WO2003082310A1

WO2003082310A1 - Composition and method for the treatment or prevention of il-1 mediated disorders

Info

Publication number: WO2003082310A1
Application number: PCT/NL2003/000250
Authority: WO
Inventors: Hobbe Friso Smit; Yvo Maria Franciscus Graus
Original assignee: Nutricia NV; GENERAL NUTRITION INVESTMENT Co
Current assignee: Nutricia NV; GENERAL NUTRITION INVESTMENT Co
Priority date: 2002-04-03
Filing date: 2003-04-03
Publication date: 2003-10-09
Anticipated expiration: 2004-10-03
Also published as: AU2003225430A1

Abstract

One aspect of the present invention relates to a composition comprising a. celery material b. an isolate from vegetable oil enriched in unsaponifiables and c. optionally a pharmaceutically acceptable carrier. The present composition is particularly suitable for use in a method for the prophylactic and/or curative treatment of interleukin-1 mediated diseases, in particular for the treatment or prevention of damage to the connective tissue and for reducing inflammation and inflammation associated complaints. Thus, another aspect of the invention concerns a method for the curative or prophylactic treatment of interleukin-1 mediated disorders in mammals, said method comprising the enteral, topical or transdermal administration to a mammal of the aforementioned composition.

Description

COMPOSITION AND METHOD FOR THE TREATMENT OR PREVENTION OF

IL-1 MEDIATED DISORDERS

FIELD OF THE INVENTION

This invention relates to compositions that contain a celery material. The present composition is particularly suitable for use in a method for the prophylactic and/or curative treatment of interleukin-l mediated diseases, in particular for the treatment or prevention of damage to the connective tissue and for reducing inflammation and inflammation associated complaints.

BACKGROUND OF THE INVENTION

Inflammatory conditions of a joint are chronic joint diseases that afflict and disable, to varying degrees, millions of people worldwide. Arthritis is one of the most common disabling diseases. It has been estimated that about 10 percent of the Western population suffers from one or many forms of arthritis.

There are over 75 types of different diseases of the joint, classified according to their specific symptoms and causes. Representatives of the more common disorders are rheumatoid arthritis and osteoarthritis. Rheumatoid arthritis is a disease of articular joints in which the cartilage and bone are slowly eroded away by a proliferative, invasive connective tissue called pannus, which is derived from the synovial membrane. Subjects suffering from rheumatoid arthritis often have insufficient or abnormal cartilage in the joints due to a chronic inflammation reaction.

Osteoarthritis is a common joint disease characterized by degenerative changes in articular cartilage and reactive proliferation of bone and cartilage called osteophytes. Degenerative changes in joint cartilage are for example caused by reduced synthesis of matrix molecules and increased degradation of articular cartilage. In order to counteract these phenomena it is essential that the cartilage formation is stimulated. Most arthritic complaints relate to decreased mobility, pain, stiffness that develops during sleep and other discomforts due to insufficient cartilage in the joints.

The effects of reduced, damaged or insufficient cartilage, in particular in the case of osteoarthritis, are most often and first seen in the hands or feet of the patient. Other body joints such as shoulder, elbow, knees, ankles, hips, spine and jawbone can also be involved.

The involvement of interleukin-l in arthritis has been implicated by two distinct lines of evidence. First, increased levels of interleukin-l (IL-1), and of the mRNA encoding it, have been found in the synovial tissue and fluid of arthritic joints. Additionally the administration of interleukin-l to healthy joint tissue has been shown on numerous occasions to result in the erosion of cartilage and bone. It is presently believed that IL-1 dominates the effects of cartilage thinning and destruction. IL-1 causes proteoglycan loss, which can prevent proper maintenance of the cartilage. Also, IL-1 has been shown to inhibit collagen synthesis, which prevents proper matrix formation with collagen and makes the cartilage matrix vulnerable to more damage. Additionally IL -1 stimulates production of inflammatory components which have adverse effects on the integrity of the joint, such as prostaglandin E2 (PGE2), nitric oxide (NO), matrix metalloproteases (MMPs), and cyclooxygenase-2 (COX-2).

