AU2015372438B9

AU2015372438B9 - Formulation and method of use

Info

Publication number: AU2015372438B9
Application number: AU2015372438A
Authority: AU
Inventors: Amanda Bernadette SMITH
Original assignee: Sanofi Aventis Healthcare Pty Ltd
Current assignee: Sanofi Aventis Healthcare Pty Ltd
Priority date: 2014-12-24
Filing date: 2015-12-22
Publication date: 2021-07-08
Anticipated expiration: 2035-12-22
Also published as: WO2016101028A1; CN107205978B; AU2015372438B2; AU2015372438A1; CN107205978A; NZ733215A

Abstract

A formulation comprising one or more omega-3 fatty acids and/or one or more derivatives thereof, willow bark or an extract, one or more components and/or derivatives thereof; and withania or an extract, one or more components and/or derivatives thereof is capable of treating or preventing pain, inflammation and/or improving mobility in a subject. A method of treating or preventing one or more diseases, disorders or conditions such as pain, inflammation and/or lack of mobility includes administration of the formulation to a subject.

Description

TITLE

FORMULATION AND METHOD OF USE TECHNICAL FIELD THIS INVENTION relates to treatment of pain and/or inflammation. More particularly, this invention relates to a formulation comprising omega-3 fatty acids, willow bark and withania that has particular efficacy in treating joint pain and/or inflammation. The formulation may assist in improving patient mobility.

BACKGROUND

Pain is a neurological response to stimulation of peripheral nerves ("nociceptive pain"), inflammatory diseases or conditions and/or damage or destruction of elements of the nervous system ("neuropathic pain") that is perceived by the sufferer. Joint pain can be particularly problematic due to the chronic, repeated episodes of pain as well as the severity of the pain. This type of pain is often associated with bone joints or other skeletal structures where bone or cartilage damage and/or inflammation have occurred. These include pain in the knee, ankle, hip, back, neck, elbow, finger, wrist and shoulder which may be caused by structural damage and/or by inflammation. In some cases, the pain is exacerbated by the reduced mobility of the sufferer, which can in turn lead to weight gain, thereby adding to the physical strain on the painful joints.

Both acute and chronic pain can be extremely discomforting and debilitating for a sufferer and hence considerable effort has been expended in developing drugs that treat pain, typically referred to as "analgesics" or "painkillers". As a result, there are many potent drugs for treating pain, whether mild pain such as associated with headaches, fever or toothache or severe pain such as following a serious injury or surgery. One particularly popular and effective class of drugs for treating joint pain are non-steroidal anti-inflammatory drugs (NSAIDs). These provides analgesic and antipyretic effects, and, in higher doses, anti-inflammatory effects.

While drugs such as NSAIDs have proven to be potent and effective for treating pain and inflammation, patients can overuse these drugs and suffer side effects that detract from their positive effects. There is also concern that drugs such as NSAIDs merely mask the pain and inflammation without any treatment of the underlying cause of the pain. Increasingly, patients have turned to natural or traditional medicines for the treatment of pain and inflammation. SUMMARY

The present invention is directed to formulations and methods of treatment to treat pain and/or inflammation that comprise natural or traditional medicine constituents which act together to prevent or treat pain and/or inflammation.

In a particular broad form, the invention relates to a formulation comprising omega-3 fatty acids, willow bark and withania for use in preventing, relieving or otherwise treating pain and/or inflammation.

In a first aspect, the invention provides a formulation comprising:

(i) one or more omega-3 fatty acids and/or one or more derivatives thereof;

(ii) willow bark or an extract, one or more components and/or derivatives thereof; and

(iii) withania or an extract, one or more components and/or derivatives thereof.

In an embodiment, the formulation further comprises at least one pharmaceutically acceptable carrier, diluent and/or excipient.

In one embodiment, the formulation is capable of treating or preventing pain in a subject.

In another embodiment, the formulation is capable of treating or preventing inflammation in a subject.

In a related embodiment, the formulation is capable of improving the mobility of a subject.

In a second aspect, the invention provides a method of producing a formulation, said method including the step of combining:

(i) one or more omega 3 fatty acids and/or one or more derivatives thereof;

(iii) withania or an extract, one or more components and/or derivatives thereof;

to thereby produce the formulation.

In an embodiment, the method further includes combining (i)-(iii) with at least one pharmaceutically acceptable carrier, diluent and/or excipient. In one embodiment, the formulation treats or prevents pain.

In another embodiment, the formulation treats or prevents inflammation.

In a related embodiment, the formulation improves the mobility of a subject.

In a third aspect, the invention provides a method of treating or preventing a disease, disorder or condition in a subject, said method including administering to the subject an effective amount of the formulation according to the first aspect or an effective amount of the formulation produced according to the second aspect to thereby treat or prevent the disease, disorder or condition in the subject.

In one embodiment, the method treats or prevents pain in the subject.

In another embodiment, the method treats or prevents inflammation in the subject.

In a related embodiment, the method improves the mobility of the subject.

In certain embodiments, the pain and/or inflammation or a lack of mobility is associated with, or caused by, a disease, disorder or condition of bones and/or bone joints.

Throughout this specification, unless otherwise indicated, "comprise", "comprises" and "comprising" are used inclusively rather than exclusively, so that a stated integer or group of integers may include one or more other non-stated integers or groups of integers.

As used in this specification the indefinite articles "a" and "an" may refer to one entity or a plurality of entities and are not to be read or understood as being limited to a single entity.

BRIEF DECSRIPTION OF THE FIGURES

Figure 1 : (A) Body weight change from Day (-22) to 0; and (B) body weight change from Day 0 to 18.

Figure 2: Ankle diameter over time;

Figure 3 : Ankle diameter with AUC calculation;

Figure 4: Final paw weight;

Figure 5 : Relative liver weight;

Figure 6: Relative spleen weight;

Figure 7: Relative thymus weight;

Figure 8: Von Frey analysis; Figure 9: Operant pain readout (seconds);

Figure 10: Operant pain with AUC calculation;

Figure 11 : Individual histopathology scores (ankle);

Figure 12: Summed histopathology scores (ankle);

Figure 13 : Periosteal bone measure;

Figure 14: Individual histopathology scores (knee); and

Figure 15: Summed histopathology scores (knee).

DETAILED DESCRIPTION

The invention is at least partly predicated on the surprising effect of omega-3 fatty acids, willow bark and withania in combination for treating and preventing pain, in particular pain associated with inflamed and/or damaged bone joints. Omega-3 fatty acids, willow bark and withania appear to act synergistically to treat or prevent pain, thereby providing a particularly effective pain remedy based on natural products. In particular, the formulation and method disclosed herein may provide a particularly effective remedy for joint pain, by synergistically targeting the prostaglandin pathway and COX-2 to reduce inflammation while also having a positive effect on the central nervous system that assist alleviating the sufferer's perception of pain. One particular benefit of the invention is an improvement in the mobility of the sufferer which can lead to a loss in weight, which in turn can lead to a reduction in pain and/or inflammation.

