A composition comprising an extract derived from fish cartilage and an extract derived from Boswellia plants

Abstract

Provided is a composition derived from natural products that enables easy intake of nutritional components capable of exhibiting excellent functionality. 【Solution means】A composition containing a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from Boswellia plants. The collagen-containing extract contains type II collagen and type XI collagen, and the mass ratio of their total content is 10:1 to 1:10, and it is preferable that their total content is 30% by mass or more. The boswellic acid-containing extract preferably contains 20% by mass or more of boswellic acid.

Description

Technical Field

[0001] The present invention relates to a composition derived from natural products that enables easy intake of nutritional components capable of exhibiting excellent functionality.

Background Art

[0002] Boswellia serrata is a deciduous tall tree of the Burseraceae family that grows wild in the dry highlands of India. In Ayurveda, an Indian traditional medicine, Boswellia serrata is said to be effective against arthritis, rheumatism, hyperlipidemia, bronchial asthma, etc., and the resin (frankincense) secreted from the tree contains boswellic acid compounds (such as 3-O-acetyl-11-keto-β-boswellic acid). There are various reports on the activity of boswellic acid-containing extracts derived from Boswellia serrata (see the background art section of Patent Document 1).

[0003] On the one hand, collagen is a protein that is widely distributed in animals in general, such as mammals, birds, and fish, and functions as a major component of the extracellular matrix. In humans, it is said to account for approximately 25 to 30% of the total protein mass. As for its molecular structure, three peptide chains with a molecular weight of about 100,000 gather and form a triple helix structure through hydrogen bonds. Also, the units of the triple helix structure are cross-linked in the telopeptide region that occupies the ends of collagen molecules, forming higher-order structures such as fibrous structures and network structures. The types of collagen that have been confirmed in humans are currently about 28, and they are also classified by function. For example, type I collagen, which is the main protein in bone, dermis, tendon, etc., forms a triple helix structure with two α1 chains and one α2 chain, and its fibrous structure has excellent tensile strength and functions as a structural protein that maintains the shape of the body and organs. Also, type II collagen forms a triple helix structure with three α1 chains, is localized in cartilage, etc., and has the same function as a structural protein as type I. On the other hand, type III and type V collagens coexist with type I in skin, etc., and type XI collagen coexists with type II in cartilage, etc., and it is considered that they are subsidiarily involved in the formation of collagen fibers adapted to each tissue (see Non-Patent Documents 1 and 2). Also, as the main molecular species composition of type XI collagen, it is considered to form a triple helix structure with three types: α1 chain, α2 chain, and α3 chain (Non-Patent Document 3).

[0004] Regarding the use of collagen materials, type I collagen, which occupies the most of the collagen in animal tissues, is widely used as a raw material for cosmetics, health foods, pharmaceuticals, etc. Also, in recent years, the use of cartilage-derived collagen materials has been increasing. For example, Patent Document 2 describes that a type II / type XI collagen-containing composition derived from fish cartilage has the functionality of promoting the hyaluronic acid-producing ability of chondrocytes and is useful for cartilage protection and knee joint protection.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2012-502901 [Patent Document 2] Japanese Patent No. 7357189 [Non-Patent Document]

[0006] [Non-Patent Document 1] Shunji Hattori, "Properties and Applications of Animal-Derived Fiber Molecules Collagen," Fibers and Industry, vol. 65, No. 12 (2009) pp453-461. [Non-Patent Document 2] Shunji Hattori, "Collagen - Molecular Assembly and Its Applications -," Polymer, Vol. 47, June issue (1998) pp394-397. [Non-Patent Document 3] Hidetoshi Yoshioka, "XI-Type Collagen α1 Chain Gene: Primary Structure of α1 Chain, Its Gene Expression and Regulatory Mechanism," Connective Tissue, 29 (1997) pp39-47. [Summary of the Invention] [Problems to be Solved by the Invention]

[0007] However, there is no report on the complementary relationship between boswellic acid and collagen in terms of their biological functionality.

