WO2021229501A1

WO2021229501A1 - Mixture and composition for the prevention and treatment of diseases of the circulatory system and of related symptoms

Info

Publication number: WO2021229501A1
Application number: PCT/IB2021/054113
Authority: WO
Inventors: Umberto DI MAIO
Original assignee: Neilos S.r.l.
Priority date: 2020-05-13
Filing date: 2021-05-13
Publication date: 2021-11-18
Also published as: IT202000010924A1

Abstract

The present invention relates to a composition comprising a combination or mixture of: (a) diosmin, (b) an extract of Betula pendula, (c) an extract of Hamamelis virginiana and (d) rutin useful in prevention and/or in the treatment of diseases associated with the circulatory system. The combination is particularly effective thanks to the synergistic action of the components thereof.

Description

Industrial invention patent application having the title:

“Mixture and composition for the prevention and treatment of diseases of the circulatory system and of related symptoms’’

DESCRIPTION

The present invention relates to a combination or mixture comprising or, alternatively, consisting of: (a) diosmin; (b) an extract of Betula pendula; (c) an extract of Hamamelis virginiana; e (d) rutin.

Furthermore, the present invention relates to a composition comprising diosmin, an extract of Betula pendula, an extract of Hamamelis virginiana and rutin useful in prevention and/or in the treatment of diseases associated with the circulatory system.

Furthermore, the present invention relates to a composition comprising said combination or mixture (a)-(d), and food or pharmacologically acceptable additives and/or excipients.

Furthermore, the present invention relates to said composition for use in a method for the preventive or curative treatment of a disorder, or of an ailment, or of a symptom, or of a disease of the circulatory system in a subject, preferably in a mammal, more preferably in a human (human being) or animal.

The combination or composition subject of the present invention is particularly effective thanks to the synergistic action of the components thereof.

Background of the invention Circulatory system

The circulatory system can be divided into organs and vessels for the transport of blood and organs and vessels for the transport of lymph: however, this division is mainly intended for illustrative purposes, given that the two types of circulation (blood and lymphatic) work in a close and synergic relationship.

The circulatory system consists of: heart (main organ), arteries, veins, capillaries, blood. The main functions of this system are: to ensure cell survival, disease prevention, to keep body temperature and pH under control, to maintain the right homeostatic levels.

The heart is the main organ of the circulatory system, that is the pump that facilitates the proper blood circulation. It is a single organ, present in the thoracic cage, consisting of four cavities: two atria and two ventricles. This organ is directly connected to the venae cavae, the pulmonary artery, the pulmonary veins, the aorta. Venae cavae (lower and upper) introduce non-oxygenated blood into the right atrium. The pulmonary artery starts from the right atrium and carries non-oxygenated blood to the lungs. The pulmonary veins carry oxygenated blood from the lungs to the left atrium. The aorta starts from the left ventricle and carries oxygenated blood to all anatomical districts. This blood transport system allows to ensure proper oxygenation in the various anatomical districts and the elimination of waste substances.

The arteries are vessels that start from the heart and carry oxygenated blood to the various anatomical districts (except for the pulmonary artery that carries non-oxygenated blood to the lungs to allow gaseous exchange at the alveolar level and ensure that blood is oxygenated). Given the function thereof which entails reaching the points farthest from the heart, the diameter of the arteries gradually shrinks the farther they are from the heart. Depending on the diameter, the arteries are classified as: large-sized arteries (diameter equal to or larger than 7 mm), medium-sized arteries (diameter comprised between 7 mm and 2.5 mm) and small-sized arteries (diameter smaller than 2.5 mm). Arteries are not simple ducts through which blood flows. As a matter of fact, they have a complex structure of the walls able to facilitate the flow of the blood and resist stresses of various types: a cross-section of the wall of an artery shows three layers (tunica intima, tunica media, tunica adventitia).

Unlike arteries, veins are vessels that carry blood from the periphery to the heart. Despite the functional similarity with the arteries, their structure is remarkably different given that the walls (beside tunica adventitia, media and intima being starkly recognisable) are thinner, less muscular and less elastic; while sharing the same variation in diameter, which increases the closer to the heart until it reaches a diameter comprised between 20 mm and 22 mm (inferior vena cava and superior vena cava).

