Composition for inhibiting amyloid beta accumulation

Abstract

The purpose of the present invention is to provide a novel composition for inhibiting the accumulation of amyloid beta. The present invention relates to a composition for inhibiting the accumulation of amyloid beta, which is characterized by containing Cyclo (His-Pro) and / or Cyclo (Gly-Pro) as an active ingredient.

Description

Technical Field

[0001] The present invention relates to a composition for inhibiting the accumulation of amyloid-β. Background Art

[0002] Amyloid-β (Aβ) is a protein produced in the brain. Aggregation of amyloid-β occurs through oligomerization or fibrillation, thereby exhibiting neurocytotoxicity. In addition, the aggregated amyloid-β does not excrete from the brain but accumulates, which is considered to be a trigger for the onset of Alzheimer's disease. Thus, it is generally considered that inhibiting the accumulation of amyloid-β is effective, for example, for the prevention of Alzheimer's disease or the inhibition of disease progression.

[0003] As a medicament for improving the symptoms of Alzheimer's disease, acetylcholinesterase inhibitors are known. Inhibition of acetylcholinesterase increases the amount of acetylcholine in the brain and is effective for inhibiting the progression of cognitive function decline. However, the accumulated amyloid-β cannot be removed by inhibiting acetylcholinesterase. Therefore, a substance that can inhibit the accumulation of amyloid-β and can be taken daily as a food or drink is pursued.

[0004] It is described in Patent Document 1 that the accumulation of amyloid-β is inhibited by combining Zn and Cyclo(His-Pro). However, the effect of Cyclo(His-Pro) alone is not disclosed.

[0005] Patent Document Patent Document 1: U.S. Patent Application Publication No. 2009 / 0004291 Summary of the Invention

[0006] An object of the present invention is to provide a novel composition for inhibiting the accumulation of amyloid-β.

[0007] The present inventors conducted in-depth research on the above problems and found that Cyclo(His-Pro) and / or Cyclo(Gly-Pro) as cyclic dipeptides are effective for inhibiting the accumulation of amyloid-β.

[0008] That is, although the present invention is not limited thereto, it relates to the following composition for inhibiting the accumulation of amyloid-β and the like. [1] A composition for inhibiting the accumulation of amyloid-β, characterized by containing Cyclo(His-Pro) and / or Cyclo(Gly-Pro) as an active ingredient. [2] The composition according to the above [1], characterized in that the amyloid-β is at least 1 selected from amyloid-β1-38, amyloid-β1-40, and amyloid-β1-42. [3] The composition according to [1] or [2] above, characterized in that it is an oral composition. [4] The composition according to any one of [1] to [3] above, characterized in that it is a food or drink or a pharmaceutical product. [5] The composition according to any one of [1] to [4] above, characterized in that it is attached with a label of one or more functions selected from "enhancing cognitive function", "inhibiting the decline of cognitive function", "maintaining good cognitive function", "enhancing memory", "inhibiting the decline of memory", "maintaining good memory", "enhancing the accuracy of memory", "preventing memory disorders", "improving memory disorders", "maintaining memory as part of cognitive function", "suitable for the function of those who care about memory decline", "enhancing the accuracy of memory or the correctness of judgment as part of cognitive function", "improving memory retention or integration", "maintaining the enhancement of cognitive function", "improving executive function", "promoting attention and concentration", "improving learning ability", "maintaining and improving orientation ability", "delaying the decline of cognitive function with age", "strengthening short-term and long-term memory", "promoting verbal and visual-spatial memory", and "preventing cognitive function disorders during the aging process". [6] An application, characterized in that Cyclo(His-Pro) and / or Cyclo(Gly-Pro) is used for the manufacture of a composition for inhibiting amyloid-β accumulation. [7] An application, characterized in that Cyclo(His-Pro) and / or Cyclo(Gly-Pro) is used for inhibiting the accumulation of amyloid-β.