[0008] In view of the above circumstances, an object of the present invention is to provide a natural product-derived composition that enables easy intake of nutritional components capable of exhibiting excellent functionality. [Means for Solving the Problems]

[0009] The present inventors have conducted intensive studies to achieve the above object and have completed the present invention.

[0010] That is, the present invention provides a composition containing a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from Boswellia plants.

[0011] In the above composition, the collagen-containing extract contains type II collagen and type XI collagen, and the mass ratio of the content of the type II collagen to the type XI collagen is 10:1 to 1:10, and the total content of the type II collagen and the type XI collagen is preferably 30% by mass or more.

[0012] In the above composition, the boswellic acid-containing extract preferably contains 20% by mass or more of boswellic acid.

[0013] In the above composition, the boswellic acid-containing extract preferably contains 10% by mass or more of 3-O-acetyl-11-keto-β-boswellic acid.

[0014] In the above composition, the collagen-containing extract contains type II collagen and type XI collagen, and the mass ratio of the content of the type II collagen to the type XI collagen is 10:1 to 1:10, and the total content of the type II collagen and the type XI collagen is 30% by mass or more, the boswellic acid-containing extract contains 10% by mass or more of 3-O-acetyl-11-keto-β-boswellic acid, and the mass ratio of the content of the 3-O-acetyl-11-keto-β-boswellic acid to the total content of the type II collagen and the type XI collagen is preferably 0.001 to 10.

[0015] In the above composition, the collagen-containing extract contains type II collagen and type XI collagen, the mass ratio of the content of the type II collagen to the type XI collagen is 10:1 to 1:10, the total content of the type II collagen and the type XI collagen is 30% by mass or more, the boswellic acid-containing extract contains 3-O-acetyl-11-keto-β-boswellic acid in an amount of 10% by mass or more, the composition is in the form of tablets, and the total content of the type II collagen and the type XI collagen contained per 300 mg of the tablets is 0.1 mg or more and 100 mg or less, and the content of 3-O-acetyl-11-keto-β-boswellic acid contained per 300 mg of the tablets is preferably 0.1 mg or more and 100 mg or less.

[0016] In the above composition, it is preferable that the collagen-containing extract derived from fish cartilage is made from salmon nasal cartilage as a raw material.

[0017] In the above composition, it is preferably in a form for oral intake.

[0018] In the above composition, it is preferably in the form of tablets.

[0019] In the above composition, it is preferably used for anti-inflammatory purposes.

Advantages of the Invention

[0020] According to the present invention, a natural product-derived composition can be provided in which nutritional components capable of exhibiting excellent functionality can be easily ingested by using a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from Boswellia plants.

Brief Description of the Drawings

[0021]

Figure 1

Modes for Carrying Out the Invention

[0022] When numerical ranges are indicated in this specification, the upper and lower limit values can be appropriately combined, and the numerical ranges obtained thereby are also considered to be disclosed. Also, when indicating the content of components, unless otherwise specified, it is the content in terms of dry matter.

[0023] The collagen-containing extract used in the present invention is not particularly limited as long as it is an edible material containing collagen derived from fish cartilage. For example, it may be a collagen-containing extract prepared from salmon nasal cartilage as a raw material. Generally, as methods for solubilizing and extracting collagen from animal tissues, there are methods such as heat denaturation treatment, solubilization treatment with acid or alkali, and solubilization by enzyme treatment. However, in heat denaturation treatment, the triple helix unfolds and randomly denatures. Also, in acid solubilization or alkali solubilization, only a very small amount elutes under normal non-denaturing conditions, resulting in poor productivity. On the other hand, it is considered that by using digestive enzymes such as pepsin, only the cross-linked structure part can be partially decomposed, and the triple helix structure units can be efficiently extracted.

