A composition containing an extract derived from fish cartilage and an extract derived from the Boswellia plant.

Abstract

To provide a natural product-derived composition that allows for the easy intake of nutrients that can exhibit excellent functionality. [Solution] The composition comprises a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from the Boswellia plant. The collagen-containing extract preferably contains type II collagen and type XI collagen, with a total mass ratio of their contents being 10:1 to 1:10, and a total content of 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] Collagen, on the other hand, is a protein widely distributed in animals such as mammals, birds, and fish, and functions as a major component of their extracellular matrix. In humans, it is said to account for approximately 25-30% of the total protein mass, and its molecular structure consists of three peptide chains with a molecular weight of about 100,000, which come together to form a triple helix structure through hydrogen bonds. Furthermore, these triple helix units cross-link at the telopeptide regions that occupy the ends of the collagen molecule, forming higher-order structures such as fibrous structures and network structures. There are currently about 28 types of collagen identified in humans, and they are classified according to their function. For example, type I collagen, the main protein in bone, dermis, and tendons, has a triple helix structure formed by two α1 chains and one α2 chain, and its fibrous structure has excellent tensile strength, functioning as a structural protein that maintains the shape of the body and organs. Type II collagen, on the other hand, has a triple helix structure formed by three α1 chains, and is localized in cartilage, etc., and functions as a structural protein similar to type I. On the other hand, type III and type V collagen coexist with type I collagen in the skin, etc., and type XI collagen coexists with type II collagen in cartilage, etc., and is thought to be involved in the formation of collagen fibers suited to each tissue (see Non-Patent Documents 1 and 2). Furthermore, type XI collagen is thought to be composed mainly of three molecular species: α1 chains, α2 chains, and α3 chains, forming a triple helix structure (Non-Patent Document 3).

[0004] Regarding the use of collagen materials, type I collagen, which accounts for the largest portion of collagen in animal tissues, is widely used as a raw material for cosmetics, health foods, and pharmaceuticals. In recent years, the use of cartilage-derived collagen materials has also been increasing. For example, Patent Document 2 describes that a composition containing type II / type XI collagen derived from fish cartilage has the functionality to promote the hyaluronic acid production ability of chondrocytes and is useful for protecting cartilage and knee joints. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Special Publication No. 2012-502901 [Patent Document 2] Patent No. 7357189 [Non-patent literature]

[0006] [Non-Patent Document 1] Shunji Hattori, "Properties and Applications of Animal-Derived Fiber Molecules: Collagen," Textiles and Industry, vol. 65, No. 12 (2009), pp. 453-461. [Non-Patent Document 2] Shunji Hattori, "Collagen - Molecular Assembly and its Applications," Polymer Science, Vol. 47, June issue (1998), pp. 394-397. [Non-Patent Document 3] Hidekatsu Yoshioka, "Type XI Collagen α1 Chain Gene: Primary Structure of the α1 Chain, Its Gene Expression and Regulatory Mechanism," Connective Tissue, 29 (1997), pp. 39-47. [Overview of the project] [Problems that the invention aims to solve]

[0007] However, there are no reports indicating that boswellic acid and collagen complement each other in terms of their respective biological functions.

[0008] In view of the above circumstances, the object of the present invention is to provide a composition derived from natural products that allows for the easy intake of nutritional components that can exhibit excellent functionality. [Means for solving the problem]

[0009] The inventors of this invention have diligently conducted research to achieve the above objectives and have now completed this invention.

[0010] In other words, the present invention provides a composition comprising a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from the Boswellia plant.

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

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

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

[0014] In the above composition, it is preferable that the collagen-containing extract contains type II collagen and type XI collagen, with a mass ratio of type II collagen to type XI collagen content of 10:1 to 1:10, and the total content of type II collagen and type XI collagen of 30% by mass or more; and that 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 3-O-acetyl-11-keto-β-boswellic acid to the total content of type II collagen and type XI collagen of 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 10% by mass or more of 3-O-acetyl-11-keto-β-boswellic acid, 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 that enables easy intake of nutritional components that can exhibit excellent functionality 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]In Test Example 1, using the production amount of TNF-α secreted by macrophage-like cell line (J774.1, derived from mouse) stimulated by LPS as an index, it is a chart showing the results of examining the effects of a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from Boswellia plants on the sensitivity of immune cells to cell stimulants.