An aspect of the invention provides a formulation comprising:

(i) one or more omega-3 fatty acids and/or one or more derivatives thereof;

Another aspect of the invention provides a method of producing a formulation by combining: one or more omega-3 fatty acids and/or one or more derivatives thereof; willow bark or an extract, one or more components and/or derivatives thereof; and withania or an extract, one or more components and/or derivatives thereof.

In one embodiment, the formulation is capable of treating or preventing pain in a subject. In another embodiment, the formulation is capable of treating or preventing inflammation in a subject.

Suitably, the formulation comprises a therapeutically effective amount each of the one or more omega-3 fatty acids and/or one or more derivatives thereof, willow bark or an extract, one or more components and/or derivatives thereof and withania or an extract, one or more components and/or derivatives thereof.

The term "therapeutically effective amounf describes a quantity or concentration of a therapeutically active substance that alone or in combination with one or more other therapeutically active substances achieves a desired effect in a subject treated with the formulation. In the context of the present invention, the desired effect is the treatment or prevention of pain and/or inflammation. Preferably, the therapeutically effective amount does not elicit or cause any undesired "side-effects", or elicits an acceptable or tolerably low level of undesired "side-effects" in the subject, when the formulation is administered to the subject.

As generally used herein, the term extract refers to a portion, fraction or component obtained, isolated or purified from a source, wherein the extract comprises one or more therapeutically active substances, compounds or molecules present in the source. The therapeutically active substances, compounds or molecules in the extract may be in a more concentrated or enriched form compared to the source.

The term "derivative" refers to a modified form of a particular therapeutically active substance, compound or molecule. In some embodiments, the derivative is a chemically modified or otherwise related form of the particular compound, substance or molecule which at least partly retains or improves the therapeutic activity of the substance, compound or molecule.

An "omega-3 fatty acid" includes and encompasses polyunsaturated fatty acids with a double bond (C=C) at the third carbon atom from the methyl (CH₃) end of the carbon chain. This numbering of the fatty acid is taken from the location of the first double bond, counted from the methyl end, that is, the omega (o ) or the n- end. By way of example, linolenic acid is an 18 carbon omega 3 fatty acid having double bonds at carbons 9, 12 and 15. The primary essential omega-3 fatty acids in the human diet include docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and a-linolenic acid (ALA) which may be converted to EPA and DHA in the mammalian body. Food sources of omega-3 fatty acids include fish such as tuna, pink salmon, mackerel, herring, cod, molluscs such as squid and octopus, crustaceans including Decapoda such as crabs, lobsters, shrimp and Euphausia such as krill, algae, plants and plant nuts and seeds, including oils or other extracts of these sources include fish oils such as menhaden oil, salmon oil and cod liver oil, krill oil, calamari oil, canola oil, soybean oil, butter fat oil, and corn oil.

It will be appreciated that formulation disclosed herein may comprise one or more isolated, purified or synthetic omega-3 fatty acids and/or may comprise sources of omega-3 fatty acids, or extracts thereof, such as fish oils, crustacean oils, plant oils or seed oils, although without limitation thereto. Preferably, the one or more omega-3 fatty acids are in the form of krill oil. A preferred source of krill oil is obtained from Antarctic krill {Euphausia superba).

Suitably, the formulation comprises 100 mg to 3 g omega-3 fatty acids, such as in the form of krill oil. Preferably, the formulation comprises 300 mg to 2 g omega-3 fatty acids. In embodiments, the formulation comprises 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500, 1520, 1530, 1540, 1550, 1560, 1570, 1580, 1590, 1600, 1610, 1620, 1630, 1640, 1650, 1660, 1670, 1680, 1690, 1700, 1710, 1720, 1730, 1740, 1750, 1760, 1770, 1780, 1790, 1800, 1810, 1820, 1830, 1840, 1850, 1860, 1870, 1880, 1890, 1900, 1910, 1920, 1930, 1940, 1950, 1960, 1970, 1980, 1990, 2000, 2010, 2020, 2030, 2040, 2050, 2060, 2070, 2080, 2090, 2100, 2110, 2120, 2130, 2140, 2150, 2160, 2170, 2180, 2190, 2200, 2210, 2220, 2240, 2250, 2260, 2270, 2280, 2290, 2300, 2310, 2320, 2330, 2340, 2350, 2360, 2370, 2380, 2390, 2400, 2410, 2420, 2430, 2440, 2450, 2460, 2470, 2480, 2490, 2500, 2510, 2520, 2530, 2540, 2550, 2560, 2570, 2580, 2590, 2600, 2610, 2620, 2630, 2640, 2650, 2660, 2670, 2680, 2690, 2700, 2710, 2720, 2730, 2740, 2750, 2760, 2770, 2780, 2790, 2800, 2810, 2820, 2830, 2840, 2850, 2860, 2870, 2880, 2890, 2900, 2910, 2920, 2930, 2940, 2950, 2960, 2970, 2980 or 2990 mg omega-3 fatty acids or the equivalent such as in the form of krill oil, or any range or amount between any of these stated amounts.

As hereinbefore described, the formulation further comprises willow bark and/or one or more extracts or derivatives thereof.

As used herein, the term willow" and "willow bark" relate to plants or bark extracts of plants of the genus Salix that contain or comprise salicin. Some Salix species are particularly high in salicin content including Salix alba L. (white willow), Salix fragilis L., Salix purpurea L. (purple willow), Salix daphnoides, and Salix pentandra L. Traditional sources for medicinal willow products are the bark of young, two to three year-old willow branches harvested during the early spring and dried. Salicin is converted to salicylic acid in the mammalian body. Methylsalicylic acid or "aspirin" is a chemically synthesized derivative of salicylic acid.

It will therefore be understood that the formulation disclosed herein may comprise willow bark or extracts or derivatives thereof such as salicin, salicylic acid and/or acetylsalicylic acid, although without limitation thereto.

The formulation may comprise willow bark such that the amount of salicin is about 10 mg to 1000 mg. Preferably, the formulation comprises 20 mg to 500 mg salicin. More preferably, the formulation comprises 60 mg to 240 mg salicin. In embodiments, the formulation may comprise willow bark such that the amount of salicin is about 10, 20, 30, 40, 50, 60, 70, 80 , 90„ 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480 , 490 , 500 , 510 , 520 , 530, 540, 550, 560,570, 580, 590 , 600, 610, 620, 630 , 640 , 650 , 660 , 670 , 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, or 990 mg salicin, or any range or amount between any of these stated amounts.

Generally, this is standardized as the amount of salicin in an ethanolic extract of willow bark. Typically, 2 g of willow bark may comprise about 20 mg total salicin. By way of example, the proprietary preparation Assalix® is standardized to 15% salicin, corresponding to about 240 mg of salicin per tablet.

In alternative embodiments, the formulation may comprise plants or plant extracts of other salicin-containing plants not in the Salix genus including Cassia acutifolia, Polygala senega L. and Dorema ammoniacum, although without limitation thereto.

As hereinbefore described, the formulation disclosed herein further comprises withania and/or one or more extracts or derivatives thereof.

