TECHNICAL FIELD he present invention relates generally to the use of a combination of a P&a!fa spp, extract and another plant extract (e.g. deri ed from Sesamum spp,} to inhibit activation of mast cells, in particular, the present invention relates to the use of a combination of a P ila spp. extract and another plant extract (e.g. derived from S&sa um spp,) to prevent and/o treat a disorder associated with mast cell activation or degranuSailon such as a allergic disorder, anaphylaxis, an autoimmune disorder, or symptoms associated with mast cell activatio disorder, mast cell clonal disorder or mast cell neoplastic disorder,

BACKGROUND

Mast cells are granulocytes derived from myeloid stem cells. Mast cells are activated by allergens throug cross-linking with IgE receptors on the surface of the cells, physical injury through damage-associated molecular pattern molecules, pathogen- related agents through pathogen-associated molecular ^' pattern molecules, various compounds with their associated G-protei coupled receptors and complement proteins. Activation of mast cells causes complex intracellular signalling pathway t occur, which results in degranulatioo of mast cells. This is wher molecules stored in the granules of the mast cells are released outside of the cell. This includes:, for example, serine proteases {e.g, tryptase). histamine, serotonin and proteoglycan (e.g. heparin). Mast cells play a role in the immune and neuroimmurse systems, wound healing, angiogenesis and blood-brain barrier function, fviast cells have thus been implicated in numerous diseases.

Therefore, there is an ongoing need for compositions that inhibit mast ceils, whic may, for example, have prophylactic and/or therapeutic eff ects on disorders associated with mast cell activation and/or degranuiaSon. SUMMARY

The present invention aims, at least In part, to provide a composition having a specific combination of active ingredients, which acts to inhibit roast eel activation, in certain embodiments, the present invention aims to provide composition having a specific combination of active ingredients, which act synerglstically, to inhibit mast ceil activation and/or mast celi deg adatio and/or release of histamine from mast ceils.

According to a first aspect, ther Is provided a composition comprising a Perifte spp, extract and another plant extract.

The other plant extract may, for ex m le, be an anti-inflammatory and/or an anil- ai!ergic plant extract The other plant extract may, for example be derived from Ntg&Ma spp., Urti sp ^ Asi g&ius pp., P$ta$ii&s spp,, Qitrm zpp,, U a spp,, Layanduia pp., M&ntha spp,, E s!ypt spp,, Matricaria spp,, Rosmarinus spp., Curcuma spp., Aflmm spp.. or S&satm pp, In certai embodiments, the other plant extract is derived from S& mum spp;

The Peeifta spp, extract and/or the other plant extract may be an oil. In certain embodiments, the composition comprises periila oil and sesame oil.

The PBril!a spp. extract and other plant extract may be present in the composition In a weight ratio ranging from about 1:10 to about 10:1, The composition may, for example, be a solid preparation such as a tablet, capsule, capiet, dragee, lozenge, granule, powder, pellet, bead, cachet or bolus. The composition may, for example, be a liquid preparation such as an elixir, syrup, suspension, spray, emulsion, lotion, solution or tincture. The composition may, for example, be a semi solid preparation such as ointment, cream, paste, gel or Jelly.

According to a second aspect, there is provided a pharmaceutical composition comprising a composition according to the first aspect and a phamiaceuticaliy- acceptable carrier, excipient, diluent and/or additive. The pharmaceutically acceptable carrier, ©xeipient and/or dteent for use in the pharmaceutical composition can be selected from the group consisting of fillers _* binders, surfactants, preservatives, colourants, flavouring substances, pH regulators, regulators of the osmotic activity and salt-forming groups.

The composition may, for example, inhibit activation of roast cells, The composition may, for example, inhibit degranuiatioo of mast cells. The composition may, for example, Inhibit release of histamine from mast ^■ ceils. Thus, according to a third aspect, there Is provided the use of a composition comprising a PeriHa spp. extract and another plant extract to inhibit activation of mast cells,

According to a fourt aspect, ther is provided a therapeutic or non-therapeutic or vitro method of inhibiting activation of mas cells, the method comprising administering a compositio comprising a Pen? fa spp. extract and another plant extract to a subject.

According to a fifth aspect, there is provided a composition comprising a P&ri spp. extract and another plant extrac for use in a therapeutic method, for example for use in a therapeutic method of Inhibiting mast cell activatio .

According to a sixth aspect, there I provided the use of Persia, spp, extrac and another plant extract in the manufacture of a medicament or pharmaceutical composition,

The composition may. for example, inhibit degranulation of mast cells. The composition may, for example, inhibit releas of histamine from mast cells.

The composition may, for example, reduce histamine release by at least about 20% or by at least about 40% or by at least about 5δ%. For example, the composition may reduce the concentration of histamine in th tissue of a subject by at least shout 20% or by at least about 40% or by at least about S0%.

The composition may, for example, prevent and/or treat a disorder associated with mast cell activation and/or mast cell degranulation. The composition may, for example, prevent and/or treat symptoms associated wit a mast ceil activation disorder (e.g, mast ceil activation syndrome (¾?CAS)} _< an allergic disorder (e.g. allergic asthma, allergic rhinitis, conjunctivitis, aogsoederoa, urticaria, eczema, food allergy, drug allergy and insect allergy), anaphylaxis, an autoimmune disorder, symptoms associated with a mast ceil clonal disorder {e.g. mastocytosis) or symptoms associated with a mast cell neoplastic disorder (e.g, .mastocytoma, mast cell sarcoma and mast cell leukemia).