Capillaries can be considered as the endings of the veins and of the arteries which - upon reaching the tissues in question - acquire microscopic dimensions that facilitate entry into the spaces between the cells and the exchange of various types of substances: gaseous molecules, carbon dioxide, ions, nutrients, waste products, water. Capillary walls are made of a single layer, consisting of endothelial cells and have a much simpler structure with respect to the veins and arteries.

Blood is the fluid that flows through the aforementioned vessels and it reaches a total volume of about 5 litres in the adult human body. It consists of 55% plasma and 45% haemocytes. Plasma can be described as an aqueous solution containing mineral salts and proteins, while haemocytes (literally "blood cells”) are red blood cells (erythrocytes), white blood cells (leukocytes) and platelets.

The lymphatic system primarily consists of: lymphatic vessels, lymph nodes, lymph.

Lymphatic vessels are ducts in which the lymph flows in a centripetal direction (i.e. from the periphery toward the centre). Unlike blood, whose flow is mainly influenced by a motor organ that is the heart, the lymph flows through the vessels thanks to the action of the smooth muscle of the vessels. The lymphatic vessels run parallel to the venous vessels until they join in two points: the right lymphatic duct joins the right subclavian vein and pours the lymph contained therein thereinto, the thoracic lymphatic duct enters the left subclavian vein, pouring the content contained therein thereinto.

Lymph nodes are basically biological filters which intercept germs, foreign substances and/or neoplastic cells present in the lymph. In addition to lymph nodes, other "lymphatic” organs are: thymus, spleen, bone marrow.

The parent substance of this system is the lymph, liquid with different degrees of viscosity (depending on the physiopathological conditions of the individual and the anatomical district concerned), with a colour varying between transparent, straw yellow and opalescent. Lymph contains sugars, proteins, salts, lipids, amino acids, hormones, vitamins, white blood cells.

Given the extension of the circulatory system, it is clear that any disorder affecting it can lead to immediate and/or long-term systemic effects. In the case of blood vessel disorders, “venous insufficiency, a pathological condition caused by the difficult return of blood from the veins to the heart, is often cited.

Venous insufficiency, preferably chronic venous insufficiency, may be organic or functional: in the case of organic venous insufficiency, preferably chronic organic venous insufficiency, the causes can be traced to pathological impairment of the veins, often related to variables such as the number of pregnancies, the use of oral contraceptives, water retention, overweight, chronic constipation; in the case of functional venous insufficiency, preferably chronic functional venous insufficiency, the causes are linked to the functional overload of the veins linked in turn to incorrect posture, impairment of the muscle pump, lymphedema.

In Italy, venous insufficiency, preferably chronic venous insufficiency, affects 30% of women and 15% of men, with variable symptoms: swelling of the legs, feeling of heaviness of the legs, formication, itching, burning sensation, pain and cramps (especially nocturnal), visible capillaries, skin changes, varicose veins. This variety of symptoms can only be the tell-tale sign of a more severe chronic venous insufficiency and which can lead to related pathological conditions such as: increased venous pressure, local inflammation, phlebitis, thrombosis, ulceration, cellulitis, haemorrhoids.

From this description of venous insufficiency (organic and functional), preferably chronic venous insufficiency (organic and functional), it can be deduced that this is a multifactorial disease, therefore it can be prevented (as far as possible) by adopting some good habits and avoiding some bad ones. Activities that assist in the prevention of venous insufficiency are: physical exercise (preferably swimming or sports with slow and harmonious movements), maintaining ideal weight, moving the legs (since sedentary lifestyle contributes to the venous stasis), wearing of elastic stockings, using a raised plane in the lower part of the bed (that of the feet). Bad habits to be avoided are: wearing shoes with very high heels, using the contraceptive pill, smoking and taking fried food, alcohol and excessive amounts of coffee.

Shifting the focus from prevention to treatment of chronic venous insufficiency, the following drug protocols are currently used: anticoagulants (for example heparan sulphate) suitable to facilitate blood circulation by decreasing blood viscosity, fibrinolytic drugs (for example streptokinase) which dissolve fibrin accumulations which inevitably form at the level of the varices and which therefore facilitate blood flow, phlebotonics (for example diosmin/oxerutin), injectable sclerosing drugs (for example tetradecyl sulphate, ethanolamine oleate) used mainly in medium-sized vessels, which cause local inflammation resulting in fibrosis in the vessel causing the flow to be diverted into a healthy vessel.

