FR3104024A1 - ANTI-ALLERGIC COMPOSITIONS BASED ON SAFFRON AND FLAVONOID - Google Patents

Abstract

The present invention relates to a composition comprising saffron or a mass fraction of saffron and at least one flavonoid for use in the prevention and / or treatment of IgE-mediated type I allergic conditions. This invention also relates to a dietetic and / or food composition comprising saffron or a mass fraction of saffron and at least one flavonoid.

Description

ANTI-ALLERGIC COMPOSITIONS BASED ON SAFFRON AND FLAVONOID

The present invention relates to a composition comprising saffron or a mass fraction of saffron and at least one flavonoid for use in the prevention and/or in the treatment of IgE-mediated type I allergic conditions. This invention also relates to a dietary and/or food composition comprising saffron or a mass fraction of saffron and at least one flavonoid.

Allergy is defined as a symptomatic reaction to a normally harmless environmental antigen. Hypersensitivity develops in the sensitizing and effector stages. Sensitization is an asymptomatic primary immune response to antigen. The effector phase is a harmful secondary response. Specifically, there are four types of allergies: Type I hypersensitivity (also known as allergy or atopy) is mediated by IgE antibodies specific for antigens that are normally non-pathogenic (allergens). In type I hypersensitivity, an allergen triggers the production of allergen-specific IgE which binds via its Fc region to mast cells and basophils FcεRs. At the early effector stage, re-exposure of allergen-sensitized mast cells triggers immediate degranulation and release of preformed mediators, as well as synthesis of inflammatory molecules. Sensitized eosinophils and basophils are involved in the late effector phase by degranulation and production of additional mediators. Local atopic responses include dermatitis, urticaria, asthma, and rhinitis, whereas anaphylaxis is a life-threatening systemic atopic response. Food allergies are atopic reactions to allergens consumed. Type II hypersensitivity occurs when IgG or IgM antibodies bind to epitopes on fixed or mobile cells, triggering antibody-dependent cell-mediated cytotoxicity, complementary activation and/or phagocytosis. Type III hypersensitivity occurs when soluble antigen binds to IgG or IgM to form large, insoluble immune complexes. Complex deposits in narrow channels like capillaries trigger inflammation. Type IV hypersensitivity occurs when effector T cells, macrophages, and other leukocytes induce delayed damage to inflammatory tissue.

Type I hypersensitivity (IgE-mediated immediate-type hypersensitivity reactions) is most commonly referred to as allergy and will be used in that sense here.

The main allergens responsible for IgE-mediated allergic reactions (type I) are inhaled allergens such as pollens (eg birch, rye, ragweed), house dust mites, mold spores, animal dander and food (eg. nuts, seafood and peanuts).

In type 1 hypersensitivity, B cells are stimulated (by CD4+TH2 cells) to produce antigen-specific IgE antibodies that coat mast cells and basophils. Mast cells play a central role in triggering and maintaining inflammation, especially in allergies and asthma. Although mast cells are found in all well-vascularized tissues, they are mostly found in tissues where the body comes into contact with the outside world. These tissues include the gastrointestinal tract, upper and lower respiratory tract, and skin. Antigens binding to IgE antibodies trigger anaphylactic degranulation of mast cells and basophils, which is the immediate and explosive release of pharmacologically active preformed mediators like histamine, platelet activating factor, IL-6, leukotrienes (LTC4 , LTD4 and LTB4), prostaglandin and other cytokines.

The main effects of these products are vasodilation and contraction of smooth muscles such as bronchoconstriction.

Histamine, the most important mediator, exerts its effects in allergic diseases mainly by interacting with histamine H1 receptors which are present in various organs like nerve endings, blood vessel walls and smooth muscle of the pathways. respiratory. Histamine has broad biological effects. Depending on where the histamine is released, its biological effects vary from mild discomfort due to itching to life-threatening bronchoconstriction. In the nose or skin, histamine induces vasodilation and increases vascular permeability leading to edema and erythema. It stimulates sensory nerve endings that cause itching or sneezing. In the lungs, histamine causes vasodilation of the bronchial airways leading to bronchoconstriction.

According to the World Allergy Organization, more than 20% of populations in most developed countries have allergies (Pawankar R, et al, 2013), of which about 20-25% eventually develop allergic asthma. This transition to allergic asthma is accompanied by a change in medication: Over-the-counter antihistamines and decongestants are replaced or supplemented with more potent topical and systemic corticosteroids, leukotriene inhibitors and sometimes even monoclonal antibodies, which inhibit the production of IgE completely.

About 60% of all asthmatics have allergic (IgE-mediated) asthma. The avoidable indirect costs of inadequate allergy treatment in the EU alone are estimated at €55-151 billion per year (European Academy of Allergy and Clinical Immunology EAACI, 2016).

The most common allergic manifestation of type 1 hypersensitivity is allergic rhinitis, mainly triggered by airborne allergens. Allergic rhinitis (AR) affects approximately 20% of the population in industrialized countries and significantly impairs daily activity, work productivity, sleep and quality of life for people of all ages.

Allergic rhinitis involves inflammation of the mucous membranes of the nose, eyes, Eustachian tubes, middle ear, sinuses and pharynx. Although the nose is always involved, the other organs are not necessarily involved in all individuals. Mucosal inflammation is characterized by a complex interplay of inflammatory mediators, but is ultimately triggered by an immunoglobulin E (IgE)-mediated response to an extrinsic protein.

In the case of bronchial asthma, a first general distinction is made between intermittent asthma and persistent asthma. Symptoms of wheezing and coughing that do not exceed 2 days per week are considered intermittent asthma. Persistent asthma is sub-categorized into mild persistent asthma (symptoms: twice a week, lung function 80% of normal), moderate persistent asthma (symptoms: daily, lung function 60 and 80% of normal), and persistent asthma severe (symptoms: daily and often, lung function less than 60% of normal).

Symptoms of severe persistent asthma consist of swelling of the throat and lungs which leads to coughing, wheezing and shortness of breath.

According to the "International European Respiratory Society (ERS)/American Thoracic Society (ATS) Guidelines on Definition, Evaluation and Treatment of Severe Asthma", severe asthma is defined as "asthma that requires treatment with high-dose inhaled corticosteroids ( ICS) plus a second controller (and/or systemic corticosteroids) to avoid becoming "uncontrolled" (Fan Chung, K; Wenzel, S. International ERS/ATS Guidelines on Definition, Evaluation, and Treatment of Severe Asthma, London 2013) Corticosteroids are synthetic forms of cortisone like prednisone, methylprednisolone, prednisolone, dexamethasone and hydrocortisone.

A severe allergy affects quality of life, and those affected often have to take sick days because the symptoms prevent them from working or they are unable to perform outdoor activities. day when pollen conditions are poor. Systemic effects, including fatigue, drowsiness and malaise, accompany the inflammatory allergic response to varying degrees. These symptoms are responsible for the deterioration in quality of life reported by most allergy sufferers (Thompson AK, Juniper E, Meltzer EO. Quality of life in patients with allergic rhinitis. Ann Allergy Asthma Immunol, 2000).

Currently, allergic pathologies such as allergic rhinitis, allergic conjunctivitis and allergic asthma are incurable. Treatment is symptom-focused and includes allergen avoidance, symptomatic drug therapy, and allergen-specific immunotherapy (ITA), commonly referred to as desensitization.

Although allergen avoidance and immunotherapy are theoretically ideal, in practice they are either impossible to achieve or immediately ineffective. Pharmacological treatment mainly focuses on

a) prevent the degranulation of mast cells and basophils,

b) Blockade of receptors for mediators such as histamine, cytokine, leukotriene,

c) counter the inflammatory aspects of allergy with potent corticosteroid agents,

d) and, as a last resort in the event of permanent severe asthma, the total suppression of the production of IgE by the action of humanized anti-immunoglobulin E (IgE) monoclonal antibodies.