According to a seventh aspect, there is provide a method for making a composition comprising a Perifla spp. extract and another plan extract, the method comprising combining the PeriHa spp. extract and the other plant extract.

Embodiments of the inventio will be further described in the detailed description, Any embodiment described herein or any combination of embodiments described herein is applicable to any one or more aspects of th present invention unless clearl contradicted by cont xt

BRIEF DESCRIPTION OF THE D AWINGS

FIGURE 1 Is a graph showing the percentages of histamine reduciion from NMC-I ceil line when treated with periiSa seed oil, sesame seed oil, cromolyn and the combination, as .compared to the respective controls,

FIGURE 2 is a graph showing th percentages of histamine reduction from ^' HMC-1 cell fine when treated with the combination and Cromolyn, compared to the averaged histamine values of perfiia and sesame oils.

DETAILED DESCRIPTION

The present invention is based on, at least In part, the surprising finding that a combination of a Perifts spp. extract and another plant extract can inhibit mast cell activation. For example, embodiments of the present invention are based on the surprising finding that a Peri!!a spp. extract and another plant extract may work synerg sticail to inhibit mast ceil activation. In certain embodiments, the combination of a Per ilia spp> extract and another plant extract can be used to treat disorders associated with mast ceil activation and/or mast cell degranulation. Hereinafter, the invention shall be described according to preferred embodiments of the present invention and by referring to the accompanying description and drawings. However, it is to be understood that limi ing th descriptio to the preferred embodiments of the invention is merely to facilitate discussion of th present invention and it is envisioned that those skilled in the art may devise various modifications without departing from the scope of the appended claim.

Th terms generall used hereinbefore and hereinafter have for preferenc th meanings indicated below, unless Indicated otherwise, whereby more specific meanings may be used independently of one another in preferred embodiments of the present inventions instead of the general definitions _* these more specific significances describing especially preferred embodiments of the invention, W ere the term "at least one" or "one or more" occurs hereinbefore and hereinafter, ibis signifies in particular one to ten, for preference one to three, and in particular one or, further, two of the features enumerated, such as components, Where ranges are indicated, such as weigh percentage ranges, these include the limi value indicated; thus, for example, "between X and Y" signifies "from and including X up to and including Y",

The term "product" or "composition may mean In particular a phanrsaoayticai product or pharmaceutical compositio in the sense of a formulation, whereby this term is not restricted to pharmaceutical products suitable for registration, or a medical product or fictitious pharmaceuticals. The ^s producf or "composition" may refer fe e notraoeutical product in the sense of a formulation.

The term ^¾ anti~ailergic" is to be understood to relate to an agent o measure that prevents, inhibits, or alleviates an allergic reaction.

The term "aniHnfiarTimatory ^e ss to be understood to relate to an agent or measure that prevents, inhibits, or alleviates an inflammatory reaction.

The term "therapeutic treatment" or 'therapeutic method", also includes prophylaxis and the alleviation of symptoms i a subject, although not cosmetic treatments. This relates to therapeutic treatment, and in particular prophylaxis or alleviation, of disorders relating to mast cells, for example disorders relating to mast cell activation and/or mast .cell degranuiafion. The components referred to hereinbefore and hereinafter are in particular selected from among those such as are listed in pharmacopoeia, e,g, in the US Pharmacopoeia National Formulary, the Pharmacopoea Europea, the Pharmacopoea Helvetica, the British Pharmacopoeia, the German Pharmacopoeia, the Japanese Pharmacopoeia, the Chinese Pharmacopoeia, or supplements, such as by wa of decrees.

The presen invention relates to a composition comprising a P&rilta spp. extract and another plant extract. The composition may, for example, comprise one or more further plant extracts, For example* the composition may comprise a third or fourth or fifth or sixth plant extract. The composition may, for example, consist essentially of or consist of a P lia zpp. extract and another plant extract. The composition may, for example, consist essentially of or consist of a P&rilf spp. extract and another plant extract and a third plant extract and optionally a fourth, fifth or sixth plant extract, The term "extract ^* encompasses aqueous extract, non-aqueou extract, aloohoiic extracts,: evaporation, distillation (e.g. by water, steam, hydro diffusion, cohobation, rectification, fractional, etc), expression / cold pressing, supercritical COs extraction, dilutions, plant concentrates, plant oils, plant macerations, plant exudates, plant resins, plant powders and plant granules and any combination of two or more thereof. However, th invention should not be construed as being limited to such embodiments..

The P$r a spp. extract may, for exampie, be derived from Perfe frutescens {also known as pertila, Chinese basil and beefsteak), P illa i dora, Periifa hirteita, P&fffla seioyenms or a combination of two or mor thereof,

The other plant extract and the optional third, fourth, fifth or sixth plant extracts may, for example, each independently be an anti-inflammatory or an anti-ailergic plant extract The other plant extract and the optional third, fourth, fifth or sixth plant extracts may, for exampie, each independently be a mast cell stabilizing extract. The other plant extract and the optional third, fourth, fifth o sixth p\ani extracts may, for example, each Independently be a medicinal or therapeutic or nutritional or health benefiting plant extract,

The other plant extract and the optional third, fourth, fifth or sixth plant extracts may, for example, each be independently derived ^{' '} from Nigella spp. (e.g. Nigella saliva which is also known as black cumin, black caraway or fennel seed), Urtica spp. (e,g, rteftie). Astragalus spp, {e.g. milk vetch), Peimlfes spp, (e,g. butterbur), Citrus spp, (e.g. lemon), UrtG&ria spp, (e.g. Cat's claw), Lavandula spp. (e.g. lavender), Mentha spp. (e.g. peppermint), -Eucalyptus- spp. (e.g. Eucalyptus globulus which is also know as Tasmanian blue gum, souther blue gum or blue gum), Matricaria spp, (e.g. German chamomile), Rosmarinus spp. (e.g. rosemary), Curcum spp. (e turmeric). Allium spp. (e.g. garlic), Sesamum spp. or a combination of any I» or more thereof. In certain embodiments, the other (second) plant extract is derived torn Sesami/mspp.