As used herein, "withania" refers to plants and extracts of plants of the species Withania somnifera, also known as Physalis somnifera and ashwagandha which is a small evergreen shrub that grows to about four or five feet tall. It is found in dry areas of India and the Middle East, as well as parts of Africa. The word ashwagandha means "odor of the horse" due to its distinctive smell. The therapeutically active constituents of withania plants and plant extracts may include alkaloids and steroidal lactones that together are called withanolides (particularly withaferin A). The whole plant, leaves and/or the long, brown tuberous roots, or extracts thereof, are typically used for medicinal purposes.

Suitably, the formulation may comprise about O. lg to 20 g of withania plant. Preferably, the formulation comprises 0.5g to 10 g withania plant. More preferably, the formulation comprises lg to 6 g withania plant. In embodiments, the formulation may comprise about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0,

I .1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 3.0, 3.1, 3.2, 3 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0. 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0. 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0. 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7,

I I .8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17.0, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18.0. 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19.0. 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8 or 19.9 g of withania plant, or any range or amount between any of these stated amounts.

Suitably, the formulation may compromise about 10 to 6000 mg of an aqueous or ethanolic extract of withania roots and/or leaves. Preferably the formulation may compromise about 60 to 2000 mg of the aqueous or ethanolic extract of withania roots and/or leaves, more preferably the formulation may compromise about 125 to 750 mg of the aqueous or ethanolic extract of Withania roots and/or leaves.

In embodiments, the formulation may comprise 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400,

410, 420, 430, 440, 450 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930. 940, 950, 960, 970, 980, 990, 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070 1080, 1090, 1100 1110, 1120, 1130 1140, 1150, 1160, 1170, 1180, 1190, 1200 1210, 1220, 1230 1240, 1250, 1260 1270, 1280, 1290, 1300, 1310, 1320, 1330 1340, 1350, 1360 1370, 1380, 1390 1400, 1410, 1420, 1430, 1440, 1450, 1460 1470, 1480, 1490 1500, 1520, 1530 1540, 1550, 1560, 1570, 1580, 1590, 1600 1610, 1620, 1630 1640, 1650, 1660 1670, 1680, 1690, 1700, 1710, 1720, 1730 1740, 1750, 1760 1770, 1780, 1790 1800, 1810, 1820, 1830, 1840, 1850, 1860 1870, 1880, 1890 1900, 1910, 1920 1930, 1940, 1950, 1960, 1970, 1980, 1990 2000, 2010, 2020 2030, 2040, 2050 2060, 2070, 2080, 2090, 2100, 2110, 2120 2130, 2140, 2150 2160, 2170, 2180 2190, 2200, 2210, 2220, 2240, 2250, 2260 2270, 2280, 2290 2300, 2310, 2320 2330, 2340, 2350, 2360, 2370, 2380, 2390 2400, 2410, 2420 2430, 2440, 2450 2460, 2470, 2480, 2490, 2500, 2510, 2520 2530, 2540, 2550 2560, 2570, 2580 2590, 2600, 2610, 2620, 2630, 2640, 2650 2660, 2670, 2680 2690, 2700, 2710 2720, 2730, 2740, 2750, 2760, 2770, 2780 2790, 2800, 2810 2820, 2830, 2840 2850, 2860, 2870, 2880, 2890, 2900, 2910 2920, 2930, 2940 2950, 2960, 2970 2980, 2990, 3000, 3010, 3020, 3030, 3040 3050, 3060, 3070 3080, 3090, 3100 3110, 3120, 3130, 3140, 3150, 3160, 3170 3180, 3190, 3200 3210, 3220, 3230 3240, 3250, 3260, 3270, 3280, 3290, 3300, 3310, 3320, 3330, 3340, 3350, 3360, 3370, 3380, 3390, 3400, 3410, 3420, 3430, 3440, 3450, 3460, 3470, 3480, 3490, 3500, 3510, 3520, 3530, 3540, 3550, 3560, 3570, 3580, 3590, 3600, 3610, 3620, 3630, 3640, 3650, 3660, 3670, 3680, 3690, 3700, 3710, 3720, 3730, 3740, 3750, 3760, 3770, 3780, 3790, 3800, 3810, 3820, 3830, 3840, 3850, 3860, 3870, 3880, 3890, 3900, 3910, 3920, 3930, 3940, 3950, 3960, 3970, 3980 or 3990 mg of an aqueous or ethanolic extract of withania roots and/or leaves, or any range or amount between any of these stated amounts.

In one embodiment, the withania extract is Sensoril®, a commercially available product sold as an ashwagandha extract.

The formulation disclosed herein may further comprise one or more other components or constituents. Non-limiting examples include components or constituents that facilitate weight loss, enhance palatability, facilitate joint repair, modulate the immune system, improve mood, promote normal sleep patterns, aid nutrition and/or enhance blood circulation, although without limitation thereto.

In embodiments relating to facilitating weight loss, the component or constituent may suppress appetite, elicit a feeling of "fullness", stimulate metabolism (e.g "fat burners"), act as a diuretic or otherwise facilitate weight loss. Particular non-limiting examples include Acai berry, African mango (Irvingia gabonensis), Amorphaphallus rivieri, Barley Grass, Beta-glucan, Bitter orange {Citrus aurantium), Bromelian, Capsaicin (Capsicum annuum), Caralluma fimbriata, Cayenne, Chitosan, cinnamon extract and/or cinnamon powder (from Cinnamomum spp.), Citrimax, Coleus forskohlii, forskolin, Fucoidin, Fucus versiculosis, galactomannans, Garcinia cambogia, glavonoid (a licorice extract from Glycyrrhiza glabra), Green coffee bean, Green tea (Camellia sinensis), Guarana (Paullina cupana), Gymnema, Hoodia gordonii, L-carnitine, Psyllium, raspberry ketone, saffron, Spirulina, Vitamin Bi₂ and/or Zantrex-3, or extracts or derivatives of these.

Appropriate dose may readily be determined by persons skilled in the art. It will be appreciated that in some cases, weight gain may exacerbate joint pain due to increased load on already painful or damaged joints. This weight gain may be at least partly attributable to a depressed or otherwise unmotivated mood experienced by the sufferer as a result of the pain experienced during movement. This leads to reduced mobility and thereby creates a negative feedback loop that adds to the pain of the sufferer. In such cases, the inclusion of one or more components that facilitate weight loss may reduce or eliminate weight gain, thereby increasing the mobility of the subject, which eventually reduces the pain experienced by the sufferer as mobility improves and weight is lost.

Non-limiting examples of further additional components or constituents may include ginger, turmeric (Curcuma longa), curcumin (from Curcuma longa), glucosamine hydrochloride, glucosamine sulfate, chondroitin sulfate, amino acids, collagen or collagen derivatives, methlysulfonylmethane (MSM) and/or bioperine (Black pepper extract), or extracts or derivatives of these. These further additional components or constituents may facilitate repair of damaged, inflamed or otherwise painful joints and/or modulate the immune system to reduce or ameliorate inflammation.

Suitably, the formulation further comprises at least one pharmaceutically acceptable carrier, diluent and/or excipient. Preferably, at least one other pharmaceutically acceptable carrier, diluent and/or excipient may be or include one or more of diluents, solvents, pH buffers, binders, fillers, emulsifiers, disintegrants, polymers, lubricants, oils, fats, waxes, coatings, viscosity-modifying agents, glidants and the like.