Many methods for the treatment of arthritic complaints are currently available, including the use of unspecifϊc non-steroidal anti-inflammatory drugs (NSAIDs). However, most of the currently available methods solely treat or prevent the symptoms of the arthritic complaints, such as joint swelling and pain. Additionally, several of the drugs used today have side effects, e.g. gastrointestinal complaints.

In contrast, herbal preparations mostly have limited side effects. However due to limited knowledge of the effects induced by herbal preparations, medicinal compositions containing herbal preparations often provide only a limited medicinal effect. Furthermore, due to the complexity of herbal preparations, research directed towards the unraveling of the effects of the herbal preparations on the disease specific mechanism is extremely cumbersome.

Celery extract has been reported to have an anti-inflammatory effect. EP708651 describes a therapeutic agent containing celery seed concentrate for the treatment of inflammatory complaints.

Still the effects of administration of celery extract alone provide insufficient relief of arthritis or related complaints. Therefore celery seed extract is often combined with other components in the hope to improve its effects.

WO0040258 describes the combined use of biologically active extracts of celery seed and non-steroidal anti-inflammatory drugs (NSAIDS) for the treatment and prevention of acute and chronic pain, inflammation and gastrointestinal irritation.

US5916565 describes a composition for the treatment of arthritis, joint stiffness, joint mobility and joint pain in vertebrates, consisting essentially of a palatability agent (e.g. hydrogenated vegetable oils), cayenne, ginger, turmeric, yucca, Devils Claw, nettle leaf, Black Cohosh, alfalfa, celery seeds, D-glucosamine HC1 and mucopolysacchrides.

CN1212128 describes a vegetable oil composition for eliminating bad breath. The oil mixture comprises celery seed oil, sunflower seed oil and peppermint camphor.

Herron's "Celery and Juniper", a nutritional supplement, is advertised to reduce inflammatory complaints. The preparation consists of Juniper (Juniperus communis), Celery (Apium graveolens) and Devil's claw.

Joint & Ligament Support Formula™ includes Curcumin (Curcuma longa), Boswellia serrata, Bromelain, Cat's Claw's Root (Uncaria tomentosa), Ginger ( Zingiber officinale), Devil's Claw (Harpagophytum procumbens), Ascorbic acid, Methyl-sulfonyl-methane, Magnesium (citrate), Green Lipped Mussel (Perna canaliculus), Celery Seed extract (Apii/rMctu.? and Apium graveolens), Withania somnifera (Ashwagandha), Asphaltum (Silajit), Vitamin E, Black pepper (Piper nigrum), Zinc monomethionine, Boron, Manganese and Selenomethionine.

Without detailed knowledge of the mechanism of action of celery it is difficult to provide advantageous combinations, which support the action of celery.

SUMMARY OF THE INVENTION

Interleukin-l (IL-1) is one of the most potent inflammatory cytokines yet discovered and a cytokine, which is thought to be a key mediator in many diseases and medical conditions, termed "interleukin-l mediated diseases". A disease or medical condition is considered to be an "interleukin-l mediated disorder" if the disease or medical condition is associated with elevated levels of IL-1 in bodily fluids or tissue or if cells or tissues taken from the body of a patient suffering from the disease or medical condition, produce elevated levels of IL-1 in culture.

The present inventors have surprisingly found that celery material is capable of mildly reducing IL-1 release in lipopolysaccharide (LPS) activated human peripheral blood monocytes (hPBMC). This result indicates that celery material can be used by mammals for reduction of IL-1 release in vivo. However, since celery material was found to be only capable of mildly reducing IL-1 release, these results show that additional agents are needed to enhance the IL-1 inhibitory effect of celery material.