The Periiia spp, extract, the other plant extract and the optional third, fourth, fifth and sixth plant extracts may, for example, each be Independently obtained from the seed roo leaf, or whole of the plant as listed in the preceding description. In certain embodiments, the plant extracts are each independently (e.g. all) derived from the seed of the pla nt in certai embodiments, the Periiia spp, extract is periiia oil and the other plant extract is sesame oil. In certain embodiments, the Periiia spp, extract Is periiia oil derived fnom the seed of the -plant of Periiia spp., and the othe plant extract Is sesame oil. In certain embodiments, the Periiia spp. extract is Periiia fmias m seed oil and the other plant extract Ϊ& Sesamum mdicum seed oil. in certain embodiments, the Periiia spp, extract Is periiia oil and the other plant extract Is nigella oil, eucalyptus oil, peppermint oil, lavender oil, lemon oil, chamomile oil, frankincense oil, basil oil, tea tree oil. clove oil or a combination of any two or more thereof.

The P&rilia spp, extract and the other plant extract may, for example, be present in the composition in a weight ratio ranging from about 1 :10 to about 10:1, For example, the Periiia spp. extract and the other plant extract may he present In the com osition in weight ratio from about 1:9 to about 9:1 or from about 1 :B to about 8:1 or from about 1:7 to about 7:1 or from about 1:8 to about 6:1, or from about 1:5 to about 5:1, or from about 1:4 to about 4:1 or from about 1:3 to about 3:1 _s or from about 1:2 to about 2:1. For example, the Peritte spp. extract and the other plant extract may be presen In a weight ratio of about 1:1. for example, the Per$a spp. extract and the other plant extfaot may be present n a weight ratio ranging from about 1 1 to about 10:1 or from about 1:1. to about 8: 1 or from about 1:1 to about 5:1 or from about 1:1 to about 2:1 ,

In certain embodiments, the PwMfe spp. extract is perjSIa oil and the other plant extract is sesame oil and th weight ratio of peria oil to sesame oil is in the range of about : 10 t 10:1, for example, from about 1:9 to about 9:1 , or from about; 1:8 to about 8:1, or from abou 1:7 to about 7:1 , or from about 1:6 to about 8:1, or from about 1 :5 to about 5:1, or from abouM:4 to about 4:1, or from about 1:3 to about 3:1, or from about 1:2 to about 2:1, or about 1:1, For example, the weight ratio of perii!a oil to sesame oil may range rom about 1 :1 to about 10:1 or from about 1 : 1 to about 8:1 o from about 1:1 to about S:1 or from about 1:1 to about .2:1,

The Pe a spp. extract and the other plant extract may, for example, be present in the composition in an amount by weight from about 5% to about 09.5% or from about 10% to about 9S% or from about 20% to about 90% or from about 30% to about β0% or from about 40% to about 70% or from abou 50% to about 60%. For example, the P&rilla spp. extract and ^' he other plant extract may, for example, be present in the composition in an amount by weight from about 20% to about 60% or from about 25% to about 55% or from about 30% to about 50% or from about 35% to about 45%,

The -composition of the present invention may be administered in the form of a composition comprising any suitable additional component. The composition may, for example, be pharmaceutica composition (medicament), suitable fo oral, nasal, topical, suppository, intravenous, intradermal, transdermal, transmucosal or inhalation administration. The composition may alternatively be a nutraoeuScai composition, for example, foodstuff, food supplement, dietary supplement, health supplement, meal replacement product beverage, beverage supplement, food additive, animal feed or feed additive. The compositions may take the form, for example, of solid preparations including tablets, capsules, eaplets, dragees, baanges, granules, powders, pellets feeads, cachets and boluses; liquid preparations including elixirs, syrups, suspensions, sprays, emulsio s, lotions, solutions and tinctures; o serai solid preparations including ointments, creams, pastes, gels and jellies. Techniques and formulations generally may be found In Remington, The Science and Practice of Pharmacy, Mack Publishing Co., Easfoa PA, latest edition.

The term "pharmaceutical composition" or "medscanien in the context of this invention means a composition comprising (a pharmaceutically effective amount of) a ΡΜ8 spp, extract and another plant extract, and additionally one or mor pharmaceutically acceptable carriers and/or excipients. The pharmaceutical composition may further contain ingredients selected from, for example, diluents, adjuvants, exeiplents, vehicles, c untehons, preserving agents, fillers, binders, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavouring agents, colouring agents, preserving agents, gelling agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents, coating agents, encapsulating agents, buffering agents, surfactant and dispersing agents, depending on th nature of the mode of administration and dosage forms.