[0009] According to the present invention, a composition for inhibiting amyloid-β accumulation can be provided. The composition for inhibiting amyloid-β accumulation of the present invention can be used as a food or drink or a pharmaceutical product for inhibiting amyloid-β accumulation. The cyclic dipeptide can be ingested by humans as a food or drink or a pharmaceutical product, and also has the advantage of high safety. Description of the Drawings

[0010] Figure 1A It is a graph showing the concentration of amyloid-β1-38 in the culture supernatant obtained by adding Cyclo(Gly-Pro) (CGP) or Cyclo(His-Pro) (CHP) to H4 glioma cells (H4-hAPP cells) overexpressing wild-type amyloid precursor protein with Swedish double mutations K595N / M596L. Figure 1B It is a graph showing the concentration of amyloid-β1-40 in the culture supernatant obtained by adding CGP or CHP to H4-hAPP cells. Figure 1CA graph showing the concentration of amyloid-β1-42 in the culture supernatant obtained by adding CGP or CHP to H4-hAPP cells. Detailed implementation mode

[0011] The composition for inhibiting amyloid-β accumulation of the present invention (also referred to as the composition of the present invention) contains Cyclo(His-Pro) and / or Cyclo(Gly-Pro) as active ingredients. The composition for inhibiting amyloid-β accumulation of the present invention contains either Cyclo(His-Pro) or Cyclo(Gly-Pro) as an active ingredient, or contains both Cyclo(His-Pro) and Cyclo(Gly-Pro) as active ingredients.

[0012] Both of the above-mentioned Cyclo(His-Pro) (cyclo-histidylproline) and Cyclo(Gly-Pro) (cycloglycylproline) are cyclic dipeptides. In this specification, the so-called "cyclic dipeptide" refers to a compound characterized by having a diketopiperazine structure formed by dehydration condensation of the amino group of the amino acid on the N-terminal side and the carboxyl group of the amino acid on the C-terminal side with amino acids as constituent units. In addition, in this specification, when the amino acid compositions of cyclic dipeptides are the same, the order of their descriptions may be either one first. For example, Cyclo(Gly-Pro) and Cyclo(Pro-Gly) (cycloprolylhistidine) represent the same cyclic dipeptide. In this specification, Cyclo(His-Pro) and Cyclo(Gly-Pro) may be respectively referred to as CHP and CGP.

[0013] CHP and CGP may be in the form of salts. The composition for inhibiting amyloid-β accumulation of the present invention preferably does not contain the zinc salt of CHP.

[0014] There are no particular limitations on the sources and manufacturing methods of CHP and CGP. CHP and CGP can be manufactured according to known methods. CHP and CGP can be derived from natural products, can be artificially synthesized, can also be manufactured by enzymatic methods or microbial fermentation methods, and can also be synthesized by dehydrating and cyclizing linear dipeptides. For example, by heating protein hydrolysates such as collagen hydrolysates, a peptide heat treatment product rich in cyclic dipeptides such as CHP and CGP can be obtained. CHP and CGP can be formulated into the composition using the protein hydrolysate containing them or its heat treatment product, or can also be formulated into the composition using a concentrate, dry powder of the protein hydrolysate or its heat treatment product, or a substance obtained by improving the refinement degree. For example, if the composition of the present invention contains a protein hydrolysate or its heat treatment product, in terms of CHP and CGP, it can also be a part of the protein hydrolysate or its heat treatment product. CHP and CGP can also use commercially available products.

[0015] Amyloid-β is a peptide sometimes referred to as amyloid-β protein, amyloid-β peptide, or amyloid-β. Amyloid-β is usually a peptide composed of about 40 amino acids, and examples include amyloid-β 1-38, amyloid-β 1-40, amyloid-β 1-42, etc. Amyloid-β is generated by the cleavage of amyloid precursor protein (APP) in two stages by β-secretase and γ-secretase.

[0016] Amyloid-β in a non-aggregated state is sometimes also referred to as amyloid-β monomer. Amyloid-β can form aggregates of two or more. For amyloid-β, for example, amyloid-β can form soluble aggregates (amyloid-β oligomers) aggregated from two or more (e.g., 2 to 50). Examples of amyloid-β oligomers include amyloid-β 1-38 oligomer, amyloid-β 1-40 oligomer, amyloid-β 1-42 oligomer, etc. The amyloid-β in the present invention includes amyloid-β in a non-aggregated state and amyloid-β forming aggregates such as oligomers.