Diluents may include one or more of microcrystailine cellulose, lactose, mannitol, calcium phosphate, calcium sulfate, kaolin, dry starch, powdered sugar, and the like. Binders may include one or more of povidone, starch, stearic acid, gums, hydroxypropylmethyl cellulose and the like. Disintegrants may include one or more of starch, croscarmellose sodium, crospovidone, sodium starch glycolate and the like. Solvents may include one or more of ethanoi, methanol, isopropanol, chloroform, acetone, methyl ethyl ketone, methylene chloride, water and the like. Lubricants may include one or more of magnesium stearate, zinc stearate, calcium stearate, stearic acid, sodium stearyl fumarate, hydrogenated vegetable oil, glyceryl behenate and the like. A glidant may be one or more of colloidal silicon dioxide, talc or cornstarch and the like. Buffers may include phosphate buffers, borate buffers and carbonate buffers, although without limitation thereto. Fillers may include one or more gels inclusive of gelatin, starch and synthetic polymer gels, although without limitation thereto. Coatings may comprise one or more of film formers, solvents, plasticizers and the like. Suitable film formers may be one or more of hydroxypropyl methyl cellulose, methyl hydroxyethyi cellulose, ethyl cellulose, hydroxypropyl cellulose, povidone, sodium carboxymethyl cellulose, polyethylene glycol, acrylates and the like. Suitable solvents may be one or more of water, ethanol, methanol, isopropanol, chloroform, acetone, methylethyl ketone, methylene chloride and the like. Plasticizers may be one or more of propylene glycol, castor oil, glycerin, polyethylene glycol, polysorbates, and the like.

Reference is made to the Handbook of Excipients 6^th Edition, Eds. Rowe, Sheskey & Quinn (Pharmaceutical Press), which provides non-limiting examples of excipients which may be useful according to the invention,

It will be appreciated that the choice of pharmaceutically acceptable carriers, diluents and/or excipients will, at least in part, be dependent upon the mode of administration of the formulation. By way of example only, the formulation may be in the form of a tablet, capsule, caplet, chew or other edible strip, an edible powder or powder concentrate for subsequent dilution or addition to food or drink, an injectable liquid, a suppository, a slow release formulation, an osmotic pump formulation or any other form that is effective and safe for administration.

In one particular embodiment, the formulation is in the form of an edible soft gel capsule. Such formulations are well known in the art, although reference is made to Encyclopaedia of Pharmaceutical Technology 3 Edition Vol. 1 Ed. Swarbrick (Informa Healthcare USA) which provides non-limiting examples of soft gel capsule formulations and methodology.

In a further aspect, the invention provides a method of treating or preventing a disease disorder or condition in a subject, said method including the step of administering to said subject an effective amount of the formulation disclosed herein or an effective amount of the formulation produced according to method disclosed herein to thereby treat or prevent the disease disorder or condition in the subject.

The terms "administration" or "administered" describe the introduction of the formulation to a subject such as by a particular route or vehicle. Routes of administration may include topical, parenteral and enteral which include oral, buccal, sub-lingual, nasal, anal, gastrointestinal, subcutaneous, intramuscular and intradermal routes of administration, although without limitation thereto. The term "subject" is used in its broadest sense to include and encompass any individual, preferably a mammal. Non-limiting examples of mammals include humans, pets such as dogs, cats and livestock such as horses, cows, sheep, goats and pigs. Preferably, a subject includes any human or non-human mammal, including for example, a primate, cow, horse, pig, sheep, goat, dog, cat, rabbit or rodent. In a preferred embodiment, the subject is a human.

By "treat, treatment or treating" is meant administration of the formulation to a subject to at least ameliorate, reduce or suppress existing signs or symptoms of the disease, disorder or condition experienced by a subject.

By "prevent, preventing" or "preventative" is meant prophylactically administering the formulation to a subject who does not exhibit signs or symptoms of a disease disorder or condition, but who is expected or anticipated to likely exhibit such signs or symptoms in the absence of prevention. Preventative treatment may at least lessen or partly ameliorate expected symptoms or signs. For example, the formulation may be administered to a subject as a preventative treatment before exercise or other activities that could lead to joint or back pain or inflammation.

In one embodiment, the method treats or prevents pain in the subject.

In a related embodiment, the method improves the mobility of the subject.

It will be appreciated that the method and formulation disclosed herein may be suitable for the prevention or treatment of pain and/or inflammation in a subject.

As generally used herein "inflammation" is used in the context of inflammatory diseases, disorders or conditions which are associated with, or caused by, acutely or chronically aberrant or pathological immune activation. Such inflammatory diseases, disorders or conditions may include autoimmune diseases, disorders or conditions. Non-limiting examples include Type I diabetes, rheumatoid arthritis, gout and inflammatory bowel disease (IBD), although without limitation thereto.

In particular embodiments, the pain and/or inflammation is associated with joints. As used herein, "joints" include bones and articulating surfaces of bones and other anatomic structures located at, near or connected to, joints. These structures include cartilage, muscles, tendons, connective tissue (e.g. collagenous tissue) and ligaments, although without limitation thereto. Joints may include knees, ankles, hips, pelvic joints, craniofacial joints such as the temporomandibular joint, spine and vertebral joints, rib joints, shoulder joints, neck joints, elbows, finger joints and wrist joints, although without limitation thereto. Joint pain may be caused by, or associated with, structural damage to the joint and/or by inflammation at or near the joint.

Particular diseases, disorders and conditions that may be prevented or treated by the formulation and/or method disclosed herein include arthritis inclusive of osteoarthritis, rheumatoid arthritis and infectious arthritis, bursitis, osteochnodritis dissecans, sacriliitis and inflammation and/or pain associated with hip dysplasia, sciatica, slipped intervertebral discs, sprains and/or joint dislocations or sub-luxations, although without limitation thereto.

The formulation and method therefore provides a synergistic approach to relieving or preventing pain symptoms and also reducing the inflammation that may be associated with painful, damaged joints. As previously described, the formulation or method may therefore be useful for improving the mobility of sufferers, thereby assisting weight loss which in the longer term reduces pain and inflammation.

In certain embodiments, the composition and treatment method disclosed herein may be particularly useful for treating pain/and or inflammation associated with other specific diseases, disorders and conditions such as metabolic syndrome, Type II diabetes and/or obesity. Subjects suffering from metabolic syndrome, Type II diabetes and/or obesity may be particularly susceptible to joint pain and/or inflammation due to multifactorial, interacting problems such as poor cardiovascular health, physical inactivity, gout and other inflammatory conditions and general weight gain (particularly abdominal adiposity) through metabolic dysregulation.

So that the invention may be readily understood and put into practical effect, reference is made to the following non-limiting Examples.

EXAMPLES

INTRODUCTION Rat collagen arthritis is an experimental model of polyarthritis that has been widely used for preclinical testing of numerous anti-arthritic agents that are either under preclinical or clinical investigation or are currently used as therapeutics in this disease (Trentham et al., 1977; Bendele, McComb et al, 1999; Bendele, 2001). The hallmarks of this model are reliable onset and progression of robust, easily measurable, polyarticular inflammation, marked cartilage destruction in association with pannus formation and mild to moderate bone resorption and periosteal bone proliferation. Therapeutic agents that inhibit interleukin-1 (IL-1) production or activity are especially active in this test system ( Bendele, McAbee et al, 1999) but other types of anti-inflammatory agents have good to excellent activity (refer to Bendele, McComb et al, 1999; Bendele, 2001).