Corticosteroids have cumulative, partly irreversible side effects, such as weight gain, water retention, hypertension, depression, hypercholesterolemia and irreversible, dose-related side effects, such as an increased disposition to diabetes, cataract formation, osteoporosis and skin atrophy.

In the treatment of IgE-mediated allergies, there is an unmet need for alternative anti-inflammatory and anti-allergic agents with similar or superior efficacy to corticosteroids, providing satisfactory long-term results and reduced side effects.

Unexpectedly, the applicant has discovered that the combination of saffron and flavonoids in the compositions according to the present invention has a synergistic and remarkable effect in the prevention and treatment of IgE-mediated type I allergic conditions, including asthma. severe allergic, significantly exceeding the effectiveness of corticosteroids. The Applicant has also identified that this combination can also be used as a dietetic and/or food composition in order to reduce the uncomfortable manifestations associated with allergies.

According to a first aspect, the invention relates to a composition comprising saffron or a mass fraction of saffron and at least one flavonoid for use in the prevention and/or in the treatment of IgE-mediated type I allergic conditions.

By "mass fraction of saffron" is understood according to the invention a part of the saffron comprising molecules which are usually found in natural saffron. These saffron fractions can be expressed as a percentage of saffron mass. These mass fractions may thus correspond to safranal, crocin, crocetin or picrocrocin initially present in total saffron.

The terms “prevention and/or treatment” designate both therapeutic treatment and prophylactic or preventive measures, the objective of which is to prevent or attenuate the condition or the pathological disorder in question.

Saffron is a spice derived from the stigmas and styles of the Crocus Sativus flower. Metabolomics analysis detects nearly 1000 compounds in saffron, of which only about 150 compounds, volatile and non-volatile, have been identified. Saffron is originally marketed for its color, flavor and aroma and includes the main metabolites that determine the quality of saffron. The main bioactive compounds are crocetin, crocin, picrocrocin, safranal and their derivatives. The chemistry of saffron is complex and there are many types of saffron, obtained from different origins and packaged differently, which are distinguished by the amount of their main components, such as safranal, crocin, crocetin, or picrocrocin.

Picrocrocin (4-(β-D-glucopyranosyloxy)-2,6,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde, C16H26O7) is the substance responsible for the characteristic bitter taste of saffron. So far, picrocrocin has not been detected in any other raw material, of vegetable or animal origin, and it is therefore considered a molecular marker of true saffron. Its concentration is generally between 1-15% (del Campo CP, et al. Picrocrocin Content and Quality Categories in Different (345) Worldwide Samples of Saffron (Crocus sativus L., 2010) although in some samples the picrocrocin content may reach 20%.

Safranal (2,6,6,6-trimethyl-1,3-cyclohexadiene-1,3-cyclohexadiene-1-carboxaldehyde, C10H14O) is the main volatile compound of saffron and the aglycone of picrocrocin. It has been detected in very few plant products and can also be generated when certain carotenoids undergo a thermal process. The concentration of safranal is much lower than that of crocin and picrocrocin, usually between 0.01 and 0.6%.

Crocin is a carotenoid that is primarily responsible for the color of saffron. Chemically, crocin is the diester formed from the disaccharide gentiobiose and the dicarboxylic acid crocetin. When isolated as a pure chemical compound, it has a dark red color and forms crystals with a melting point of 186°C. The term crocins can also refer to members of a series of related hydrophilic carotenoids which are either monoglycosyl or diglycosyl esters of crocetin.

Crocetin is the substance responsible for the coloring properties of saffron which is a glycosidic ester of the carotenoid dicarboxylic crocetin (8,8'-diapocarotenedioic acid). The glycosides linked to crocetin are gentiobiose, glucose, neopolitanose and triglycose, which are found in saffron in their trans (mainly) and cis (minority) forms. All of these compounds are referred to as crocins in the literature, although crocin is actually just an ester of trans-crocetin di(β-D-gentiobiosyl). The coloring strength of saffron depends on the concentration of these compounds, which normally varies from 10 to 30% in the dried stigma of Crocus sativus L., and rarely reaches up to 36% in exceptional years.

Crocin and crocetin can be obtained in large quantities from the fruits of Gardenia Jasminoides, an evergreen plant in the Rubiaceae family.

The concentrations of safranal, crocin, crocetin, picrocrocin vary depending on the source of the saffron. An exhaustive analysis of 16 saffron samples from Greece, Iran and Spain by the Institute of Pharmaceutical Biology of the University of Münster yielded, on average, 12.04% Picrocrocin and 20.06% Crocin in Greek Saffron, 6.9% Picrocrocin and 15.49% Crocin in Iranian Saffron, 8.03% Picrocrocin and 18.14% Crocin in Spanish Saffron (M. Lechtenberg et al, Quality and Functionality of saffron: Quality Control, Species Assortment and Affinity of Extract and Isolated Saffron Compounds to NMDA and σ1 Receptors, 2009).

An analysis of 22 Greek saffron samples yielded 15.3% ± 3.1% Picrocrocin and 33.5% ± 4.2% Crocetin sugar esters, with an average Picrocrocin/Crocetin ratio of 0. 5 ± 0.1 (Chrysanthou A, Pouliou E, et al. Sensory Threshold Studies of Picrocrocin, the Major Bitter Compound of Saffron, 2015).

In a Moroccan study, saffron from different origins was analyzed, yielding 4%-28% Picrocrocin, 18-36% Crocins and 0%-0.5% Safranal (Lage M, Cantrell C. Quantification of saffron ( Crocus sativus L.) crocin, picrocrocin and safranal metabolites for quality determination of the spice grown under different environmental Moroccan conditions. 2009).

Preferably, the mass fraction of saffron comprises at least one compound from natural or synthetic sources chosen from safranal, crocin, crocetin, picrocrocin, derivatives thereof and/or mixtures thereof.

Even more preferably, said mass fraction of saffron comprises only the fraction from a natural or synthetic source corresponding to safranal, to crocin, to crocetin, or to picrocrocin.

According to one embodiment of the present invention, the saffron can be in the form of one of its extracts. The extracts can be obtained by means of extraction methods well known to those skilled in the art, such as steam distillation, maceration, solvent extraction (for example H2O, ethanol, methanol, hexane and propane/butane), counter-current extraction, supercritical fluid extraction, microwave-assisted extraction, phytonic processes, ultrasonic-assisted extraction, etc.

Flavonoids are a class of secondary phenolic metabolites found especially in the leaf epidermis and skin of fruits, vegetables and grains, and are therefore common substances in the daily diet (Hollman & Katan , 1999; Middleton et al., 2000). There are more than 15,000 natural flavonoid compounds (Xiao, J. Dietary flavonoid aglycons and their glycosides: Which show better biological significance? Crit. Rev.Food Sci. Nutr.2017). Their general structure consists of a skeleton with 15 carbon atoms, forming two aromatic rings (A and B) and a heterocyclic ring (C), abbreviated C6-C3-C6. The carbon skeleton of flavonoids is:

Depending on their respective degree of oxidation of the heterocyclic C ring, flavonoids are subdivided into different subclasses, of which flavonols, flavones, flavan-3-ols, anthocyanidins, flavanones and isoflavones are the most common.

Preferably according to the invention, the IgE-mediated type I allergic conditions are allergic asthma and/or allergic bronchitis and/or rhinitis and/or conjunctivitis and/or skin rash and/or dermatitis atopic and/or clogged ears.

These allergic conditions can be intermittent, persistent, mild, moderate or severe.

Preferably, according to the invention, said at least one flavonoid is at least one flavonol, from a natural or synthetic source.

Flavonols such as quercetin, isoquercetin, quercitrin, isoquercitrin (quercetin-3-O-β-D-glucoside), kaempferol, fisetin, isorhamnetin, hyperoside, rutin, l Astragalin, avicularin, and myricetin are a distinct class of flavonoids that possess a 3-hydroxyflavone backbone. These are the most common flavonoids. The distribution and structural variations of flavonols are extensive and have been well documented. The main dietary flavonols, quercetin, kaempferol, isorhamnetin (3-O' methylated quercetin) and myricetin, are most often O-glycosides. Conjugation most often occurs at the 3 position of the C ring, but substitutions can also occur at carbons 5, 7, 4', 3', and 5'. Although there is obviously only one aglycone per individual flavonol, there are many flavonol conjugates, for example over 200 different glycosidic conjugates of quercetin alone.