The present invention provides novel compositions, comprising celery material and an isolate from vegetable oil enriched in unsaponifiables. The combination of celery material and an isolate from vegetable oil enriched in unsaponifiables produces a synergistic effect, which can be explained from the individual actions of these compositions on the effects of IL- 1. Celery material mildly inhibits the release of IL- 1 , while an isolate from vegetable oil enriched in unsaponifiables decreases the deleterious effects of IL-1 on synoviocytes and chondrocytes. Deleterious effects of IL-1 on synoviocytes and chondrocytes include for example, reduction of proteoglycan synthesis (Mauviel, A. et al. 1991, Rev. Phum. Mai. Osteoartic. 58(4):241-5) and inhibition of collagenase production (Henrotin, Y.E. et al. 1998, Clin. Rheumatol. 17(l):31-9). A combination of celery material and an isolate from vegetable oil enriched in unsaponifiables will effectively prevent/reduce the detrimental effects of IL-1, particularly in the joints, including one or more of proteoglycan loss, inhibition of collagen synthesis and production of prostaglandin E2 (PGE2), nitric oxide (NO), matrix metalloproteases (MMPs) and cyclooxygenase-2 (COX-2).

Hence, in a further aspect the present invention provides a method for the treatment or prevention of IL-1 mediated disorders, preferably a method for the repair, treatment or prevention of connective tissue damage, said method comprising the enteral, topical or transdermal administration of celery material and an isolate from vegetable oil enriched in unsaponifiables.

It is especially advantageous to use unsaponifiable material from vegetable oil, since this fraction is deemed to have additional beneficial effects besides inhibiting the effects of IL-1. Unsaponifiable fractions of vegetable oil have been described to stimulate chondrocyte expression of transforming growth factor (TGF) beta 1, beta 2 and plasminogen activator inhibitor 1. (Boumediene K., et al.,"Avocado/soya unsaponifiables enhance the expression of transforming growth factor beta 1 and beta 2 in cultured articular chondrocytes, "Arthritis Rheum. 42 (1): 148-56 (Jan. 1999)).

Additionally, an important advantage of the present composition, particularly when used in a method wherein enteral administration is employed, resides in the fact that the present composition does not produce significant side effects. DETAILED DESCRIPTION

The present invention provides a composition suitable for use in a method for the treatment or prevention of IL-1 mediated diseases, particularly the repair, treatment and prevention of damage to the connective tissue, the composition comprising celery material and an isolate from vegetable oil enriched in unsaponifiables.

Celery material

The celery (Apium graveolens) material used in the present invention can be obtained from a variety of celery species, however is preferably obtained from one or more species selected from the group of A. graveolens var. dulce, A. graveolens var. rapaceum, A. graveolens var. graveolens and A. graveolens var. secalinum, more preferably the celery material is obtained from A. graveolens var. graveolens.

The celery material can be obtained from whole plants or from one or more parts thereof, for example stems, stalks, roots, shoots, rhizomes, tubers, fruits, foliage, kernels, husks, hulls, seeds or mixtures thereof. According to a preferred embodiment the celery material is obtained from the seeds of celery.

Celery isolate In a particularly preferred embodiment of the present invention the celery material is a celery isolate. The term "isolate" as referred to in here, preferably encompasses any fraction that can be obtained from a plant material by means of isolation techniques known in the art, e.g. extraction, distillation, squeezing etc. and that displays the desired functional properties described herein before. Hence, the celery isolate is preferably prepared by subjecting the celery plant or part thereof to one or more isolation processes selected from the group of solvent extraction, cold pressing, hot pressing, distillation, chromatography and filtering. The celery isolate is suitably prepared using steam distillation, solvent extraction or essential oil distillation. Celery extract

The celery material is preferably provided in the form of a celery extract. The term "extract" as used in the present invention refers to an isolate that has been obtained by means of solvent extraction. The celery extract is preferably prepared by the following procedure:

A. Reducing celery raw material to yield a composition of particulate matter

B. Contacting the particulate matter obtained in A with a suitable extraction solvent

C. Incubation: keeping the particulate matter in contact with the solvent for a sufficiently long time to allow the transfer of soluble components from the particulate matter into the solvent.

D. Separating the insoluble particulate matter from the extraction solvent containing the extracted soluble components

E. Optionally, removing at least part of the solvent from the extraction solvent containing the extracted soluble components.

Reduction of the celery material can be achieved by a variety of methods of commuting well known in the art, including crushing or grinding in a mill. Preferably, soon after celery has been converted to particulate matter having a sufficiently small average diameter, e.g. below 1.5 mm in diameter, the celery particular matter is contacted with a suitable solvent.