[0017] The composition for inhibiting amyloid-β accumulation of the present invention preferably inhibits the amyloid-β whose accumulation is inhibited and is at least one selected from amyloid-β 1-38, amyloid-β 1-40, and amyloid-β 1-42. It is considered that the composition for inhibiting amyloid-β accumulation of the present invention inhibits the accumulation of amyloid-β by inhibiting the generation of amyloid-β, promoting the degradation of the generated amyloid-β, or both.

[0018] The composition for inhibiting amyloid-β accumulation of the present invention can be used to inhibit the accumulation of amyloid-β in the brain.

[0019] In one aspect, the composition of the present invention can be used for the prevention or improvement of conditions or diseases related to the accumulation of amyloid-β, and can preferably be used for the prevention or improvement of conditions or diseases related to the accumulation of amyloid-β in the brain. Examples of such conditions or diseases include conditions or diseases accompanied by or caused by the accumulation of amyloid-β. Examples of conditions or diseases accompanied by or caused by the accumulation of amyloid-β in the brain include Alzheimer's disease (including Alzheimer's type dementia), mild cognitive impairment, etc. The composition of the present invention can be used for the prevention or improvement of the above-mentioned conditions or diseases. In this specification, the prevention of a condition or disease includes: preventing the onset, delaying the onset, reducing the incidence rate, reducing the risk of onset, etc. The improvement of a condition or disease includes: restoring the subject from the condition or disease, alleviating the symptoms of the condition or disease, improving the symptoms of the condition or disease, delaying or preventing the development of the condition or disease, etc. Restoration includes partial restoration.

[0020] In Alzheimer's disease and mild cognitive impairment, a decrease in cognitive function is known. As symptoms of the decrease in cognitive function in Alzheimer's disease and mild cognitive impairment, for example, a decrease in memory, memory impairment (forgetfulness), aphasia (difficulty naming objects), apraxia, agnosia (getting lost in a familiar place, etc.), a decrease in orientation ability (the ability to correctly recognize one's own situation such as place, time, names of people, etc.), a decrease in language and non-verbal learning ability, a decrease in auditory and visual processing, a decrease in executive function, executive dysfunction (becoming unable to make a plan and execute it), a decrease in concentration, a decrease in attention, a decrease in judgment, a decrease in spatial recognition ability, a decrease in cognitive flexibility, a decrease in information processing speed, etc. can be cited. By inhibiting the accumulation of amyloid-β, it is expected to obtain a preventive effect on the decrease in cognitive function or an improvement effect on cognitive function, for example, a preventive or improvement effect on the above symptoms.

[0021] The composition for inhibiting amyloid-β accumulation of the present invention is applicable to either therapeutic use (medical use) or non-therapeutic use (non-medical use). The so-called non-therapeutic does not include the concept of medical acts, that is, surgery, treatment, or diagnosis of humans. As an example, the composition for inhibiting amyloid-β accumulation of the present invention can be provided in the form of an agent, but is not limited to this form. The agent can be provided directly as a composition or as a composition containing the agent. In one mode, the composition for inhibiting amyloid-β accumulation of the present invention can also be referred to as an amyloid-β accumulation inhibitor.

[0022] The composition for inhibiting amyloid-β accumulation of the present invention can be either oral or non-oral. The composition of the present invention is preferably an oral composition. The composition of the present invention can be made into forms such as food and drink products, pharmaceuticals, quasi-drugs, feeds, etc., and is preferably food and drink products or pharmaceuticals. The composition of the present invention can also be added to food and drink products, pharmaceuticals, quasi-drugs, feeds, etc. and used. The form of the composition of the present invention is not particularly limited and can be any of solid (for example, powdery, granular, tablet-like, etc.), liquid, paste-like, etc.

[0023] The composition of the present invention contains CHP and / or CGP, and various diluents, acidulants, antioxidants, stabilizers, preservatives, fragrances, emulsifiers, pigments, flavoring agents, pH regulators, nutritional fortifiers, etc. that are allowed as additives to food and drink products or pharmaceuticals, etc. can further be added.

[0024] For example, when the composition of the present invention is made into food or drink products, ingredients that can be used in food or drink products (e.g., food materials, food additives used as needed, etc.) can be formulated into CHP and / or CGP to make various food or drink products. The food or drink products are not particularly limited, and examples include general food or drink products, health foods, foods with functional claims, foods for specified health uses, health supplements, food or drink products for patients, etc. The above-mentioned health foods, foods with functional claims, foods for specified health uses, health supplements, etc. can be used in various preparation forms such as liquid preparations, fine granules, tablets, granules, powders, capsules, chewable tablets, dry syrups, liquid foods, etc.

