Composition with synergistic effects

Abstract

The present invention relates to a novel composition comprising two or more different sources of ω-3 fatty acids, wherein the composition comprises 5% (w / w) of hokial in powder or oil form and green-lipped mussel in powder or oil form. The composition is particularly useful as a supplement and / or animal or human food. Additionally, the present invention may relate to the maintenance or treatment of human and veterinary conditions, such as use in the improvement of inflammation or joint-related disorders that may be associated with inflammation.

Description

Technology Field

[0001] The present invention relates to a novel composition comprising a combination of two different blended marine-source components comprising at least two components, including at least hokial (HR) powder or oil, or preferably a combination with green-lipped mussel (GLM) powder or oil.

[0002] Furthermore, the use of the composition of the present invention is extended to human and veterinary applications, particularly as a supplement or functional food, and provides nutritional support for health, and especially for joint health. Background Technology

[0003] Functional foods, foods, and health supplements are often used to improve or support joint structures in mammals, including humans and animals, and to maintain joint mobility.

[0004] Biological inflammatory pathways are directly related to joint health, including, for example, joint mobility. Lipids and bioactive peptides, including potentially pro-resolving lipid mediators, may also play a role, but they may act through different pathways.

[0005] The effects were shown to be induced by essential fatty acids (EFAs). It is thought that various enzymes involved in biological inflammatory pathways are affected by the presence or absence of these EFAs, and also vary depending on the type of EFA.

[0006] Significant levels of omega (ω) polyunsaturated fatty acids, particularly ω-3, can be useful as they are likely to support the body's natural anti-inflammatory processes. In particular, these EFAs are metabolized into anti-inflammatory eicosanoids. The presence of ω-3 enhances existing anti-inflammatory pathways because it acts preferentially in the process, whereas in their absence, natural substrates, including other EFAs such as arachidonic acid, are metabolized to produce pro-inflammatory agents.

[0007] GLM powder can be used to provide an ω-3 source within supplement compositions, but the quality and content of ω-3 fatty acids are not always reliable and may not always be sufficient for products intended to maintain joint health in humans or animals.

[0008] However, whether it is a long-term maintenance product such as food or nutritional supplement or a short-term dietary solution, it is still desirable to provide a composition effective for joint health use. In particular, the ability to provide a high-specification source of GLM powder that correlates with the desired technical specifications sufficient to meet these applications is becoming increasingly difficult.

[0009] Accordingly, the present invention was conceived out of a continuing need to provide a novel composition useful for the aforementioned commercial application. Prior art literature

[0010] Japanese Patent No. 4101876

[0011] Bligh and Dyer, A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology 1959, 37: 911-917 The problem to be solved

[0012] The present invention relates to a composition comprising a combination of two powder or oil components, comprising a green-lipped mussel (GLM) powder or oil component and a hoki roe (HR) powder or oil component in an amount of at least 5% (w / w).

[0013] The present invention also relates to a composition comprising a blend of at least two different powder or oil components, each containing an ω-3 fatty acid, wherein the first component among the at least two different powder or oil components comprises 5% (w / w) or more of HR powder or oil, and the blend of the at least two different powder or oil components together provides a total ω-3 fatty acid of 3.3% (w / w) or more in the composition. Preferably, the second component among the at least two powder components is GLM powder or oil.

[0014] In the first case, the applicant has found that the selection of the ω-3 fatty acid source, i.e., the source, is very important when providing a composition for the application described above. The ω-3 fatty acids in powder sources available on the market vary considerably. In the applicant's preliminary investigation, it was found that some GLM powder specifications do not provide sufficient ω-3 fatty acids to meet the technical criteria for the intended purpose compared to higher specification products that are rare and not easily available on the market.

[0015] However, GLM remains an important primary source of ω-3 fatty acids even in low-grade cases, and the inventors further investigated other ways in which GLM-based compositions can nevertheless provide a solution. By doing so, they confirmed that including a minimum amount of other specially selected ω-3 fatty acids in combination with lower-grade GLM powder raises the ω-3 content to an acceptable level. Tests revealed that the use of at least 5% HR powder blended with lower-grade GLM powder can achieve this solution.

[0016] In vitro analysis further revealed that unexpectedly enhanced levels of inhibitory or regulatory activity can be observed in the composition by introducing specific selections or combining sources of ω-3 content. Such activity is highly relevant to supporting inflammatory response pathways and maintaining optimal joint health. The anti-inflammatory activity of the blend of HR powder and GLM powder is preferably measured with a coefficient of 0.05 to 0.25, as measured according to the method described herein.