The current study was done to determine the potential efficacy of krill oil, willow bark, and Sensoril® (withania extract), dosed alone or in combination, administered twice daily (BID) by oral (PO) gavage for 21 days prior to collagen immunization and then for the 18 days post-immunization (study days -21 to 18) in inhibiting the inflammation (paw swelling), cartilage destruction, and bone resorption that occurs in developing type II collagen arthritis in rats.

MATERIALS & METHODS

Animals

Female Lewis rats (n = 54) were obtained from Charles River Laboratories, Inc., Wilmington, MA. Rats weighed 139-156 grams (mean approx. 147 g) on study day -22. Animals were identified by a distinct number at the base of the tail delineating group and animal number. After randomization, all cages were labeled with protocol number, group number and animal numbers with a specific color-coding.

Upon arrival, animals were housed 3-4/cage in shoe-box polycarbonate cages with wire tops, wood chip bedding, and suspended food and water bottles. Animal care including room, cage, and equipment sanitation conformed to the guidelines cited in the Guide for the Care and Use of Laboratory Animals (refer to Guide) and the applicable standard operating procedures of Bolder BioPATH, Inc. Animals were acclimated for 9 days prior to enrolment. An attending Veterinarian was on site or on call during the live phase of the study. No concurrent medications were given. During the acclimation and study periods, animals were housed in a laboratory environment with temperatures ranging from 67-76°F and relative humidity of 30%-70%. Automatic timers provided 12 hours of light and 12 hours of dark. Animals were allowed access ad libitum to Harlan Teklad Rodent Chow and fresh municipal tap water.

Experimental Design

Animals (10 rats/group for arthritis) were randomized by body weight and enrolled into treatment groups on study day minus 21. Dosing began upon enrolment (day -21 ; three weeks prior to collagen injection) and continued through study day 18. Animals were anesthetized with Isoflurane and injected intradermally/subcutaneously (ID/SC) with 400 μΐ of Freund's Incomplete Adjuvant (Sigma Aldrich) containing 2 mg/ml porcine type II collagen (Chondrex) at 2 sites (200 μΐ per site) at the base of the tail on day 0 and then 100 μΐ in one site at the base of the tail on day 7. Rats were euthanized on day 18.

Experimental groups were as follows:

Group N Dose mg kg Treatment Dose mg/ml

1 4 Olive oil

control

2 10 - Vehicle -

3 10 1000 Krill oil 400

4 10 60* Willow bark 24

5 10 500 Sensoril 200

(Withania

extract)

6 10 1000, 60*, Krill oil, 400, 24, 200

500 Willow bark &

Sensoril

(Withania extract)

*Equivalent to about 9 mg/kg salicin

All routes were PO at a dosage volume of 2.5 ml/kg. All Regimens were BID (- 12)-17.

Observations, Measurements, and Specimens

Rats were weighed on study days -22, -21, -18, -14 -11, -7, -4, 0, 3, 7, and 9-18. Caliper measurements of right and left ankle diameters were taken every day beginning on day 9 (day -1 of arthritis) and continuing through day 18. Ankle caliper measurements were made with a Digitrix II micrometer (Fowler & NSK). Baseline measurements were taken using one ankle with values rounded to one-thousandth of an inch. Measurements were confirmed as clinically normal (0.260-0.264 in) by comparison with historical values for rats based on a range of body weights. Baseline measurements were then applied to both ankles, and these values remained with the animal so long as the ankle was clinically normal with good definition of all the ankle bones and no evidence of inflammation.

Pain testing was performed as follows: Von Frey testing was performed on days -4, 10, 12, and 14, and Operant read-out testing was performed on days -3, - 1, 11, 13, and 18.

On study day 18, rats were anesthetized with Isoflurane, bled to exsanguinate, and euthanized by cervical dislocation. Hind paws were transected at the level of the medial and lateral malleolus, weighed, and collected with knees into 10% neutral buffered formalin (NBF) for histopathologic evaluation. Livers, spleens, and thymuses were collected, trimmed of extraneous tissue, weighed and then discarded.

Pain Testing Methods

Von Frey Testing Methods

Rats are habituated to the animal colony for one week and handled four times for five minutes each after the week of habituation. Rats are habituated to the testing rack three times during this process. A Von Frey (VF) testing kit for our purposes consists of a set of hairs that range from 3.61 to 5.18 absolute threshold. Each researcher uses the same set of hairs since there may be some variability between kits.

The purpose of testing is to determine which stiffness hair the rat responds to 100% of the time.

Data is entered into a spreadsheet, and a separate program (Psychofit) is used to translate this 100% response rate into an absolute threshold by fitting s- shaped psychometric functions to the data using a maximum-likelihood technique. The absolute threshold, which is used in reporting results, is the threshold at which a stimulus will be detected 50% of the time.

Testing begins with three applications of the 4.31 hair.

• If the rat responds two or three times in a row to 4.31, then testing is dropped down to the 3.61 hair and continued until the rat responds three times in a row.

· If there is no response or one response, then testing continues upwards until the rat responds three times.

• If the rat does not respond at 5.18, any abnormalities in behaviour are recorded.

Responses are recorded on a template as either a 0 (no response) or a 1 (response). Often times the rat will respond once or twice to a certain hair and then not a third time. This is recorded as a 110 or 101 on the template.

When testing, the hair is placed on the surface of the hind paw (forepaw may work as well, but we have not tested this) and pushed smoothly until the hair has a significant bend in it. The hair is kept as steady as possible since wiggling increases the likelihood of getting a response. The hair is kept pressed against the left paw for eight seconds and then the right paw for eight seconds.

• If the hair slips off the paw or skids along the bottom of the paw without bending then at least 10 seconds are allowed to pass before retesting.

• If the hair slips off several times or in a way that startles the rat, testing moves on to a different rat and then comes back to the original rat.

Testing is done on the hind portions of the hind paw - the heel tends to give a more reliable and sensitive response.

A response is recorded when the rat has an obvious reaction to the pressure of the hair. Usually this manifests itself as a lifting of the hind paw from the grate to relieve the pressure. Sometimes responses are very obvious, sometimes more subtle. Some rats jerk their leg away and quiver, while others respond by walking away from the offending hair.

• If the tester is unsure whether or not the rat is responding, the rat is retested.

• If it is the hair that they were responding to, and not general agitation, then they will continue to respond.

The tester should be aware of hyper-responding. Rats quickly learn that something is annoying them and may put undue attention on their paws. This may result in over responding to a stimulus that they would not normally respond to. Freezing is also a large concern during testing, because if rats become too agitated or frightened, they may be feeling the hair, but not responding because of their freezing instinct. If this occurs, the rat is left alone or distracted to help them calm down.

Operant Read-out Testing

Rats are given 30% sweet milk for approximately 12 hours prior to conditioning them to the operant conditioning apparatus. Rats are then conditioned to the apparatus by letting them individually explore the dark box at the top of the ramp, which contains a bowl of sweet milk, for 3 minutes.