According to a preferred embodiment the solvent is capable of solubilising the lipophilic components present in celery. Preferred solvents include acetone, ethanol, chloroform, hexane, mixtures thereof and mixtures of any one of the previous mentioned solvents with water. Alternatively and equally preferred are solvents used in supercritical extraction processes such as carbondioxide and nitrogen. According to a particularly preferred embodiment celery particulate matter is suspended in 80 - 100% ethanol, or a solvent used in supercritical extraction procedure. Suitable methods for the preparations of supercritical fluid celery extracts are given in WO0040258, the entire content of which is hereby incorporated by reference. The weight ratio solvent to celery particulate matter is preferably between 0.1 and 100, more preferably between 1 and 50, even more preferably between 2 and 25. The celery particulate matter is preferably suspended in the solvent for a sufficiently long time to extract soluble components therefrom, preferably for at least 5 minutes. The suspension of the solvent and particulate matter may for example be subjected to agitation.

After incubation, the non-dissolved particulate matter is removed from the suspension, for example by centrifugation or filtration. The extract may, for example, be filtered with one or more filters having a pore size between 0.2-10 microns. Subsequently the liquid phase is concentrated by removal of a part or all of the solvent. According to a preferred embodiment, at least 95% of the solvent is removed from the preparation. The solvent may for example be removed through distillation and/or by applying heat or vacuum. The concentration ratio of the celery extract used in accordance with the present invention, expressed as celery derived dry matter in the extract divided by the amount of dry celery particulate material that was extracted, is preferably 0.002 and 0.6, more preferably between 0.01 and 0.25, even more preferably between 0.013 and 0.2, especially between 0.02 and 0.1.

Active ingredients

The celery isolate preferably contains phthalides. Preferably the amount of phthalides in the isolate exceeds 4%, more preferably 7%, even more preferably 10% by weight of dry matter. Although celery isolate may consist essentially of phthalides, preferably the content of phthalides does not exceed 60 % by weight of dry matter, more preferably is below 40% by weight of dry matter. According to a preferred embodiment, the celery isolate comprises at least 0.5%, preferably at least 1% by weight of dry celery isolate of at least three phthalides selected form the group of butylphthalides3-n-butyl, phthalide3- n-butylidenephthalide, 3-iso-butylidene phthalide, 3-n-butyl-4,5-dihydrophthalide (also referred to as sedanenolide or senkyunolide), 3-n-butylidene-4,5-dihydrophthalide (also referred to as ligustilide), 3-isobutylidene-3a, 4-dihydrophthalide, 3-isoval(er)idene phthalide, 3-isoval(er)idene-3a,4-dihydrophthalide, sedanolide and sedanonic anhydride. The celery isolate preferably contains terpenes, which may include monoterpenes and sesquiterpenes. Preferably the amount of terpenes in the isolate exceeds 2%, more preferably exceeds 4%, even more preferably exceeds 5% by weight of dry matter. Although celery isolate may comprises high weight percentages of terpenes, preferably the content of terpenes does not exceed 90%, more preferably it is below 50 % by weight of dry matter. According to a preferred embodiment, the celery isolate comprises at least 0.5 % by weight of dry matter of at least three terpenes selected from the group consisting of the monoterpenes beta-caryophyllene, p-cymene, D-carvone, caryophyllene oxide, beta-elemene, limonene, limonen-oxide, linalool, myrcene, alpha-pinene, beta-pinene and the sesquiterpenes alpha-selinene and beta-selinene.

The celery isolate preferably contains coumarins. Preferably the amount of coumarins in the isolate exceeds 0.2 %, more preferably 1 %, even more preferably 3 % by weight of dry matter. According to a preferred embodiment, the celery isolate comprises at least 0.1%), preferably at least 0.5% by weight of dry matter of at least two coumarins selected form the group of nodakenetin, rutaretin, celereoin and celerin.

Celery functionality

According to a further preferred embodiment, a celery material is used which significantly reduces IL-1 production in hPBMC. Preferably the celery isolate is capable of reducing the IL-1 production of LPS activated hPBMC by 50% at a concentration of below 12.5 μg dry celery material/ml, even more preferably at a concentration below 10 μg/ml, most preferably at a concentration below 5 μg/ml.