In solid dosage forms of th invention {e.g. for oral or topical administration), the active ingrecfient(s} may be mixed with one or more phar iaceu ically acceptable carriers, such as dicslciofTi phosphate, and or any of the following: diluents, fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, microcrystaiiin cellulose, maifodexirine and/or silicic acid _s binders, such as, for example, hydroxypropyScellulose, hypromellose, hydroxypropyl methylceliuiose, carhoxymethyfcellulose, gelatine, polyvinyl pyrrolldones, polyvinyl acetate, sucrose and/or acacia; disintegrating agents, such as starch, for example, potato or tapioca starch, starch derivatives such as sodium starch glycotaie, crospoSyvinyipyroliidone, calcium carbonate, croscarmelfose sodium, alginlc acid, and certain silicates; lubricants, such as talc, calcium stearate,. magnesium sfearaie, stearic acid, sodium sulfate stearyl fumarate, solid polyethylene glycols, soiubiiiser such as sodium lauryi sulfate, g!idants such as silicon dioxide, flavouring and colouring agents and mixtures thereof. Tablets, and other solid dosage forms of the compositions of the invention, may optionally e prepared with coatings and shells, such as enteric coatings and other coatings well known in the phaFmaoeutical-formuSation art. They may also be formulated so as to rovde stew or controlled release of the active sngredient{s) therein S using, for exampl , natural and synthetic polymers such as hydroxypropyl methyl cellulose triethaerylates, methacrylic acid copolymers {e.g< methyl aorylate-meihscryilc acid copolymer and methyl me hacrylate-methacryile acid copolymers), shellac, eiby!cellulose, cellulose acetate phthalate, cellulos acetat Ifimetlitate, polyvinyl acetate pothalate, cellulose acetate succinate, hydroxy! propyl methyl cellulose acetate

10 succinate, sodium alginate, waxes, fatty acids , .zein, respectively, in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres may also used. These compositions may also optionally contain colourant and/or opacifying agents and may he of a compositio such ^' that they release the active ingredients) only, or preferentially, In a certain portion of the

I S gastrointestinal tract, optionally, in a delayed manner.

The compositions ma comprise no mor than a out SO % w/w of pharmaceutically acceptable carrier and/or excipieni, for example, no more than about 45 % w w of pharmaceuiloally acceptable carrier and/or exoiplents, or no more than about 40 % of20. w/w pharmaceutically acceptable carrier and/or excipients, o no more than about 35 % w/w of pharmaceutically acceptable carrier and/or excipients. For example, the pharmaceutical composition may comprise at least about 1 % w/w, or at least about 1.0 % w w, or at least about 15 w/w o at least about 2D % w/w, o at least about 25 % w/w, or at least about 30 % of phamiaceutica!iy acceptable carrier and/or

25 excipients.

Liquid ©r semi-solid form preparations include solutions, suspensions, and emulsions, fo example, water or waier-propyiene glycol solutions (e,g> for oral or topical administration). Liquid or semi-solid preparations can also be formulated In solution In30 aqueous polyethylene glycol solution or an ointment base as known In the art (e.g. petrolatum). In certain embodiments, the active ingredients), I.e., plant extracts, may be mixed with one or more pharmaceutically acceptable carriers, such as water and/or an of the following: solvent such a propylene glycol, !sopropyi myrlstate, alcohol; humeciant such .as glycerol; sweeteners such as liquid glucose, corn syrup and

35 sucrose; artificial sweeteners such as aspartame, stevia and syeralose; preservatives such as' phehox ethaho!, capryiyi glycol _* ethylhexylglycehn, benzalkonium chloride, potassium sorbate, disodium -EOTA, benzoates and parabens; viscosity modifiers/thickeners such as. gums, soybean oil, polyacrylaies (e.g. sodium po!yaorylaie. Carfcopoi 980 polymer) and alginates; buffering agents such as monopofassium phosphate, dt-potasslum drogen phosphate and glycerin; going or emulsifying agents such as bentooite, xanthan gum, carrageenan, potysorbate 80, poiysorbate 20, gelatin, cetearyi alcohol, lecithin, glycerol monostaarate, flavouring •agents In aqueous or in oil forms such as vanilla oil; cooling agents suo as menthol; soothing or moisturising agents such as Aloe vera, hyaluronic acid; antioxidants such as tocopherols and citric acid; colouring agents; pH regulators and surfactants.

Also included are solid form preparations, for example, tablets, capsules, granules and powder, which are intended to be converted, shortl before use, to liquid -form preparations for oral administration. Such liquid .forms, include solutions, suspensions, and emulsions. These particular solid form preparations are most convenientl provided In unit dose form and as such are used to provide a single liquid dosage unit. Alternatively, sufficient solids may be provided so that multiple individual liquid dose may be reconstituted when required, by measuring pr¾determfned volumes of the solid for preparation: as with a spoon, o other measuring device. The solid form preparations intended to be converted to liquid form may contain, in addition t the active material, flavourings, colourants, stabilizers, buffers, artificial and natural sweeteners, dispersants:, thickeners, solubiiising agents, and the like. The liquid utilized for preparing the i! uid form preparation may be water, isotonic water, juices, milk, eihanol, and the like as welt as mixtures thereof .

The terms "food", "foodstuff', food- supplement", "dietary supplement, "health supplement", "meal replacement producf , "beverage ^* and "beverage supplemenf used herein have the normal meanings fo those terms, and are not restricted to pharmaceutical preparations. Other composition forms are also included within the present invention. These may, fo example, include, a foodstuff precursor such as a rehydratafela powder or a beverage precursor such as a powder dispersible In water, milk or other liquid.