[0024] The above collagen-containing extract preferably has a collagen content of 30% by mass or more, more preferably 40% by mass or more, still more preferably 50% by mass or more, and even more preferably 60% by mass or more. On the other hand, there is no particular limitation on the upper limit value of the collagen content, but typically, for example, it can be 100% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less, etc. Further, among collagens, it is preferable to contain type II collagen and type XI collagen, and the mass ratio of the content of type II collagen to type XI collagen is 10:1 to 1:10, and the total content thereof is preferably 30% by mass or more, and more preferably 40% by mass or more. The mass ratio of the content of type II collagen to type XI collagen can be, for example, 9:1 to 1:9, 8:1 to 1:8, 7:1 to 1:7, 6:1 to 1:6, 5:1 to 1:5, etc. In addition, as collagen-containing extracts derived from fish cartilage, "SCP Complex-LS", "SNC Complex-LS" (both are product names, manufactured by RINACE Co., Ltd.) and the like are commercially available, so such commercially available products may be used. Further, Japanese Patent No. 7138873 discloses a preparation method in which fish cartilage is used as an extraction raw material and type II collagen and type XI collagen can be efficiently recovered in a non-denatured state. That is, for example, the extraction raw material is subjected to enzymatic treatment with an acidic protease in a dilute acidic solvent to solubilize the collagen, the liquid part containing the collagen is recovered, the pH of the liquid part is neutralized, and then salting out is performed with sodium sulfate or the like having a salt concentration of 4M or more, and the salt is removed from the salting-out precipitate by ultrafiltration, dialysis, etc., to obtain the collagen containing type II collagen and type XI collagen in a non-denatured state. Therefore, it may be prepared and used by such a known method. Further, the collagen in the above collagen-containing extract may be an atelo type that has been partially decomposed by an acidic protease such as pepsin or lysopeptidase.

[0025] The boswellic acid-containing extract used in the present invention is not particularly limited as long as it is an edible material containing boswellic acid derived from Boswellia plants. For example, it may be a boswellic acid-containing extract prepared from resin (frankincense) secreted from the tree of Boswellia serrata as a raw material.

[0026] The above-mentioned boswellic acid-containing extract preferably has a boswellic acid content of 20% by mass or more, more preferably 30% by mass or more, still more preferably 40% by mass or more, and even more preferably 50% by mass or more of the material. On the other hand, there is no particular limitation on the upper limit of the boswellic acid content, but it can typically be, for example, 100% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less, etc. Further, it preferably contains 3-O-acetyl-11-keto-β-boswellic acid among boswellic acids, and its content is preferably 10% by mass or more, and more preferably 20% by mass or more. Incidentally, as the boswellic acid-containing extract derived from Boswellia plants, "Boswellin Super" (trade name, manufactured by Sabinsa Japan Corporation), "Boswellia serrata extract 20%" (trade name, manufactured by Bioactives Japan), "Boswellia extract AKB50%" (trade name, manufactured by Bioactives Japan), etc. are commercially available, so such commercial products may be used. Further, Japanese Patent Publication No. 2002-543125 discloses a method for efficiently recovering boswellic acid using Boswellia serrata gum as an extraction raw material. Therefore, it may be prepared and used by such a known method. That is, for example, after extraction with an organic solvent such as isopropyl alcohol, the organic solvent is removed by heating and concentration, an alkaline aqueous solution such as 5% KOH is added thereto to dissolve the total organic acids in the aqueous phase. After washing the remaining organic solvent and impurities with ethyl acetate, concentrated hydrochloric acid is slowly added dropwise to the aqueous phase to recover the total organic acids as a precipitate, and water is added to the precipitate to recover boswellic acid as an aqueous solution. Further, the above-mentioned boswellic acid-containing extract can also be prepared by a subcritical extraction or supercritical extraction method using liquid carbon dioxide with, for example, Boswellia serrata gum as an extraction raw material.

[0027] The present invention provides a composition comprising the collagen-containing extract as described above and the boswellic acid-containing extract as described above in combination and containing them as edible materials.