The carbon skeleton of flavonols is:

Preferably according to the invention, said at least one flavonol is chosen from quercetin, myricetin, kaempferol, rutin, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin, their metabolites, their analogues, their chemically modified analogues, their derivatives or their mixtures.

Myricetin is commonly derived from vegetables, fruits, nuts, berries, tea, and is also found in red wine. Its structure is similar to that of fisetin, luteolin and quercetin.

Kaempferol (3,4′,5,7-tetrahydroxyflavone) is found in a wide range of plants and is common in Pteridophyta, Pinophyta and Angiospermae.

Rutin, also called rutoside, quercetin-3-O-rutinoside and sophorin, is a glycoside combining quercetin and the disaccharide rutinose (α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranose). It is a citrus flavonoid found in a wide variety of plants, including citrus fruits.

Fisetin (7,3′,4′-flavon-3-ol) is a polyphenol found in many plants, where it serves as a dye. It is also found in many fruits and vegetables, such as strawberries, apples, persimmons, onions and cucumbers.

Luteolin is the main yellow dye compound obtained from the Reseda luteola plant, which has been used as a source of the dye since at least the first millennium.

Apigenin is a natural product found in many plants, in the form of a yellow crystalline solid that historically has been used to dye wool.

Taxifolin, also known as dihydroquercetin, is the 2,3-dihydrogen derivative of quercetin. It is found in acai (Euterpe oleracea), Siberian larch (Larix sibirica) and silymarin extracted from milk thistle seeds. It is also present in small amounts in red onions.

Dihydromyricetin, or ampelopsin, is found in Ampelopsis japonica, megalophylla, and grossedentata species; Cercidiphyllum japonicum; Hovenia dulcis; Rhododendron cinnabarinum; certain species of Pinus; and some species of Cedrus, as well as in Salix sachalinensis.

Laricitrin is formed from myricetin by the action of the enzyme myricetin O-methyltransferase. This compound is present in red grapes and in Vaccinium uliginosum (bog bilberry).

Seringetin is formed from laricitrin by the action of the enzyme laricitrin 5'-O-methyltransferase. This compound is present in red grapes and in Vaccinium uliginosum (bog bilberry).

Preferably according to the invention, said at least one flavonol comprises at least one aglycone chosen from the aglycones of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof.

Preferably according to the invention, said at least one aglycone comprises quercetin aglycone.

Quercetin aglycone (non-glycosylated) is rarely found in nature. But in its glycosylated forms, quercetin is the most abundant flavonoid in plants, fruits and vegetables, but most often bound to one or more sugar molecules in the form of β-glycosides.

The carbon skeleton of quercetin is:

Preferably according to the invention, said at least one flavonol comprises at least one glycoside chosen from the glycosides of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof.

Preferably according to the invention, said composition comprises at least one quercetin glycoside.

A quercetin glycoside is formed by placing a glycosyl (a sugar such as glucose, rhamnose or rutinose) in place of one of the OH groups (usually in position 3).

Preferably according to the invention, said at least one quercetin glycoside is chosen from isoquercetin, isoquercitrin, at least one isoquercetin alpha-glycoside, at least one isoquercitrin alpha-glycoside and/or mixtures thereof .

Isoquercitrin and isoquercitrin are terms that are sometimes used interchangeably, as these two molecules have a very similar structure. The difference between the two is that isoquercitrin has a glucofuranose ring structure while isoquercetin has a glucopyranose ring structure. Functionally, the two molecules are nearly indistinguishable. Isoquercitrin and isoquercetin have been shown to be much more bioavailable and absorbed faster than quercetin aglycone or quercetin glycosides, such as rutin (quercetin rutinoside). Enzymes and transport mechanisms active in cells lining the small intestine (enterocytes) interact with the glucose moiety of isoquercitrin and isoquercetin molecules, accelerating their absorption and transformation into quercetin aglycone. Without this glucose moiety, absorption of quercetin from the small intestine is the slowest and least efficient passive diffusion process. Isoquercitrin and isoquercetin therefore provide quercetin, with all its benefits, in a more bioavailable form.

Alpha-glycosyl isoquercitrin (alpha-oligoglucosyl quercetin 3-O-glucoside, abbreviation AGIQ) is for example produced by the Japanese manufacturer San-Ei Gen F.F.I. of Osaka and marketed under the name Enzymatically Modified Isoquercitrin (Enzymatically Modified Isoquercitrin) under the trademark EMIQ. EMIQ contains 30% AGIQ, declared as 18-20% Rutin trihydrate.

Alpha-glycosyl Isoquercitrin is a mixture of the monoglucoside isoquercitrin and its alpha-glycosyl derivatives which, independently of each other, comprise from 2 to 11 linear glucose fragments (Figure 1).

It is made by enzymatic hydrolysis of rutin followed by transglycosylation using dextrin and cyclodextrin glucanotransferase (CGTase). Conjugation of multiple glucose moieties confers increased water solubility and 3 times greater bioavailability than isoquercitrin and 17.5 times that of quercetin aglycon. Because it is actively and rapidly absorbed, alpha-glycosyl isoquercitrin reaches peak plasma levels within 15 minutes (C _max and AUC _0-12h : Enzymatically Modified Isoquercitrin, a-Oligoglucosyl Quercetin 3-O Glucoside, Is absorbed more easily than other Quercetin Glycosides or Aglycone after oral Administration in Rats; T. MAKINO, R. SHIMIZU, M. KANEMARU, Y. SUZUKI, M. MORIWAKI, H. MIZUKAMI).

Table 1: Significantly equally effective dosages of quercetin aglycone, isoquercetin/isoquercitrin, alpha-glycosyl isoquercitrin

_Cmax C _max rel. with quercetin AUC _{0-12 hrs} AUC _{0-12 hr} rel. with quercetin Bioequivalent to 1000 mg quercetin aglycone Quercetin aglycon 0.26 100% 2.6 100% 1000mg Isoquercetin
Isoquercitrin 2.66 1000% 15.8 600% 170mg α-glycosyl Isoquercitrin 10.7 4100% 46 1800% 50mg

Preferably according to the invention, the composition also comprises quercetin metabolites such as the O-methylated, O-glucuronidated and O-sulphated quercetin metabolites, chemically modified analogues of quercetin or mixtures thereof.

Preferably according to the invention, said composition comprises for a daily dose between 0.01 mg and 20 mg, preferably between 0.02 mg and 15 mg, more preferably between 0.03 mg and 10 mg of saffron or a fraction mass of saffron per kilogram of body weight, and between 0.01 mg and 15 mg, preferably between 0.05 mg and 12 mg per kilogram of body weight of one or more flavonoids, preferably quercetin aglycone and/or isoquercetin and/or isoquercitrin and/or alpha-glycosyl isoquercitrin or mixtures thereof.

Preferably according to the invention, the saffron contains from 5 to 45% crocin.

Preferably according to the invention, the saffron contains from 1 to 20% picrocrocin.

Preferably according to the invention, the saffron comprises from 0.0001% to 0.7% of safranal.

Preferably according to the invention, the mass fraction of saffron comprises from 5 to 45% of saffron, corresponding to its part of crocin, and/or from 1 to 20% of saffron, corresponding to its part of picrocrocin and/or 0.0001% to 0.5% saffron, corresponding to its saffron part.

The specific weight ratio of saffron to isoquercetin/isoquercitrin depends on parameters such as, for example, the composition of the saffron and the bioavailability of the quercetin compound used. The higher the bioavailability of the quercetin compound, the lower the amount required for a therapeutic effect.

Preferably according to the invention, said saffron or said mass fraction of saffron and said at least one flavonoid are administered simultaneously.