Also included are solid form preparations which are intended to be combined with a food or foodstuff before oral consumption. The solid form preparations may e mixed. Into the food or foodstuff or applied to the food or foodstuff, e.g., by sprinkling onto the food or foodstuff. Such solid forms include powders, granules, pellets and the Ike, in certain embodiments, the food or foodstuff , and the .like, comprises from about 0,1 wt % to about 50 wt % of the composition of the invention described herei , based on the total weight of the food or foodstuff, for example, from about 0.1 wt, % to about 40 wt, or from about 0.1 wt. % to about 30 wt , o from; about Q ^: .t wt:, % to about .20 wt, %, or from about 0.1 wt. % to about i ^' § wt or from about 0.1 wt, % to about ^" 10 wt. %, o from mhaut 0.1 wt. % to about 8 wt. Si, or from about 0.1 wt. % to about 8 wt, %, or from abou 0. wt, % to about 4 wt, %, or from about 0.1 wt. % to about 2 wt. % of the composition of the Invention described herein, i certain embodiments, the food or foodstuff, and the iike. comprise at least about 0. 2 wt. % of the compositions of the invention described herein, based on the total weight of the food or foodstuff, for example, at least about 0,5 wt, %, or at least about 1 wt. %: _> or at least about 5 wt % of the composition of ^' the invention described herein.

The composition may further comprise other biologicall active agents, for example, bioiogieal active agents suitable for Inhibiting mast ceil activation or suitable: for preventing or treating a disease or disorder associated with mast ceil activation or mast ceil degranulation. For example, the composition may further comprise other anti- allerg active agents.

The composition or pharmaceutical composition may further comprise a nutrient ingredient selected from the group consisting of vitamins and minerals, and combinations thereof. The vitamin may be any one or more of vitamin A, vitamin D, vitamin £, vitami K, thiamine, riboflavin; pyridoxin©, cyanocobalaroln, carotenoids (including be!a-carotefte, aeaxanthin;. Lutein and lycopene}, niacin, folic acid, pantothenic acid, biotin, vitamin€, choline, inositol, and salts and derivatives thereof. The mineral may b any one or more of calcium, phosphorous, magnesium, Iron, zinc, manganese, copper, cobalt, boron, iodine, sodium, potassium, molybdenum, selenium, ^• chromium, fluorine and chloride.

The composition or pharmaceutical composition may further comprise dietary fibre of plant and/or non-piant origin. The term "dietary fibre ^* used herein has Its normal meaning for this term. It is generally regarded as the indigestible portion of food derived from plants. Typically, there are two mam components of dietary fibre; soluble fibre, which dissolves in water, and Insoluble fibre, which does not dissolve in water. Soluble fibres _. Include in lln, chitosan, gym acacia, guar gum, low-metboxy and hlgb- methaxy pectin, oat and/or barley beta giucans, carrageenan, psyllium, cyetodextfin, and derivatives thereof. Insoluble fibres include oat hull fibre, pea hull fibre, so hull fibre, soy cotyledon fibre, sugar beet fibre, cellulose, corn bran and derivatives thereof.

The compositions and pharmaceutical compositions described herein may be used In _. , various therapeutic, non4herapeutic and m viirv applications. For example, the .compositions ^' and pharmaceutical com ositions ^{' '} described herein ^{' '} ma be used to inhibit mast cell activation and/or mast cell degranulation and/or release of histamine from mast cells. For example, the compositions and pharmaceutical compositions describe herein may be used In an vitm method or in a in mo method. The methods may comprise administering the composition or. pharmaceutical composition described herein to a subject.

According to one aspect of the present invention, there is provided a composition •according to any aspect or embodiment described herein for use in a therapeutic or non-therapeutic method for .Inhibiting, mast cell activation (a therapeutic or non- therapeutic method for mast cell stabilization). The method may, for example^ comprise contacting the mast ceils with a composition according to any aspect or embodiment described herein.

In certain embodiments, the plant extract components have a synergistic effect on Inhibiting mast cell activation and/or mast cell degranulation and/or release of histamine from mast cells, ast ceils are activated by allergens through cross-linking with IgE receptors on the surface of the cells, physical injury through damage-associated molecula pattern molecules, pathogen-related agents through pathogen associated molecular pattern molecules, various compounds with their associated G-pro!ein coupled receptors and complement proteins. Activation of a mast cell causes a complex intracellular signalling pathway to occur, which results in degranulation of the mast cells.- Thi is where molecules stored in the granules of the mast cell are released outside of the cell. This Includes* for example, serine proteases (e.g. tryptase) _* histamine, serotonin and proteoglycans (e.g. heparin). Inh bi ing activation of mast cells may, for example, include any on© or more of these steps. For example, the compositions disclosed herein may be useful in inhibiting mast ceil degranuSation, For example, the compositions disclosed herein may be useful in inhibiting release of histamine from the mast cel.

Thus, in certain embodiments, the composition according to any aspect or embodiment may reduce histamine release by at least about 20 , For example, the compositio may reduce histamine release by at least about 30% or at least about 40% or at least abou S0 or at least about 60% or a least about 70% or at least about 75% or at least about 8:0 or at least about 85% or a least about 90% or at leas about 95%, For example, the composition may reduce histamine release by Lip to 100%, for example up to 99% or u to 98%. This may, fo example, he determined by measuring histamin content In cell culture medium, for example as described in the examples below,

According to further aspects, there is provided a composition according to any aspect or embodiment of th present Invention for use in preventing and/or treating a disorder associated with mast ceils, for example a disorder associated with mast cell activation and/or mast cell degranuiation. for example, there is provided a composition according to an aspect for use in preventing and/or treating an allergi disorder and/or anaphylaxis and/or an autoimmune disease, or symptoms associated with mast ceil activation disorder, a mast ceil clonal disorder and/or a mast cell neoplastic disorder. In certain embodiments, the disorder is not a nasal disorder. In certain embodiments, the disorder i not an allergic disorder o the nasal cavity, in certain embodiments, the disorder is not seasonal allergic rhiniti or perennial allergic rhinitis.