The inhibition of IL-1 production in LPS activated hPBMC by celery isolate can be determined according to the method as given in example 1.

A suitable celery material, showing an inhibition of IL-1 production in LPS activated hPBMC of 50%) at a concentration of below 5 μg/ml can be obtained by extracting celery seed with wateπethanol mixture (weight ratio 2:98), wherein the weight ratio celery seed to dry celery seed extract is about 16. Dosages of celery material

The celery material needs to be administered in a sufficient quantity to inhibit the release of IL-1 in vivo. Celery material is preferably administered in a daily amount between 0.5 and 250 mg, based on dry weight, of celery material per kg body weight. More preferably, a daily dosage comprises an amount between 1 and 150 mg, even more preferably between 1.5 and 100 mg, most preferably between 2 and 25 mg, based on dry weight, of celery material per kg of body weight.

For a human subject a daily dose of the composition according to the invention preferably provides an amount between 25 mg and 3 g, more preferably between 50 mg and 2 g, even more preferably between 100 mg and 1.5 g, based on dry weight, of celery seed isolate on a daily basis.

In a further aspect, the present invention provides a discrete dosage unit containing celery material and unsaponifiables from vegetable oil. The term "discrete dosage unit" refers to a dosage unit suitable for unitary administration to human subjects and other mammals, and wherein the unit contains a predetermined quantity of the present active principles and optionally, a pharmaceutically acceptable carrier. The discrete dosage unit according to the invention may. for example, comprise one or more tablets that are to be administered together, i.e. within a brief time interval (e.g. of less 15 minutes, more particularly of less than 5 minutes). In a particularly preferred embodiment the dosage unit is an enteral dosage unit, more preferably a solid or semi-solid dosage unit. Most preferably the dosage unit is designed for oral administration. Preferably the discrete unit dosage preferably contains between 10 mg and 4 g, more preferably between 50 mg and 1 gram, even more preferably between 100 mg and 750 mg celery seed material, preferably celery isolate, even more preferably celery seed isolate.

Unsaponifiable fraction of vegetable oil The unsaponifiable matter content of vegetable oil is a measure of the proportion of vegetable lipid material which does not react with alkali to form soap. Preferably the unsaponifiables employed in the present invention are non- volatile at 100°C to 105°C. The unsaponifiable matter content of vegetable oil enriched in unsaponifiables can be suitably determined according to the method described in "European Pharmacopoeia - Supplement 2001 (page 56). The unsaponifiable organic materials may include sterols, tocopherols, carotenoids, higher aliphatic hydrocarbons, squalene and lipidic furans. Preferably the vegetable oil enriched in unsaponifiables comprises a mixture of at least 3 of the aforementioned components. Even more preferably the vegetable oil enriched in unsaponifiables is enriched in at least three of the aforementioned components when compared to the crude, unfiltered vegetable oil from which it is obtained. The unsaponifiable matter content is usually expressed as a percentage of the total sample. Compositions from vegetable oil enriched in unsaponifiables have an increased percentage of unsaponifiable matter compared to the filtered crude oil. Vegetable oil generally contains below 3.5 % unsaponifiables, e.g. soy oil contains 0.7 % unsaponifiables and avocado oil contains 3 % unsaponifiables (Thiers, M. H., 1972 J. Med. Lyon 53 (222): 195-8). Suitable methods for the isolation of unsaponifiables from vegetable oil are well known to the art and have been already been described in the 1960's. Suitable methods are for example described in GB1142804, WO0151596 and WOO 170046, which are herein incorporated by reference.

Content of unsaponifiables

According to a preferred embodiment the isolate from vegetable oil enriched in unsaponifiables comprises at least 10 wt.% unsaponifiables based on the total weight of the vegetable oil isolate, more preferably at least 25 wt.%>, more preferably at least 50 wt.%, even more preferably at least 90 wt.%. The content of unsaponifiables in the vegetable oil enriched therein preferably is below 100 wt.%, more preferably below 98 wt.%, even more preferably below 95 wt.%.