Preferably according to the invention, the composition comprises saffron and at least one flavonol chosen from quercetin aglycone and its glycosides such as isoquercetin, isoquercitrin, alpha-glycosyl isoquercitrin or mixtures thereof.

Preferably according to the invention, said composition further comprises at least one pharmaceutically acceptable compound, preferably chosen from anti-inflammatory compounds and/or for the treatment of IgE-mediated type I allergic conditions, and/or for the treatment of allergic asthma and/or allergic bronchitis and/or rhinitis and/or conjunctivitis and/or skin rashes and/or atopic dermatitis and/or ear plugging.

Preferably, the compound for the prevention and/or treatment of IgE-mediated type I allergic conditions and/or for the treatment of allergic asthma, allergic bronchitis, rhinitis, conjunctivitis, atopic dermatitis and/or blocked ears includes corticosteroids, antihistamines, fluticasone propionate, salmeterol xinafoate, prednisolone, injections of steroids such as triamcinolon, ectoin, loratadine, azelastine, ipratropium bromide, fenoterol, phlorizine hydrolase, glucosidase or mixtures thereof.

Preferably, the composition further comprises pharmaceutically acceptable excipients and carriers.

The compositions covered by the present invention may comprise all the pharmaceutical forms normally used for any route of administration (orally, by injection or topical application, for example) to obtain the desired therapeutic effect. In one case, the composition of the present invention is formulated in a solid form which is chosen from among tablets, granules, dragees or capsules. According to one embodiment, the composition is formulated in the form of a hard capsule. According to one embodiment, the composition is formulated in the form of a liposome. According to one embodiment, in particular in the case of administration of quercetin metabolites, injection or inhalation can be used.

The administration of saffron and the administration of one or more flavonoids can be done in any order, i.e. first the saffron and then the flavonoid(s), or first the flavonoid(s). flavonoids and then saffron.

In one case, the administrations of saffron and one or more flavonoids are carried out in a period not exceeding 6 hours, preferably not exceeding 4 hours, more preferably not exceeding 2 hours, even more preferably not not exceeding 1 hour and even more preferably not exceeding 30 minutes.

Preferably according to the invention, said saffron or said mass fraction of saffron and said at least one flavonoid are administered simultaneously.

The composition according to the present invention can be administered to a patient daily, once a day, twice a day, three times a day or even more times a day. In the case of more than one administration per day, the single doses of saffron and one or more flavonoids are adjusted so as to possibly administer the total daily dose required for a period of 24 hours. In one embodiment, the composition of the present invention is administered to a patient twice daily and every 12 hours, the daily dose of saffron and one or more flavonoids being divided equally between the two administrations.

To optimize the filling of the capsules and the dosage of the composition according to the present invention, the following classes of known excipients could preferably be used: anti-caking agents, trehalose, pullulan, sweeteners, surfactants (cationic, anionic or nonionic) , diluents, aggregating or binding agents, lubricants, slip agents, stabilizers, solubilizers, emulsifiers, humectants, flavors, coating agents, coloring agents, flavor correctors or mixtures thereof.

According to a second aspect, the present invention relates to a dietary and/or food composition comprising saffron or a mass fraction of saffron and at least one flavonoid.

By dietetic and/or food composition is understood according to the invention a composition which can be ingested, such as a food supplement, intended to supplement the normal diet and which constitute a concentrated source of nutrients or other substances having a nutritional effect. or physiological.

Preferably according to the invention, the mass fraction of saffron comprises at least one compound from natural or synthetic sources chosen from safranal, crocin, crocetin, picrocrocin, their derivatives and/or their mixtures.

According to one embodiment of the present invention, the saffron can be in the form of one of its extracts.

Preferably according to the invention, said at least one flavonoid is at least one flavonol, from a natural or synthetic source.

Preferably according to the invention, said at least one flavonol is chosen from quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin, their metabolites, analogues, chemically modified analogues, derivatives or mixtures thereof.

Preferably according to the invention, said at least one flavonol comprises at least one aglycone chosen from the aglycones of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof.

Preferably according to the invention, said at least one aglycone comprises quercetin aglycone.

Preferably according to the invention, said at least one flavonol comprises at least one glycoside chosen from the glycosides of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof.

Preferably according to the invention, said flavonol comprises at least one quercetin glycoside.

Preferably according to the invention, said at least one quercetin glycoside is chosen from isoquercetin, isoquercitrin, at least one isoquercetin alpha-glycoside, at least one isoquercitrin alpha-glycoside and/or mixtures thereof .

Preferably according to the invention, the composition also comprises quercetin metabolites such as the O-methylated, O-glucuronidated and O-sulphated quercetin metabolites, chemically modified analogues of quercetin or mixtures thereof.

Preferably according to the invention, said composition comprises for a daily dose between 0.01 mg and 20 mg, preferably between 0.02 mg and 15 mg, more preferably between 0.03 mg and 10 mg of saffron or a fraction mass of saffron per kilogram of body weight, and between 0.01 mg and 15 mg, preferably between 0.05 mg and 12 mg per kilogram of body weight of one or more flavonoids, preferably quercetin aglycone and/or isoquercetin and/or isoquercitrin and/or alpha-glycosyl isoquercitrin or mixtures thereof.

Preferably according to the invention, the saffron contains from 5 to 45% crocin.

Preferably according to the invention, the saffron contains from 1 to 20% picrocrocin.

Preferably according to the invention, the saffron comprises from 0.0001% to 0.7% of safranal.

Preferably according to the invention, the mass fraction of saffron comprises from 5 to 45% of saffron, corresponding to its part of crocin, and/or from 1 to 20% of saffron, corresponding to its part of picrocrocin and/or 0.0001% to 0.5% saffron, corresponding to its saffron part.

The specific weight ratio of saffron to isoquercetin/isoquercitrin depends on parameters such as, for example, the composition of the saffron and the bioavailability of the quercetin compound used. The higher the bioavailability of the quercetin compound, the lower the amount required for a therapeutic effect.

Preferably according to the invention, said composition further comprises at least one dietary and/or food-acceptable compound, preferably chosen from anti-caking agents, trehalose, pullulan, sweeteners, surfactants, diluents, anti-caking agents. aggregators or binders, lubricants, slip agents, stabilizers, solubilizers, emulsifiers, humectants, flavors, coating agents, coloring agents, acidity regulators or mixtures thereof.

The compositions according to the present invention can comprise all dietary and/or food acceptable forms normally used.

According to one embodiment, the composition according to the present invention is formulated in a solid form which is chosen from tablets, granules, dragees or capsules. According to another embodiment, the composition is formulated in the form of a hard capsule. According to one embodiment, the composition is formulated in the form of a liposome.

The administration of saffron and the administration of one or more flavonoids can be done in any order, i.e. first the saffron and then the flavonoid(s), or first the flavonoid(s). flavonoids and then saffron.

In one case, the administrations of saffron and one or more flavonoids are carried out in a period not exceeding 6 hours, preferably not exceeding 4 hours, more preferably not exceeding 2 hours, even more preferably not not exceeding 1 hour and even more preferably not exceeding 30 minutes.

Preferably according to the invention, said saffron or said mass fraction of saffron and said at least one flavonoid are administered simultaneously.

According to one embodiment, saffron or a mass fraction of saffron and one or more flavonoids are taken orally.

The composition according to the present invention can be ingested daily, once a day, twice a day, three times a day or even more times a day. In the case of more than one administration per day, the single doses of saffron and one or more flavonoids are adjusted so as to possibly administer the total daily dose required for a period of 24 hours. In one embodiment, the composition of the present invention is administered to a patient twice daily and every 12 hours, the daily dose of saffron and one or more flavonoids being divided equally between the two administrations.

Preferably according to the invention, the dietary and/or food composition comprises saffron and at least one flavonol chosen from quercetin aglycone and its glycosides such as isoquercetin, isoquercitrin, alpha-glycosyl isoquercitrin or mixtures thereof.