[0028] As shown in the examples described below, boswellic acid and collagen complement each other's functionality. More specifically, these components actively contribute to each other in the mechanism of action of alleviating sensitivity when immune cells such as macrophages respond to cell stimulants. This mechanism of action of immune tolerance can bring about an anti-inflammatory effect in the body. Therefore, it can be a functional composition for improving and preventing symptoms such as discomfort and pain in the knee joint during walking, discomfort and pain in the knee joint during ascending and descending stairs, and discomfort and pain in the knee joint during sitting and standing. The subject may be a healthy person, and in particular, it may be a composition for healthy persons aged 50 or above. Also, it may be a composition for animals such as pet animals.

[0029] In the composition provided by the present invention, there is no particular limitation on the inclusion of other components in addition to the above collagen-containing extract and / or the above boswellic acid-containing extract, as long as the object is not impaired. Examples of other components include proteoglycan, vitamin C, imidazole peptide, collagen peptide, salmon ovary outer skin peptide, β-hydroxy-β-methylbutyric acid (HMB), and the like.

[0030] In the composition provided by the present invention, boswellic acid and collagen may be used together so as to be administered to a living body, and there is no particular limitation on the form of use. For example, in the case of an oral intake form, it may be in the form of tablets (tablets, tablets, chewable tablets, orally disintegrating tablets), liquids (liquids), syrups (syrups), powders (granules, fine granules), capsules (capsules), soft capsules (soft capsules), solids, semi-liquids, creams, pastes, etc.

[0031] Regarding the intake amount, it may be appropriately set according to the health condition or disease state of the human or animal to which it is applied, the purpose, etc., and there is no particular limitation. For example, as the intake amount per day, for example, in terms of the amount of 3-O-acetyl-11-keto-β-boswellic acid conversion, it may be 10 mg or more and 200 mg or less, it may be 20 mg or more and 200 mg or less, it may be 30 mg or more and 100 mg or less, it may be 40 mg or more and 100 mg or less. In terms of collagen amount conversion, it may be 5 mg or more and 100 mg or less, it may be 5 mg or more and 40 mg or less, it may be 5 mg or more and 20 mg or less, it may be 10 mg or more and 20 mg or less, it may be 10 mg or more and 15 mg or less. As the intake period, it is preferably used so as to be ingested over 12 weeks or more. As the intake period, for example, it may be 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, etc., and it may also be used so as to be ingested continuously or intermittently over these periods.

[0032] From the perspective of the above intake amount, in the composition provided by the present invention, the mass ratio of the content of the above-mentioned 3-O-acetyl-11-keto-β-boswellic acid to the total content of the above-mentioned type II collagen and type XI collagen is preferably 0.001 to 10. The mass ratio may be 0.01 to 10, may be 0.1 to 10, or may be 0.1 to 1. Further, when the form of the composition provided by the present invention is a tablet, from the perspective of the above intake amount, the total content of the above-mentioned type II collagen and type XI collagen contained per 300 mg of the tablet is 0.1 mg or more and 100 mg or less, and the content of the above-mentioned 3-O-acetyl-11-keto-β-boswellic acid contained per 300 mg of the tablet is preferably 0.1 mg or more and 100 mg or less. The total content of the above-mentioned type II collagen and type XI collagen contained per 300 mg of the tablet may be 1 mg or more and 75 mg or less, may be 2 mg or more and 50 mg or less, may be 3 mg or more and 40 mg or less, or may be 3 mg or more and 30 mg or less. The content of the above-mentioned 3-O-acetyl-11-keto-β-boswellic acid contained per 300 mg of the tablet may be 1 mg or more and 75 mg or less, may be 2 mg or more and 50 mg or less, may be 3 mg or more and 40 mg or less, or may be 3 mg or more and 30 mg or less.

[0033] The composition provided by the present invention can be used, for example, for foods such as health foods and foods with functional claims, supplements, pharmaceuticals, medical materials, etc., and can be particularly preferably used as a raw material. Further, as described above, it can also be used for animals such as pet animals as well as humans.