[0025] When the composition of the present invention is made into pharmaceuticals or quasi-drugs, pharmacologically acceptable carriers, additives added as needed, etc. can be formulated into CHP and / or CGP to make pharmaceuticals or quasi-drugs in various dosage forms. Such carriers, additives, etc. only need to be pharmacologically acceptable substances that can be used in pharmaceuticals or quasi-drugs. For example, one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, etc. can be cited. As the administration (ingestion) methods of pharmaceuticals or quasi-drugs, oral or non-oral (transdermal, transmucosal, transintestinal, injection, etc.) administration methods can be cited. When the composition of the present invention is made into pharmaceuticals or quasi-drugs, it is preferably made into oral pharmaceuticals or oral quasi-drugs. As dosage forms for oral administration, for example, liquid preparations, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewable tablets, etc. can be cited. As dosage forms for non-oral administration, for example, injections, drip infusions, ointments, lotions, patches, suppositories, nasal preparations, pulmonary preparations (inhalants), etc. can be cited. The pharmaceuticals can also be pharmaceuticals for non-human animals.

[0026] When the composition of the present invention is made into feed, CHP and / or CGP can be formulated into the feed. The feed also includes feed additives. As the feed, for example, livestock feeds for cattle, pigs, chickens, sheep, horses, etc.; small animal feeds for rabbits, rats, mice, etc.; pet foods for dogs, cats, birds, etc. can be cited.

[0027] When the composition of the present invention is made into, for example, food or drink products, pharmaceuticals, quasi-drugs, feed, etc., its manufacturing method is not particularly limited, and CHP and / or CGP can be used and manufactured by general methods.

[0028] In one embodiment, the composition of the present invention is preferably a liquid composition, more preferably a beverage. The beverage can be, for example, a functional beverage. The form of the beverage is not particularly limited and can be a container-packed beverage. The container for the container-packed beverage is not particularly limited, and any form and material of container can be used. For example, metal containers such as aluminum cans and steel cans; resin containers such as PET bottles; paper containers such as paper boxes; glass containers such as glass bottles; wooden containers such as wooden barrels and other commonly used containers can be used. By filling the beverage into such a container and sealing it, a container-packed beverage can be obtained.

[0029] When the composition of the present invention contains CHP, its content is not particularly limited and can be set according to its form and the like. In one embodiment, the content of CHP in the composition of the present invention can be, for example, 1.0×10 -6 wt% or more, preferably 1.0×10 -5 wt% or more, more preferably 0.001 wt% or more. In addition, it is preferably 10 wt% or less, more preferably 1 wt% or less, further preferably 0.5 wt% or less, and particularly preferably 0.1 wt% or less. In one embodiment, the content of CHP is preferably 1.0×10 -6 ~10 wt% in the composition of the present invention. More preferably 1.0×10 -5 ~10 wt%, further preferably 1.0×10 -5 ~1 wt%, even more preferably 1.0×10 -5 ~0.5 wt%, and particularly preferably 0.001~0.1 wt%.

[0030] When the composition of the present invention contains CGP, its content is not particularly limited and can be set according to its form and the like. In one embodiment, the content of CGP in the composition of the present invention can be, for example, 1.0×10 -6 wt% or more, preferably 1.0×10 -5 wt% or more, more preferably 0.001 wt% or more, further preferably 0.01 wt% or more. In addition, it is preferably 50 wt% or less, more preferably 10 wt% or less, further preferably 5 wt% or less, and particularly preferably 2 wt% or less. In one embodiment, the content of CGP is preferably 1.0×10 -6 ~50 wt% in the composition of the present invention. More preferably 1.0×10 -5 ~10 wt%, further preferably 0.001~5 wt%, and even more preferably 0.01~2 wt%.