[0017] Furthermore, the applicant was able to further confirm that, provided the source remains the same, using the HR component and / or GLM component in oil form can produce a composition having equivalent technical effects, thereby providing an additional solution based on the fundamental concept of the present invention.

[0018] During a similar analysis, as performed on the powder components, the effect of the combination of these two specific oil components on the inflammatory pathway was found to be greater than expected from the sum of the individual component parts.

[0019] Therefore, the combination of these two components in powder or oil form is superior to what is predicted from levels disclosed in the art in terms of the level of inhibition or control of inflammation, as measured by relative COX2 activity. In other words, the applicant has demonstrated that when the combination of components is selected from either the powder or oil form, there is a useful synergistic effect in maintaining a better level of support for the anti-inflammatory system.

[0020] The in vitro inhibitory or regulatory activity observed by the applicant is considered to be much greater than the simple additive effect expected from the combination of ω-3 EFA levels alone. Accordingly, the applicant has confirmed that the most unexpected synergistic effect can be achieved by the present invention. That is, a specific selection of the powder or oil components of the present invention involving a specific introduction of HR at a minimum level and / or a lower specification GLM extract maintains a higher level of competitive enzymatic activity than expected based purely on the ω-3 EFA content. Accordingly, in specific embodiments, the present invention relates to a food, supplement, or functional food comprising a composition having the components cited in the claims and described herein. means of solving the problem

[0021] Such compositions will meet or exceed the required technical standards in terms of content and / or activity, and thus enable consistent and improved efficacy of GLM-based products for the purpose of supporting joint health. The present invention is useful for providing foods or supplements comprising compositions as described herein. Foods or supplements may further include one or more desirable useful excipients that may make the supplement more palatable or provide other benefits. The excipients may include, but are not limited to, one or more of the following excipients: glucosamine, hyaluronic acid, and chondroitin sulfate.

[0022] In a preferred embodiment of any aspect of the present invention, the HR powder or oil is in the range of 5% to 95% (w / w) of the total mass of the composition. In an embodiment, the HR powder or oil component contains a total fat content of 14% to 40% (w / w), but may importantly contain a total ω-3 fatty acid content of 2% to 20% (w / w).

[0023] With respect to the GLM content, the GLM powder or oil contains a total fat content of 7% to 13% (w / w). In a specific example, the GLM powder or oil is expected to contain a total ω-3 fatty acid content in the range of 2.0% to 7.0% (w / w), ideally 2.0% to 5.0% (w / w).

[0024] The compositions of the present invention are particularly useful as supplements for human or veterinary products and may be used to prevent inflammation, maintain inflammation at low levels, and / or treat inflammation. The compositions of the present invention themselves and / or foods or supplements containing said compositions may be intended for use in the treatment and / or maintenance of the health of humans or animals, preferably joint health. Additionally, treatment and / or maintenance involves support of anti-inflammatory pathways in humans or animals.

[0025] In addition, the present invention provides a method for maintaining joint health, the method comprising administering a composition of the present invention to an animal in need. In a specific embodiment, the maintenance of health comprises supporting an anti-inflammatory pathway in the animal, including the prevention of inflammation.

[0026] The present invention also provides a method for treating a subject, the method comprising administering a composition as disclosed in the present invention to a subject, the subject being a human or an animal. In a specific embodiment, the present invention treats inflammation in a subject or maintains inflammation at a low level, the subject being an animal. In a specific embodiment, the present invention maintains the health or joint health of a subject. Brief explanation of the drawing

[0027] Figure 1 is a graph showing the dose-response curve for the gold standard specifications of GLM. Figure 2 is a graph showing various GLM quality sample operations. Figure 3 is a graph illustrating the ω-3 / COX2 ratio data. Figure 4 is a graph showing the screening work of marine samples comparing GLM, Paua, blue mussels, and HR. Figure 5 is a graph showing the effects at specific % combinations, representing the predicted and actual effects on the inflammatory process. Figure 6 is a graph showing the concentration-dependent inhibition or regulation of COX2 of three different sample powders. Figure 7 is a graph showing the anti-inflammatory activity versus % powder blend compared to the expected trend lines of H powder and X powder. Figure 8 is a graph showing the anti-inflammatory activity of the x+H blend compared to the level of total ω-3 fatty acids compared to the expected trend lines of H powder and X powder. Figure 9 is a graph showing that the combination of these two components in oil form (at various blend ratios) is more effective in regulating COX2 activity than expected from the trend line. Specific details for implementing the invention

[0028] GLMs are used to provide EFAs to the body. They act preferentially in the anti-inflammatory system to support the anti-inflammatory process by generating anti-inflammatory mediators. In the absence of these ω-3 EFAs, other EFAs, particularly arachidonic acid, are used as substrates, resulting in pro-inflammatory mediators.

