Composition for inhibiting the degradation of collagen type IV, as well as foods or food supplements containing this composition

Abstract

Composition for inhibiting the degradation of collagen type IV, characterized in that it comprises: Green coffee extract, olive fruit extract; wherein, based on parts by weight of the raw materials, the green coffee extract amounts to 100-300 parts and the olive fruit extract amounts to 15-167 parts.

Description

Technical field

[0001] The present utility model belongs to the technical field of functional compositions and specifically relates to a composition for inhibiting the degradation of collagen type IV, as well as food or health foods containing this composition. State of the art

[0002] With increasing age and due to environmental influences, collagen loss is one of the key factors leading to skin aging. Current anti-aging research for the skin focuses primarily on type I collagen in the dermis and pays little attention to collagen in the basal membrane zone.

[0003] The basal membrane zone represents the second physical barrier of human skin and, through the basal membrane between the epidermis and dermis, forms an optimal "epidermis-basal membrane-dermis" circulation, allowing the free exchange of substances and information and thus maintaining the healthy vitality of the skin. Collagen type IV, as one of the main collagen components that constitute the basal membrane, provides structural support and mechanical strength to the basal membrane through self-organization into complex network structures, supports cells and tissues, and is involved in various physiological processes such as filtration, cell adhesion, cell differentiation, and tissue regeneration.

[0004] Currently, US patent 20160175217A1 discloses an anti-aging composition for topical application, specifically a topical composition containing at least one substance from catechins and flavonols of green tea extract as an active ingredient. This substance can reduce the biosynthesis of matrix metalloproteinases (MMP-2, MMP-9), increase the biosynthesis of collagen type IV, collagen type VII, and laminin, inhibit the degradation of the dermal-epidermal junction, and promote its formation, thereby protecting the dermal-epidermal junction. The composition also discloses a composition containing catechins and theanine. However, this is a topical preparation and not an ingestible composition.

[0005] Currently, there are no reports on the combination of green coffee extract and olive fruit extract for inhibiting the degradation of collagen type IV components. Content of the present utility model

[0006] The purpose of this section is to outline some aspects of the embodiments of this utility model and to briefly describe some preferred embodiments.

[0007] In view of the problems described above and / or existing in the prior art, the present utility model is proposed.

[0008] Therefore, the aim of the present utility model is to overcome the shortcomings of the prior art and to provide a composition for inhibiting collagen type IV degradation.

[0009] To solve the aforementioned technical problems, the present utility model provides the following technical solution: A composition for inhibiting collagen type IV degradation, comprising green coffee extract and olive fruit extract; wherein, based on parts by weight of the raw materials, the green coffee extract comprises 100-300 parts and the olive fruit extract comprises 15-167 parts.

[0010] As a preferred embodiment of the composition described in the utility model, wherein: the active ingredients in the green coffee extract comprise chlorogenic acid.

[0011] As a preferred embodiment of the composition described in the utility model, wherein: the active ingredients in the olive fruit extract comprise hydroxytyrosol and olive polyphenol.

[0012] As a preferred embodiment of the composition described in the utility model, wherein: the mass fraction of the active component chlorogenic acid in the green coffee extract is ≥0.1%

[0013] As a preferred embodiment of the composition described in the utility model, wherein: the mass fraction of the active component hydroxytyrosol in the olive fruit extract is ≥0.1% and the mass fraction of the active component olive polyphenol in the olive fruit extract is ≥0.1%.

[0014] As a preferred embodiment of the composition described by the utility model, wherein: based on parts by weight of the raw materials, the green coffee extract comprises 100-200 parts and the olive fruit extract comprises 15-67 parts.

[0015] As a preferred embodiment of the composition described by the utility model, wherein: based on parts by weight of the raw materials, the green coffee extract is 200 parts and the olive fruit extract is 34 parts.

[0016] Another purpose of the utility model is to overcome the shortcomings of the prior art and to provide a food or health food containing a composition to inhibit the degradation of collagen type IV.