On testing days, rats are placed at the bottom of the ramp and a timer is started the moment the rat is released. The timer is stopped once all four paws are inside the dark box, and the time is recorded. This process is repeated three times, consecutively, for each rat. The mean of all three values is treated as a single data point for statistical purposes.

Morphologic Pathology Methods

Preserved and decalcified (5% formic acid) ankle and knee joints were cut in half longitudinally (ankles) or in the frontal plane (knees), processed through graded alcohols and a clearing agent, infiltrated and embedded in paraffin, sectioned, and stained with Toluidine Blue (T. blue) by Bolder BioPATH, Inc. associated personnel (HistoTox Labs, Inc.). Tissues from all animals were examined microscopically by a board certified veterinary pathologist (Dr. Alison Bendele) and observations were entered into a computer-assisted data retrieval system. Scoring of Joints

Collagen arthritic ankles and knees are given scores of 0-5 for inflammation, pannus formation and bone resorption according to the following criteria:

Knee and/or Ankle Inflammation

0 = Normal.

0.5 = Minimal focal inflammation.

1 = Minimal infiltration of inflammatory cells in synovium/periarticular tissue.

2 = Mild infiltration.

3 = Moderate infiltration with moderate edema.

4 = Marked infiltration with marked edema.

5 = Severe infiltration with severe edema.

The inflammatory infiltrate in mice and rats with type II collagen arthritis consists of neutrophils and macrophages with smaller numbers of lymphocytes when the lesions are in the acute to subacute phase. Tissue edema and neutrophil exudates within the joint space are common in the acute to subacute phase. As the inflammation progresses to chronic, mononuclear inflammatory cells (monocytes, lymphocytes) predominate and fibroblast proliferation, often with deposition of metachromatic matrix, occurs in synovium and periarticular tissue. Exudate is less common in the joint space. Unless indicated in the comments area, the inflammation type is acute to subacute.

Ankle Pannus

0 = Normal.

0.5 = Minimal infiltration of pannus in cartilage and subchondral bone, affects only marginal zones and affects only a few joints.

1 = Minimal infiltration of pannus in cartilage and subchondral bone, primarily affects marginal zones.

2 = Mild infiltration (<l/4 of tibia or tarsals at marginal zones). 3 = Moderate infiltration (1/4 to 1/3 of tibia or small tarsals affected at marginal zones).

4 = Marked infiltration (1/2 to 3/4 of tibia or tarsals affected at marginal zones).

5 = Severe infiltration (>3/4 of tibia or tarsals affected at marginal zones, severe distortion of overall architecture).

Knee Pannus

0 = Normal.

1 = Minimal infiltration of pannus in cartilage and subchondral bone, approximately 1%-10% of cartilage surface or subchondral bone affected.

2 = Mild infiltration (extends over up to 1/4 of surface or subchondral area of tibia or femur), approximately l l%-25% of cartilage surface or subchondral bone affected.

3 = Moderate infiltration (extends over >l/4 but < 1/2 of surface or subchondral area of tibia or femur) approximately 26%-50% of cartilage surface or subchondral bone affected.

4 = Marked infiltration (extends over 1/2 to 3/4 of tibial or femoral surface) approximately 51%-75% of cartilage surface or subchondral bone affected.

5 = Severe infiltration approximately 76%-100% of cartilage surface or subchondral bone affected.

Ankle Cartilage Damage (emphasis on small tarsals)

0 = Normal

0.5 = Very minimal = Minimal decrease in T blue staining, affects only marginal zones and affects only a few joints.

1 = Minimal = Minimal to mild loss of toluidine blue staining with no obvious chondrocyte loss or collagen disruption.

2 = Mild = Mild loss of toluidine blue staining with focal mild (superficial) chondrocyte loss and/or collagen disruption. 3 = Moderate = Moderate loss of toluidine blue staining with multifocal moderate (depth to middle zone) chondrocyte loss and/or collagen disruption, smaller tarsals affected to 1/2 to 3/4 depth with rare areas of full thickness loss.

4 = Marked = Marked loss of toluidine blue staining with multifocal marked (depth to deep zone) chondrocyte loss and/or collagen disruption, 1 or 2 small tarsals surfaces have full thickness loss of cartilage.

5 = Severe = Severe diffuse loss of toluidine blue staining with multifocal severe (depth to tide mark) chondrocyte loss and/or collagen disruption affecting more than 2 cartilage surfaces.

Knee Cartilage Damage

0 = Normal

0.5 = Veryminimal = Minimal decrease in T blue staining, affects only marginal zones.

2 = Mild = Mild loss of toluidine blue staining with focal mild

(superficial) chondrocyte loss and/or collagen disruption, may have few small areas of 50% depth of cartilage affected.

3 = Moderate = Moderate loss of toluidine blue staining with multifocal to diffuse moderate (depth to middle zone) chondrocyte loss and/or collagen disruption, may have 1-2 small areas of full thickness loss affecting less than 25% of the total width of a surface and not more than 25% of the total width of all surfaces.

4 = Marked = Marked loss of toluidine blue staining with multifocal to diffuse marked (depth to deep zone) chondrocyte loss and/or collagen disruption or 1 surface with near total loss and partial loss on others, total overall loss less than 50% of width of all surfaces combined.

5 = Severe = Severe diffuse loss of toluidine blue staining with multifocal severe (depth to tide mark) chondrocyte loss and/or collagen disruption on both femurs and/or tibias, total overall loss greater than 50% of width of all surfaces combined.

Ankle Bone Resorption 0 = Normal

0.5 = Very minimal = Minimal resorption affects only marginal zones and affects only a few joints.

1 = Minimal = Small areas of resorption, not readily apparent on low magnification, rare osteoclasts.

2 = Mild = More numerous areas of resorption, not readily apparent on low magnification, osteoclasts more numerous, < 1/4 of tibia or tarsals at marginal zones resorbed.

3 = Moderate = Obvious resorption of medullary trabecular and cortical bone without full thickness defects in cortex, loss of some medullary trabeculae, lesion apparent on low magnification, osteoclasts more numerous, 1/4 to 1/3 of tibia or tarsals affected at marginal zones.

4 = Marked = Full thickness defects in cortical bone, often with distortion of profile of remaining cortical surface, marked loss of medullary bone, numerous osteoclasts, 1/2 to 3/4 of tibia or tarsals affected at marginal zones.

5 = Severe = Full thickness defects in cortical bone, often with distortion of profile of remaining cortical surface, marked loss of medullary bone, numerous osteoclasts, >3/4 of tibia or tarsals affected at marginal zones, severe distortion of overall architecture.

Knee Bone Resorption

0 = Normal

0.5 = Very minimal = Minimal resorption affects only marginal zones.

1 = Minimal = Small areas of resorption, not readily apparent on low magnification, approximately 1%-10% of total joint width of subchondral bone affected.

2 = Mild = More numerous areas of resorption, definite loss of subchondral bone, approximately l l%-25% of total joint width of subchondral bone affected.

3 = Moderate = Obvious resorption of subchondral bone approximately 26%-50% of total joint width of subchondral bone affected.