[0029] The dose-response effect is observed depending on the amount of GLM used, that is, as the concentration increases, the activity of the inflammatory process (COX2 activity) decreases (see Fig. 1).

[0030] The inventors confirmed that when GLM is used at the same concentration but has a lower EFA specification, a decisively reduced effect on the inflammatory process is observed (see Fig. 2).

[0031] In addition, the inventors confirmed the correlation between ω-3 EFA and the activity of the inflammatory process, namely that as ω-3 EFA increases, the activity of the inflammatory process decreases (see Fig. 3).

[0032] Furthermore, despite having similar levels of total EFA, certain marine species were found to be more effective in lowering the level of inflammatory processes, suggesting that biological activity is associated with specific EFAs compared to all species tested (see Fig. 4).

[0033] HR was identified as a source in which specific EFA profiles similar to GLM appear higher in some EFAs. Therefore, the potential to utilize HR as a substitute or supplement for reduced concentrations of reduced EFA-specific GLM and full-specific GLM was further investigated.

[0034] Based on the relative effects of GLM and HR on the inflammatory process, multiple combinations were tested by adding reduced concentrations of GLM / HR or GLM / HR with reduced EFA. The expected additional effect was calculated based on the inflammatory performance of the original GLM and HR samples to maintain the expected effect at the full concentration and full specification GLM (see Fig. 5).

[0035] In fact, the inventors have confirmed that the combination has a greater effect on the inflammatory process than is expected from the sum of the individual components. The present invention exhibits a distinct synergistic effect, or synergy, that can be used to maintain a better level of support for the anti-inflammatory system.

[0036] As mentioned above, EFA is considered important as a component in all compositions intended to help support the natural anti-inflammatory process associated with human and animal joints.

[0037] In particular, ω-3 EFA appears to act as a dual competitor for arachidonic acid oxygenation via both the cyclooxygenase (COX) and lipoxygenase (LOX) pathways, and is therefore useful for the above application.

[0038] In particular, EFAs and ω-3 fatty acids are found in green-lipped mussels (GLM) or green-shelled mussels (GSM). Perna canaliculus It can be found in ). As mentioned above, due to variations in the specifications of GLM powder and the fact that many existing GLM powder sources did not necessarily provide sufficient specifications for ω-3 fatty acid content, alternatives or additives were sought for investigation.

[0039] Various marine sources have been examined as potential sources of desirable ω-3 fatty acid components. Generally, marine-derived sources of high ω-3 fatty acids are oils from shellfish or fish organs.

[0040] A range of oils of this type was selected as potential substitutes for parts of compositions containing GLM to improve the total ω-3 fatty acid content. Although it is a good source of ω-3 fatty acids in itself, it was found that blending GLM powder with oil is difficult to provide a homogeneous powder suitable for formulation into applicable products.

[0041] However, such a single source of ω-3 fatty acids is Macruronus novaezelandiae , or dried egg powder from white fish hoki. So, hoki egg (HR) powder was tested to determine whether the addition of hoki egg (HR) powder to GLM powder improves the content of EFAs, especially ω-3 fatty acids.

[0042] To test whether the ω-3 level could be raised to the specifications required by the combination, the suitability of the HR powder was confirmed by measuring the levels of ω-3 fatty acids in the GLM powder before and after the addition of the HR powder. The required specifications were provided by the GLM powder, which is known to provide a commercially desired therapeutic effect with a relatively high ω-3 fatty acid specification.

[0043] In order to confirm whether the novel powder blend exhibits the desired anti-inflammatory activity (despite the inclusion of GLM with a lower ω-3 content), in vitro Enzyme activity was tested and compared with a standard product of specified specifications used by the applicant.

[0044] commonly used to measure anti-inflammatory activity in vitro The process was used. That is, as mentioned above, the COX2 enzyme, known as a pathway significantly involved in the inflammatory process in vitroAnti-inflammatory activity was measured by measuring the level of competition.