In the most severe cases of disorders of the circulatory system, there arises the need for medical and surgical intervention: valvuloplasty, removal of the diseased venous portion, radio-frequency ablation, and/or laser therapy.

DE 20 2007 006374 U1 illustrates a preparation for venous therapy comprising extracts of Gingko biloba, Hamamelis virginiana and Vitis vinifera, combined with a hydroxymethyl and/or hydroxyethyl derivative of rutin (or rutoside).

WO 98/51291 A1 illustrates a pharmaceutical composition for treating and/or preventing an ischaemia or energy deficit, preferably chronic venous insufficiency.

CORSALE ITALO ET AL: "Flavonoid mixture (diosmin, troxerutin, rutin, hesperidin, guercetin) in the treatment of HM degree hemorroidal disease: a double-blind multicenter prospective comparative stud y", INTERNATIONAL JOURNAL OF COLORECTAL DISEASE, SPRINGER VERLAG, BERLIN, DE, vol. 33, no. 11, pages 1595-1600) is a comparative, prospective, multicentre, double-blind study aimed at establishing the efficacy of a flavonoid mixture in the treatment of grade l-lll haemorrhoidal diseases. HOSTETTMANN K ET AL: "ZU INHALTSSTOFFEN UND PHARMAKOLOGIE PFLANZLICHER VENENMITTELI/COMPONENTS AND PHARMACOLOGY OF PHYTOPHARMACEUTIC PREPARATIONS FOR VARICOSE VEINS", ACTA CRYSTALLOGRAPHICA SECTION E, JOHN WILEY & SONS, GB, vol. 23, no. 3, pages 71-77, is an article illustrating several natural substances (coumarins, flavonoids, saponins) that could be used in the treatment of chronic venous insufficiency. RASTOGI SUBHA ET AL: "Medicinal plants of the genus Betula— Traditional uses and a phytochemical- pharmacological review", JOURNAL OF ETHNOPHARMACOLOGY, ELSEVIER IRELAND LTD, IE, vol. 159, pages 62-83, is a study aimed at providing information on botany, traditional uses, phytochemistry as well as pharmacological and toxicological research on species of Betula.

US 6565850 B2 illustrates compositions in the form of gel for use in the treatment of anorectal disorders such as haemorrhoids.

The object of the present invention, consisting of the synergistic combination (or mixture) of diosmin, extract of Betula pendula, extract of Hamamelis virginiana and rutin, is to solve the problem related to the prevention and/or treatment of diseases of the circulatory system both in humans and animals.

In particular, the combination (or mixture) of the present invention is particularly effective in the prevention and/or treatment of venous insufficiency, having the main advantage of including substances of natural origin and not giving rise to problems that are usually typically observed in conventional drug protocols: therapeutic competition between various drugs (polytherapy) and side effects.

GLOSSARY

The terms used in the present description are as generally understood by the man skilled in the art, unless otherwise indicated.

In the context of the present description, the expression "extract” is used to indicate any product related to a material or plant drug including all products derived from mechanical treatments (pulverisation, crushing, mixing and/or other methods) and / or from extract-based treatments (solvent extraction, distillation, and/or other specific methods) conducted on said material or drug.

In the present description, the expression “in an amount comprised between ... and ..." is used to indicate any amount comprised between the end points of the ranges specified from time to time, including the end points of the ranges in question.