4 = Marked = Obvious resorption of subchondral bone approximately 51%-75% of total joint width of subchondral bone affected. 5 = Severe = Distortion of entire joint due to destruction approximately 76%-100% of total joint width of subchondral bone affected.

Periarticular Matrix Deposition (only scored if an increase is seen in any treated group relative to disease controls)

0 = Normal.

1 = Faint, multi-focal metachromatic staining, no excessive expansion of periarticular tissue.

2 = Darker, diffuse metachromatic staining, no excessive expansion of periarticular tissue.

3 = Darker, diffuse metachromatic staining, mild expansion of periarticular tissue.

4 = Darker, diffuse metachromatic staining, moderate expansion of periarticular tissue.

5 = Darker, diffuse metachromatic staining, severe expansion of periarticular tissue.

Periosteal New Bone Formation (Ankles-measure on 16x)

Studies that go beyond the acute inflammatory stage often show varying degrees of periosteal new bone formation. In order to convey the degree of periosteal new bone formation the following scores are applied based on the distribution of periosteal bone proliferation and a measurement of width of periosteal new bone formation at the widest location.

0 = Normal, no periosteal proliferation.

0.5 = Minimal focal or multifocal proliferation, measures less than 127 μιη width (1-2) at any location.

1 = Minimal multifocal proliferation, width at any location measures 127- 252 μιη (3-4 units).

2 = Mild multifocal on tarsals, diffuse in some locations, width at any location 253-441 μιη (5-7 units).

3 = Moderate multifocal on tarsals, diffuse in most other locations, width at any location measures 442-630 μιη (8-10 units). 4 = Marked multifocal on tarsals, diffuse at most other locations, width at any location measures 630-819 μιη (11-13 units).

5 = Severe, multifocal on tarsals, diffuse at most other locations, width at any location measures >819 μιη (>13 units).

Statistical Analysis

Clinical data for ankle joint diameter were analyzed by determining the area under the dosing curve (AUC). For calculation of AUC, the daily measurement of ankle joints (using a caliper) for each rat were entered into Microsoft Excel and the area between the treatment days after the onset of disease to the termination day was computed. Means for each group were determined and % inhibition from arthritis controls was calculated by comparing values for treated and normal animals. Test groups were compared to disease controls using a one-way analysis of variance (1-way ANOVA) for measured parameters or a Kruskal-Wallis test for scored parameters (non-parametric) along with the appropriate multiple comparison post-test. The model was validated by comparing normal controls to disease controls using a Student's two-tailed ί-test. Unless indicated, Bolder BioPATH, Inc. performs statistical analysis on raw (untransformed) data only. Statistical tests make certain assumptions regarding the data's normality and homogeneity of variance, and further analysis may be required if testing resulted in violations of these assumptions. Significance for all tests was set at p < 0.05, with p values rounded to the third decimal place. Percent inhibition of paw weight and AUC is calculated using the following formula:

% Inhibition = B/A x 100

A = Mean Normal - Mean Disease Control

B = Mean Treated - Mean Disease Control

RESULTS

Live Phase and Necropsy Parameters

Vehicle control rats had disease-induced body weight loss of -12.3 g from day 0 to day 18 (collagen injection to termination). Rats treated with krill oil/willow bark/Sensoril (combination therapy) had body weight gain that was significantly (70%) increased toward normal compared to vehicle controls. Body weight loss was not significantly affected by treatment with krill oil, willow bark, or Sensoril dosed alone. Body weight gain prior to collagen injection was similar across groups, which indicates that the treatments were well tolerated and had no observed toxicity (see Figures 1 A & B).

Daily ankle diameter measurements differed significantly from vehicle controls over time. Rats given combination therapy had significantly reduced

(toward normal) ankle diameter measurements on days 12-13 and 15-18 compared to vehicle controls. Daily ankle diameter measurements were not significantly affected by treatment with krill oil, willow bark, or Sensoril^® dosed alone (Figure 2).

Ankle diameter AUC was significantly reduced toward normal in rats given combination therapy (64% reduction) compared to vehicle controls. Ankle diameter AUC was not significantly affected by treatment with krill oil, willow bark, or Sensoril dosed alone (Figure 3).

Final paw weights were significantly reduced toward normal in rats given combination therapy (62% reduction) compared to vehicle controls. Final paw weights were not significantly affected by treatment with krill oil, willow bark, or

Sensoril dosed alone (Figure 4).

Liver weights relative to body weight were significantly increased toward normal in rats given willow bark, Sensoril, or combination therapy compared to vehicle controls (Figure 5). Spleen weights relative to body weight were similar across groups (Figure 6).

Thymus weights relative to body weight were significantly increased in rats given combination therapy compared to vehicle controls, which is consistent with efficacy. Relative thymus weights were not significantly affected by treatment with krill oil, willow bark, or Sensoril^® dosed alone compared to vehicle controls (Figure 7).

Von Frey analysis revealed that absolute pain thresholds were significantly increased on day 14 in rats given combination therapy or willow bark compared to vehicle controls, indicating a reduction in average hind foot allodynia (Figure 8).

Operant read-out times were significantly reduced on day 18 in rats given combination therapy compared to vehicle controls, which is an indication of increased mobility and reduced pain (Figure 9). Operant readout times AUC were significantly (49%) reduced in rats given combination therapy compared to vehicle controls. Operant readout times AUC were not significantly affected by treatment with krill oil, willow bark, or Sensoril dosed alone compared to vehicle controls (see Figure 10).

Morphologic Pathology

All vehicle-treated disease control rats had severe synovitis and periarticular inflammation in at least one and usually both ankle joints with none to moderate pannus formation, bone resorption, and periosteal bone formation, and none to marked cartilage damage. Mean periosteal bone width was 242.55 μπι.

Ankle inflammation was significantly reduced toward normal in rats given combination therapy (55% reduction) compared to vehicle controls. Ankle histopathology parameters were not significantly affected by treatment with krill oil, willow bark, or Sensoril^® dosed alone compared to vehicle controls (see Figures 11-13). All vehicle control rats had severe synovitis and periarticular inflammation in at least one knee joint with none to mild pannus formation and bone resorption, and none to marked cartilage damage.

All knee histopathology parameters were significantly reduced toward normal in rats given combination therapy (68% reduction of summed knee scores) compared to vehicle controls.

Knee histopathology parameters were not significantly affected by treatment with krill oil, willow bark, or Sensoril^®dosed alone compared to vehicle controls (see Figures 14-15). DISCUSSION & CONCLUSIONS

Female Lewis rats with developing type II collagen-induced arthritis (CIA) were treated twice daily (BID) by the oral (PO) route for 21 days prior to collagen immunization and then for 18 days post-immunization (study days -21 to 18) with vehicle (olive oil), krill oil (1000 mg/kg), willow bark (60 mg/kg), Sensoril® (withania extract: 500 mg/kg), or combination therapy (krill oil/willow bark/Sensoril, 1000/60/500 mg/kg). Rats were euthanized on study day 18. Efficacy evaluation was based on animal body weights, daily ankle caliper measurements, ankle diameter expressed as area under the curve (AUC), terminal hind paw weights, pain testing (Von Frey and operant read-out), and histopathologic evaluation of ankles and knees.