Mode for Carrying Out the Invention

[0022] When indicating a numerical range in this specification, the upper limit value and the lower limit value can be appropriately combined, and the numerical range obtained thereby is also considered to be disclosed. Also, when indicating the content of a component, 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 a method 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 strands unwind and denature randomly. 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 a digestive enzyme 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 collagen-containing extract described above preferably has a collagen content of 30% by mass or more, more preferably 40% by mass or more, even more preferably 50% by mass or more, and even more preferably 60% by mass or more. On the other hand, there is no particular upper limit on the collagen content, but it can typically be, for example, 100% by mass or less, 90% by mass or less, 80% by mass or less, or 70% by mass or less. Furthermore, it is preferable that the collagen contains both type II collagen and type XI collagen, with a mass ratio of type II collagen to type XI collagen of 10:1 to 1:10, and a total content of 30% by mass or more, and more preferably 40% by mass or more. The mass ratio 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. Furthermore, commercially available collagen-containing extracts derived from fish cartilage, such as "SCP Complex-LS" and "SNC Complex-LS" (both trade names, manufactured by Linace Co., Ltd.), may be used. In addition, Japanese Patent Publication No. 7138873 discloses a preparation method that allows for the efficient recovery of type II and type XI collagen in an undenatured state using fish cartilage as the raw material. Specifically, for example, the raw material is enzymatically treated with an acidic protease in a dilute acidic solvent to solubilize the collagen, the liquid portion containing the collagen is recovered, the pH of the liquid portion is neutralized, and then salted out with sodium sulfate or the like at a salt concentration of 4M or higher. The salt is then removed from the salted-out precipitate by ultrafiltration, dialysis, etc., to obtain collagen containing type II and type XI collagen in an undenatured state. Therefore, it may be prepared and used using such a known method. Moreover, the collagen in the above collagen-containing extract may be in the ateloform form, which has undergone partial decomposition by acidic proteases such as pepsin or lysopepsin.

[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 the Boswellia plant. For example, it may be a boswellic acid-containing extract prepared using the resin (frankincense) secreted from the Boswellia serrata tree as a raw material.

[0026] The above-mentioned boswellic acid-containing extract preferably contains 20% by mass or more of boswellic acid, more preferably 30% by mass or more, even more preferably 40% by mass or more, and even more preferably 50% by mass or more. On the other hand, there is no particular upper limit to 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, or 70% by mass or less. Furthermore, it is preferable to contain 3-O-acetyl-11-keto-β-boswellic acid among the boswellic acid, with a content of 10% by mass or more, and more preferably 20% by mass or more. As for boswellic acid-containing extracts derived from Boswellia plants, commercially available products such as "Boswellin Super" (product name, manufactured by Sabinsa Japan Corporation), "Boswellia serrata extract 20%" (product name, manufactured by Bioactives Japan), and "Boswellia extract AKB 50%" (product name, manufactured by Bioactives Japan) can be used. Furthermore, Japanese Patent Publication No. 2002-543125 discloses a method for efficiently recovering boswellic acid using Boswellia serrata gum as the extraction 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, and an alkaline aqueous solution such as 5% KOH is added 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. In addition, the above boswellic acid-containing extract can also be prepared by subcritical or supercritical extraction using liquid carbon dioxide, for example, using Boswellia serrata gum as the extraction material.

[0027] The present invention provides a composition comprising a collagen-containing extract as described above, combined with a boswellic acid-containing extract as described above, and containing these as edible materials.

[0028] As shown in the examples described later, boswellic acid and collagen complement each other's functionality. More specifically, these components mutually contribute positively to a mechanism of action that mitigates sensitivity when immune cells such as macrophages respond to cellular stimulants. This immune tolerance mechanism can produce anti-inflammatory effects in the body. Therefore, it can be used as a functional composition for improving or preventing symptoms such as discomfort and pain in the knee joint when walking, when climbing stairs, and when sitting down or standing up. The target population may be healthy individuals, and in particular, the composition may be formulated for healthy individuals aged 50 and over. It may also be formulated as a composition for animals such as pets.

[0029] In the compositions provided by the present invention, there are no particular restrictions on the inclusion of other components in addition to the collagen-containing extract and / or boswellic acid-containing extract, as long as the purpose is not impaired. Examples of other components include proteoglycans, vitamin C, imidazole peptides, collagen peptides, salmon ovary membrane peptides, and β-hydroxy-β-methylbutyrate (HMB).

[0030] In the composition provided by the present invention, boswellic acid and collagen can be administered to a living organism together, and there are no particular restrictions on the form of use. For example, in the form for oral administration, it may be in the form of tablets (tablets, chewable tablets, orally disintegrating agents), liquid (liquid), syrup (syrup), powder (granules, fine granules), capsules (capsules), soft capsules (soft capsules), solid, semi-liquid, cream, paste, etc.

[0031] Regarding the intake amount, it can be set appropriately according to the health condition, disease state, and purpose of the person or animal to whom it is applied, and there are no particular restrictions. For example, the daily intake amount may be 10 mg to 200 mg in terms of 3-O-acetyl-11-keto-β-boswellic acid, 20 mg to 200 mg, 30 mg to 100 mg, or 40 mg to 100 mg. In terms of collagen amount, it may be 5 mg to 100 mg, 5 mg to 40 mg, 5 mg to 20 mg, 10 mg to 20 mg, or 10 mg to 15 mg. It is preferable that the intake period be 12 weeks or longer. The intake period may be, for example, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, etc., and it may be used to be taken continuously or intermittently over these periods.