The present composition preferably contains between 1 and 99 wt.%, more preferably between 2 and 75 wt.%, even more preferably between 5 and 50 wt.%, most preferably between 10 and 50 wt.% unsaponifiables based on the total dry weight of the composition. Sources of unsaponifiables

According to a preferred embodiment the isolate from vegetable oil enriched in unsaponifiables is obtained from a vegetable oil selected from the group consisting of avocado oil, soy oil, palm oil, corn germ oil, sunflower oil, canola oil and mixtures thereof. More preferably the unsaponifiables of avocado oil, soy oil or a mixture of these unsaponifiables are used, even more preferably the unsaponifiables of avocado oil are used, most preferably a mixture of unsaponifiables from soy oil and avocado oil is used. When a mixture of isolate from soy oil enriched in unsaponifiables and isolate from avocado oil enriched in unsaponifiables is used, the weight ratio soy oil unsaponifiables to avocado oil unsaponifiables is preferably between 1:10 and 10:1, more preferably between 5:1 and 1:1.

Dosage of unsaponifiables The vegetable oil enriched in unsaponifiable matter needs to be administered in an amount sufficient to decrease the effects of IL-1 in vivo. Preferably the isolate from vegetable oil enriched in unsaponifiables is administered in a daily amount equivalent to between 0.1 and 250 mg unsaponifiables per kg body weight on a daily base. More preferably, a daily dosage comprises an amount between 0.5 and 150 mg, even more preferably between 1 and 100 mg, most preferably between 2 and 25 mg unsaponifiables per kg of body weight.

For a human subject a daily dose of the composition according to the present invention preferably contains between 10 and 5000 mg, more preferably between 50 and 4000mg, even more preferably between 200 and 2000 mg unsaponifiables from vegetable oil. Preferably, the unsaponifiables are provided by oil enriched in unsaponifiables from vegetable oil.

The aforementioned discrete unit dosage preferably contains between 10 and 2500 mg, more preferably between 50 and 1000 mg, even more preferably between 100 mg and 1000 mg unsaponifiables from vegetable oil, preferably unsaponifiables from avocado oil. Preferably, the unsaponifiables from vegetable oil used in the discrete unit dosage is a mixture of at least 3, preferably at least 4 unsaponifiables. More preferably it is a mixture of at least 3, more preferably at least 4 unsaponifiables selected from the group consisting of sterols, tocopherols, carotenoids, higher aliphatic hydrocarbons, squalene and lipidic furans

Additional components

Further components may be coadministered in the method according to the present invention. Preferably these components contribute to the repair, treatment and/or prevention of connective tissue damage. Preferably these components are selected from the group consisting of glucosamine, chondroitin, collagen type II, S- adenosylmethionine, methylsulfonylmethane (MSM), hydroxyproline and mixtures thereof, more preferably form the group consisting of glucosamine and chondroitin. Glucosamine is a building block for glycosaminoglycans in cartilage and other connective tissue. When supplied exogenously, glucosamine stimulates connective tissue repair and synthesis. Glucosamine can thus be used advantageously in the present method, i.e. both celery and the composition enriched in unsaponifiable matter from vegetable oil reduce the effects caused by IL-1, while glucosamine provides the building blocks for the repair and/or maintenance of a healthy cartilage. The composition used in the present method can thus be advantageously combined with glucosamine, which can have a synergistic effect on the repair, treatment or prevention of connective tissue damage. Glucosamine and/or salts thereof are preferably administered in an amount equivalent to between 1 and 100 mg glucosamine per kg body weight on a daily basis, even more preferably in an amount equivalent to between 2 and 25 mg glucosamine. Chondroitine, similar to glucosamine, also provides building blocks for the in vivo production of cartilage and/or other connective tissue. The composition used in the present method can advantageously be combined with chondroitine, which is deemed to have a synergistic effect on the treatment or prevention of connective tissue damage. Chondroitine and/or a salt thereof is preferably administered in a daily amount equivalent to between 1 and 100 mg chondroitine per kg body weight , even more preferably in an amount equivalent to between 2 and 25 mg chondroitine per kg body weight. According to a particularly preferred embodiment the present composition includes glucosamine and chondroitine, making use of the synergistic effects provided by the combination of glucosamine and chondroitine.