According to a third aspect, the invention relates to a kit for the manufacture of a dietary and/or food composition according to the invention comprising:
- saffron or a mass fraction of saffron;
- at least one flavonoid.

Preferably according to the invention, the mass fraction of saffron comprises at least one compound from natural or synthetic sources chosen from safranal, crocin, crocetin, picrocrocin, their derivatives and/or their mixtures.

According to one embodiment of the present invention, the saffron can be in the form of one of its extracts.

Preferably according to the invention, said at least one flavonoid is at least one flavonol, from a natural or synthetic source.

Preferably according to the invention, said at least one flavonol is chosen from quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin, their metabolites, analogues, chemically modified analogues, derivatives or mixtures thereof.

Preferably according to the invention, said at least one flavonol comprises at least one aglycone chosen from the aglycones of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof.

Preferably according to the invention, said at least one aglycone comprises quercetin aglycone.

Preferably according to the invention, said at least one flavonol comprises at least one glycoside chosen from the glycosides of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof.

Preferably according to the invention, said flavonol comprises at least one quercetin glycoside.

Preferably according to the invention, said at least one quercetin glycoside is chosen from isoquercetin, isoquercitrin, at least one isoquercetin alpha-glycoside, at least one isoquercitrin alpha-glycoside and/or mixtures thereof .

Preferably according to the invention, said at least one flavonol further comprises quercetin metabolites such as the O-methylated, O-glucuronidated and O-sulphated quercetin metabolites, chemically modified analogues of quercetin or mixtures of those -this.

Preferably according to the invention, said kit allows the manufacture of a daily dose of dietary and/or food composition according to the invention, comprising between 0.01 mg and 20 mg, preferably between 0.02 mg and 15 mg , more preferably between 0.03 mg and 10 mg of saffron or a mass fraction of saffron per kilogram of body weight, and between 0.01 mg and 15 mg, preferably between 0.05 mg and 12 mg per kilogram of body weight one or more flavonoids, preferably quercetin aglycone and/or isoquercetin and/or isoquercitrin and/or alpha-glycosyl isoquercitrin or mixtures thereof.

Preferably according to the invention, the saffron contains from 5 to 45% crocin.

Preferably according to the invention, the saffron contains from 1 to 20% picrocrocin.

Preferably according to the invention, the saffron comprises from 0.0001% to 0.7% of safranal.

Preferably according to the invention, the mass fraction of saffron comprises from 5 to 45% of saffron, corresponding to its part of crocin, and/or from 1 to 20% of saffron, corresponding to its part of picrocrocin and/or 0.0001% to 0.5% saffron, corresponding to its saffron part.

The specific weight ratio of saffron to isoquercetin/isoquercitrin depends on parameters such as, for example, the composition of the saffron and the bioavailability of the source of quercetin used. The higher the bioavailability of the quercetin compound, the lower the amount required for a therapeutic effect.

Preferably according to the invention, said kit further comprises at least one dietary and/or food-acceptable compound, preferably chosen from anti-caking agents, trehalose, pullulan, sweeteners, surfactants, diluents, anti-caking agents. aggregators or binders, lubricants, slip agents, stabilizers, solubilizers, emulsifiers, humectants, flavors, coating agents, coloring agents, acidity regulators or mixtures thereof.

Figure 1 shows a high performance liquid chromatography (HPLC) chromatogram of alpha-glycosyl isoquercitrin powder (commercial product EMIQ® R 20 from San-Ei Gen FFI, Osaka, Japan) used in the examples herein. invention. The distribution of isoquercitrin and its alpha glycosides is given in percentage by weight, according to the total weight of the EMIQ®-R-20 composition. The concentrations of alpha oligo-glycosides n = 7 to 11 are consolidated into a single value.

EXAMPLES

Example 1: Composition comprising saffron and quercetin aglycone

Quercetin aglycone (Zein Pharma), made from Sophora Japonica flowers, was orally administered to 6 adult volunteers, with severe allergies but otherwise healthy, in the midst of the annual pollen allergy season.

All subjects had a multi-year history of allergic rhinitis and additionally suffered from mild to severe allergic asthma, confirmed by positive skin tests to various tree and grass pollens (all subjects), specific radioallergen IgE tests (3 subjects ), spirometric respiratory function tests and clinical history (all subjects).

All subjects were taking antiallergic drugs, three of them had received inhaled corticosteroids and two had received systemic corticosteroids. Initially, despite their respective medications, the subjects presented with strong symptoms of nasal and/or ocular seasonal allergic rhinitis and mild, moderate and severe allergic asthma, reducing their quality of life.

The efficacy of the protocol was assessed by daily recording of the total symptom score in an allergy diary, supplemented by spirometric lung function tests and changes in consumption of the respective usual antiallergic drugs.

At 250 mg twice daily (500 mg/day), quercetin aglycone alone did not produce a significant reduction in allergic symptoms in any of the 6 study subjects, according to the Total Symptom Score Diary, Flow Meter and the usual need to continue anti-allergic treatment, and no improvement in quality of life. Due to extremely weak or no responses, the dose was increased to 500 mg twice daily (1000 mg daily), again with no noticeable improvement.

100-150 mg of pure, unaltered saffron (200-300 mg/day), was then added to quercetin aglycone, and allergic symptoms including allergic asthma showed significant, rapid and unexpected remission, making the use of obsolete systemic and/or inhaled corticosteroids.

Suppression of asthmatic symptoms began 40-50 minutes after oral administration in all subjects and lasted an average of 12 hours.

Nasal allergy symptoms were significantly reduced, but still required occasional treatment with topical antihistamines, although at a much lower level than before treatment with the flavonoid/saffron composition.

The suppression of eye symptoms was less pronounced than the suppression of asthma and nasal symptoms. The suppressive effect of nasal and ocular symptoms was approximately 12 hours in all subjects at the height of allergen exposure, and 16 to 24 hours towards the end of the allergy season.

Taken alone, neither saffron nor quercetin aglycon produced noticeable relief of allergy symptoms in any of the people in the trial.

In a systematic trial and error approach, it quickly became apparent that the therapeutic action of the saffron aglycone/quercetin combination depends on the dose administered, hence the need to calibrate the saffron aglycone/quercetin combination according to the patient's body weight. subject.

Accordingly, subjects were treated with 2 capsules per day, each containing:
- 1.5 mg of saffron per kg of body weight
- 2.5 mg quercetin aglycon per kg body weight

Conclusion: The saffron/quercetin aglycone formulation resulted in a very significant reduction in allergic asthma symptoms: approximately 60-70% reduction in symptom scores and 70-80% reduction in the use of systemic and/or inhaled corticosteroids . Nasal symptoms such as sneezing, runny nose, nasal obstruction were also significantly reduced: about 50% reduction in symptom scores and 60% reduction in nasal spray use. Ocular symptoms such as itchy eyes, watery eyes, and conjunctival congestion all showed slightly less, but still significant, reduction: about 30-40% reduction in symptom scores and 40-50% reduction in eye symptoms. use of topical allergy eye drops.

Negative side effects consisted of two subjects reporting mild stomach discomfort, while continuing treatment due to its antiallergic efficacy.

The positive side effects were:

Mild hypotension in 2 subjects, considered a welcome side effect

Improvement in symptoms of prostatitis in 1 subject

None of the subjects normally taking antihistamines reported sedation, drowsiness, fatigue, or decreased concentration while using the saffron/quercetin aglycone composition.

Example 2: Composition comprising saffron and isoquercetin

The composition of saffron/quercetin aglycone was changed to saffron/isoquercetin (Merck KG), to assess whether the significantly higher bioavailability of isoquercetin (compared to quercetin aglycone) improves the efficacy of symptomatic treatment of allergies IgE, including asthma. The test was carried out on 14 adult volunteers, suffering from acute allergies but otherwise in good health, over the course of 6 months (March-August).

The saffron dose remaining unchanged (1.5 mg/kg of body weight, twice a day), the composition containing isoquercetin (instead of quercetin aglycon) proved to be significantly more effective at a rate of 1-1.2 mg per kg of body weight, twice a day, which is 40-48% of the dosage required in the case of quercetin aglycone.