Diosmin

Diosmin is a molecule of plant origin, belonging to the family of flavonoid glycosides. It consists of a carbohydrate part (rutinoside) and a flavonoid part with the benzopyrone group (diosmetin) responsible for pharmacological activities: anti-oedematous activity, anti-inflammatory activity and anti-varicose activity. Thanks to these actions thereof mentioned above, it is a useful remedy for haemorrhoids, cellulitis, varicose veins, capillary fragility. The mechanism of action of this molecule mainly consists in a decrease in the catabolism of noradrenaline, with consequent increase in venous tone and therefore vasoconstriction: vasoconstriction in turn averts venous stasis, which is among the main factors determining phenomena such as oedemas and varicose veins. The study conducted on a rat artery tissue “Diosmin-induced increase in sensitivity to Ca 2+ of the smooth muscle contractile apparatus in the rat isolated femoral vein", shows that the administration of diosmin results in a higher sensitivity of the tissue to the vasoconstrictive action of Calcium. Another distinctive characteristic of diosmin lies in the anti inflammatory action, mediated by the reduction of the production and release of phlogosis mediators (cyclooxygenase, prostaglandins, thromboxanes), and by the inactivation of free radicals. The characteristics listed above indicate diosmin as the preferred substance in the treatment and prevention of disorders of the circulatory system. The in vivo study “Medicamentous treatment of chronic venous insufficiency using semisynthetic diosmin - a prospective stud ’ shows that the use of diosmin for 30 days in patients with chronic venous insufficiency leads to a significant decrease in swelling, pain, feeling of heaviness and tightness of the legs: this obviously accompanied by a physical, social and psychological improvement in the quality of life. In another study “Flavonoids mixture (diosmin, troxerutin, hesperidin) in the treatment of acute hemorrhoidal disease: a prospective, randomized, triple-blind, controlled trial’, the argument showing diosmin to be useful against disorders linked to venous insufficiency is corroborated by positive results in the treatment of symptoms of acute haemorrhoidal disease.

Betula pendula is a type of Birch commonly found in the Balkans all the way to the Alps and across Atlantic Europe and Asia. In particular, in Italy it is very common in Piedmont and Lombardy. However, specimens can often be found in some areas of Abruzzo, Campania and Sicily. The genus Betula, belonging to the family of Betulaceae, comprises about 40 species variously scattered in the northern hemisphere. This variety of species justifies the wide extension of this kind of plants that fit well to a wide range of conditions: as a matter of fact, Birch proves to adapt well to all temperatures, it can vegetate up to 2000 meters of altitude, it bears pH values of the soil up to 3.30 and shows polymorphism of the roots depending on the conditions of the soil (whether it is rich or poor in water). All of these characteristics make the Birch a colonising plant. Betula pendula usually reaches a height of 30 metres (more generally the family of the Betulaceae has a height varying between 15 and 30 metres), it has a relatively bulky foliage and tends to the pyramidal shape. The leaves are sharp triangular with a width varying between 3 and 4 cm and a length between 4 and 6 cm. The name Betula pendula is currently accepted universally. However, in the past this plant used to be known as Betula alba and Betula verrucosa : names derived from some of its morphological characteristics such as the light colour and rough surface of the bark. Betula pendula is significantly used in the nutraceutical field due to its capacity to facilitate diuresis, due to its organism purifying properties (claims recognised even by the Ministry of Health and of the EMA) and due to its anti-inflammatory and antioxidant properties. The EMA monograph shows the Birch's ability to increase the amount of urine and its adjuvant function in resolving disorders of the urinary tract, while the study “Potassium -sodium ratio for the characterization of medicinal plant extracts with diuretic activit ’ shows that this diuretic derives from the sodium and potassium concentrations present in the leaves of this plant. The arguments that attribute diuretic and anti-inflammatory properties to Birch are also reflected in the study “Studies of the anti-inflammatory and diuretic properties of the tablet with dense extract ofbetula pendula leaves", in which the tests are conducted on rat paw with carrageenin-induced inflammation.

Hamamelis virginiana

Witch-hazel ( Hamamelis virginiana L.) is a plant of the family of hamamelidaceae, native to the eastern regions of the United States and Canada. Despite its origins, it is also widely cultivated in Europe and it is a shrub that can reach up to 6 meters high. The deciduous leaves with serrated edge and sharp apex, vary in colour from brownish green (spring-summer) to yellowish (autumn), they are oval or obovate- shaped, they are up to 15 centimetres long and up to 10 centimetres wide. The flowers, which usually blossom between September and October, are joined in two or three-element axillary glomeruli. The fruits are woody with two loculi, between 10 and 14 millimetres long, which at maturity open releasing one or two dark and shiny seeds. The therapeutic properties of the witch-hazel are now widely recognised by the scientific community; as a matter of fact, the EMA monograph mentions local anti-inflammatory actions linked to the use of extracts of witch-hazel. These properties are also recognised in the study “Medicinal trees and shrubs introduced and cultivated in Iran: a pharmacological approach". Furthermore, annex 1 of the decree dated 9 July 2012 regulating the use of plant substances and preparations in dietary supplements attributes to this plant the ability to normalise the functionality of venous circulation (at the level of the haemorrhoidal plexus) and antioxidant properties. In particular, the antioxidant properties of the witch-hazel are also highlighted in the study conducted on cells “Antioxidant and potential anti inflammatory activity of extracts and formulations of white tea, rose and witch hazel on primary human dermal fibroblast cells".