[0017] In a preferred embodiment, wherein said food and health foods comprise tablets, beverages, capsules, powders, confectionery and jellies.

[0018] Advantageous effects of the present utility model: (1) The present utility model provides a composition for inhibiting the degradation of type IV collagen which exhibits a significant synergistic effect compared to individual materials for inhibiting the degradation of type IV collagen. (2) The composition provided by the present utility model for inhibiting the degradation of collagen type IV exhibits a high level of safety, with all raw material components belonging to the legally permitted ordinary food raw materials. Brief description of the characters

[0019] To clarify the technical solutions of the embodiments of this utility model, the figures necessary for describing these embodiments are briefly introduced below. Obviously, the figures shown in the following description represent only some embodiments of this utility model. Those skilled in the field can easily derive other figures based on these without any creative effort. The following applies: Fig.Figure 1 shows a comparison diagram of the collagen type IV content after treatment with compositions of different mixing ratios in the embodiment of the present utility model. Fig. Figure 2 shows a diagram of the degradation inhibition rate of collagen type IV of the comparison model group in the embodiment of the present utility model. Detailed descriptions

[0020] In order to make the aforementioned purposes, features and advantages of the present utility model clearer and more understandable, the specific design of the present utility model will be explained in detail below with reference to the exemplary embodiments in the description.

[0021] The raw materials used in the production of the composition of this utility model are obtained through the following steps: Green coffee extract can be obtained through an extraction process that includes the following steps: 1) Crush the green coffee beans, grind; 2) Extraction with water as solvent, continuous countercurrent extraction, water supply: 1200±10kg / h, extraction temperature: 50-75°C, extraction time: 1-2h or alternatively extraction with 5-10 times the amount of water twice, each with an extraction time of 1-2 hours; 3) Performing fine filtration using a ceramic membrane or plate filter, retaining the filtrate; 4) Concentration with the following parameters: vacuum -70.0 kPa to -95.0 kPa, temperature 75-80°C, Brix 30-35%; 5) Spray drying: Inlet temperature 195±15°C, outlet temperature 95±15°C; 6) Packaging for the production of a commercial green coffee extract; 7) Carrying out the identification and determination of the content of chlorogenic acid in the green coffee extract, wherein the mass fraction of chlorogenic acid may be ≥0.1%; in embodiments 1-10 and comparative examples 1-5, green coffee extract with a chlorogenic acid mass fraction of 20% is used.

[0022] Olive fruit extract can be obtained through an extraction process that includes the following steps: 1) Fruit harvest; 2) Wash, press and grind to 10-80 mesh; 3) Reflux extraction in an extraction vessel using water or ethanol as solvent, extracting twice with 5-10 times the amount, each time for 1-3 hours, or continuous countercurrent extraction with water supply: 1200±10kg / h, extraction temperature: 50-75°C, extraction time: 1-2 hours; 4) Filtration using a ceramic membrane or plate filter with retention of the filtrate; 5) Concentration at the following parameters: Temperature: 60-80°C, Vacuum: -70.0kPa to -95.0kPa; Relative density 1.03-1.12kg / m³ 3 ; 6) Addition of maltodextrin (maltodextrin content 5%-40%) and subsequent spray drying after mixing, inlet temperature 160-220°C, outlet temperature 60-110°C; 7) After packaging, commercial olive fruit extract is obtained; 8) Carrying out the identification and determination of the hydroxytyrosol content in olive fruit extract, wherein the mass fraction of hydroxytyrosol and the mass fraction of olive polyphenol are to be ≥0.1%. In embodiments 1-10 and comparative examples 1-5, olive fruit extract with a hydroxytyrosol mass fraction of 1.5% and an olive polyphenol mass fraction of 2% is used. Examples 1-10 and comparative examples 1-5

[0023] The extract powders are mixed uniformly according to Table 1 below in order to obtain the compositions of the respective exemplary embodiments and comparative examples.

