KR20090103918A - An extract - Google Patents

Abstract

The invention relates to compositions that include an oil extract rich in fatty acid amide compounds including N-acylethanolamine (NAE) compounds such as N-palmitoylethanolamide (PEA) and N-arachidonoylethanolamide (AEA). The composition may be derived from marine materials including mussel meat.

Description

Extract {AN EXTRACT}

The present invention relates to extracts. More specifically, the present invention is rich in fatty acid amide compounds including N-acylethanolamide (NAE) compounds such as N-palmitoylethanolamide (PEA) and N-arachidonoylethanolamide (anadamide, AEA) An oil extract derived from marine based raw materials.

N-palmitoylethanolamide (PEA) is an N-acylethanolamine (NAE's; or N-acylethanolamide), which includes 'cannabis-like' compounds such as N-arachidonoylethanolamide (anandamide, AEA). Endogenous fatty acid amides belonging to the family known as. PEA and AEA have the ability to treat a series of human / animal conditions involving abnormal inflammatory and / or immune responses and associated pain (Lambert et al., 2002).

Overview of the historical and scientific literature of PEA and other NAE's

a) origin

PEA is a naturally occurring lipid found in many diverse cells of animal, marine and plant origin (Lambert et al., 2002, homologous). In living organisms, PEA synthesis is rapidly induced in response to cellular stressors, such as tissue damage or pathological injury, often accompanied by inflammation and pain (Darmani, et al., 2005). Although the biological function of human PEA is not fully understood, it is assumed that PEA constitutes one of a variety of natural anti-inflammatory and analgesic drugs (Darmani, et al., 2005).

Crude lipids extracted from freeze-dried pulp of green mussels (chlorous mussels) using chloroform and methanol are mixed mixtures of triglycerides, sterol esters, sterols, polar lipids, free fatty acids and their derivatives such as NAEs. (Sepe et al., 1998; Murphy et al .; Murphy et al., 2003). PEA and N-stearoylethanolamide contain much smaller amounts of N-myristoyl (C14: 0)-, N-oleoyl (C18: 1)-, N-linoleyl (C18: 2)-, and N Arachidonoyl (C20: 4; Anandamide / AEA) -ethanolamide is the most abundant NAEs in blue mussel lipid extracts. The nature and quantity of NAE's present in the crude or purified extracts may have therapeutic significance after the presence of complex synergistic interactions between NAE's in whole animals and animal tissues has been reported in the prior art. For example, the results obtained from animal studies indicate that some action of PEA may be mediated through AEA and possibly other NAEs. In addition, evidence of synergistic effects between NAEs has been reported (described in detail below).

b) chemical

PEA is saturated 16 carbon fatty acid ethanolamide (C16: 0) having the structure:

AEA is a polyunsaturated 20 carbon fatty acid ethanolamide (C20: 4) having the structure:

c) historical views

The anti-inflammatory properties of PEA were first identified in the early 1950s by Coburn et al. Who discovered breeding guinea pigs in a high yolk diet to protect them from experimental allergies. Subsequent studies have isolated and purified PEA from egg yolk and characterized anti-inflammatory properties in animals (Lambert et al., 2002). Research on NAEs resumed in the 1990s with the discovery of another endogenous NAE, AEA, which had properties similar to the active ingredients of cannabis. Cloning of cannabinoid receptors (named CB1 and CB2) and generation of selective CB receptor ligands provided a mechanism to further promote research activity (Devane et al., 1992). Subsequent evidence of evidence obtained from animal and human studies showed that PEA has anti-inflammatory and analgesic properties when given through different routes of administration.

d) pre-clinical studies

Many studies have shown that synthetic PEA has anti-inflammatory and analgesic properties in various animal models. Typically, inflammatory components, such as carrageenan, collagen or povol esters, are injected under the skin so that pathological and behavioral changes are measured within hours (acute model) or days (chronic model). To date, the vast majority of these studies have been conducted in acute animal models of inflammation (eg, Aloe et al., 1993; Mazzari et al., 1996; Conti et al., 2002; Costa et al., 2002).

e) clinical trials

In comparison to animal studies, only a few human clinical trials were conducted to investigate the anti-inflammatory effects of PEA. During the early 1970s several experiments were carried out in Czechoslovakia using an oral formulation provided under the tradename Impulsin® (N-2-hydroxyethyl palmitamide, SPOFA United Pharmaceutical Works). The experiment in Article 1 evaluated the effect of PEA (3 times / day / 12 days) on reducing the frequency and severity of respiratory tract infections in 1345 adult volunteers (young male soldier or Skoda Car Co.) Employees of; Masek et al., 1974). The results indicated that when impulsine® is given before an infectious contingency, it helps to prevent viral infection but does not reduce the duration of infectious symptoms. An additional series of similar experiments was conducted between 1973 and 1975 in a total of 1864 young male soldiers (same dose regimen), and prophylactic impulse® was used to determine the frequency of acute respiratory infections in these populations. Significant reduction (Kahlich et al., 1979). The frequency of adverse events was particularly low during the 12-week experiment (only a few percent; Kahlich et al., 1979). The apparent success of these experiments has led to the use of impulsin® in previous Slovakians for acute respiratory disease. After several years on the market, the drug was withdrawn due to unknown reasons that seemed not to be associated with toxicity (Lo Verme et al., 2005b). Two subsequent clinical trials were initiated to investigate the effects of PEA on chronic lumbosciatalgia and multiple sclerosis (Lambert et al., 2002).

Creams comprising “structured natural lipids” with PEA for administration have also been developed (Physiogel® A.I., Stiefel Laboratories) and evaluated in two small clinical trials. The first was an observational study in which 19 adult patients diagnosed with anal eczema were instructed to apply the cream to the affected area between 6 and 63 days (Rohde and Ghyczy, 2003). After 4 weeks of experiment, 68% of patients reported reductions in pain, burning and itching, and 21% reported worsening of symptoms. This cream has been repeatedly well adapted to 95% of patients. In a second experiment, the cream was applied twice daily for three weeks to 21 adult patients suffering from end stage renal failure and suffering from urinary itch (Szepietowski et al., 2005). At the end of the three-week trial, there was a statistically significant decrease in itching (the itch disappeared completely in less than 40% of patients) and 81% of patients reported a decrease in dryness in the injured area (dryness). This cream was well tolerated in all patients without reported adverse effects. These results provide an encouraging signal that PEA-containing creams may provide an alternative therapeutic option in the treatment of inflammatory skin conditions.

f) mode of action

The exact mechanism (s) by which PEA exerts its anti-inflammatory and analgesic effects is not fully understood. It has been found that PEAs effects are generally not mediated through traditional cannabinoid receptors, as it has been discovered that psychotropic effects may be accounted for. In contrast, most AEAs action appears to be mediated through CB1 and / or CB2 receptors in the brain and periphery. One problem associated with the use of AEA is its psychotropic side effects, which are thought to be mediated through the CB1 receptor. However, there is evidence that such unwanted effects may be reduced or eliminated by co-administration of low dose (sub-therapeutic) AEA and other NAEs (Calignano et al., 1998; Di Marzo et al., 2001; De Petrocellis et al. , 2001; Lo Verme et al., 2005b).