According to a further aspect, there Is provided the use of a Pmiia spp, extract and another plant extract in th manufacture of a medicament or pharmaceutical composition. The medicament or pharmaceutical composition may, for example, be for preventing and/o treating an allergic disorder and/or anaphylaxis and/or an autoimmune disease, or symptoms associated with mast ceil activation disorder, a mast ceil clonal disorder and/or a mast cell neoplastic disorder.

Ivlasf cell activation disorders encompass disorders caused b or associated with Inappropriate .activation of mast ceils.: The classification of mast cell activation disorders was laid out in Akin ef ai. ^* ast-ceif activation syndrome: Proposed diagnostic criteria", J. Allergy Gtin. Immunol, (2010), 126 (8): 1099-1104. For example, . the mast cell activation disorder may be mast cell activation syndrome (MGAS). Allergic disorders encompass disorders caused by hypersensitivity of the Immune system. Generally, allergic disorders are caused by hypersensitivity to an allergen that causes littl reaction in most subjects. Allergens thai may be related to allergic disorders may be derived from, for example, foods, latex, medications, pollen and other animals fe,g. animal hair, insect stings). Allergic disorders encompass allergic disorders of the nasal cavity, including seasonal allergic rhinitis and perennial allergic rhinitis. Allergic disorders also include allergic conjunctivitis, allergic contact dermatitis, atopic dermatitis and allergic asthma. Anaphylaxis is a serious allergic reaction, in certain embodiments, the allergic disorder is not an allergic disorder of the nasal cavity (e.g. is not seasonal allergic rhinitis and/or Is not perennial allergic rhinitis).

Autoimmune disorder encompass disorders caused by or associated with an abnormal immune response against a substance that is normall present in the body. Autoimmune disorders Include, fo example, allergic asthma, Addison disease, celiac disease, derrn tomyositis, tSravas disease, Hashimoto thyroiditis, multiple sclerosis, myasthenia gravis, pernicious anemia, reactive arthritis, rheumatoid arthritis, Sjogren syndrome, systemic lupus erythematosus, bullous pemphigoid and type I diabetes. For .example, autoimmune disorders include autoimmune, inflanin atory disorders of the Joints and skin.. Mmi cell clonal disorders encompass disorders caused by or associated with the presence of an increased number of mast celis in comparison to a normal (healthy} subject For example, mast ceil clonal disorders include mastocytosis.

Mast ceil neoplastic disorders encompass disorders caused by or associated with abnormal growth of mast cells, For example, mast cell neoplastic disorders includes mastocytomas, mast call sarcoma and mast cell leukemia.

The composition according to any aspect or embodiment may reduce histamine concentration in he Issue of a subject by at least about 20%, For example, the composition may reduce histamine concentration in the tissue of a subject by at least about: 30% or at least about 40% or at least about 50 or at least about 60% or at least about 70% or at least about 75% or at least about 80% or at least about. SS% or at leas! about 90% or at least about 95%·. For example, the composition may reduce histamine concentration in the tissue of a subject by up to 100%, for example up to 99% or up to 98%,

Histamine concentration in the tissue of a subject may be measured by any method known to a person skilled in the art. For example, histamine concentration may be determined by microdialysis sampling followed by qyantificatson with commercially availabl ELiSA kits or by HPtC, For example, histamine concentration at a localised site may be determined using tissue samples such as tissue homoganates. For example, histamine concentration ma be determined using tissue from a site where an allergen has come into contact with mast ceils or a sampling site ( g. skin, mucosal lining (respiratory tract, gastrointestinal), blood, organs, bone marrow, etc),

The composition according to the first aspec or a pharmaceutical composition according to the second aspect of the in ention can further comprise- an anti-al!ergie drug or pharmaceutically active anti-allergic substance, for example an antihistamine or an additional mast cell stabilizer (e.g. cromoglicate, etc). In certain embodiments, the composition or the pharmaceutical composition can further comprise an antihistamine, including H anfihist.am!ne (e.g. a elastine, olopatadine, t iproSidine, bilastine, cetiriEine, pyrilamioe etcX. Hj-antinisiamsne (e.g. cimetidine, famotidine:, etc) and Hs-antihlstamine (a.g, fhio eramide, etc). The antihistamine opposes the activit of histamine receptors in a subject nd: further enhances the inhibitory effect of mast ceil activation of the anti allergic oil and oily component (e.g. pehlia oil and sesame oil) In the composition or the pharmaceutics! composition. I certain embodiments, azelastine ca he used as the antihistamine In the composition of the invention* as I has a triple mode of action, i.e. antihistamine effect, mast -cell stabilizing effec and antl nfiammatory effect. Fo example, the composition or the pharmaceutical composition may comprise from about 0.1 wt. % to about 50 wt %, or from about 0,01 wt % to about 10 wt %, or from about 10 wt, to about 40. wt. %, or from about 20 wt % to about 30 wt. % of antihistamine by weight of the composition of the invention described herein.