Application

In the present method, the composition may be administered enterally, topically or transdermally, however is preferably administered enterally in the form of an edible composition, more preferably in the form of a pharmaceutical composition or a nutritional supplement. The terms pharmaceutical composition and nutritional supplement within the spirit of the present invention refer to compositions that include a pharmaceutically acceptable carrier. Pharmaceutical acceptable carriers are well known and described in the art.

For topical application, formulations in the form of gels, ointments, fatty ointments, creams, pastes, powders, milk, colloidal suspensions and tinctures are preferred. Additionally, compositions suitable for use with transdermal patches are encompassed. The topical product should contain between 0.1 wt.% and 80 wt.% of the combination of celery material and unsaponifiable matter from vegetable oil to achieve adequate pharmacological action.

According to a preferred embodiment the present method comprises the enteral administration of a composition comprising celery material and an isolate from vegetable oil enriched in unsaponifiables. Examples of compositions suitable for enteral administration, and particularly oral administration, include meals, bars, pills, capsules, gels, biscuits, drinks etc. According to a further preferred embodiment the present method comprises the administration of the composition in a solid or semisolid dosage form, more preferably in the form of one or more pills, capsules, tablets, caplets, microparticles and microspheres. The single solid or semisolid dosage unit form preferably has a weight between 0.1 and 30 grams, more preferably between 0.2 and 10 gram. When a pill is used to provide the celery material and unsaponifiable matter from vegetable oil, the pill preferably has a weight between 0.2 and 4 grams, even more preferably between 0.5 and 3 grams. A daily dosage may include one or more pills, however, preferably a daily dosage consists of 1 to 10 pills.

Treatments

According to a preferred embodiment the preparation according to the present invention is used in a method for the treatment of a interleukin-l mediated disorders selected from the group of: inflammatory conditions of a joint, including osteoarthritis, psoriatic arthritis and rheumatoid arthritis; pain and joint inflammatory condition resulting from strain, sprain, cartilage damage, trauma, orthopedic surgery, infection or other disease processes, more preferably from the group consisting of inflammatory conditions of a joint. The composition is especially useful in a method for the treatment, repair or prevention of damage to the connective tissue.

The present invention thus makes available effective therapeutic and prophylactic methods for restoring cartilage function and integrity. Such methods are useful, in particular for the repair of defects or lesions in cartilage tissue which are the result of degenerative wear such as observed in osteoarthritis, as well as other mechanical derangements which may be caused by trauma to the tissue.

The present method for the treatment, repair or prevention of damage to the connective tissue may be manifested in the form of products, such as nutritional supplements, advertised to treat or prevent undesirable conditions of the joint and/or the treatment or prevention of symptoms thereof. Nutritional supplements labeled with terminology pointing to the above method are encompassed by the present method. Such terminology includes for example "joint health", "mobility", "cartilage plus", "artri-cure", "joint pain", "flexibility", "joint-repair", "cartila", "osteobiflex" or "joint-flex". The term "symptoms" includes pain, tenderness of the joints, swelling, stiffness, morning stiffness and loss of cartilage. EXAMPLES

Example 1: Inhibition of IL-1 release by Celery seed extract.