[0024] Table 1. Composition formulations of embodiments 1-10 and comparative examples 1-5 group Material composition (parts by weight) Composition of the basic material components (parts by weight) Green coffee extract (contains 20% chlorogenic acid) Olive fruit extract (contains 1.5% hydroxytyrosol) Chlorogenic acid Hydroxytyrosol Example 1 100 15 20 0.225 Example 2 300 167 60 2.505 Example 3 100 167 20 2.505 Example 4 300 15 60 0.225 Example 5 200 100 40 1.5 Example 6 150 134 30 2.01 Example 7 100 67 20 1.005 Example 8 150 15 30 0.225 Example 9 200 67 40 1.005 Example 10 200 34 40 0.51 Comparative example 1 50 34 10 0.51 Comparative example 2 350 100 70 1.5 Comparative example 3 250 10 50 0.15 Comparative example 4 200 0 40 0 Comparative example 5 0 67 0 1.005

[0025] In Table 1, comparison examples 4~5 are presented as two components, where comparison example 4 does not contain olive fruit extract and comparison example 5 does not contain green coffee extract.

[0026] In comparison example 1, the content of green coffee extract and olive fruit extract is lower; in comparison example 2, the content of green coffee extract and olive fruit extract is higher. Experimental setup:

[0027] The determination of the collagen type IV content in HaCaT cells was carried out on all compositions obtained from the preceding exemplary embodiments and comparative examples, as described below: Cell line: Immortalized keratinocytes HaCaT (ZQ0044\Zhongqiao Xinzhou); Instruments and consumables: incubator, enzyme labeling device, 96-well plates, 24-well plates, various types of pipettes and pipette tips, and other cell culture consumables; Reagents: DMEM basic culture medium, DMEM complete culture medium (DMEM+15% serum+1% dual antibiotic), CCK-8 reagent (C0039\Beyotime), 0.25% trypsin, human collagen type IV (Col IV) enzyme immunoassay kit (CSB-E17116h\Wuhan Huamei); Experimental procedure steps:

[0028] 1. HaCaT cells are cultured in DMEM culture medium with 15% fetal calf serum at 37°C and 5% CO2 in a cell culture incubator; HaCaT cells in the logarithmic growth phase are extracted and treated with 1×10 5 Cells / wells seeded in 24-well plates, 500 µL per well, at 5% CO₂ 2 and cultivated at 37°C in an incubator for 24 hours.

[0029] 2. The culture medium is discarded, washed once with PBS; 200 µL of PBS is added to each well.

[0030] 3. The entire experiment is divided into three groups: an empty control group, a model group, and treatment groups with different sample mixtures (i.e., groups treated with the compositions of the respective exemplary implementations and comparison examples, hereinafter referred to accordingly). a) Empty control group: DMEM medium is co-incubated with HaCaT cells without receiving UVA irradiation (i.e., the empty control group contains in this well: 500 µL cell culture medium + no UVA irradiation); b) Model group: Placement in the UV crosslinker (25 J / cm²)2 UVA) for irradiation treatment of the cells, treatment for 87.5 minutes (i.e. the model group contains in this well: 500 µL cell culture medium + receives UVA irradiation); c) Sample treatment group: Placement in UV crosslinker (25 J / cm²) 2UVA) for irradiation treatment of the cells, after 87.5 minutes of treatment the culture medium is aspirated, a total concentration of 250 µg / mL of various mixing-ratio composition solutions is added (the various mixing-ratio composition solutions are combined according to the weight part ratios of green coffee extract and olive fruit extract in Table 1 for the preparation of the composition solution, the solvent for the preparation of the composition solution being pure water) and co-incubated with HaCaT cells for 24 h (i.e. the sample treatment group contains in each well: 500 µL of composition solution with a total concentration of 250 µg / mL + receives UVA irradiation).

[0031] 4. The perforated plate is repeatedly frozen and thawed three times at -80°C, and all liquids in the perforated plate are collected; at 4°C, centrifugation is performed for 20 minutes at 5000 rpm, and the supernatant is collected.