In certain embodiments, the subject is a human, in other embodiments, the subject ^' is a mammal other than a human, such as non-human primates (e.g. apes, monkeys and lemurs), companion animals such as cats or dogs, working and sporting animals such as dogs, horses and ponies, farm animals such as pigs, sheep, -goats, deer, oxen and cattle, and laboratory animals such as rodents (e.g, rabbits, rats, mice, hamsters, gerbils or guine pigs).

In certain embodiments, the composition is administered daily to the subject (e.g. orally or topically administered daily to the subject). The amount of composition administered may be varied depending upon the requirements of the subject For both therapeutic and non-therapeutie applications, the amount of composition administered ma b varied depending upon the desired results, the requirements of the subject and the severity of the condition being treated. Determination of the proper amount/dosage for a particular situation is within th skill of the art. Fo example, fo therapeutic applications physician o veterinarian having ordinary skill in the art can readil determine and prescribe the effective amount of the pharmaceutical com position required , The total daily amount dosage may be divided and administered in portions during th day if desired .

In general, a suitable daily close of active agents i the composition according to the invention will be that amount which is the lowest dose effective to produce the desired effect, for example, a therapeutic effect, and/or to inhibit mast cell activation and/or to inhibit mast ceil degranulatlon and/or to inhibit release of histamine from mast ceils. It is contemplated that a wide range of doses may be used, due to the non-toxic nature of the composition. person of ordinary skill in the art will understand that a suitable dos or dosage will typicall vary from subject to subject and will be dependent on factors such as the severity of health conditions of the subject at the outset of administration of the composition. For example, the dose of active agents (i.e. plant extracts) in the composition ma be up to 15 § per day, for example,, tip to about 10 g per day, or up to about 5 g pe day. In certain embodiments, the doses of active agents In the compositio is i the range of 100 mg to about 3 g per day, which may be administered as two or three or more sub-dose administered separately at appropriate Intervals throughout the day, optionally in unit dosage forms. In certain embodiments,, the dose of active agents In the composition may be from about 10 mg to about 3 g of each extract component per day, for example, from about 500 mg to about g of each component per day, or from about 750 mg to about 2.S g of each component per day, or from about 1000 mg to about 2008 mg of each component per day. In certain embodiments, the composition may be administered two or three times a day, optionally before, with, or after a meal. In certain embodiments, each dose of active agents is no more than about S g, for example, no more than about 3 g, for example, no more than about 2.5 g. Each dose of the plant extracts in the composition may toe combined with other conventional agents for inhibiting mas cell activation.

Use for the manufacture of a product for the therapeutic treatment may comprise in particular the appropriat procurement of th produc (e,g. as a pharmaeeuHca roduct i the broader or narrower sense); in other words, the manufacture of the preparation, its introductio i partioslar into any device.' for its administration, its packing, and the corresponding provision of instructions for use I therapeutic- treatment, for example by way of a package insert and/or printings on the package.

The compositions and pharmaceutical compositions described herein may be prepared by combining a Psrilfa spp, extract and a second plant extract and optionally any one or more of the other ingredients described herein, such as one ©r more further piani extracts, dietary fibre, nutrients, biologically active: agents and pharmaceutical exeipiants end/o earners and/or diluents. The components are combined in suitable amounts to obtain ^' a composition having the desired quantity of each component. Each component may be combined! with one or more other components in any order and combination suitable to obtain the desired product, Such methods are well know In the art, for example, methods known in the food industr (e.g. those used in the preparation of health food bars and the like) and methods teown in the pharmaceutical industry. The composition may be prepared in th dry solid form, for example, powder form,, and subject to further processing step depending on the types of the formulation for the intended finished products. The methods may further comprise forming step, wherein the mixture is moulded, pressed, spray dried or otherwise formed into a shape (e.g. bar, bail, pellet, clusters, tablet, beads), preferably with dimensions and/or textures suitable for consumption by a human or other mammalian animal of the types described herein. In one of the embodiments, the composition or pharmaceutical composition may be produced in the form of pills, including gel capsules.

The preparations can be manufactured according to conventional and known methods, which include in particular the mixing of the components, if required step by step and/or under movement and/or heating of the fluid, for example by stirring. The Invention will now be ^■■ described in detail byway of reference only to the following non-limiting examples. A PLES

■l amate 1: Μ&$ί C®$tJMt

Pas ceil lines, i.e. HMQi cells {provided by Dr Joseph H. Butterlieid; -Mayo Clinic:, Rochester, MH _} USA} _> were cultured in Isocove ^' s Modified DuSbecco ^' s Medium (t M) containing 10 % Fetal Bovine Seum (FBS), 2 mU l-giutamine, SO μΜ 2~ mercapiQethanoi, 100 U/ml penicillin and 100 pgimL streptomycin. Cells were maintained at 37 ^* C in 5% CG ₃ in a humidified atmosphere.