PBMCs from human donors (hPBMC) stored in liquid nitrogen were thawed, washed and counted. In parallel a visual viability check is performed using trypan blue. 20 μl celery seed extract solutions (Celery seed ext dry cone 1:10, Mediherb, Warwick, Australia), with concentrations of 0; 3.5; 17.5; 35; and 70 μg celery seed extract/ml were added to the wells of a 96-well culture plate (Falcon-BD, Franklin Lakes, NJ, USA). Subsequently 150 μl hPBMC suspension was pipetted into the wells, 1.5 x 10⁵ cells per well. After 1 hour pre-incubation at 37 °C in a humidified atmosphere 30 μl LPS was added (10 ng/ml). Supernatants were collected 20 hours after incubation and human IL- lβ was measured using the human IL-lβ cytoset from Biosource (Camarillo, CA, USA). EIA/RIA plates from Corning Costar (cat.# 9018, Acton, MA, USA) were coated with coating antibodies 1: 1000 overnight from the Biosource cytoset. Plates were washed and blocked with 1% BSA in PBS. 100 μl 1 : 10 diluted supernatant was pipetted into the wells. 50 μl biotinylated antibody was added (1:1250) and the plates were incubated at room temperature on a shaker for 2 hours. Plates were washed 5 times and 100 μl streptavidine-HRP was added (1 :20000) and incubated at room temperature for 30 minutes. Subsequently the plates were washed 5 times. 3,3',5,5'-tetramethylbenzidine substrate solution was added. The color reaction was stopped by adding 100 μl 10% H₂SO₄ and the plates were measured at 450nm in a microplate reader.

It was found that the celery seed extract inhibited IL-1 production in LPS activated hPBMC by 50% at a concentration of at least 3.5 μg celery seed extract/ml. Example 2: Composition reducing the effects of IL-1 in humans. Nutritional supplement providing on a daily base:

• 800 mg celery seed extract comprising 10 wt.%> phtalides (Celery seed ext dry cone 1:10, Mediherb, Warwick, Australia)

• 800 mg avocado soy unsaponifiable material (Piascledine™, weight ratio unsaponifiables of avocado:unsaponifiables of soy 1 :2, (Pharmascience, Courbevoie, France)

• 600 mg tablet excipient

Example 3: Composition preventing damage to the connective tissue Nutritional supplement in the form of a soft gel providing on a daily base:

• 250 mg celery seed extract containing 10 wt.% phtalides (Celery seed ext dry cone 1:10, Mediherb, Warwick, Australia) • 250 mg avocado soy unsaponifiable material (Piascledine™, Pharmascience, Courbevoie, France)

• 375 mg glucosamineHCl

• 300 mg chondroitin sulfate

• Excipients: soybean oil, gelatin, glycerin, soya lecithin, yellow beeswax, Red 40 and titanium dioxide.

Claims

1. A composition comprising a. celery material b. an isolate from vegetable oil enriched in unsaponifiables; and c. optionally a pharmaceutically acceptable carrier

2. Composition according to claim 1, wherein the isolate from vegetable oil comprises at least 10 wt.% unsaponifiables based on the total weight of the isolate.

3. Composition according to any one of the preceding claims, wherein the celery material is a celery seed isolate.

4. Composition according to any one of the preceding claims, wherein the celery material is capable of inhibiting interleukin- 1 (IL- 1 ) production of lipopolysaccharide

LPS activated human peripheral blood monocytes (hPBMC) by 50% at a concentration below 12.5 μg dry celery material/ml.

5. Composition according to any one of the preceding claims, wherein the isolate from vegetable oil is obtained from oil selected from the group consisting of avocado oil, soy oil or mixtures thereof.

6. Composition according to any one of the preceding claims, wherein the composition further comprises at least one component selected from the group consisting of glucosamine, chondroitin, collagen type II, S-adenosylmethionine, methylsulfonylmethane (MSM) and hydroxyproline.

7. A composition for use in a method for the curative or prophylactic treatment of interleukin-l mediated disorders in mammals, said method comprising the enteral, topical or transdermal administration to a mammal of a composition according to any one of claims 1 to 6.

8. A composition according to claim 7, for use in a method for the prophylactic or curative treatment of damaged connective tissue.

9. A composition according to claim 7 or 8, wherein the method comprises the daily administration of between 0.5 and 250 mg, based on dry weight, of celery material per kg body weight.

10. A composition according any one of the claims 7-9, wherein the isolate from vegetable oil is administered in an amount equivalent to between 0.5 and 250 mg unsaponifiables per kg body weight on a daily base.

11. A discrete dosage unit comprising a. between 10 mg and 4 g celery material b. between 10 and 2500 mg unsaponifiables from vegetable oil.

12. The dosage unit according to claim 11, wherein the celery material is a celery isolate contains at least 4 wt.% phthalides.

13. The dosage unit according to claim 12, wherein the vegetable oil is avocado oil