Rutin

Rutin is aflavonoid glycoside present in plants of the genera Citrus and Sophora, in the buckwheat, in red wine, and in less significant amounts in other plants such as Mint and Eucalyptus. The structure of this glycoside consists of flavanol quercetin (aglycone) and rutinose disaccharide. Thanks to its ability to modulate the vessel tone, rutin is used in the treatment of disorders of the circulatory system. As a matter of fact, the study “Pharmacodynamics and pharmacokynetics of Veliten (rutine, alpha-tocopherol and ascorbic acid) in patients with chronic venous insufficiency’ states that rutin (although not acting on blood parameters) determines an increase in vessel tone. Increased vasoconstriction and anti-inflammatory action are also cited in the study “Evidence for a protective and antioxidant properties of rutin, a natural flavone, against ethanol induced gastric lesions". The prominent role of this glycoside can also be deduced from the fact that it is cited (together with the diosmin) as a functional active component useful in the treatment of disorders of the circulatory and lymphatic systems in the study “Pharmacological targets of drugs employed in chronic venous and lymphatic insufficiency. Being useful in circulatory system disorders system disorders, rutin is also a useful remedy against haemorrhoids: “Treatment of acute symptoms ofhemomhoid disease with hugh-dose oral 0-(beta-hydroxyethyl)-rutosides”.

Thus, forming an object of the present invention is a combination or mixture comprising or, alternatively, consisting of:

(a) diosmin;

(b) an extract of Betula pendula;

(c) an extract of Hamamelis virginiana; e

(d) rutin.

Furthermore, forming an object of the present invention is a combination of (a) diosmin, (b) an extract of Betula pendula, (c) an extract of Hamamelis virginiana and (d) rutin useful in prevention and/or in the treatment of diseases related to the circulatory system such as venous insufficiency and symptoms thereof.

Preferably, said rutin in said mixture is not:

- a hydroxymethyl and/or hydroxyethyl derivative of rutin;

- hydroxyethyl-rutin;

- hydroxymethyl-rutin;

- an extract of rutoside.

Said rutin is preferably present in said mixture as an extract of Fagopyrum esculentum.

Furthermore, forming an object of the present invention is a composition comprising said mixture (a)-(d), and food or pharmacologically acceptable additives and/or excipients.

In a preferred embodiment, diosmin (a) is present in said composition in an amount comprised from 1 mg to 5000 mg, more preferably in an amount comprised from 10 mg to 3000 mg, more preferably in an amount comprised from 20 mg to 2000 mg, even more preferably in an amount comprised from 200 mg to 700 mg.

The extract of Betula pendula (b) is preferably present in said composition in an amount comprised from 0.1 mg to 2000 mg, more preferably in an amount comprised from 1 mg to 1000 mg, even more preferably in an amount comprised from 5 mg to 800 mg, further preferably in an amount comprised from 6 mg to 100 mg.

The extract of Hamamelis virginiana (c) is preferably present in said composition in an amount comprised from 1 mg to 2500 mg, more preferably in an amount comprised from 5 mg to 1500 mg, even more preferably from 10 mg to 1000 mg, further preferably in an amount comprised from 11 mg to 200 mg.

Rutin (d) is preferably present in said composition in an amount comprised from 1 mg to 3000 mg, more preferably in an amount comprised from 5 mg to 2000 mg, even more preferably in an amount comprised from 10 mg to 1200 mg, further more preferably in an amount comprised from 15 mg to 300 mg.

The composition subject of the present invention is preferably a pharmaceutical composition, or a dietary supplement composition, or a composition for medical device, or a composition for food for special medical purposes including or comprising the aforementioned active ingredients (a)-(d) mixed with each other.