[0031] The total content of CHP and CGP contained in the composition of the present invention is not particularly limited and can be set according to its form and the like. In one embodiment, the total content of CHP and CGP in the composition of the present invention can be, for example, 1.0×10 -6 % by weight or more, preferably 1.0×10 -5 % by weight or more, more preferably 0.0001% by weight or more, further preferably 0.001% by weight or more, particularly preferably 0.01% by weight or more. In addition, it is preferably 50% by weight or less, more preferably 10% by weight or less, further preferably 5% by weight or less, and particularly preferably 3% by weight or less. In one embodiment, the total content of CHP and CGP can be, for example, 1.0×10 -6 to 50% by weight, preferably 1.0×10 -5 to 10% by weight, more preferably 0.0001 to 5% by weight, further preferably 0.001 to 5% by weight, and particularly preferably 0.01 to 3% by weight. CHP and CGP can be quantified, for example, by liquid chromatography mass spectrometry (LC / MS) in a known method.

[0032] The composition of the present invention is preferably taken orally (oral administration). The dosage (also referred to as the intake amount) of the composition of the present invention is not particularly limited. The dosage of the composition of the present invention only needs to be an amount that can obtain the cumulative inhibitory effect of amyloid-β, and can be appropriately set according to the administration form, administration method, body weight of the subject, etc.

[0033] In one embodiment, when the composition of the present invention is taken or administered to a human (adult) subject, its dosage, as the total dosage of CHP and CGP, is preferably 0.001 mg or more per day, more preferably 0.01 mg or more per day, further preferably 0.1 mg or more per day. In addition, it is preferably 5000 mg or less, more preferably 3000 mg or less, and further preferably 2000 mg or less. In one embodiment, when the composition of the present invention is taken or administered to a human (adult), as the total dosage of CHP and CGP, it is preferably 0.001 to 5000 mg per day, more preferably 0.01 to 3000 mg per day, and further preferably 0.1 to 2000 mg per day. It is preferably divided into one or more times a day, for example, once or multiple times a day (such as 2 to 3 times) for intake or administration. In one embodiment, it is preferred to orally intake or administer the above amount of CHP and / or CGP. In the case of a human (adult), per day, for every 60 kg of body weight, it is preferred to intake or administer the above amount of CHP and / or CGP. In one embodiment, the composition of the present invention can be an oral composition for a human to intake or be administered with the above amount of CHP and / or CGP per 60 kg of body weight per day.

[0034] The composition of the present invention is preferably continuously ingested or administered. Higher effects can be expected by continuously ingesting or administering CHP and / or CGP. In one mode, the composition of the present invention is preferably continuously ingested or administered for 1 week or more, more preferably continuously ingested or administered for 4 weeks or more, and further preferably continuously ingested or administered for 8 weeks or more. CHP and CGP can be ingested as food and beverages, etc. From the viewpoint of safety, it is considered that there are few problems even if they are ingested for a long time, for example.

[0035] The subject to which the composition of the present invention is ingested or administered (also referred to as the administration subject) is not particularly limited. It is preferably a human or a non-human mammal, and more preferably a human. In one mode, as the administration subject of the composition of the present invention, preferably a subject who needs or desires to inhibit the accumulation of amyloid-β, a subject who needs or desires the prevention or improvement of a condition or disease related to the accumulation of amyloid-β, etc. In one mode, as the administration subject in the present invention, the middle-aged and elderly can be cited. The middle-aged and elderly include the elderly. The middle-aged and elderly can be, for example, humans 40 years old or older. In one mode, among the middle-aged and elderly, the elderly are preferably the subjects. The elderly can be, for example, humans 60 years old or older or 65 years old or older. In one mode, the administration subject of the composition of the present invention can also be a healthy subject. For example, it can also be used for healthy subjects for the purpose of inhibiting the accumulation of amyloid-β in the brain, preventing the accumulation of amyloid-β in the brain, preventing Alzheimer's disease or mild cognitive impairment, etc.