[0034] In addition, the analysis of the amount of collagen described in this specification can be measured by an analysis method well-known in the food analysis field. For example, it can be measured by a method of quantifying hydroxyproline, which is an acid hydrolyzate, and converting it from the characteristics of the amino acid composition.

[0035] In addition, the amount of boswellic acid compounds such as 3-O-acetyl-11-keto-β-boswellic acid can be measured by HPLC analysis using a standard product of boswellic acid compounds prepared at a specific concentration as an index.

Example

[0036] The present invention will be described in more detail with reference to the following examples. However, these examples do not limit the scope of the present invention.

[0037] [Test Example 1] [1-1. Materials] · ELISA kit: Mouse TNF-α ELISA Kit (Registered trademark: Revis) · Cells: Macrophage-like cell line (J774.1, derived from mouse) · Cell stimulant: LPS (Lipopolysaccharide) · Liquid medium: D-MEM (containing 1% or 10% FBS) · Collagen-containing extract derived from fish cartilage: (1) "SCP Complex-LS" (Trade name, manufactured by RINICE Co., Ltd., non-denatured proteoglycan, mixture of non-denatured type II & XI collagen) (2) "SNC Complex-LS" (Trade name, manufactured by RINICE Co., Ltd., mixture of non-denatured type II & XI collagen) (Hereinafter, the collagen contained in the above collagen-containing extract may be simply referred to as "RSC".) · Boswellic acid-containing extract derived from Boswellia plant: (1) "Boswellin Super" (Trade name, manufactured by Sabinsa Japan Corporation) (2) "Boswellia serrata extract 20%" (Trade name, manufactured by Bioactives Japan) (Hereinafter, 3-O-acetyl-11-keto-β-boswellic acid contained in the above boswellic acid-containing extract may be simply referred to as "boswellic acid".)

[0038] [1-2. Test groups] (#1)LPS: Not added Boswellic acid: Not added RSC: Not added (#2)LPS: 1 μg / mL Boswellic acid: Not added RSC: Not added (#3)LPS: 1 μg / mL Boswellic acid: 0.001 mg / mL RSC: Not added (#4)LPS: 1 μg / mL Boswellic acid: Not added RSC: 0.01 mg / mL (#5)LPS: 1 μg / mL Boswellic acid: Not added RSC: 0.1 mg / mL (#6)LPS: 1 μg / mL Boswellic acid: Not added RSC: 1 mg / mL (#7)LPS: 1 μg / mL Boswellic acid: 0.001 mg / mL RSC: 0.01 mg / mL (#8)LPS: 1 μg / mL Boswellic acid: 0.001 mg / mL RSC: 0.1 mg / mL (#9)LPS: 1 μg / mL Boswellic acid: 0.001 mg / mL RSC: 1 mg / mL

[0039] [1-3. Cell culture] - Suspend the cells in 10% FBS / D-MEM medium and adjust the cell concentration to approximately 2×10 4 cells / mL - Inoculate the cell suspension (approximately 100 μL) into each well of a 96-well plate to achieve 2.0×10 3 cells per well - Incubate overnight (37 °C, 5% CO2) - Remove the 10% FBS / D-MEM medium and replace it with 1% FBS / D-MEM medium containing the test substance at a predetermined concentration - Incubate for 2 hours (37 °C, 5% CO2) - Add LPS to each well (final concentration 1 μg / mL) - Incubate for 24 hours (37 °C, 5% CO2) - Collect the culture supernatant