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

[0033] The compositions provided by the present invention can be used, for example, in foods such as health foods and functional foods, supplements, pharmaceuticals, and medical materials, and are particularly suitable as raw materials. Furthermore, as mentioned above, they can be used not only for humans but also for animals such as pets.

[0034] The amount of collagen described herein can be measured using analytical methods well known in the field of food analysis. For example, it can be measured by quantifying hydroxyproline, an acid hydrolysate, and then converting it based on the characteristics of its amino acid composition.

[0035] Furthermore, 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 sample of the boswellic acid compound prepared to a specific concentration as an indicator. [Examples]

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

[0037] [Test Example 1] [1-1. Materials] • ELISA kit: Mouse TNF-α ELISA Kit (Registered Trademark: Rebis) • Cells: Macrophage-like cell line (J774.1, mouse-derived) • Cell stimulant: LPS (Lipopolysaccharide) • Liquid culture medium: D-MEM (containing 1% or 10% FBS) • Collagen-containing extract derived from fish cartilage: (1) "SCP Complex-LS" (product name, manufactured by Linice Co., Ltd., a mixture of undenatured proteoglycan and undenatured type II & type XI collagen) (2) "SNC Complex-LS" (product name, manufactured by Linice Co., Ltd., a mixture of undenatured type II and type XI collagen) (Hereafter, the collagen contained in the above collagen-containing extract may be simply referred to as "RSC.") • Boswellic acid-containing extract derived from the Boswellia plant: (1) "Boswelline Super" (product name, manufactured by Sabinsa Japan Corporation) (2) Boswellia serrata extract 20% (product name, manufactured by Bioactives Japan Co., Ltd.) (Hereafter, 3-O-acetyl-11-keto-β-boswellic acid contained in the above-mentioned 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 cells in 10% FBS / D-MEM medium and adjust the cell concentration to approximately 2 × 10⁻⁶ 4 Adjust to cells / mL - 2.0 × 10⁶ per well of a 96-well plate 3 Inoculate the cell suspension (approximately 100 μL) to form cells. - 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 the specified concentration. - 2 hours incubation (37℃, 5%CO2) - Add LPS to each well (final concentration 1 μg / mL) - 24 hour culture (37℃, 5%CO2) - Collect the culture supernatant.

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

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

[0042] [Table 1]

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

[0044] Therefore, it became clear that boswellic acid and collagen mutually contribute positively to the mechanism by which immune cells such as macrophages mitigate their sensitivity to cellular stimulants in response to them.

[0045] (Prescription example 1) Tablets containing 300 mg each were manufactured using a conventional method with the formulations shown in Table 2.

[0046] [Table 2]

[0047] As a result, we were able to create tablets without any problems with tableting, such as capping or sticking.

[0048] (Prescription example 2) Tablets containing 300 mg each were manufactured using a conventional method with the formulations shown in Table 3.

[0049] [Table 3]

[0050] As a result, we were able to create tablets without any problems with tableting, such as capping or sticking.

[0051] <Survey conducted> Eighteen healthy individuals aged 50 to 80 with knee pain symptoms were given three tablets of Prescription Example 1 per day for 10 weeks. The results of a questionnaire showed that, in general, their pain symptoms disappeared or improved.

Claims

1. A composition comprising a collagen-containing extract derived from fish cartilage and a boswellic acid-containing extract derived from the Boswellia plant.

2. The composition according to claim 1, wherein the collagen-containing extract contains type II collagen and type XI collagen, the mass ratio of the content of type II collagen to 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.

3. The composition according to claim 1, wherein the boswellic acid-containing extract contains 20% by mass or more of boswellic acid.

4. The composition according to claim 3, wherein the boswellic acid-containing extract contains 10% by mass or more of 3-O-acetyl-11-keto-β-boswellic acid.

5. The composition according to claim 1, wherein the collagen-containing extract contains type II collagen and type XI collagen, the mass ratio of the content of type II collagen to 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, and 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 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.

6. The composition according to claim 1, wherein the collagen-containing extract contains type II collagen and type XI collagen, the mass ratio of the content of type II collagen to 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 10% by mass or more of 3-O-acetyl-11-keto-β-boswellic acid, the composition is in the form of a tablet, the total content of type II collagen and type XI collagen contained in 300 mg of the tablet is 0.1 mg to 100 mg, and the content of 3-O-acetyl-11-keto-β-boswellic acid contained in 300 mg of the tablet is 0.1 mg to 100 mg.

7. The composition according to claim 1, wherein the collagen-containing extract derived from fish cartilage is made from salmon nasal cartilage.

8. The composition according to claim 1, in a form for oral intake.

9. The composition according to claim 1, in the form of a tablet.

10. A composition according to claims 1 to 9, used for anti-inflammatory purposes.

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