[0032] The concentration of collagen type IV (Col IV) in the collected solution is determined according to the human collagen type IV (Col IV) enzyme immunoassay kit.

[0033] Based on the results of the collagen type IV content, the degradation inhibition rate of collagen type IV is calculated in comparison to the model group. the calculation method is as follows: Collagen type IV degradation inhibition rate = Collagen type IV content of the sample group − Collagen type IV content of the model group / Collagen type IV content of the model group 100% Test results:

[0034] The corresponding formulations of the compositions obtained in all the preceding exemplary embodiments and comparative examples, as well as the data on the degradation inhibition rate of collagen type IV, are shown in Table 2. Table 2 Summary of the corresponding formulations of the compositions obtained in the exemplary embodiments and comparative examples, as well as data on the degradation inhibition rate of collagen type IV group Material composition (parts by weight) Composition of the basic material components (parts by weight) Collagen type IV, pg / mL Degradation inhibition rate of collagen type IV, % Green coffee extract (contains 20% chlorogenic acid) Olive fruit extract (contains 1.5% hydroxytyrosol) Chlorogenic acid Hydroxytyrosol Empty control group 0 0 0 0 123.21550 / Model group 0 0 0 0 43.19189 / Example 1 100 15 20 0.225 56.03871 29.74% Example 2 300 167 60 2.505 56.45864 30.72% Example 3 100 167 20 2.505 57.56786 33.28% Example 4 300 15 60 0.225 58.57338 35.61% Example 5 200 100 40 1.5 59.79902 38.45% Example 6 150 134 30 2.01 59.46408 37.67% Example 7 100 67 20 1.005 70.29293 62.75% Example 8 150 15 30 0.225 70.64613 63.56% Example 9 200 67 40 1.005 69.49147 60.89% Example 10 200 34 40 0.51 76.66509 77.50% Comparative example 1 50 34 10 0.51 43.93217 1.71% Comparative example 2 350 100 70 1.5 44.59555 3.25% Comparative example 3 250 10 50 0.15 43.04499 -0.34% Comparative example 4 200 0 40 0 49.67564 15.01% Comparative example 5 0 67 0 1.005 44.38740 2.77%

[0035] Table 2 shows that the composition prepared using green coffee extract and olive fruit extract in specific ratios can effectively inhibit the degradation of collagen type IV. According to embodiments 1-10, the degradation inhibition rate of collagen type IV is 29.74% to 77.50% when the composition has the specific ratio of 100-300 parts green coffee extract to 15-167 parts olive fruit extract. The highest degradation inhibition rate of collagen type IV, 77.50%, is achieved with 200 parts by weight of green coffee extract and 34 parts by weight of olive fruit extract, representing a significant increase compared to the other embodiments.

[0036] In comparative example 1, the weight fraction of the green coffee extract is not in the range of 100-300 parts (insufficient amount of green coffee extract), which reduces the rate of degradation inhibition of collagen type IV and prevents the synergistic enhancing effect of the two substances.

[0037] In comparative example 2, the weight fraction of the green coffee extract is not in the range of 100-300 parts (overdosed amount of green coffee extract), which reduces the rate of degradation inhibition of collagen type IV and prevents the synergistic enhancing effect of the two substances.

[0038] In comparative example 3, where the weight parts of the olive fruit extract are not in the range of 15-167 parts (olive fruit extract in insufficient quantity) and the weight parts of the green coffee extract are in the range of 100-300 parts, the rate of degradation inhibition of collagen type IV decreases, and the synergistic enhancing effect of the two substances becomes ineffective.

[0039] Comparative example 4 serves as a single-substance control, meaning that with an exclusive content of green coffee extract, the rate of degradation inhibition of collagen type IV is significantly higher than in the model group, but significantly lower than in embodiments 1 to 10.

[0040] Comparative example 5 serves as a single-substance control, meaning that with an exclusive content of olive fruit extract, there is virtually no inhibitory effect on the degradation of collagen type IV.