[0040] [1-4. ELISA] - Pipette 25 μL of buffer and 25 μL of the collected culture supernatant into each well of the ELISA plate - After stirring, leave it to stand at room temperature (20 - 25 °C) for 2 hours - Discard the liquid from each well and wash 4 times with the washing solution - Dispense 50 μL of biotin-conjugated anti-TNF-α antibody into each well - After stirring, leave it to stand at room temperature (20 - 25 °C) for 1 hour - Discard the liquid from each well and wash 4 times with the washing solution - Dispense 50 μL of peroxidase-avidin conjugate into each well - After stirring, leave it to stand at room temperature (20 - 25 °C) for 30 minutes - Discard the liquid from each well and wash 4 times with the washing solution - Dispense 50 μL of the chromogenic solution into each well - After stirring, leave it to stand at room temperature (20 - 25 °C) for 20 minutes - Dispense 50 μL of the reaction stop solution into each well and stir - Measure the absorbance (450 nm) using a microplate reader

[0041] 〔1-5. Evaluation〕 The results are shown in Table 1 and Figure 1

[0042]

Table 1

[0043] As a result, compared with the results of test group #3 in which the boswellic acid-containing extract was acted alone in addition to the stimulation by LPS, and the results of test groups #4 - #6 in which the collagen-containing extract was acted alone in addition to the stimulation by LPS, in test groups #7 - #9 in which the boswellic acid-containing extract and the collagen-containing extract were acted together in addition to the stimulation by LPS, the TNF-α production amount was significantly decreased with a significant difference of p < 0.005 (indicated by "**" in Table 1 or Figure 1.) or p < 0.001 (indicated by "***" in Table 1 or Figure 1.)

[0044] Therefore, it was revealed that boswellic acid and collagen positively contribute to each other in the mechanism of action that alleviates the sensitivity when immune cells such as macrophages respond to cell stimulants.

[0045] (Formulation Example 1) Tablets of 300 mg per tablet were produced by a conventional method with the formulation shown in Table 2.

[0046]

Table 2

[0047] As a result, there were no problems with tableting obstacles such as capping and sticking, and it was possible to tabletize.

[0048] (Formulation Example 2) Tablets of 300 mg per tablet were produced by a conventional method with the formulation shown in Table 3.

[0049]

Table 3

[0050] As a result, there were no problems with tableting obstacles such as capping and sticking, and it was possible to tabletize.

[0051] <Questionnaire Implementation> When 18 healthy people in their 50s to 80s with symptoms of knee pain were given 3 tablets of the formulation example 1 per day for 10 weeks, questionnaire results were obtained indicating that the pain symptoms generally disappeared or improved.

Claims

1. A composition for inhibiting TNF-α production, comprising a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from Boswellia plants, wherein the collagen-containing extract contains type II collagen and type XI collagen, the mass ratio of the content of type II collagen to the content of type XI collagen is 10:1 to 1:10, and the total content of type II collagen and type XI collagen is 30% by mass or more, the boswellic acid-containing extract contains 3-O-acetyl-11-keto-β-boswellic acid in an amount of 10% by mass or more, and the mass ratio of the content of 3-O-acetyl-11-keto-β-boswellic acid to the total content of type II collagen and type XI collagen is 0.001 to 10.

2. A composition for inhibiting TNF-α production, comprising a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from Boswellia plants, wherein the collagen-containing extract contains type II collagen and type XI collagen, the mass ratio of the content of type II collagen to the content of type XI collagen is 10:1 to 1:10, and the total content of type II collagen and type XI collagen is 30% by mass or more, the boswellic acid-containing extract contains 3-O-acetyl-11-keto-β-boswellic acid in an amount of 10% by mass or more, the composition is in the form of tablets, the total content of type II collagen and type XI collagen contained per 300 mg of the tablets is 0.1 mg or more and 100 mg or less, and the content of 3-O-acetyl-11-keto-β-boswellic acid contained per 300 mg of the tablets is 0.1 mg or more and 100 mg or less. The composition according to claim 1.

3. The composition according to claim 1 or 2, wherein the collagen-containing extract derived from fish cartilage is prepared from salmon nasal cartilage as a raw material.

4. The composition according to claim 1 or 2, which is in a form suitable for oral administration.

5. The composition according to claim 1 or 2, which is in the form of tablets.

6. The composition according to claim 1 or 2, which is used for anti-inflammatory purposes.

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