For example, sub- analgesic doses of PEA or AEA provide analgesia when given in combination at the same low dose (Calignano et al., 2001).

At the molecular and behavioral levels, PEA interacts with a number of important targets in the body. Evidence collected from in vitro and in vivo studies indicates that PEA reduces edema, mast cell proliferation, neutrophil infiltration and numerous endogenous mediators, including: mast cell degranulation (inhibition of histamine and serotonin secretion), cyclo-jade Cigenase-2 (COX-2) activity, endothelial nitric oxide synthesis activity, nitric oxide production from macrophages and lipid peroxidation during acute hypoxia (Gulaya et al., 1998). PEA also reduces hyperalgesia in animal models of inflammatory pain (Jaggar et al., 1998; Farquhar & Smith, 2001; Conti et al., 2002).

Recent results suggest that peroxide growth factor-activated receptor alpha (PPAR-α) is very important for the anti-inflammatory action of PEA (Lo Verme et al., 2005a).

Standard Therapeutic Approaches to the Control of Inflammatory Conditions

A. Diseases Including Chronic Inflammation

Inflammation is characterized by typical signs of local redness, swelling, fever and pain as part of the body's normal response to wounds and infections. Normal inflammatory response is an acute process that resolves the initiation of tracer elimination and repair of tissues and tissue healing. In some situations, acute inflammation can progress to chronic inflammation, which is an important component of numerous pathologies acting on bones and joints, respiratory system, skin, gastrointestinal tract, cardiovascular system and nervous system. Importantly, the processes underlying the acute and chronic inflammation are distinct.

The popularity of non-steroidal anti-inflammatory drugs for the treatment of inflammation and pain is mainly due to their more favorable risk profile compared to other anti-inflammatory drugs.

However, there are some major limitations to long-term NSAID use, including: their propensity to cause gastric injury (digestive ulcers and gastrointestinal bleeding), kidney failure and increased risk of bleeding. In addition, because NSAIDs do not alter disease progression, those who suffer suffer from additional classes of drugs, disease modifying anti-inflammatory drugs (DMARDs) such as methotrexate, etanercept, Corticosteroids). Unfortunately, most of these agents have a narrow range of stability and are characterized by frequent unwanted effects that negatively affect the quality of life.

B. Atopic Dermatitis / Eczema

Atopic dermatitis / eczema (AD) is a chronic skin condition that traditionally results in the formation of blisters that may cause redness, itching, and moistening and stiffness. AD affects more than 15% of developing countries and is often associated with other forms of allergies such as asthma and hay fever (Lee, Y-A., Et al., 2000).

Topical corticosteroids are the gold standard for drug treatment for AD. However, one of the major problems with long-term topical corticosteroid use is the risk of side effects such as skin contraction, oral and allergic contact dermatitis, acne, reduced skin pigmentation and excessive hair growth within the treatment site. Although numerous non-steroidal products are available (eg, pimecrolimus, tacrolimus, antibiotics, cyclosporine, methotrexate, etc.), they are often less effective and none of these are sometimes serious It does not have side effects (see Abramovits, 2005) for review. For example, Pimecrolimus and Tacrolimus have recently received FDA Black Box warnings regarding possible cancer risks. Clearly, there is a need for an effective anti-inflammatory product with less adverse effects.

Anti-inflammatory Effects of Green-lipped Mussels

The "stabilized" lipid extract of New Zealand Green-lipped mussel (NZGLM) has a useful effect that has been demonstrated in several other animal models as well as in chronic inflammatory conditions in humans (Whitehouse et al., 1997; Shiels & Whitehouse, 2000; Tenikoff et al., 2005; Gibson & Gibson, 1998; Emelyanov et al., 2002; Cho et al., 2003; Gruenwald, et al., 2004). Stabilization typically involves the addition of organic acids, such as tartaric acid, to reduce oxidation of PUFAs prior to processing (WO 85/05033; New Zealand Patent 211928). However, not all human studies in which stabilized mussel lipid extracts have been used to treat inflammation have reported positive results (Lau et al., 2004; see Cobb and Ernst, 2006, for review) and inflammation in healthy volunteers. It was not possible to demonstrate a reduction in blood markers in humans (Murphy et al., 2006).

Mussel patent

Mussel patents in the art relate to mussel extract formulations from various aspects, for example:

New Zealand Patent No. 211928 describes a formulation for stabilizing green skin mussel extracts by harvesting organic acids (acetic acid, citric acid, tartaric acid, lactic acid) and / or metal salts and then adding to salted mussels.

New Zealand patent 270754 describes a combination of finely ground mussel extracts suspended in fish oil.

New Zealand Patent No. 314867 describes a green bark hop-derived protein extract mixed with glycosamino-glycans.

New Zealand patent 514389 describes the delivery of green bark mussel extracts at a rate of 0.18 to 114 mg per kg body weight of animal per day in pet food.

Other patents describe extraction methods for concentrating an extract with selected ingredients, for example:

New Zealand Patent No. 329018 describes a method for extracting glycans from mussels that is treated with proteolytic enzymes in water, separating solid residues and recovering glycans from aqueous solutions.

New Zealand patent 510407 describes extracts of green husk mussels containing carbohydrates and lipids and having a protein fraction removed.

New Zealand Patent No. 328489 discloses protein extracts of green skin mussels, as well as stirring the flesh in a phenol solution for 45 minutes, then sucking the top layer with a centrifuge / inhaler and precipitating the protein containing product using ethanol to extract the extract. Describe the method of preparation.

None of the above patents refer to the preparation of concentrated NAE compound extracts or recognize the utility of these compounds in a variety of treatments.