Replacing quercetin aglycone with isoquercetin in the composition resulted in a faster manifestation of the antiallergic effect, 25-30 minutes after taking the formulation. Additionally, the efficacy in suppressing the symptoms of asthma, allergic rhinitis, itchy eyes and allergic conjunctivitis further increased significantly.

Isoquercetin administered alone did not produce satisfactory antiallergic effects.

Conclusion: The remission of allergic symptoms, in particular asthma, was significantly stronger compared to the composition composed of saffron/quercetin aglycone: approximately 70-80% reduction in symptom scores and 70-85% reduction in the use of inhaled corticosteroids. Asthma relief was perceived as 'more comprehensive' by trial participants, especially for those with severe asthma. Nasal symptoms: Approximately 60-75% reduction in symptoms and 65-80% reduction in nasal spray use. Allergic conjunctivitis, itchy eyes and watery eyes showed improved efficacy over the saffron/quercetin aglycone formulation, including sun sensitivity being greatly reduced in virtually all subjects: approximately 60-70% reduction in eye symptom scores and 70-80% reduction in the use of topical allergy eye drops.

Negative side effects consisted of a single test subject reporting rather mild stomach discomfort but continuing treatment.

The positive side effects were:

Mild hypotension, improvement in prostatitis symptoms in one subject, absence of sedation, absence of drowsiness/fatigue or loss of concentration, improvement in mood.

For the preparation of the saffron/isoquercetin composition, grade I Iranian saffron was used, with spectrophotometric readings of crocin 266, picrocrocin 95 and safranal 42 (according to ISO 3632-2).

Example 3: Composition comprising saffron and alpha-glycosyl isoquercitrin.

The saffron/isoquercetin composition is in this example replaced by the saffron/alpha-glycosyl isoquercitrin (AGIQ) composition. AGIQ was obtained from Japanese manufacturer San-Ei Gen F.F.F.I. from Osaka, marketed as Enzymatically Modified Isoquercitrin under the trademark EMIQ. EMIQ contains 30% AGIQ as the active ingredient, the remaining 70% being dextrin. The saffron dose remaining unchanged (1.5 mg/kg body weight, twice a day), the composition containing alpha-glycosyl isoquercitrin (instead of isoquercetin) proved to be significantly more effective even at doses of 0.4 to 0.6 mg per kg of body weight, twice a day, which is 50% of the intake required with isoquercetin. Alpha-glycosyl isoquercitrin (AGIQ) dosing is the result of a systematic trial-and-error approach, with each subject reporting changes in total symptom score and reduction in medication, especially systemic corticosteroids and/or inhalation.

Alpha-glycosyl isoquercitrin taken alone did not produce a noticeable anti-allergic effect.

Conclusion: Of 12 study subjects, one subject reported unspecified gastric discomfort but continued treatment. In all 12 subjects, the use of alpha-glycosyl isoquercitrin significantly accelerated the onset of anti-allergic effects: 10-15 minutes after oral administration of the formulation. Remission of asthma symptoms was significantly stronger than that of saffron/isoquercetin, and was perceived to be significantly better than systemic or inhaled corticosteroids by all subjects with asthma: approximately 80-95% reduction in symptoms and 80-100% reduction in the use of inhaled corticosteroids. Subjects reported an antidepressant effect from the formulation due to the rapid asthma relief and, possibly, the antidepressant effect of saffron. Nasal decongestion and almost complete drying of the mucous membranes occurred within 15 to 30 minutes, accompanied by a decrease in facial swelling and sensitivity to light in practically all subjects: reduction of 75 to 95% for symptom scores and 70-100% for nasal spray use. Allergic conjunctivitis, eye itching and tearing were significantly reduced and only rarely required the use of topically applied eye drops: about 70-90% reduction for eye symptoms and 70-90% reduction in the use of eye drops for local allergy.

Negative side effects: rather mild gastric discomfort (bloating), reported by one study subject, who nevertheless continued treatment.

The positive side effects were:

Mild hypotension, improvement in symptoms of prostatitis, remission of gastritis, no sedation, drowsiness, fatigue or loss of concentration, improvement in mood.

For the preparation of the saffron/alpha-glycosyl isoquercitrin composition, grade I Iranian saffron, with spectrophotometric readings of crocin: 217, picrocrocin: 88 and safranal: 46 (according to ISO 3632-2) was used. Quantitative analysis of this saffron revealed the presence of crocin: 11.3%, picrocrocin: 3.1%, safranal: 0.07% (Laboratory of Chemistry and Food Technology, Aristotle University of Thessaloniki).

Example 4: summary of examples 1 to 3

Table 2 summarizes the results described in Examples 1 to 3.

Table 2: Summary of the results of examples 1 to 3

Example Example 1 Example 2 Example 3 Composition Saffron - Quercetin aglycone Saffron - Isoquercetin Saffron-alpha-Glycosyl Isoquercitrin Carbohydrate parts 0 1 1-11 Test subjects 6 14 12 % Allergic Asthma Symptom Relief 60-70% 70-80% 80-95% % Allergic Rhinitis Symptom Relief 50% accompanied by drying of the nasal mucosa 60-75% accompanied by pronounced drying of the nasal mucosa 75-95% + accompanied by very pronounced drying of the nasal mucosa % Relief of Allergic Eye Symptoms 30-40% 60-70% 70-90% Subject side effects in % Gastric disorders: 17%
Mild hypotension: 33%
Gastric disorders: 7%
Mild hypotension: 14%
Gastric disorders: 8%
Mild hypotension: 25%
Positive effects in % of subjects Improvement of prostatitis
No sedation, drowsiness, fatigue or impaired concentration Improvement of prostatitis
No sedation, drowsiness, fatigue or impaired concentration improved mood Improvement of prostatitis
Remission of gastritis
No sedation, drowsiness, fatigue or impaired concentration Improved mood
Appearance of therapeutic effect 45-50 mins. 25-30 mins. 10-15 mins. Effective therapeutic duration, in hours 10-14 hours 10-14 hours 10-14 hours Dosing Frequency 12 hours 12 hours 12 hours

Even when used for 4 consecutive years, treatment with the composition according to the present invention did not produce a habituation effect over consecutive allergy seasons.

Example 5: Comparison of the antiallergic efficacy of the aglycone forms of the flavonols kaempferol, myricetin, luteolin, apigenin and fisetin with that of quercetin aglycone/isoquercetin/alpha-glycosyl-isoquercitrin (AGIQ)-saffron as described above

To compare the antiallergic efficacy of the flavonols kaempferol aglycon and myricetin aglycon with quercetin/isoquercetin/alpha-glycosyl-isoquercitrin (AGIQ)-saffron aglycon as described in Examples 1-3, a composition containing kaempferol aglycone and myricetin aglycone at 5 mg/kg body weight and saffron at 1.5 mg body weight was administered twice daily to 4 allergic but otherwise healthy consenting adult volunteers.

The efficacy of these two formulations was significantly inferior to the quercetin aglycone/isoquercetin/alpha-glycosyl-isoquercitrin (AGIQ)/saffron composition in treatment efficacy and duration.

The aglycone forms of luteolin, apigenin and fisetin at 5 mg per kg body weight, combined with saffron at 1.5 mg per kg body weight, were tested twice daily for 2-3 days per 10 allergic subjects who participated in the alpha-glycosyl isoquercitrin/saffron composition study.

The compositions all showed some antiallergic effect, but were significantly less effective in the extent of symptom reduction and more importantly of much shorter therapeutic duration than the quercetin aglycone/isoquercetin/alpha-glycosyl-isoquercitrin composition. (AGIQ)-saffron as described above.

