to lactoferrin hydrolysates, other ingredients commonly used in formulations, such as excipients, pH adjusters, colorants, and flavorings, can be used. Furthermore, other pharmacoactive ingredients, or known or potentially discovered ingredients with IFN production-promoting effects, can be used in combination. In addition, formulation can be carried out by known methods as appropriate, depending on the dosage form. When formulating, carriers commonly used in formulation may be added as appropriate. Such carriers include excipients. Examples include agents, binders, disintegrants, lubricants, stabilizers, and flavor / odor-correcting agents.

[0044] Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, carboxymethylcellulose, and carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

[0045] Examples of binders include, in addition to the above-mentioned excipients, gelatin, polyvinylpyrrolidone, macrogol, and the like.

[0046] Examples of disintegrants include, in addition to the above-mentioned excipients, chemically modified starches or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and cross-linked polyvinylpyrrolidone.

[0047] Examples of lubricants include talc; stearic acid; metal stearate salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and gayl wax; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylate salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as anhydrous silicic acid and silicic acid hydrate; and starch derivatives.

[0048] Examples of stabilizers include para-hydroxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

[0049] Examples of flavoring and odor-modifying agents include sweeteners, acidulants, and flavorings. In the case of liquid formulations for oral administration, examples of carriers used include solvents such as water.

[0050] The timing of taking the pharmaceutical product of the present invention is not particularly limited, for example, before meals, after meals, between meals, or before going to bed. [Examples]

[0051] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0052] [Example 1] (1) Preparation of bovine lactoferrin hydrolysates The bovine lactoferrin hydrolysate was prepared as follows: Bovine lactoferrin (manufactured by Morinaga Milk Industry Co., Ltd.) was dissolved in purified water to a concentration of 5% by mass, and the pH was adjusted to 3 with hydrochloric acid. Heat, then add 3% by mass of pepsin (porcine pepsin; mixed with the mass of bovine lactoferrin). Hydrolysis was carried out by adding a solution manufactured by Kojun Pharmaceutical Co., Ltd. (optimal pH 2-3) and stirring for 4 hours. After the hydrolysis reaction was complete, the reaction solution was heated to 80°C and kept warm for 10 minutes to inactivate the enzyme. After cooling the reaction solution, sodium hydroxide solution was added to adjust the pH of the reaction solution to 6, and then the reaction solution was freeze-dried to obtain bovine lactoferrin hydrolysate. It was confirmed by conventional methods that this hydrolysate contained peptides containing the amino acid sequence shown in SEQ ID NO: 2, and in particular peptides consisting of the amino acid sequence shown in SEQ ID NO: 1.

[0053] (2) Evaluation test of the interferon production promoting effect of bovine lactoferrin hydrolysates To compare and evaluate the interferon production-promoting effects of bovine lactoferrin and bovine lactoferrin degradation products, we used single-stranded RNA (ssRNA) that induces interferon production and bovine lactoferrin. Interferon, or bovine lactoferrin degradation products, was cultured in a culture dish with peripheral blood mononuclear cells (PBMCs), and the amount of interferon released by the PBMCs into the culture supernatant was quantified.

[0054] Specifically, PBMCs were recovered from peripheral blood samples taken from collaborators (n=3). ssRNA40 / LyoVec (registered trademark, Invitrogen) was added to all culture dishes to a final concentration of 2.5 μg / mL. It was added. In addition, bovine lactoferrin (Morinaga Milk Industry Co., Ltd.) was added to a culture dish containing ssRNA40 / LyoVec at a final concentration of 100 μg / mL, or the bovine lactoferrin degradation product prepared in (1) was added. The solution was added to a concentration of 100 μg / mL. Bovine lactoferrin, bovine lactoferrin Each solution was pre-dissolved in sterile water. Only sterile water was added to the control group. PBMC was dissolved in 5% human serum (Sigma-Aldrich), Glutamax (Thermo Fisher Scientific), and 1 Each culture dish was created by suspending the cells in RPMI1640 medium (Sigma-Aldrich) supplemented with % Penicillin Streptomycin (Fujifilm Wako Pure Chemical Industries, Ltd.) and adding it to the respective culture dishes. PBMCs were placed in one culture dish. 5 x 10 5 The cells were cultured at 37°C under 5% CO2 conditions. After 24 hours of culture, the culture supernatant was collected, and the concentration of interferon-α was measured using a commercially available ELISA kit (PBL Assay Science) according to the manufacturer's specified protocol.