[0036] The composition of the present invention can also be labeled with a function exerted by inhibiting the accumulation of amyloid-β. Such a label is also referred to as a functional label. The above label is not particularly limited. As such a label, for example, "improve cognitive function", "inhibit the decline of cognitive function", "maintain good cognitive function", "improve memory", "inhibit the decline of memory", "maintain good memory", "improve the accuracy of memory", "prevent memory impairment", "improve memory impairment", "maintain memory as part of cognitive function", "suitable for the function of those who care about memory decline", "improve the accuracy of memory or the correctness of judgment as part of cognitive function", "improve memory retention or integration", "maintain enhanced cognitive function", "improve executive function", "promote attention and concentration", "improve learning ability", "maintain and improve orientation ability", "delay the decline of cognitive function with age", "strengthen short-term and long-term memory", "promote verbal and visuospatial memory", and "prevent cognitive dysfunction during the aging process" and labels or functional labels that can be regarded as equivalent thereto can be cited. In one aspect of the present invention, the composition of the present invention is preferably a food or drink product attached with one or more of the above-mentioned labels. In addition, the above-mentioned label may also be a label indicating the use of the above-mentioned composition for obtaining the above-mentioned function. This label may be attached to the composition itself, or to the container or packaging of the composition.

[0037] The present invention also includes the following methods and applications. A method for inhibiting the accumulation of amyloid-β, characterized by administering Cyclo(His-Pro) and / or Cyclo(Gly-Pro). An application, characterized by using Cyclo(His-Pro) and / or Cyclo(Gly-Pro) for inhibiting the accumulation of amyloid-β. The above method may be a therapeutic method or a non-therapeutic method. The above application may be a therapeutic application or a non-therapeutic application.

[0038] In the above methods and applications, it is preferred to administer or give CHP and / or CGP to the subject once or more a day, for example, once to multiple times (e.g., 2 to 3 times) a day. The above application is preferably for humans or non-human mammals, and more preferably for humans. In one aspect, CHP and / or CGP can be used for preventing or improving conditions or diseases related to the accumulation of amyloid-β. The method for preventing or improving conditions or diseases related to the accumulation of amyloid-β by administering CHP and / or CGP is also included in the present invention.

[0039] In the above methods and applications, it is sufficient to use an amount (also referred to as an effective amount) of CHP and / or CGP that can obtain an inhibitory effect on the accumulation of amyloid-β. The preferred dosage, administration method, administration subject, etc. of CHP and / or CGP are the same as those of the composition of the present invention described above. CHP and / or CGP can be directly ingested or administered, or can be ingested or administered as a composition containing them. For example, the composition of the present invention can also be ingested or administered.

[0040] CHP and / or CGP can be used for manufacturing food or drink products, pharmaceuticals, quasi-drugs, feeds, etc. for inhibiting the accumulation of amyloid-β. In one aspect, the present invention also includes the application of CHP and / or CGP in the manufacture of a composition for inhibiting the accumulation of amyloid-β.

[0041] In this specification, a numerical range represented by a lower limit value and an upper limit value, that is, "lower limit value to upper limit value", includes these lower limit values and upper limit values. For example, the range represented by "1 to 2" means 1 or more and 2 or less, and includes 1 and 2. In this specification, the upper limit and the lower limit can be set as ranges based on any combination. Examples

[0042] Hereinafter, the present invention will be further described in detail by way of examples, but the scope of the present invention is not limited thereby.

[0043] <Examples 1-2> Using in vitro assays, the effects of Cyclo(Gly-Pro) (CGP) and Cyclo(His-Pro) (CHP) on the accumulation of amyloid-β were investigated. H4 glioma cells (H4-hAPP cells) overexpressing wild-type amyloid precursor protein with the Swedish double mutation K595N / M596L were used in the assays. To evaluate the effects of CGP and CHP on the accumulation of amyloid-β in H4-hAPP cells, untreated cells (Vehicle Control) were compared with CGP-treated cells and CHP-treated cells.

[0044] (Materials and Methods) The following substances were used as reagents and cells. Gibco Opti-MEM TM I Reduced Serum Medium (Opti-MEM) (cell culture medium), fetal bovine serum (FCS), penicillin / streptomycin (P / S), hygromycin B, and blasticidin S hydrochloride were products of Thermo Fischer Scientific. MSD (registered trademark), 96-well MULTISPOT (registered trademark), 6E10 Abeta Triplex Assay kit (amyloid-β detection kit) were products of Meso Scale Discovery. N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine tert-butyl ester (DAPT, γ-secretase inhibitor) was a product of Calbiochem. CGP and CHP were products of Bachem. H4-hAPP cells were obtained by stable transfection of H4 glioma cells based on the pAG3 vector containing human amyloid precursor protein 695 (APP695) with the Swedish double mutation K595N / M596L. The pAG3 vector is a modified pcDNA3 plasmid containing a hygromycin B resistance expression cassette and having the transgene under the control of the fused cytomegalovirus (CMV) and chicken β-actin promoters.