Example 6: Monitoring a patient's treatment

Individual 1, male, weighing 94 kg, suffering from severe allergy since early childhood, presents the symptoms listed in Table 3. Very sensitive to tree pollen (hazel, birch, alder, cedar, cypress ) and grass pollen (e.g., timothy, Bermuda), as evidenced by medical history, skin prick test, pulmonary function tests, immuno-allergen diagnosis in phase solid (ISAC) and total IgE >2000. Person 1 has had four hospital emergency room visits over 20 years.

Table 3: Allergic manifestations before treatment – Individual 1.

Symptoms Gravity Conjunctivitis: Itching and burning sensation in the eyes; excessive tearing; eye pain, redness of the eyes, sensitivity to light Severe persistent during allergy season
Rhinitis: swelling of the mucous membranes of the nose, itchy nose, sneezing serious persistent in period of allergy Allergic asthma: Chest tightness, coughing, wheezing, airflow limitation Persistent severe through allergy season

The allergy treatment protocol in the years preceding the trial consisted of inhaled steroids (e.g. fluticasone propionate, salmeterol xinafoate), prednisolone tablets (5 mg/day), steroid injections (triamcinolon 40 mg) before and during the allergic season. The trial using the quercetin/saffron composition began in April 2016, in the height of allergy season, with minimal conventional treatment (occasional salmeterol xinafoate) and therefore a full manifestation of allergic symptoms, in particular severe allergic asthma.

Individual 1 was treated as described in Table 4, for four consecutive years.

The saffron/quercetin aglycone/isoquercetin/AGIQ compositions were effective in treating the symptoms suffered. Given its superior efficacy to conventional treatment before the trial and its tolerability (e.g., no sedation, no side effects as with corticosteroids), treatment with saffron/quercetin aglycon/isoquercetin/AGIQ continued through subsequent allergy seasons. During this treatment, person 1 stopped taking inhaled steroids, steroid tablets and steroid injections and only used eye drops occasionally (eg ectoin formulations).

Table 4: Individual 1- Treatments 2016 – 2019

Allergy Season Composition of the dose
Dosing frequency Symptom relief (%) 2016 Saffron: 150mg; quercetin aglycon: 250 mg Every 12 hours Conjunctivitis: 40-60% after 45-50 minutes Rhinitis: 50-70% after 45-50 minutes Allergic asthma: 60-80% after 45-50 minutes 2017 Saffron: 150mg; Isoquercetin: 100 mg Every 12 hours Conjunctivitis: 50-80% after 30-40 minutes Rhinitis: 70-80% after 30-40 minutes Allergic asthma 70-80% after 30-40 minutes 2018 Saffron: 150mg; AGIQ : 15 mg Every 12 hours Conjunctivitis 80-95% % after 20-30 minutes Rhinitis: 80-100% after 20-30 minutes Allergic asthma: 80-100% after 20-30 minutes 2019 Saffron: 150mg; AGIQ: 15mg Every 12 hours Conjunctivitis 80-100% % after 20-30 minutes Rhinitis: 80-100% after 20-30 minutes Allergic asthma: 80-100% after 20-30 minutes

Three years after the first treatment, Individual 1 is still reactive, in the same way, to the saffron/quercetin aglycon/isoquercetin/AGIQ treatments described in this example.

Example 7: Follow-up of the treatment of individual 2

Individual 2, male, weighing 107 kg, has multiple grass pollen sensitivities as evidenced by clinical history, skin prick testing, Immuno Solid-phase Allergen Chip (ISAC) IgE diagnosis and measurement forced expiratory flow rate (PEFR), and exhibits the symptoms shown in Table 5. azelastine, H1 receptor antagonist administered topically and berodual-aerosol (ipratropium bromide, fenoterol).

Table 5: Individual 2 - Allergy (before administration)

Symptoms Gravity Conjunctivitis: Itching and burning sensation in the eyes; swollen eyelids Severe persistent during allergy season Rhinitis: Post-nasal drip, sneezing Moderate to severe persistent during allergy season Allergic asthma: Lung obstruction, wheezing Moderately persistent during allergy season

Person 2 was treated as described in Table 6. The saffron/isoquercetin/AGIQ compositions were effective in treating the symptoms.

Table 6: Individual 2 – Treatments 2017-2019

Allergy Season Composition of the dose
Dosing frequency Symptom relief (%) 2017 Saffron: 180mg; Isoquercetin: 120 mg Every 12 hours Rhino-Conjunctivitis: 70%-80% after 30-40 minutes Facial swelling: 70-80% after 30-40 minutes Allergic asthma: 70-80% after 30-40 minutes 2018 Saffron: 180mg; AGIQ: 20 mg Every 12 hours Rhino-Conjunctivitis: 80%-90% after 20-30 minutes Facial swelling: 70-90% after 20-30 minutes Allergic asthma: 80-90% of respiratory capacity restored 30-40 minutes

Individual 2 reported significantly better efficacy of isoquercetin/IQFA/saffron compositions compared to pre-trial treatment, particularly rapid improvement of airway obstruction, preservation of mental acuity, absence of adverse sedative effects typical of traditional treatment.

Example 8: Monitoring another patient's treatment

Individual 3, female, weighing 67 kg, has house dust mite allergies, as evidenced by medical history, skin prick tests and the ImmunoCap-ISAC IgE test, and exhibits recurrent allergic manifestations and particularly serious during visits to relatives living in a 250-year-old farmhouse in Burgundy, France. Symptoms are listed in Table 7.

Table 7: Individual 3 - Allergy (before administration)

Symptoms Gravity Conjunctivitis: Itching and burning sensation in the eyes; swollen eyelids Severe persistent during stay in an old farmhouse Rhinitis: Post-nasal drip, sneezing Moderate to severe persistent during stay in an old farm house Allergic asthma: Lung obstruction, wheezing Severe and persistent during stay in an old farmhouse

Symptoms develop within minutes of arrival and are exacerbated by asthmatic episodes. Conventional antihistamine treatment showed slow and unsatisfactory effect.

Person 3 was treated as described in Table 8. The saffron/isoquercetin/AGIQ compositions were effective in treating the symptoms suffered for a period of ten days.

Table 8: Individual 3-Treatment 2017 – 2018

Allergy Season Composition of the dose
Dosing frequency Symptom relief (%) 2017 Saffron: 120mg; Isoquercetin: 80 mg Every 12 hours Conjunctivitis: 60-80% after 30-40 minutes Rhinitis: 80% after 30-40 minutes Allergic asthma 70-80% after 30-40 minutes 2018 Saffron: 120mg; AGIQ: 12 mg Every 12 hours Conjunctivitis 80-100% % after 20-30 minutes Rhinitis: 90-100% after 20-30 minutes Allergic asthma: 80-100% after 20-40 minutes

Two years after the first treatment, Individual 3 is still sensitive, in the same way, to the saffron/isoquercetin/AGIQ treatment described in this example.

Example 9: Preparation of the Galenic Formulation

When saffron was used alone or in combination with quercetin aglycone or isoquercetin, it was ground in a planetary ball mill to obtain a particle size between 10 and 150 microns, measured by laser diffraction using the LA -960 Laser Particle Size Analyzer from Horiba. The saffron powder obtained was then mixed with quercetin aglycon (Zein Pharma, Germany) or isoquercetin (Merck).

When saffron was used in combination with alpha-glycosyl isoquercitrin (AGIQ), trehalose was added to the pre-ground saffron powder in a weight ratio of 1:2. This mixture was placed in a sealable stainless steel jar to be cooled to -190°C in a liquid nitrogen bath prior to grinding to prevent oxidation and loss of volatiles. This mixture of powdered saffron and trehalose was mortared with alpha-oligoglucosyl isoquercitrin (AGIQ) powder using EMIQ® R 20 from San-Ei Gen F.F.I., Osaka, Japan, containing 30% by weight of AGIQ, expressed as Rutin trihydrate 18-20% by weight).

The compositions thus obtained were finally filled into size 0 HPMC (hydroxy propylmethylcellulose) hard capsules.

Example 10: Analysis of the composition of saffron

Different types of saffron were used during the experiments.