Commercial mussel products

Two widely available commercial preparations of NZGLM are Seatone® and Lyprinol®. Cytones consist of stabilized freeze-dried powders obtained from whole mussel flesh, while ripriol is an oil extracted from stabilized freeze-dried (via supercritical CO ₂ fluid) flesh with olive oil and vitamin E. Formulated (Pharmalink International Ltd., Cayman Islands). Ripriol includes five major lipids: free fatty acids, triglycerides, sterol esters, sterols and phospholipids (Sinclair et al., 2000; Wolyniak et al., 2005). Weighted rich free fatty acids in ripriol include: palmitic acid (C16: 0), linoleic acid (C18: 2n-6), EPA (C20: 5n-3), DHA (C22: 6n-3), palmitoleic acid (C16: 1n-7), C16: 1n-9,7,5 and myristic acid (C14: 0) (Sinclair et al., 2000; Wolyniak et al., 2005).

Although numerous other fatty acids have also been identified (approximately 91 in total), individually they are present only in small amounts (5% w / w or less of total lipids; Wolyniak et al., 2005).

Omega-3 PUFA's account for 40% of total fatty acids and are the most abundant in EPA and DHA (Wolyniak et al., 2005). Evidence suggests that most of the anti-inflammatory activity of ripriol is present in the fatty acid fraction (Whitehouse et al., 1999; Treschow et al., 2007).

Following the process of ripriol, human subjects reduced several proinflammatory compounds, including thromboxane B2, prostaglandin E2, and interleukin-1β, similar to those observed by pollination with low-dose omega-3 hyperunsaturated fatty acid supplements. Levels (Sinclair et al., 2000). This suggests that an important component of ripriol activity may be due to its omega-3 content, which is consistent with recent in vitro evidence (McPhee et al., 2007; Treschow et al., 2007). A hypothesized mode of ripriol action is through inhibition of both 5-lipoxygenase and COX pathways.

Assuming that the above evidences describe the therapeutic effects of PEA and other NAE compounds, it should be appreciated that products rich in these compounds may be beneficial.

It is an object of the present invention to solve the above-mentioned problems or to at least provide a useful choice for the public.

All references, including any patents or patent applications cited herein, are incorporated herein by reference. No reference is made to constitute prior art. The discussion of these references states what their authors claim, and Applicants reserve the right to challenge the accuracy and adequacy of the cited documents. Although numerous prior art publications are mentioned herein; It will be apparent that this reference does not constitute an admission that any of these documents form part of the well-known techniques common in the art, in New Zealand or in any other country.

It is appreciated that the term 'comprises', under various jurisdictions, may be considered to have an exclusive or inclusive meaning. For the purposes of this specification and unless otherwise indicated, the term 'comprises' means inclusive meaning, that is, the inclusion of cited components as well as other non-specific components or elements to which it refers directly. Will have a meaning. This theoretical interpretation will also be used when the term “included” or “comprising” is used in connection with one or more steps in a method or process.

Other aspects and advantages of the invention will be apparent from the following description, given by way of example only.

Subsequent aspects of the present invention will become apparent from the following figures, given by way of example only by reference to the accompanying figures and alone:

1 is a PEA LC / MS chromatogram of a representative green lipped mussel extract showing a peak including PEA (arrowhead, bottom trace) at time = 10 minutes. The relative magnitude of the peaks demonstrates that PEA compensates for a significant portion of the total ion current (top trace) suggesting that a significant amount of PEA is present in the extract.

For purposes of this specification, the term 'ocean based' refers to crustacean organisms that live in or near sea water or fresh water.

The term 'N-acylethanolamine' (NAE) is used to include 'N-acylethanolamide' or other names conventional in the art for compounds of this group, including fatty acid ethanolamide.

According to a first aspect of the invention, there is provided a composition comprising an oil extract derived from a marine based organism rich in fatty acid amide compounds.

According to another aspect of the present invention, there is provided a composition comprising an oil based marine extract derived from an N-acylethanolamine (NAE) compound.

Preferably, the extract is rich in N-palmitoylethanolamide (PEA).

It should be understood by those skilled in the art that PEA also has numerous other names, including but not limited to palmidol, and N- (2-hydroxyethyl) -hexadecanamide.

According to another aspect of the present invention there is provided a composition comprising an oil extract from a marine based organism comprising at least 0.10 μg / g PEA as measured in the dry weight of marine organism flesh prior to extraction.

According to another aspect of the present invention there is provided a composition comprising a marine based organism derived oil extract rich in NAE compound comprising at least 0.10 μg / g PEA as measured in the dry weight of marine organism flesh prior to extraction.

As indicated above, the composition is rich in a range of NAE compounds.

By way of example, the composition includes, but is not limited to, having a large amount of NAEs: N-stearoylethanolamide (18: 0), 70 ng / g in dry flesh prior to extraction; N-oleoylethanolamide (C18: 1), 5 ng / g; N-linoleoylethanolamide (C18: 2), 5 ng / g; And N-arachidonoylethanolamide (C20: 4, anandamide), 8 ng / g. It should not be limited to that the composition may comprise other NAEs and may also include NAEs in various amounts described above and below.

Preferably, the marine based organism is a bivalve mollusk. More preferably, the organism is a type of mussel. Most preferably, the organism is a mussel of species Perna or Mytilus (green or blue mussels, respectively). It should not be limited to that other species may also include, for example, clams and oyster species.

Preferably, the fatty acid amide compound comprises a NAE compound.

Preferably, the NAE compound is: N-myristoylethanolamide, C14: 0; N-palmitoylethanolamide (PEA), C16: 0; N-stearoylethanolamide, C18: 0; N-oleoylethanolamide, C18: 1; N-linoleoylethanolamide, C18: 2; N-arachidonoylethanolamide (anandamide, AEA), C20: 4; N-eicosaenoylethanolamide, C20: 1, and combinations thereof.

In a preferred embodiment, the PEA level in the oil extract is at least 0.10 μg / g as measured in dry tissue of marine organisms prior to extraction. More preferably, the level is at least 0.50 μg / g. In selected embodiments, the level of PEA may be at least 3.0 μg / g.

Preferably, the level of AEA in the oil extract is at least 0.008 μg / g dry tissue of the marine organism before any extract. More preferably, the level is at least 0.01 μg / g dry tissue of marine organisms before any extract. In selected embodiments, the AEA level may be at least 0.05 μg / g.

Preferably, the composition comprises PEA in the oil extract at a level of at least 1.0 μg / g as measured in the dried tissue of the marine organism before any further extraction.