[0045] method

[0046] New Zealand green-lipped mussel (GLM) powder and New Zealand HR powder were obtained from commercial sources, and lipid extracts were obtained from individual powders or blends of powders using the Bligh and Dyer method. This method is described in the following publication (see Bligh and Dyer, A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology 1959, 37: 911-917). The fatty acid content of the lipid extracts was measured using AOAC method 963.22.

[0047] result

[0048] The first set of data was generated by measuring and comparing the amount of ω-3 fatty acids in a standard reference source of GLM product (X) known to the applicant in order to determine the required amount of ω-3 fatty acids that are suitable and desirable for use with acceptable specifications. An additional source of GLM powder (x) known to have lower specifications was also measured.

[0049] GLMX powder alone was found to be insufficient as it was measured to have significantly less ω-3 fatty acid than the reference powder (X) due to low COX2 activity or regulation, and thus fell short of the necessary specifications required for the commercially desirable use mentioned above.

[0050] A new combination was tested to determine how the ω-3 fatty acid levels are affected when product X of the following specifications is supplemented with a certain amount of another powder known to be rich in ω-3.

[0051] HR powder (H) was previously selected from various sources based on the fact that it has a high potential to increase total ω-3 fatty acid content and can be easily blended with GLM powder.

[0052] In order to determine whether the low-specification GLM powder is improved to the required specifications by the addition of HR powder, it was theoretically confirmed that at least 5% (w / w), preferably 8% (w / w), of a specific HR powder (BN:HR114) added to the low-specification GLM powder increases the total ω-3 content score (measurement of the percentage of fatty acids in the total fat of the product) to a level that meets the required specifications.

[0053] It was confirmed by measurement that by including 8% (w / w) of HR powder (H) in a blend with GLM powder (X) of the following specifications, the ω-3 fatty acid content increased to 3.5% (w / w), which is very close to the level shown by standard powder X (BN32132) as shown in Table 1.

[0054] composition Batch number GLM powder ingredients (g) HR Powder Ingredients (g) Total ω-3 score (g / 100g) GLM Powder - Below Specifications (X) 100 0 3.1 HR powder (H) BN:HR114 0 100 5.1 GLM Powder - Specifications Below + HR(X+H) 95 5 3.3 GLM Powder - Specifications Below + HR(X+H) BN32127 92 8 3.5 GLM Powder - Commercially Acceptable Specifications (X) BN32132 100 0 3.6

[0055] Biological activity and graph analysis

[0056] Various quality specifications of GLM (different ω-3 fatty acid levels) were combined with HR powder in various ratios. Then, the total ω-3 fatty acids of the combined products and the COX2 inhibition or regulation scores of these powders were measured by comparing them with source GLM (x) and Hoki powder (H).

[0057] Next, the anti-inflammatory activity of the above examples was measured and compared with the expected activity based on the new combination of ω-3 fatty acids obtained to confirm the potential for the biological effect of the combination.

[0058] COX2 inhibition or regulation was measured by incubating lipid extracts of the powder blend combination with a commercially available mammalian COX2 enzyme, and the enzyme activity was measured by the co-oxidation of N,N,N,N'-tetramethyl-p-phenylenediamine (TMPD) by prostaglandin G2 (PGG2) to produce oxidized TMPD, which was blue and easily detectable at 611 nm.

[0059] The inhibitory or regulatory activity of the powder blend combination on COX2 is measured from the concentration-dependent inhibition or regulation of the lipid extract of the COX2 enzyme, or by multiplying the COX2 enzyme by 50% (IC) of the total weight of the lipids extracted from the powder blend combination. 50 It was measured by the anti-inflammatory (AI) activity score of the powder sample, calculated as the reciprocal of the concentration of the lipid extract required to inhibit or regulate it.

[0060] As can be seen in the figure, the concentration-dependent inhibition or regulation of COX2 by GLM powder (x) of the following specifications blended with 8% (w / w) HR powder (H) to produce x+H (BN32127) was much greater with GLM powder X (BN 32132) alone and was estimated to be similar as it exhibited similar ω-3 fatty acid levels (as shown in Table 1).

[0061] More surprisingly, specifically as shown in Fig. 6, the concentration-dependent inhibition or regulation of COX2 by x+H (BN 32127) was much higher than that of 100% HR powder H (BN: HR114), which exhibited significantly higher ω-3 levels than x+H (see Table 1).