[0055] Table 1 shows the concentrations of interferon-α in the culture supernatant when bovine lactoferrin and bovine lactoferrin hydrolysates were added. In PBMCs isolated from the peripheral blood of all participants, higher concentrations of interferon-α were detected when bovine lactoferrin was added in addition to ssRNA40 / LyoVec compared to the control group. Furthermore, even higher concentrations of interferon-α were detected when bovine lactoferrin hydrolysates were added compared to when bovine lactoferrin was added.

[0056] [Table 1]

[0057] [Example 2] (1) Synthesis of lactoferricin The bovine lactoferricin (LFcinB), specifically the 26-residue peptide shown in Sequence ID No. 1, was synthesized by Toray Research Center, Inc.

[0058] (2) Evaluation of the interferon production promoting effect of LFcinB To compare and evaluate the interferon production-promoting effects of bovine lactoferrin degradation products and LFcinB, we used interferon-inducing ssRNA and bovine lactoferrin degradation products, In (1), LFcinB synthesized in (1) was cultured with peripheral blood mononuclear cells (PBMCs) in a culture dish, and the interferon released by the PBMCs into the culture supernatant was quantified.

[0059] Specifically, PBMCs were recovered from peripheral blood samples taken from collaborators (n=3). ssRNA40 / LyoVec (registered trademark, Invitrogen) was added to all culture dishes to a final concentration of 2.5 μg / mL. It was added. In addition, bovine lactoferrin hydrolysate was added to a culture dish containing ssRNA40 / LyoVec to a final concentration of 100 μg / mL, or LFcinB to a final concentration of 4 μg / mL. The bovine lactoferrin hydrolysate and LFcinB were pre-dissolved in sterile water. PBMCs were added to RPMI1640 medium supplemented with 5% human serum (Sigma-Aldrich), Glutamax (Thermo Fisher Scientific), and 1% Penicillin Streptomycin (Fujifilm Wako Pure Chemical Industries). The PBMCs were suspended in (Sigma-Aldrich) and added to each culture dish. tari 1 x 10 5 The cells were cultured at 37°C under 5% CO2 conditions. After 24 hours of culture, the culture supernatant was collected, and the concentration of interferon-α was measured using a commercially available ELISA kit (PBL Assay Science) according to the manufacturer's specified protocol.

[0060] To compare and evaluate the interferon-producing ability of bovine lactoferrin hydrolysate and LFcinB, the amount of interferon-α produced per unit concentration was calculated by dividing the quantitative value of interferon-α in the culture supernatant by the concentration of the added bovine lactoferrin hydrolysate or LFcinB. The results are shown in Table 2. In PBMCs recovered from the peripheral blood of all participants, the addition of LFcinB in addition to ssRNA40 / LyoVec showed a higher interferon-α production induction ability compared to the addition of bovine lactoferrin degradation products.

[0061] [Table 2]

Claims

1. A composition for promoting interferon-α (IFN-α) production, comprising a lactoferrin hydrolysate, wherein the lactoferrin hydrolysate comprises a peptide having the amino acid sequence shown in SEQ ID NO:

1.

2. A composition for promoting interferon-α (IFN-α) production, comprising a peptide having the amino acid sequence shown in Sequence ID No.

1.

3. A composition for promoting interferon-α (IFN-α) production, comprising a pepsin-mediated degradation product of bovine lactoferrin.

4. The composition according to claim 3, wherein the bovine lactoferrin degradation product by pepsin contains a peptide having the amino acid sequence shown in SEQ ID NO:

2.

5. The composition according to claim 4, wherein the peptide containing the amino acid sequence shown in SEQ ID NO: 2 is a peptide consisting of the amino acid sequence shown in SEQ ID NO:

1.

6. A composition for promoting interferon-α (IFN-α) production, comprising a bovine lactoferrin hydrolysate, wherein the bovine lactoferrin hydrolysate comprises a peptide having the amino acid sequence shown in Sequence ID No.

1.

7. A composition for promoting interferon-α (IFN-α) production, comprising a lactoferrin degradation product by pepsin, wherein the lactoferrin degradation product by pepsin comprises a peptide having the amino acid sequence shown in Sequence ID No.

1.

8. A composition according to any one of claims 1 to 7, which is a food or beverage.

9. The composition according to claim 8, which is taken to maintain, improve, or strengthen immune function.

10. A composition according to any one of claims 1 to 7, which is a pharmaceutical product.

11. The composition according to claim 10, administered for diseases and / or conditions that can be prevented, treated, or improved by promoting IFN-α production.

12. The composition according to claim 11, wherein the disease and / or condition is one or more selected from the group consisting of multiple sclerosis, psoriasis, melanoma, demyelinating polyradiculitis, proliferation and / or metastasis of cancer cells, and diseases and / or complications thereof caused by viral proliferation.

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