Preferably, the composition comprises AEA in the oil extract at a level of at least 0.09 μg / g as measured in the dried tissue of the marine organism prior to any further extraction.

At least the above levels of PEA and AEA are significantly higher in the oil extracts of the present invention than in the prior art, which teach neither PEA levels nor the levels described. In the closest prior art, Sepe et al. Only teach the production of 0.053 ± 0.0039 μg / g PEA and 0.0018 ± 0.003 μg / g AEA in dry tissue derived from Mediterranean mussels.

It is emphasized that these values refer to PEA and AEA concentrations in dry flesh. It is to be understood that methods of reducing moisture weight (such as drying) also concentrate levels of active compounds including PEA and AEA. The extract of the present invention is substantially more concentrated already before this moisture removal process is terminated.

According to another aspect of the present invention, substantially the fatty acid amide levels in the marine raw material as described above may be concentrated.

For purposes of this specification, the term 'concentrated' refers to an increase in the concentration / amount of fatty acid amide compounds in marine material before any concentration occurs through the drying process.

In one preferred embodiment, the concentration is carried out by recovering, pulverizing and then keeping the marine material ground between 4 and 10 ° C. for at least 24 hours. In one embodiment, the material may be maintained for up to 144 hours.

We have found that by terminating this concentration step, the concentration / amount of fatty acid amide compound increases. It is believed that this is due to the biological reaction that occurs afterwards in the marine material.

Unexpectedly, this process resulted in substantial enrichment of mussel flesh fatty acid amide levels. Also in anticipation, the inventors found that using the described parameters, microbial contamination was not caused to a deleterious level for the extract suitable for human consumption.

As may be appreciated, a general manipulation process for marine based materials requires steps that are taken as quickly as possible to prevent microbial growth, such as freezing or drying. Allowing the material to remain at 100 ° C. for several days is against current practice, but the level of microbial contamination was unexpectedly low at this time and temperature.

As indicated above, the composition is rich in fatty acid amides including PEA and AEA, but may also include other compounds (both fatty acid amides and other compounds). In fact, based on prior art experience with NAEs (published literature), it is highly desirable to have a plurality of NAE compounds present in addition to PEA, as synergism between them is common and therefore, of a product comprising a plurality of NAEs The inventors understand that the potential may be substantially increased rather than including only one NAE.

In a further embodiment, the composition also comprises at least one polyunsaturated fatty acid (PUFA) compound. More preferably, the PUFA compound or compound comprises omega-3 PUFAs. It may be appreciated that it is desirable to have an oil extract that includes both fatty acid amide compounds and PUFA compounds. It may also be recognized that, because fatty acid amide compounds and PUFA compounds have different chemical properties, it is unexpected to obtain extracts with significant levels of both types of compounds. However, it should be appreciated that PUFA compounds are not essential to the compositions of the present invention and should not be considered as limiting.

Preferably, the oil extract comprises at least one PUFA compound.

More preferably, the PUFA or PUFAs are omega-3 type. Preferably, the PUFAs or PUFAs are: 4,7,10,13,16,19-docosahexaenoic acid (DHA; 22: 6n3), 5,8,11,14,17-eicosatetraenoic acid ( EPA, 20: 5n3), 6,9,12,15-octadecatetraenoic acid (OTA, 18: 4n3), 9,12,15-octadecatetrienoic acid (ALA, 18: 3n3), 7,10 , 13,16,19-docosapentaenoic acid (DPA, 22: 5n3), 11,14,17-eicosatrienoic acid (ETA, 20: 3n3), 8,11,14,17-eicosatetra Enoic acid (20: 4n3) and combinations thereof. In a preferred embodiment, the DHA content is at least 3 g / 100 g as measured in the extract.

Preferably, the EPA content is at least 5 g / 100 g as measured in the extract.

Preferably, the composition is formulated as a powder, solution, suspension, suspension, oil, tablet or capsule administered orally. The composition may alternatively be formulated for topical administration, for example creams, lotions, ointments or oils. In a further embodiment, the composition is a solid or liquid food for administration as a 'functional food'.

According to another aspect of the present invention there is provided a formulation for oral or topical administration comprising a therapeutically effective amount of an oil extract from a marine based organism rich in fatty acid amide compounds.

According to another aspect of the present invention there is provided a formulation for oral or topical administration comprising a therapeutically effective amount of an oil extract from a marine based organism enriched in NAEs.

According to another aspect of the present invention there is provided a formulation for oral or topical administration comprising a therapeutically effective amount of an oil extract from a PEA-rich marine based organism.

For oral or topical administration containing a therapeutically effective amount of an oil extract from a marine based organism comprising at least 0.10 μg / g PEA as measured in the dry weight of the marine organism prior to extraction according to another aspect of the present invention. Formulations are provided.

According to another aspect of the invention oral containing a therapeutically effective amount of an oil extract from a marine based organism rich in NAE compounds comprising at least 0.10 μg / g PEA as measured in the dry weight of the marine organism prior to extraction Or formulations for topical administration are provided.

Preferably, the oil extract also contains at least 0.008 μg / g AEA by dry weight from marine organism flesh prior to extraction.

Preferably, the formulation as described above also comprises at least one PUFA. More preferably, the PUFAs or PUFAs are omega-3 fatty acids.

In embodiments contemplated by the inventors, the formulation may comprise a foodstuff-grade antioxidant that includes a carrier material and is also acceptable to aid the long term stability of the extracted active compound.

In one preferred embodiment, the formulation is a capsule for oral administration in which the capsule is filled with an extract of mussel flesh derived oil.

In an alternative preferred embodiment, the formulation is a cream or lotion for oral administration wherein the cream / losion is filled with an oil extract of mussel flesh.

It should be appreciated that such formulations are provided by way of example only and should not be viewed as limiting as it should be appreciated that other formulations may be produced without departing from the scope of the invention as described.

According to another aspect of the present invention there is provided a method of treating inflammation and related pain substantially by oral or topical administration of a composition or formulation as described above.

In one embodiment, the inflammation is of chronic nature and not acute. By way of example, disease states that include chronic inflammation include: eczema / atopic dermatitis, asthma, inflammatory bowel disease (including Crohn's disease and ulcerative colitis), rheumatoid and osteoarthritis, glomerulonephritis, atherosclerosis, Alzheimer's disease and adult respiratory syndrome It includes. As a second example, disease states including chronic pain include neuropathy and arthritis pain.