[0041] The comparison chart of collagen type IV content is in Fig. Figure 1 shows the following, where a~f are used to mark significance between the individual groups: a. The blank control group shows significant differences from all other groups; b. Implementation 10 shows significant differences from all other groups; c. Implementation 8 shows no significant differences from Implementation 7 / 9, but each shows significant differences from the remaining groups; d. Implementation 5 shows no significant differences from Implementation 1 / 2 / 3 / 4 / 6, but significant differences from all other groups; e. Comparison example 4 shows no significant differences from Comparison example 1 / 2 / 5, but significant differences from all other groups; f. The model group shows no significant differences from Comparison examples 1 / 2 / 3 / 5, but significant differences from all other groups.

[0042] The diagram of the collagen type IV degradation inhibition rate of the comparison model group is in Fig. 2 shown.

[0043] The embodiments 1-10 exhibit a degradation inhibition rate of 29.74%-77.50%, which is significantly higher than that of the comparison examples 1~5 with a degradation inhibition rate of -0.34%~15.01%, proving that the composition has a synergistic effect within the specific ratio range.

[0044] The embodiments 7-10 exhibit a degradation inhibition rate of 60.89%-77.50%, which is significantly higher than that of embodiments 1-6 with a degradation inhibition rate of 29.74%-38.45%.

[0045] In embodiment 10, the degradation inhibition rate was 77.50%, significantly higher than in embodiments 1-9 and comparative examples 1-5, which is why embodiment 10 represents the optimal synergistic ratio of the composition.

[0046] When comparing embodiment 10 (degradation inhibition rate 77.50%) with the sum of the degradation inhibition rates of comparison examples 4~5 (i.e. 15.01%+2.77%=17.78%), it is evident that the degradation inhibition rate of embodiment 10 is far higher than the sum of the degradation inhibition rates of comparison examples 4~5, which proves the synergistic effect of the composition as a non-additive effect.

[0047] It should be noted that the foregoing embodiments serve only to illustrate the technical solution of the present utility model and are not to be understood as limiting. Although the present utility model has been described in detail with reference to preferred embodiments, the person skilled in the art should understand that modifications or equivalent substitutions may be made to the technical solution of the present utility model without departing from the spirit and scope of protection of the technical solution of the present utility model, all of which are intended to be covered by the scope of protection of the present utility model. QUOTES INCLUDED IN THE DESCRIPTION

[0000] This list of documents cited by the applicant was automatically generated and is included solely for the reader's convenience. The list is not part of the German patent or utility model application. The DPMA accepts no liability for any errors or omissions. Cited patent literature

[0000] US 20160175217A1

[0004]

Claims

[1] Composition for inhibiting the degradation of collagen type IV, characterized by that it includes: Green coffee extract, olive fruit extract; wherein, based on parts by weight of the raw materials, the green coffee extract amounts to 100-300 parts and the olive fruit extract amounts to 15-167 parts. [2] Composition according to claim 1, characterized by that the active ingredients in the green coffee extract include chlorogenic acid. [3] Composition according to claim 1 or 2, characterized by that the active ingredients in the olive fruit extract include hydroxytyrosol and olive polyphenol. [4] Composition according to claim 1 or 2, characterized by that the mass fraction of the active component chlorogenic acid in the green coffee extract is ≥0.1%. [5] Composition according to claim 3, characterized by, that the mass fraction of the active component hydroxytyrosol in the olive fruit extract is ≥0.1% and the mass fraction of the active component olive polyphenol in the olive fruit extract is ≥0.1%. [6] Composition according to one of claims 1, 2 or 5, characterized by , that, in terms of parts by weight of the raw materials, the green coffee extract comprises 100-200 parts and the olive fruit extract 15-67 parts. [7] Composition according to claim 6, characterized by , that, in terms of parts by weight of the raw materials, the green coffee extract is 200 parts and the olive fruit extract is 34 parts. [8] Food or health food containing the composition according to any one of claims 1 to 7. [9] Food or health food according to claim 8, characterized by that the food and health food products include tablets, drinks, capsules, powders, confectionery and jellies.

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