Two of them were analyzed by RP-HPLC-DAD to assess crocin and picrocrocin concentrations, and UV-VIS to assess safranal concentrations. The results are shown in Table 9.

Table 9: Crocin, picrocrocin and safranal content of two rudders

Saffron 1
Saffron 2
Result, % Result, % Unit Method Total crocin content 11.3 18.6 %, dry weight RP-HPLC-DAD (external standard method) Picrocrocin content 3.1 4.5 %, dry weight RP-HPLC-DAD (external standard method) Result mg/g saffron Result mg/g saffron Unit Method Safranal content, mg 0.70 0.54 mg/g, dry weight UV-VIS (external standard method)

Example 10: Efficacy of alpha-glycosylated isoquercitrin alone

The effects of alpha-glycosyl isoquercitrin on the immune response were assessed in-vivo in two double-blind studies of similar design in Japan (Yamada T, et al. Present state of Japanese Cedar pollinosis: The national affliction. 2014 ), where the Japanese cedar flowering season produces endemic outbreaks of pollinosis each year, with up to 40% of the population suffering from allergic symptoms (Hirano T, Kawai M, et al. Preventative effect of a flavonoid, enzymatically modified isoquercitrin on ocular symptoms of Japanese cedar pollinosis Allergol Int. 2009 - Kawai M, Hirano T, Arimitsu J, et al Effect of enzymatically modified isoquercitrin, a flavonoid, on symptoms of Japanese cedar pollinosis: a randomized double-blind placebo-controlled trial. Int Arch Allergy Immunol. 2009).

In both studies, subjects took 100 mg of alpha-glycosylated isoquercitrin or a placebo for 8 weeks. Subjective measures, Activities of Daily Living (ADL) scores and medication use were recorded daily. Quality of life (QOL) scores were obtained every 4 weeks. In both studies, taking alpha-glycosylated isoquercitrin was found to be effective in supporting eye comfort. Nasal airflow, however, remained unchanged.

CONCLUSION: Alpha-glycosyl isoquercitrin is only effective in suppressing ocular symptoms of IgE-mediated cedar pollinosis, and is not effective in relieving allergic rhinitis or allergic asthma. Other studies also demonstrate that the effects of quercetin and/or its metabolites and derivatives are much weaker than those of pharmacological steroidal and non-steroidal anti-inflammatory drugs (Rajendran et al. The vascular endothelium and human diseases, 2013).

Claims (28)

A composition comprising saffron or a mass fraction of saffron and at least one flavonoid for use in the prevention and/or treatment of IgE-mediated type I allergic conditions. A composition according to claim 1, wherein the IgE-mediated type I allergic conditions are allergic asthma and/or allergic bronchitis and/or rhinitis and/or conjunctivitis and/or skin rash and/or dermatitis atopic and/or clogged ears. Composition according to any one of the preceding claims, in which the mass fraction of saffron comprises at least one compound from natural or synthetic sources chosen from safranal, crocin, crocetin, picrocrocin, their derivatives and/or their mixtures. Composition according to any one of the preceding claims, in which the said at least one flavonoid is at least one flavonol, from a natural or synthetic source. Composition according to the preceding claim, characterized in that the said at least one flavonol is chosen from quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin, their metabolites, their analogues, their chemically modified analogues, their derivatives or their mixtures. Composition according to either of Claims 4 and 5, in which the said at least one flavonol comprises at least one aglycone chosen from the aglycones of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof. A composition according to claim 6, wherein said at least one aglycone comprises quercetin aglycone. Composition according to any one of Claims 4 to 7, in which the said at least one flavonol comprises at least one glycoside chosen from the glycosides of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof. Composition according to Claim 8, comprising at least one quercetin glycoside. Composition according to Claim 9, in which the said at least one quercetin glycoside is chosen from isoquercetin, isoquercitrin, at least one alpha-glycoside of isoquercetin, at least one alpha-glycoside of isoquercitrin and/or mixtures thereof . Composition according to any one of Claims 1 to 10, comprising for a daily dose between 0.01 mg and 20 mg, preferably between 0.02 mg and 15 mg, more preferably between 0.03 mg and 10 mg of saffron or a mass fraction of saffron per kilogram of body weight, and between 0.01 mg and 15 mg, preferably between 0.05 and 12 mg per kilogram of body weight of one or more flavonoids, preferably quercetin aglycone and/or isoquercetin and/or isoquercitrin and/or alpha-glycosyl isoquercitrin or mixtures thereof. Composition according to any one of Claims 1 to 11, characterized in that the said saffron or the said mass fraction of saffron and the said at least one flavonoid are administered simultaneously. Composition according to any one of Claims 1 to 12, additionally comprising at least one pharmaceutically acceptable compound, preferably chosen from anti-inflammatory compounds and/or for the treatment of IgE-mediated type I allergic conditions, and/or for the treatment of allergic asthma and/or allergic bronchitis and/or rhinitis and/or conjunctivitis and/or skin rashes and/or atopic dermatitis and/or ear plugging. Composition according to any one of Claims 1 to 13, characterized in that the composition is formulated in the form of a liposome. Dietetic and/or food composition comprising saffron or a mass fraction of saffron and at least one flavonoid. Dietetic and/or food composition according to Claim 15, in which the mass fraction of saffron comprises at least one compound from natural or synthetic sources chosen from safranal, crocin, crocetin, picrocrocin, their derivatives and/or their mixtures. Dietetic and/or food composition according to either of Claims 15 and 16, in which the said at least one flavonoid is at least one flavonol, from a natural or synthetic source. Dietetic and/or food composition according to Claim 17, characterized in that the said at least one flavonol is chosen from quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin , laricitrin, syringetin, their metabolites, their analogues, their chemically modified analogues, their derivatives or their mixtures. Dietetic and/or food composition according to either of Claims 17 and 18, in which the said at least one flavonol comprises at least one aglycone chosen from the aglycones of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof. Dietary and/or food composition according to claim 19, wherein said at least one aglycone comprises quercetin aglycone. Dietetic and/or food composition according to any one of Claims 17 to 20, in which the said at least one flavonol comprises at least one glycoside chosen from the glycosides of quercetin, myricetin, kaempferol, rutin, fisetin, luteolin, apigenin, taxifolin, dihydromyricetin, laricitrin, syringetin or mixtures thereof. Dietetic and/or food composition according to Claim 21, comprising at least one quercetin glycoside. Dietetic and/or food composition according to Claim 22, in which the said at least one glycoside of quercetin is chosen from isoquercetin, isoquercitrin, at least one alpha-glycoside of isoquercetin, at least one alpha-glycoside of isoquercitrin and/or mixtures thereof. Dietetic and/or food composition according to any one of Claims 15 to 23, comprising for a daily dose between 0.01 mg and 20 mg, preferably between 0.02 mg and 15 mg, more preferably between 0.03 mg and 10 mg of saffron or a mass fraction of saffron per kilogram of body weight, and between 0.01 mg and 15 mg, preferably between 0.05 mg and 12 mg per kilogram of body weight of one or more flavonoids, preferably quercetin aglycone and/or isoquercetin and/or isoquercitrin and/or alpha-glycosyl isoquercitrin or mixtures thereof. Dietetic and/or food composition according to any one of Claims 15 to 24, characterized in that the said saffron or the said mass fraction of saffron and the said at least one flavonoid are administered simultaneously. Dietetic and/or food composition according to any one of Claims 15 to 25, additionally comprising at least one acceptable dietetic and/or food compound, preferably chosen from anti-caking agents, trehalose, pullulan, sweeteners, surfactants , diluents, aggregating agents or binders, lubricants, slip agents, stabilizers, solubilizers, emulsifiers, humectants, flavors, coating agents, coloring agents, acidity regulators or mixtures thereof. Dietetic and/or food composition according to any one of Claims 15 to 26, characterized in that the composition is formulated in the form of a liposome. Kit for the manufacture of a dietetic and/or food composition according to any one of Claims 14 to 27 comprising:

• Saffron or a mass fraction of saffron;
• At least one flavonoid.