In an alternative embodiment, said inflammation is of acute nature and comprises the treatment of soft tissue by topical application outside the upper body region of a cream comprising a composition or formulation of the invention.

According to another aspect of the invention there is provided a method of treating a disease associated with inflammation by oral or topical administration of a composition or formulation substantially as described above.

According to another aspect of the present invention there is provided a method of improving the development of inflammation by oral or topical administration of a composition or formulation substantially as described above.

According to another aspect of the invention there is provided a method of treating a skin disease by oral or topical administration of the composition or formulation substantially as described above.

According to another aspect of the present invention there is provided a method of treating a skin disease by oral or topical administration of a composition or formulation substantially as described above.

According to another aspect of the present invention there is provided a method of ameliorating the symptoms of a skin disease by oral or topical administration of a composition or formulation substantially as described above.

In a preferred embodiment, the skin diseases treated above include atopic dermatitis / eczema and contact dermatitis.

In a preferred embodiment, the symptoms to be improved include itching, dryness, edema and reduced proliferation of human keratin.

In one embodiment, the substantial treatment as described above may be introduced into at least one non-steroidal anti-inflammatory drug (NSAID). The extract is even found to enhance the activity of low (therapeutic-sub) doses of NSAIDs, thereby reducing the severity of side effects. In addition, the compositions and formulations of the present invention may also act to enhance NSAID activity to allow for the benefit of reduced efficacy and / or better potency of the NSAID required.

According to another aspect of the present invention, there is provided the use of an oil extract substantially as described above in the preparation of a therapeutic formulation as described above.

From the foregoing, it should be understood that oil extracts are provided which, according to the art, comprise at least substantially elevated levels of active compounds that are beneficial for joint movement. Related oral and topical formulations are also provided for the treatment for various inflammation-related disorders and symptoms thereof.

There is an urgent need for an effective anti-inflammatory product that can be used for a long time with less unwanted effects compared to current pharmaceuticals. Extracts of the present invention can address this need. Daily administration of foods or formulations comprising NAE-enriched marine material extracts is believed to represent an ideal mode of treatment for the prevention or treatment of inflammatory symptoms.

In the following an example is provided which represents a formulation produced and grasped by the present invention relating to an NAE concentrated oil extract.

Example 1

Experiments were performed to determine the PEA levels in blue mussels (pearl mussels) and green lipped mussels (green mussels) commercially harvested from New Zealand water systems on a laboratory scale (test 1). A second experiment was conducted to determine the PEA and AEA concentrations in green lipped mussels on a virtual industrial scale (test 2).

Way

PEA and AEA levels were measured by LC / MS in crude lipids extracted from dried flesh of freshly harvested blue and green mussels and reported in μg / g in dry flesh (Table 1; Giuffrida et al., 2000).

The presence of PEA in the lipid extract of green mussels was confirmed by LC / MS as shown in FIG. 1, in which the arrowheads show PEA peaks.

result

Table 1-NAE Concentrations in Three Different Mussel Types

Dry Mussel Homogenizer (μg / g) PEA AEA M. Galloprovincialis Sepe et ¹ 0.021-0.053 0.0018-0.003 M. Edulis Exam 1 ¹ 0.15-0.39 P. canaliculus Exam 1 ¹ 0.3-0.61 Exam 2 ² 0.54-0.97 0.008-0.054

1. NAE levels were determined by back-calculating from lipid extract results.

2. NAE levels were estimated using conversion factors based on Test 1 results (Table 1).

Conclusion-test 1 and 2

1) PEA levels in green mussels were higher than those of two types of pearl mussels. Since green mussels contain slightly more flesh on the same weight basis and therefore contain a slightly higher portion of the dry matter, this at least partially accounts for higher PEA levels compared to blue mussels.

2) Even though the absolute levels were low, AEA was also present in higher levels in green mussels compared to M. galloprovincialis.

Example 2

A method for concentrating the endogenous level of NAEs in the flesh of green mussels prior to extraction is described. In this example, commercially harvested homogenized mussel flesh derived from mussels was incubated in a laboratory under various conditions, as described below.

Incubation times ranged from 0 to 144 hours at 4 or 10 ° C. in aerobic or anaerobic conditions at atmospheric pressure. Tissue pH was observed at intervals throughout the test. At the end of the incubation period, the tissue was frozen and then freeze-dried. The experimental conditions were tightly controlled to minimize possible confounding factors such as differences between mussel batches, freezing or freeze-drying conditions, and the like. NAE and omega-3 levels were measured in freeze-dried mussels.

The results obtained from the above experiments were 5-10 times more PEA compared to dried flesh made from the same batch of mussels made from homogenized mussels and incubated at 10 ° C. under aerobic conditions for up to 144 hours immediately after homogenization. And AEA.

We also unexpectedly found that the levels of main PUFA omega-3 (DHA, EPA and ALA) in the same dry mussel samples were maintained even after 144 hours incubation.

In addition, the microbial levels in the dried product were unexpectedly low given the time and temperature involved.

Example 3

As described above in the specification, it is understood that the oil extract of the present invention is rich in NAE compounds including, but not limited to, PEA. As an example, the extract of the present invention comprises PEA, AEA and / or PUFAs. Concentration profiles in lipid extracts are: 3.0 to 57.0 μg / g or more PEA; It is expected to be at least 0.1-5.2 μg / g anandamide and / or at least 3 g / 100 g DHA and at least 5 g / 100 g EPA.

Example 4

In one embodiment of the invention, oral formulations are described. As an example, typical oral formulations include mussel lipid extracts (oils) of the invention, with or without carrier lipids and antioxidant (s), contained within gel capsules. Dosages envisaged for adult humans are about 1 to 4 capsules taken 1-2 times a day. It is expected that this dosing regimen will provide relief of pain and edema due to chronic inflammation, particularly including joints (arthritis).

Example 5

In a further embodiment, oil extracts of the invention are used in topical formulations. In one embodiment, the topical formulation may comprise a cream or lotion comprising the mussel lipid extract (oil) of the present invention, with or without carrier lipid and antioxidant (s). Other materials that may be used in the creams and / or lotions include, but are not limited to: propylene glycol, water, glycerin, glycerides, hydrogenated lecithin, betaine, hydroxyethylcellulose, sodium carbomer, squalane, xanthan gum Include.

The cream is applied topically to the affected area one to three times a day for relief of dry skin, erythema, edema, itching and reduction of dermal hypertrophy.