[0045] (Cell Culture and Treatment) H4-hAPP cells were maintained in Opti-MEM supplemented with 10% FCS, 1% P / S, 200 μg / mL of hygromycin B, and 2.5 μg / mL of blasticidin S hydrochloride. Cells were seeded at a density of 25,000 cells / well into 96-well plates and cultured overnight at 37 °C in a humidified gas chamber containing 5% CO 2 . The next day, H4-hAPP cells were treated with 7.5 mg / mL of CGP (Example 1) or 31 μg / mL of CHP (Example 2). As a positive control, H4-hAPP cells were treated with 400 nM of DAPT. After 24 hours of treatment, the cell culture supernatants for amyloid-β analysis were collected.

[0046] (Amyloid-β ELISA assay) The collected cell culture supernatants were diluted 1:10, and human amyloid-β1-38 (Aβ1-38), human amyloid-β1-40 (Aβ1-40), and human amyloid-β1-42 (Aβ1-42) were analyzed using the 6E10 Abeta Triplex Assay kit. The multiplex assay was performed according to the manufacturer's instructions, and the plates were read using a Sector Imager 2400 (manufactured by Meso Scale Discovery). The concentration of amyloid-β was calculated by referring to the standard curve. The sensitivity of the multiplex kit was <5 pg / mL.

[0047] (Statistical analysis) Statistical analysis was performed using Prism version 5.0 (manufactured by GraphPad). All results were expressed as mean ± standard deviation. Statistical analysis was performed by analysis of variance (ANOVA), followed by post hoc Tukey's multiple comparison test. Data were considered significant when the p-value was p < 0.05.

[0048] Figure 1A A graph showing the concentration of amyloid-β1-38 in the culture supernatants of H4-hAPP cells treated with CGP or CHP. Figure 1B A graph showing the concentration of amyloid-β1-40 in the culture supernatants of H4-hAPP cells treated with CGP or CHP. Figure 1C A graph showing the concentration of amyloid-β1-42 in the culture supernatants of H4-hAPP cells treated with CGP or CHP. Values are expressed as mean ± standard deviation (n = 6). *** indicates a significant difference with p < 0.001 compared to the untreated vehicle control. Control is the vehicle control, and DAPT is the positive control.

[0049] In Example 1 and Example 2, both CGP and CHP significantly inhibited the accumulation of amyloid-β1-38, amyloid-β1-40, and amyloid-β1-42 in H4-hAPP cells relative to the excipient control.

Claims

1. A composition for inhibiting amyloid-β accumulation, characterized in that, it contains Cyclo(His-Pro) and / or Cyclo(Gly-Pro) as active ingredients.

2. The composition according to claim 1, characterized in that, the amyloid-β is at least one selected from amyloid-β1-38, amyloid-β1-40, and amyloid-β1-42.

3. The composition according to claim 1 or 2, characterized in that, it is an oral composition.

4. The composition according to claim 1 or 2, characterized in that, it is a food or drink or a pharmaceutical product.

5. The composition according to claim 1 or 2, characterized in that, it is labeled with one or more functions selected from "improve cognitive function", "inhibit decline of cognitive function", "maintain good cognitive function", "improve memory", "inhibit decline of memory", "maintain good memory", "improve memory accuracy", "prevent memory disorders", "improve memory disorders", "maintain memory as part of cognitive function", "suitable for those who care about memory decline", "improve accuracy of memory or correctness of judgment as part of cognitive function", "improve memory retention or integration", "maintain enhanced cognitive function", "improve executive function", "promote attention and concentration", "improve learning ability", "maintain and improve orientation ability", "delay age-related decline of cognitive function", "strengthen short-term and long-term memory", "promote verbal and visual-spatial memory", and "prevent cognitive dysfunction during the aging process".

6. An application, characterized in that, Cyclo(His-Pro) and / or Cyclo(Gly-Pro) is used for the manufacture of a composition for inhibiting amyloid-β accumulation.

7. An application, characterized in that, Cyclo(His-Pro) and / or Cyclo(Gly-Pro) is used for inhibiting the accumulation of amyloid-β.

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