[0062] As can be seen in Figure 7, it was further established that the anti-inflammatory (AI) activity represented by x+H in various ratios of %H blends (diamond data points) is greater than that expected from the anti-inflammatory (AI) activity when plotting the linear trend lines showing the activity of GLM powder X (far left square data point) and HR powder H alone (far right square data point). The linear trend line between the two indicates what can be expected from a simple additive effect of the two powders, suggesting that when the two powders are combined, there is an enhanced effect that far exceeds the addition of the individual effects.

[0063] This trend was further observed in Figure 8, as the anti-inflammatory (AI) activity of x+H in various % powder blends (diamond data points) was greater than expected from the total omega-3 fatty acid levels in H and X alone (as shown in Figure 3).

[0064] The large square data points display a linear trend line relationship representing what can be expected from the simple additive effect achieved by combining the two powders. However, as can be seen from the figure, the blended powder significantly exceeded the expected AI activity, indicating that the novel combination provides an enhanced synergistic effect rather than a simple cumulative or additive effect.

[0065] The inventors further tested the oil-base combinations of these components and confirmed that the anti-inflammatory activity observed in H blends of various ratios (original data points) was greater than expected from the additional activity of the oil components. This confirmed that the oil-base component combination is also effective in generating enhanced bioactivity compared to the individual components.

[0066] Table 2 shows the test results of the oil-based components below.

[0067] HR oil GLM oil Inflammation levels (COX2 IC 50 μg / mL) 0.0% 100.0% 28.50 100.0% 0.0% 51.50 5.0% 95.0% 22.80 10.0% 90.0% 25.90 25.0% 75.0% 23.50 50.0% 50.0% 41.60 75.0% 25.0% 29.70 90.0% 10.0% 32.40 95.0% 5.0% 40.80

[0068] Figure 9 plots the relative COX2 activity expected when either 100% GLM oil (far left square) or 100% HR oil (far right square) is used alone, illustrating the significance of these results (see Table 2) in a graph format. The linear trend line between the two plotted data square points indicates what can be expected from the simple addition effect of the two oils.

[0069] As can be seen in Table 2 above, the results plotted from the actual activity observed in various combinations (indicated by circular data points) suggest that the combination of oil components has an enhanced synergistic effect beyond the additive effect of the individual oil components.

Claims

Claim 1 A composition comprising a combination of two different powder or two different oil components, wherein the composition comprises green-lipped mussel (GLM) powder or oil and at least 5% (w / w) of hochial (HR) powder or oil, and wherein the composition synergistically inhibits COX2 activity. Claim 2 A composition according to claim 1, wherein the combination of the two different powders or two different oil components together provides at least 3.3% (w / w) of total ω-3 fatty acids in the composition. Claim 3 A composition according to claim 1 or 2, wherein the HR powder or oil is present in a range of 5% to 95% (w / w) of the total composition. Claim 4 A composition according to claim 1 or 2, wherein the GLM powder or oil contains a total fat content of 7% to 13% (w / w). Claim 5 A composition according to claim 1 or 2, wherein the GLM powder or oil contains a total ω-3 fatty acid content of 2.0% to 5.0% (w / w). Claim 6 A composition according to claim 1 or 2, wherein the HR powder or oil contains a total fat content of 14% to 40% (w / w). Claim 7 A composition according to claim 1 or 2, wherein the HR powder or oil contains a total ω-3 fatty acid content of 2% to 20% (w / w). Claim 8 A composition according to claim 1 or 2 for use in the treatment or maintenance of joint health in humans or animals. Claim 9 In claim 8, a composition wherein the treatment or maintenance comprises supporting an anti-inflammatory pathway in humans or animals. Claim 10 A food comprising the composition of claim 1 or 2 and one or more excipients selected from glucosamine, hyaluronic acid, and chondroitin sulfate. Claim 11 A functional food comprising the composition of claim 1 or 2 and one or more excipients selected from glucosamine, hyaluronic acid, and chondroitin sulfate. Claim 12 A supplement comprising the composition of claim 1 or 2 and one or more excipients selected from glucosamine, hyaluronic acid, and chondroitin sulfate. Claim 13 delete Claim 14 delete Claim 15 delete Claim 16 delete Claim 17 delete Claim 18 delete Claim 19 delete Claim 20 delete Claim 21 delete

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