Example 6

In a further embodiment, oral or topical formulations, as described above, are used for the treatment of acute inflammation. In one embodiment, the extract can be administered within 4 hours of the onset of inflammation (eg, soft tissue damage) to reduce the severity of the symptom and possible recovery period.

Example 7

In a further embodiment, the oil extract of the present invention is used in functional foods. In one such embodiment, the food may be taken to reduce the risk of developing an inflammatory condition, including those described above.

In one example, the product may be an oil extract of the invention that is ready for use in a state where it is added to (eg mixed with) a variety of food products by a consumer or by a product manufacturer.

In one embodiment, the oil extract of the present invention is ground to produce microspheres and added to a wide range of food products by the manufacturer.

Example 8

The oil extracts of the invention can be co-administered with NSAIDs. For example, low doses of NSAIDs and oral doses of the extracts of the present invention treat chronic inflammation (e.g., rheumatoid or osteoarthritis: adult dose: 1 to 4 capsules taken at the same time and frequency as NASID) To be used. The extract of the present invention then enhances the anti-inflammatory effects of low doses of NSAIDs and thereby reduces the risk of unwanted effects.

Example 9

The extracts of the present invention were further investigated in comparison with conventional commercial products (green mussel products; described in US Pat. Nos. 6,083,536 and 6,346,278). Extracts of the present invention are rich in fatty acid amides, including NAE compounds such as PEA and AEA. Extracts of the present invention may also comprise PUFA compounds such as EPA and DHA. The relative amounts of each key ingredient in the extracts of the present invention were compared in a commercial green mussel product (Table 2 below).

Table 2-Comparison Results

Inspection substance NAEs (μg / g) Omega 3 Fatty Acids (g / 100 g Oil) ² PEA AEA C22: 6n-3 docosahexanoic acid (DHA) C22: 5n-3 eicoshexahexanoic acid (EPA) C18: 4n-3 stearic acid (OTA) C18: 3n-3 alpha linoleic acid (ALA) Invention Extract 57.0 5.2 12.1 19.4 1.7 1.1 Commercial P. canaliculus products ³ 3.3 0.2 3.1 4.9 0.7 0.4

¹ ethanolamide was quantified by liquid chromatography and mass spectrometry (LC / MS; triple quadruple MS).

² omega-3 fatty acids were quantified by gas chromatography following conversion to their FAME followed by a proven method (AOAC # 963.22).

³ Each capsule contains: 50 mg green mussel extract, 100 mg olive oil and 0.225 mg d-alpha-tocopherol.

The results shown in Table 2 indicate that the extract of the present invention contains significantly higher levels of NAEs (17 times higher PEA and 26 times higher AEA) and omega-3 (particularly DHA and EPA) compared to commercial green mussel products. .

It is to be understood that the formulations using the compositions of the present invention may be simply undiluted extracts or may be mixed with other ingredients to different degrees than in a commercial green mussel product, thus diluting the components.

It is to be understood that aspects of the invention have been described by way of example only and that modifications and additions may be made thereto without departing from the scope of the invention as defined in the appended claims.

references

Calignano, A., La Rana, G., Giuffrida, A. & Piomelli, D. (1998) Control of pain initiated endogenous cannabinoids. Nature , 394: 277-281.

2. Calignano, A., La Rana, G., & Piomelli, D. (2001). Antinociceptive activity of the endogenous fatty acid amide, palmitoylethanolamide. European Journal of Pharmacology , 419 (2-3), 191-198.

3. Cho, SH, Jung, YB, Seong, S. C, Park, HB, Byun, KY, Lee, DC, et al. (2003). Clinical efficacy and safety of Lyprinol, a patented extract from New Zealand green-lipped mussel (Perna Canaliculus) in patients with osteoarthritis of the hip and knee: a multicenter 2-month clinical trial. Allergie et Immunologies , 35 (6), 212-216.

4. Cobb, CS, & Ernst, E. (2006). Systematic review of a marine nutriceutical supplement in clinical trials for arthritis: the effectiveness of the New Zealand green-lipped mussel Perna canaliculus. Clinical Rheumatology , 25 (3), 275-284.

5. Coburn, AF, CE Graham, and J. Haninger. 1954.The effect of egg yolk in diets on anaphylactic arthritis (passive Arthus phenomenon) in the guinea pig. Journal of Experimental Medicine , 100: 425-35.

6. Conti, S., Costa, B., Colleoni, M., Parolaro, D., & Giagnoni, G. (2002). Antiinflammatory action of endocannabinoid palmitoylethanolamide and the synthetic cannabinoid nabilone in a model of acute inflammation in the rat. British Journal of Pharmacology , 735 (1), 181-187.

7.Darmani, NA, Izzo, AA, Degenhardt, B., Valenti, M., Scaglinone, G., Capasso, R., Sorrentini, I., Di Marzo, V. (2005) Involvement of the cannabimimetic compound, N -palmitoylethanolamine, in inflammatory and neuropathic conditions: review of the available pre-clinical data, and first human studies. Neuropharmacology , 48: 1154-1163.

8. De Petrocellis, L., Davis, JB & Di Marzo, V. (2001) Palmitoylethanolamide enhances anandamide stimulation of human vanilloid VR1 receptors. FEBS Letters , 506 (3): 253-256.

9. Devane, WA, Hanus, L., Breuer, A., Pertwee, RG, Stevenson, LA, Griffin, G., Gibson, D., Mandelbaum, A, Etinger, A, Machoulam, R. (1992) Isolation and structure of a brain constituent that binds to the cannabinoid receptor. Science , 258 (5090): 1946-1949.

10. Di Marzo, V., Melck, D., Orlando, P., Bisogno, T., Zagoory, O., Bifulco, M., Vogel, Z. & De Petrocellis, L. (2001). Palmitoylethanolamide inhibits the expression of fatty acid amide hyrolase and enhances the anti-proliferative effect of anandamide in human breast cancer cells. Biochemical Journal , 358 (pt 1): 249-255.

Emelyanov, A., Fedoseev, G., Krasnoschekova, O., Abulimity, A., Trendeleva, T., & Barnes, PJ (2002). Treatment of asthma with lipid extract of New Zealand green-lipped mussel: a randomised clinical trial. European Respiratory Journal , 20 (3), 596-600.

12. Farquhar-Smith, WP, & Rice, AS (2001). Administration of endocannabinoids prevents a referred hyperalgesia associated with inflammation of the urinary bladder. Anesthesiology , 94 (3), 507-513.