Percentage of living cells was determined by trypan blue staining, A 1:10 dilution with 50 I cell suspension and 4S0 μΙ trypan blue was prepared and the number of living and dead cells was counted in a hemocyiometer. 10 ^s living cells were transferred into safe-lock Eppendorf tubes. Cell suspensions were oentrifuged (200 g, 5 min) and superna ant was discarded. The effect of perilla Oil, sesame oil and combination of eiilia oil and sesame oil (1:1 ratio) i the vehicle oliiphor E P (polyoxyl-35 hydrogenated castor oil) was determined. The positive control Cromolyn in phosphate buffe saline (PSS), the vehicles PBS and ELP were also tested as controls. After adding 100 μ\ IgE (100 ng/mi in PBS o ELP) to the cell pellets, cells were incubated for 12 hours over night, at 37 ^* 0. Cells were then incubated for 2 hours, at 3 with two different concentrations of compounds, i.e. 25Q/ g/mi and 600 £¾/mi. After cenfrlfugation (200 g, 5 min) supernatant was discarded and cell pellet was again incubated for 1 hour, at 37 ^* 0 with 100 μ\ anti-!gE (1 //g/rni In PBS or ELP). For the final incubation time of 6 hours, at 37X, 300 μ\ HMC1 media was added. Cells were again eeotrifuged (200 g, 5 min) and supernatant was coilected and stored as 300 1 aiiquots at -20°C for EtlSA measurements. Cell pellets were resuspended In 100≠. HMC1 media for determining cell viability. Viability of the cells (percentage of living cells) was determined by trypan blue staining (1:10 dilution wtih SO μ cell suspeftsiors and 450 trypan blue was prepared and the number of living and dead ceils was counted in a hemocytometer). The concentration of histamine In the collected supernatant was determined by ELSSA, using a kit obtained from !Bt internationai according to the manufacturer's guidelines. Each sample was measured twice. Due to expected measuring limits, vehicle {PBS, ELP) treated sample were diluted 1:2, Statistical analysis wa performed wit GraphPad Prism {Version 5.01) software (GraphRad Software inc., San Diego, C A, USA) using the unpaired two-tailed Student f test A P value of less than 0.05 was considered statistically significant

The results are shown in Figures 1 and 2, as well as Tables 1 and 2 below.

Tafole 1.

Summary of histamin mediator release from H&3C-1 ceiis. Signif leant difference compared to negative control was at P<0,G5.

Table 2.

Histamin reduction compared to averaged histamine mediaior releas of perllla oil and sesa me oil from H C-1 ceils, showing enhancement effect of the combination. Significant difference compared to averaged values of pe lla oil and sesame oil was at P<0,05.

Comparing the average histamine release of perii!a oil and sesame oil to th combination, the combination showed significantly reduced stamine- releas and therefor mast call stabilization at 250 and 500 /g ml (p~0,0185 and p~0,OQ92) respectively.

Wh n H C-1 call were treated individually with penlla oil or sesame oil, tie percentage of histamine reduction achieved was lower than that of Cromolyn, Whe treated with the combination, the percentage of histamine reduction was higher than treatment with Cromolyn (Figure 1 }. Treatment with the combination increased the percentage of histamine reduction by 18,7 - 22.5% compared to treatment with individual compounds, while treatment with Cromolyn only differed by 6.8— 8,4% of histamine reduction compared to treatment with individual compounds.

Ther Is an approximately 2-fold and 3~fold differance between t e combination and Cromolyn at concentrations of 2S0 pg/ l and 500 #g mJ respectively. It can be conducted that the combination had enhanced mast ceils stabilizing effect, which increased the percentage of histamine reductio compared to treatment with ndividual compounds, and reached a final effect that was better than Cromolyn,

Exampte 2: Antisifergie Fwmuiatmn

Tables 3 - 5 show anti-allergic pill formulations In a hard or soft shell capsule,

Table 3,

: Ingreilien Percent ge (%) by weight

Periila oil 49.75

Sesame oil 49,75

Tocopherol 0.5 Table 4

ingredient Percentage {%) by weight

Peniia oif 20 5

Uilica extract 13.8

Astragalus extract 12.2

lge !a oil 18.5

Soybean oil 30

scitte 5

Table S.

Table 6.

Ingredient Percentage (%) by weight

49.0

Sesame oil 49.0

Cetirizine hydrochloride 1.0

Citric add 1.0

Tables 7 and 8 show an anfrallergic topical gel formulation.

Table ?,

ingredient Percentage { } I jy weight

; Periiia oil 15

Sesame oil

Disodium EDTA 0.1:

; Menthol 0.5

Propylene glycol 12

I Carbopoi 980 polymer 0,85

Polysorbate 80 1.5

royrisiate 2.0

; Citric acid 0.1

Wate 62,95

Table .8.

in redient Percentage (%) by weight

Peniia oil 15

Sesame oil ^■ 5

Tables 9 and 10 stow an a f-aSlerg loiion formulation Table 9,

Table 10,

Table 11 shows a anti-allergic ointment formulation. able 11 , Menthol

Ointment base

Table 12 shows an anii-ailerg;c gel paich formulation.

Table 12,

ί Ingre ient Percentage {% by weight

I Refills oil

> Sesame oil 5

^; Water 20

; Glycerin 20

^■ Propylene glycol 10

i Hyaluronic acid 7

Potassium sorbaie 1

Sodium poiyacrylate 2

Tables 13, 14 d 15 show anti-allergic pill formulations in a tablet

Table 13.

Perilla extract 38.5

Astragalus extract 15.4

Croscamieilose sodium 7 7

f Cellulose 6.8

Silicon dioxide 0.5

Magnesium stearate 0.3

Table 14.

Tabl 15.

Ingredient Percentage {%) by weight

Perilla e tract 25.3

Urtica extract 25.3

Astragalus extract 25.3

Croscarmellos© sodium 1.5 Cellulose microcrystaSiine 20.1

Silicon dioxide 0,5

Maonesium siearaie 1.5

0.5