The preferred route of administration is oral or topical.

Said composition is preferably in the form of a tablet, granulate, or oral solution.

Said tablet preferably comprises:

(a) diosmin: from 300 mg to 700 mg, more preferably from 400 mg to 550 mg, even more preferably 500 mg;

(b) an extract of Betula pendula : from 6 mg to 70 mg, more preferably from 7 mg to 50 mg, even more preferably 10 mg;

(c) an extract of Hamamelis virginiana'. from 11 mg to 150 mg, more preferably from 12 mg to 70 mg, even more preferably 20 mg;

(d) rutin: from 20 mg to 200 mg, more preferably from 25 mg to 150 mg, even more preferably 50 mg.

Preferably said granulate, more preferably a granulate dispersible in water, comprises:

(a) diosmin: from 200 mg to 500 mg, more preferably from 210 mg to 500 mg, even more preferably 300 mg; (b) an extract of Betula pendula: from 6 mg to 80 mg, more preferably from 7 mg to 50 mg, even more preferably 20 mg;

(c) an extract of Hamamelis virginiana : from 11 mg to 180 mg, more preferably from 12 mg to 120 mg, even more preferably 40 mg;

(d) rutin: from 40 mg to 250 mg, more preferably from 50 mg to 200 mg, even more preferably 100 mg.

Said oral solution preferably comprises:

(a) diosmin: from 250 mg to 650 mg, more preferably from 300 mg to 600 mg, even more preferably 450 mg;

(b) an extract of Betula pendula : from 6 mg to 90 mg, more preferably from 10 mg to 45 mg, even more preferably 15 mg;

(c) an extract of Hamamelis virginiana'. from 11 mg to 180 mg, more preferably from 15 mg to 120 mg, even more preferably 30 mg;

(d) rutin: from 16 mg to 200 mg, more preferably from 17 mg to 100 mg, even more preferably 25 mg.

Furthermore, forming an object of the present invention is said composition for use in a method preventive or curative treatment of a disorder, or of an ailment, or of a symptom, or of a disease of the circulatory system in a subject, preferably in a mammal, more preferably in a human (that is in a human being) or animal.

Preferably, said disorder, or ailment, or disease is venous insufficiency, preferably chronic venous insufficiency, or wherein said symptom is associated with said venous insufficiency, preferably chronic venous insufficiency.

Even more preferably, venous insufficiency, preferably chronic venous insufficiency, is functional venous insufficiency, or organic venous insufficiency.

Suitable food or pharmacologically acceptable additives and/or excipients, to be used in said composition comprising said combination or mixture, may be selected from those normally known in the prior art and they include, though not limited to: diluents (for example dibasic calcium phosphate, lactose, microcrystalline cellulose and cellulose derivatives), thickeners (for example gums, hydroxypropyl methyl cellulose and other cellulose derivatives), sweeteners (for example sorbitols, mannitol and other polyols, acesulfame K, aspartame, cyclamates, saccharin, sucralose), lubricants (for example magnesium stearate, stearic acid, waxes), dispersants, surfactants (for example sodium lauryl sulfate and polysorbate), flavour-enhancement agents, adsorbents (for example silica gel, talc, starch, bentonite, kaolin), glidants and anti-adherents (for example talc, colloidal silica, maize starch, silicon dioxide), dyes (for example iron oxides), opacifiers (for example titanium oxide), antioxidants, binders (for example gums, starch, gelatine, cellulose derivatives, sucrose, sodium alginate), disaggregating agents (starch, microcrystalline cellulose, alginic acid, crospovidone), plasticisers (for example ethyl cellulose and other cellulose derivatives, acrylates and methacrylates, glycerol and sorbitol), preservatives (for example parabens, sulfur dioxide), viscosifiers, emulsifiers, humectants, wetting agents, chelating agents and mixtures thereof.

The following examples are given purely by way of illustration and they do not limit the scope of protection of the invention as defined in the attached claims. Changes or variations to the embodiments exemplified herein, obvious to the person skilled in the art, are encompassed by the attached claims.