13. Fowler, CJ. (2003) Plant-derived, synthetic and endogenous cannabinoids as neuroprotective agents non-psychoactive cannabinoids, 'entourage' compounds and inhibitors of N-acyl ethanolamine breakdown as therapeutic strategies to avoid psychotropic effects. Brain Research Reviews , 41: 26-43.

14. Gibson, SLM, and RG Gibson (1998). The treatment of arthritis with a lipid extract of Pema canaliculus: a randomized trial. Complementary Therapies in Medicine , 6: 122-26.

15. Gruenwald, J., Graubaum, H.-J., Hansen, K., & Grube, B. (2004). Efficacy and tolerability of a combination of Lyprinol and high concentrations of EPA and DHA in inflammatory rheumatoid disorders. Advances in Therapy , 21 (3), 197-201

16. Gulaya, NM, Kuzmenko, AI, Margitich, VM, Govseeva, NM, Melnichuk, SD, Goridko, TM & Zhukov, AD (1998). Long-chain N-acylethanolamines inhibit lipid peroxidation in rat liver mitochondria under acute hypoxic hypoxia. Chemistry & Physics of Lipids , 97 (1): 49-54.

17. Jaggar, SI, Hasnie, FS, Sellaturay, S., & Rice, AS (1998). The anti-hyperalgesic actions of the cannabinoid anandamide and the putative CB2 receptor agonist palmitoylethanolamide in visceral and somatic inflammatory pain. Pain , 76 (1-2), 189-199.

18.Kahlich, R., Klima, J., Cihla, F., Frankova, V., Masek, K., Rosicky, M., Matousek, F. & Bruthans, J. (1979) Studies on prophylactic efficacy of N -2-Hydroxyethyl palmitide (Impulsin) in acute respiratory infections. Serologically controlled field trials. Journal of Hygiene, Epidemiology, Microbiology and Immunology , 23 (1): 11-24.

19. Lambert, DM, Vandevoorde, S., Jonsson, EN. & Fowler, CJ. (2002) the palmitoylethanolamide family: a new class of anti-inflammatory agents? Current Medicinal Chemistry , 9 (6): 663-674.

20. Lau, CS, Chiu, PKY, Chu, EMY, Cheng, IYW, Tang, WM, Man, RYK, et al. (2004). Treatment of knee osteoarthritis with Lyprinol, lipid extract of the green-lipped mussel-a double blind placebo-controlled study. Progress in Nutrition , 6 (1), 17-31.

21. Lee, YA, Wahn, U., Kehrt, R., Tarani, L., Businco, L., Gustafsson, D., et al. (2000). A major susceptibility locus for atopic dermatitis maps to chromosome 3q21. Nature Genetics , 26 (4), 470-473.

Lo Verme, J., Fu, J., Astarita, G., La Ranna, G., Russo, R., Calignano, A. & Piomelli, D. (2005a) The nuclear receptor peroxisome proliferator-activated receptor- alpha mediates the anti-inflammatory actions of palmitoylethanolamide. Molecular Pharmacology , 67: 15-19.

23.Lo Verme, J., La Ranna, G., Russo, R., Calignano, A. & Piomelli, D. (2005b). The search for the palmitoylethanolamide receptor. Life Sciences , 77: 1685-1698.

24. Masek, K., Perlik, F., Klima, J. & Kahlich, R. (1974) Prophylactic efficacy of N-2-Hydoxyethyl palmitamide (Impulsin) in acute respiratory tract infections. European Journal of Clinical Pharmacology , 7: 415-419.

25. McPhee, S., LD Hodges, PFA Wright, PM Wynne, N. Kalafatis, DW Harney, and TA Macrides (2007). Anti-cyclooxygenase effects of lipid extracts from the New Zealand green-lipped mussel, Perna canaliculus. Comparative Biochemistry & Physiology Part B, Biochemistry & Molecular Biology , 146: 346-56.

26. Murphy, KJ. Et al., (2002) Lipid, FA and sterol composition of New Zealand green lipped mussel (Perna canaliculus) and Tasmanian blue mussel (Mytilus edulis). Lipids , 37 (6): 587-595.

27. Murphy, KJ. Et al., (2003) Fatty acid and sterol composition of frozen and freeze-dried New Zealand green lipped mussel (Perna canaliculus) from three sites in New Zealand. Asia Pacific Journal of Clinical Nutrition , 12 (1): 50-60.

28. Murphy, KJ, Galvin, K., Kiely, M., Morrissey, PA, Mann, NJ, & Sinclair, AJ (2006). Low dose supplementation with two different marine oils does not reduce pro-inflammatory eicosanoids and cytokines in vivo. Asia Pacific Journal of Clinical Nutrition , 15 (3), 418-424.

29. Rohde, H. & Ghyczy, M. (2003). Behandlung des chronischen analekzems miteiner endocannabinoidhaltigen pflegecreme. Haut , 7 (Band XIV): 281-282.

30.Sepe, N., L. De Petrocellis, F. Montanaro, G. Cimino, and Di Marzo, V. (1998) Bioactive long chain N-acylethanolamines in five species of edible bivalve molluscs: possible implications for mollusc physiology and sea food industry. B iochemica et Biophysica Acta , 1389: 101-111.

31. Shiels, IA, and MW Whitehouse (2000). Lyprinol: anti-inflammatory and uterine-relaxant activities in rats, with special reference to a model for dysmenorrhea. Allergie et Immunologies , 32: 279-283.

32. Sinclair, AJ, Murphy, KJ & Li, D. (2000). Marine Lipids: overview "news insights and lipid composition of Lyprinol. Allergie et Immunologie , 32 (7): 261-71.

33. Szepietowski, JC, Szepietowski, T. & Reich, A. (2005). Efficacy and tolerance of the cream containing structured physiological lipids with endocannabinoids in the treatment of uremic pruritis: a preliminary study. Acta Dermatovenerol Croat , 13 (2): 97-103.

34. Tenikoff, D., KJ Murphy, M. Le, PR Howe, and GS Howarth (2005). Lyprinol (stabilised lipid extract of New Zealand green-lipped mussel): a potential preventative treatment modality for inflammatory bowel disease. Journal of Gastroenterology , 40: 361-5.

35. Treschow, AP, LD Hodges, PFA Wright, PM Wynne, N. Kalafatis, and TA Macrides (2007). Novel anti-inflammatory omega-3 PUFAs from the New Zealand green-lipped mussel, Perna canaliculus. Comparative Biochemistry & Physiology Part B, Biochemistry & Molecular Biology , 147: 645-56.