All animals survived to the scheduled termination. Combination therapy with krill oil/willow bark/Sensoril showed significant and synergistic beneficial effect in the developing model of rat CIA as determined by evaluation of clinical and histopathologic parameters. Treatment with krill oil, willow bark, or Sensoril® alone resulted in disease parameters that were mostly similar to vehicle controls. Body weight gain was significantly (47%) increased toward normal in rats given combination therapy compared to vehicle controls. It is common that animals lose a significant amount of weight in this animal model because they are suffering inflammatory trauma. The combination treatment prevented such a large weight loss which is a strong signal of well-being in the animals that suggests the combination treatment is having positive effects against the inflammatory condition.

Daily ankle diameter measurements were significantly reduced toward normal on days 12-13 and 15-18 in rats given combination therapy compared to vehicle controls. Combination therapy significantly reduced ankle diameter AUC (64%) reduction) and final paw weights (62%>) compared to vehicle controls. Von Frey analysis revealed that absolute pain thresholds were significantly increased on day 14 in rats given combination therapy or willow bark compared to vehicle controls, indicating a reduction in average hind foot allodynia. Operant read-out times were significantly reduced on day 18 in rats given combination therapy, which is an indication of the brain's processing of decreased pain, increased mobility and well-being. More particularly, this is a measure of the animal's conscious processing of pain signals and also its well-being. In operant pain readings the animal is making a conscious decision on whether it thinks it is worth moving or not even though it is in discomfort or severe pain, and therefore is a more powerful reading of analgesia, the brain's processing of pain and also the animal's general well-being.

Operant readout times AUC were also significantly (49%>) reduced in rats given combination therapy compared to vehicle controls.

Evaluation of histopathology confirmed the clinical findings. Ankle inflammation was significantly (55%>) reduced toward normal, and all knee histopathology parameters were significantly reduced toward normal (68%> reduction of summed knee scores) in rats given combination therapy compared to vehicle controls. In general, knee lesions progress more slowly than ankle lesions, thus effective agents tend to be more effective on knee parameters. The test articles were well tolerated under the conditions of this study. Rats given combination therapy had thymus weights relative to body weights that were significantly increased toward normal compared to vehicle controls, which is consistent with efficacy. Liver weights relative to body weight were significantly increased toward normal.

Combination therapy with krill oil/willow bark/Sensoril showed significant and synergistic beneficial effect in the developing model of rat CIA as determined by evaluation of clinical and histopathologic parameters. Treatment with krill oil, willow bark, or Sensoril alone resulted in disease parameters that were mostly similar to vehicle controls. The test articles were well tolerated under the conditions of this study. All animals survived to the scheduled termination.

It is therefore proposed that formulations comprising krill oil, willow bark and withania extracts such as Sensoril^® may synergistically treat or prevent pain, such as joint pain, while providing the benefits of comprising natural products in contrast to typical point pain relief products such as NSAIDs. Although not wishing to be bound by theory, it is proposed that omega-3 fatty acids (such as EPA) and willowbark affect different targets in the prostaglandin pathway to reduce inflammation. Willowbark inhibits the COX enzyme inhibiting prostaglandin synthesis. Furthermore, omega-3 fatty acids (such as EPA) are a substrate for resolvins and protectins which have anti-inflammatory effects in disease and injury. Withania may have effects on the peripheral nervous system and/or the CNS effects. CNS effects may include anti-anxiolytic effects such as through activating the GABAA receptor or modulating cholinergic neurotransmission. It may be that by altering how the brain interprets or registers a painful stimulus and making the brain less responsive to pain (e.g. using a mild anti-anxiety or stress agent for relaxation together with one or more anti- inflammatory agents) the formulation disclosed herein has a "multi-pronged", synergistic effect on pain.

Throughout the specification the aim has been to describe the preferred embodiments of the invention without limiting the invention to any one embodiment or specific collection of features. It will therefore be appreciated by those of skill in the art that, in light of the instant disclosure, various modifications and changes can be made in the particular embodiments exemplified without departing from the scope of the present invention.

All patent, scientific and technical literature as well as web addresses, software and statistical packages referred to in this specification are incorporated herein by reference.

REFERENCES

Trentham DE, Townes AS, Kang AH. Autoimmunity to type II collagen: an experimental model of arthritis. J Exp Med 1977; 857-868.

Bendele AM, McComb J, Gould T, McAbee T, Sennello G, Chlipala E, Guy M. Animal models of arthritis:relevance to human disease. Toxicologic Pathol 1999; 27: 134-142.

Bendele AM. Animal Models of Rheumatoid Arthritis. J Musculoskel Interact 2001; 377-385.

Bendele AM, McAbee T, Sennello G, Frazier J, Chlipala E, McCabe D. Efficacy of sustained blood levels of interleukin-1 receptor antagonist in animal models of arthritis. Arthritis Rheum 1999; 42:498-506.

Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources National Academy Press, Washington D.C.; 1996.

Claims

1. A formulation comprising:

(i) one or more omega-3 fatty acids and/or one or more derivatives thereof;

2. The formulation of Claim 1 which is capable of treating or preventing pain and/or inflammation in a subject.

3. The formulation of Claim 1 or Claim 2, which is capable of improving mobility in a subject.

4. The formulation of any preceding claim, which further comprises one or more additional components.

5. The formulation of any preceding claim, which further comprises one or more additional components that facilitate weight loss.

6. The formulation of any preceding claim, which further comprises at least one pharmaceutically acceptable carrier, diluent and/or excipient.

7. The formulation of any preceding claim which is in the form of a soft gel capsule.

8. A method of producing a formulation, said method including the step of combining:

(i) one or more omega 3 fatty acids and/or one or more derivatives thereof;

to thereby produce the formulation.

9. The method of Claim 8, wherein the formulation treats or prevents pain and/or inflammation.

10. The formulation of Claim 8 or Claim 9, which improves mobility.

11. The method of any one of Claims 8-10, which further comprises combining one or more additional components with (i)-(iii).

12. The method of Claim 11, wherein the one or more additional components facilitate weight loss.

13. The method of any one of Claims 8-12, which further includes combining (i)-(iii) with at least one pharmaceutically acceptable carrier, diluent and/or excipient.

14. The method of any one of Claims 8-13 which produces a formulation in the form of a soft gel capsule.

15. A formulation produced according to the method of any one of Claims 8- 14.

16. A method of treating a disease, disorder or condition in a subject, said method including the step of administering to said subject an effective amount of the formulation according to any one of Claims 1-7 or Claim 15, to thereby treat or prevent the disease, disorder or condition in the subject.

17. The method of Claim 16, wherein the disease, disorder or condition is, or is associated with, joint pain and/or inflammation.

18. The method of Claim 16 or Claim 17, wherein the disease, disorder or condition is arthritis.

19. The method of Claim 18, wherein arthritis is rheumatoid arthritis or osteoarthritis.

20. The method of Claim 16, Claim 17 or Claim 18, which improves the mobility of the subject.

21. The method of any one of Claims 16-20, wherein the subject suffers from Type II diabetes, metabolic syndrome and/or obesity.

22. The method of any one of Claims 8-21, wherein the subject is a human.