36. Whitehouse, MW, TA Macrides, N. Kalafatis, WH Betts, DR Haynes, and J. Broadbent (1997). Anti-inflammatory activity of a lipid fraction (Lyprinol) from the NZ Green-lipped mussel. Inflammopharmacology , 5: 237-246.

37. Whitehouse, MW, Roberts, MS & Brooks, PM (1999). Over the counter (OTC) oral remedies for arthritis and rheumatism: how effective are they? Inflammopharmacology , 7 (2): 89-105.

38. Wolyniak, CJ. Et al., (2005) Gas Chromatography-chemical ionization-mass spectrometry fatty acid analysis of a commercial supercritical carbon dioxide lipid extract from New Zealand Green-Lipped mussel (Perna canaliculus). Lipids , 40: 355-360.

Statement of the corresponding application

This application is based on a provisional application filed with respect to New Zealand Patent Application No. 552238, which is hereby incorporated by reference in its entirety.

Claims (43)

A composition containing an oil extract from a marine based organism rich in fatty acid amide compounds. The composition of claim 1 wherein said fatty acid amide compound comprises an N-acylethanolamine (NAE) compound. A composition comprising an oil based marine derived organism rich in N-acylethanolamine (NAE) compounds. The composition of any one of the preceding claims, wherein said extract is rich in N-palmitoylethanolamide (PEA). A composition comprising a marine based organism derived oil extract rich in NAE compounds comprising at least 0.10 μg / g PEA as measured by the dry weight of marine organism flesh prior to extraction. The composition of claim 1, wherein the composition is selected from lauroylethanolamide, C12: 0; N-myristoylethanolamide, C14: 0; N-palmitoylethanolamide (PEA), C16: 0; N-stearoylethanolamide, C18: 0; N-oleoylethanolamide, C18: 1; N-linoleoylethanolamide, C18: 2; N-arachidonoylethanolamide (AEA), C20: 4; A composition comprising NAEs selected from N-Eicosaenoylethanolamide, C20: 1, and combinations thereof. The composition of claim 1, wherein the composition comprises PEA at a level in excess of 0.20 μg / g as measured by dry weight of marine organism flesh prior to extraction. The composition of claim 1, wherein said composition comprises AEA at a level above 0.008 μg / g as measured by dry weight of marine organism flesh prior to extraction. The composition of claim 1, wherein the composition comprises PEA at a level in excess of 1.0 μg / g as measured in the dried marine organism prior to extraction. The composition of any one of the preceding claims, wherein the composition comprises AEA at a level above 0.09 μg / g as measured in dried marine organisms prior to extraction. The composition of any one of the preceding claims, wherein said marine based organism is a bivalve mollusk. The composition of claim 1, wherein said organism is a mussel derived from the genus Perna or Mytilus. The composition of any one of the preceding claims wherein the marine based organism is subjected to a concentration step prior to extraction to increase fatty acid amide content. The composition of claim 13, wherein the step of concentrating comprises forming a pulverized mass and maintaining the mass at a temperature between 4 and 10 ° C. for a period of at least 24 hours. The composition of claim 1, wherein said composition also comprises at least one polyunsaturated fatty acid (PUFA) compound. The composition of claim 15 wherein the PUFA compound or compounds is an omega-3 type fatty acid. The method according to claim 15 or 16, wherein the PUFA compound or compounds are: 4,7,10,13,16,19-docosahexaenoic acid (DHA; 22: 6n3), 5,8,11,14, 17-Eicosatetraenoic acid (EPA, 20: 5n3), 6,9,12,15-octadecatetraenoic acid (OTA, 18: 4n3), 9,12,15-octadecatetrienoic acid (ALA1 18 : 3n3), 7,10,13,16,19-docosapentaenoic acid (DPA, 22: 5n3), 11,14,17-eicosatrienoic acid (ETA, 20: 3n3), 8,11, 14,17-eicosatetraenoic acid (20: 4n3) and combinations thereof. 18. The composition of claim 17, wherein said DHA content is at least 4 g / 100 g in the crude extract. 18. The composition of claim 17, wherein said EPA content is at least 6 g / 100 g in crude extract. The composition of any one of the preceding claims wherein the composition is formulated for oral administration. The composition of claim 1, wherein the composition is formulated for topical administration. A formulation for oral or topical administration containing a therapeutically effective amount of a composition according to any one of claims 1 to 19. 20. A formulation for oral or topical administration according to any one of claims 1 to 19 containing a composition comprising at least 0.10 μg / g PEA as measured in the dry weight of the marine organism flesh prior to extraction. The formulation of claim 22 or 23, wherein the formulation is a capsule for oral administration. The formulation of claim 22 or 23, wherein the formulation is a cream for topical administration. Use of a composition according to any one of claims 1 to 19 in the manufacture of a medicament for oral or topical administration in the treatment of inflammation and diseases involving inflammation. Use of a formulation according to any one of claims 22 to 25 in the manufacture of a medicament for oral or topical administration in the treatment of inflammation and diseases involving inflammation. Use according to claim 26 or 27, wherein the inflammation is chronic. Use according to claim 26 or 27, wherein said disease / pathology is asthma. Use according to claim 26 or 27, wherein said disease / pathology is an inflammatory bowel disease. Use according to claim 26 or 27, wherein the disease / pathology is rheumatoid arthritis. Use according to claim 26 or 27, wherein the disease / pathology is osteoarthritis. Use according to claim 26 or 27, wherein said disease / pathology is glomerulonephritis. Use according to claim 26 or 27, wherein said disease / pathology is atherosclerosis. Use according to claim 26 or 27, wherein the disease / pathology is Alzheimer's disease. Use according to claim 26 or 27, wherein the disease / pathology is neuropathic pain. The use of claim 26 or 27, wherein said disease / pathology comprises skin. 38. The use of claim 37, wherein the skin disease to be treated comprises atopic dermatitis / eczema or contact dermatitis. Use according to claim 26 or 27, wherein said inflammation is acute. 20. A method of treating inflammation in a non-human animal by oral or topical administration of the composition according to any one of claims 1 to 19. A method of treating inflammation in a non-human animal by oral or topical administration of the formulation according to any one of claims 22 to 25. 42. The method of claim 40 or 41, wherein said inflammation is chronic inflammation. 42. The method of claim 40 or 41, wherein said inflammation is acute inflammation.