EXAMPLES. Example 1: tablet

Example 2: granulate to be dispersed in water

Example 3: oral solution

Example 4: EXPERIMENTAL PART

The evaluation of the effectiveness of the combination subject of the present invention is based on the concept of synergy, the effectiveness must therefore be evaluated based on the benefits of the complex with respect to the benefits obtained by the individual active components.

In order to proceed with the evaluation described above, it is proposed to consider two aspects shared by all active components that form the mixture and which play a predominant role in the pathological (or at least non-homeostatic) phenomena affecting the circulatory and lymphatic system:

Anti-inflammatory action;

Antioxidant action.

For the evaluation of the antioxidant efficacy of the composition subject of the present invention, some in vitro assays are useful, such as: DPPH test, radical scavenging activity on nitric oxide or on peroxynitrite radical, TEAC (Total radicaltrapping antioxidant parameter), FRAP (Ferric reducing-antioxidant power), HORAC (Hydroxyl radical averting capacity), ORAC (Oxygen radical absorbance capacity) and the like. These tests are carried out both on the mixture of substances and on the individual elements.

For the evaluation of the anti-inflammatory efficacy there are various experimental methods that analyse the inflammatory phenomenon from different points of view: the cellular inflammatory mechanism and the inflammatory symptoms.

Thus, as regards the action of the combination of diosmin, extract of Betula pendula, extract of Hamamelis virginiana and rutin on the endogenous phenomena that regulate inflammation, in vitro assays are suitable in primary cultures of human cells (e.g. macrophages, chondrocytes and fibroblasts) which evaluate the ability to inhibit the release of pro-inflammatory cytokines such as IL-1, IL-6, TNF and/or to inhibit the expression of pro-inflammatory enzymes such as COX-2 and IL-1 b-induced metal protease-13.

Another study that can be conducted for the purposes of the evaluation of the anti-inflammatory activity is to be conducted on animals (for example mice, rats) to which the active agents under examination are administered (taken individually and combined) after having induced the inflammatory phenomenon by means of techniques and/or substances known to the person skilled in the art (for example carrageenan oedema).

Preferred embodiments F(n)of the present invention are illustrated below:

F1. A composition comprising a combination of dioxmin, an extract of Betula pendula, an extract of Hamamelis virginiana and rutin.

F2. The composition according F1, wherein diosmin is present in an amount comprised from 1 mg to 5000 mg, preferably in an amount comprised from 10 mg to 3000 mg, more preferably in an amount comprised from 10 mg to 3000 mg, even more preferably in an amount comprised from 20 mg to 2000 mg.

F3. The composition according to any one of F1-F2, wherein the extract of Betula pendula is present in an amount comprised from 0.1 mg to 2000 mg, more preferably in an amount comprised from 1 mg to 1000 mg, even more preferably in an amount comprised from 5 mg to 800 mg.

F4. The composition according to any one of F1-F3, wherein the extract of Hamamelis virginiana is present in an amount comprised from 1 mg to 2500 mg, preferably in an amount comprised from 5 mg to 1500 mg, even more preferably in an amount comprised from 10 mg to 1000 mg.

F5. The composition according any one of F1-F4, wherein rutin is present in an amount comprised from 1 mg to 3000 mg, preferably in an amount comprised from 5 mg to 2000 mg, even more preferably in an amount comprised from 10 mg to 1200 mg.

F6. The composition according to any one of F1-F5, wherein rutin is present as an extract of Fagopyrum esculentum.

F7. The composition according to any one of F1-F6, wherein the composition is a medical device, a dietary supplement, a cosmetic, a nutraceutical, dietetic and nutritional composition, a food product, a beverage, a nutraceutical product, a medicament, a medicated food, a pharmaceutical composition, a food for special medical purposes, a complementary feed.

F8. The composition according to any one of F1-F7, effective in the prevention and/or in the treatment of diseases associated with the circulatory system and related symptoms both in humans and animals. F9. The composition according to any one of F1-F8, wherein the diseases associated with the circulatory system is venous insufficiency.

Further embodiments E(n) of the present invention are illustrated below:

E1. A combination or mixture comprising or, alternatively, consisting of:

(a) diosmin;

(b) an extract of Betula pendula;

(c) an extract of Hamamelis virginiana; e

(d) rutin.

E2. The combination or mixture according to E1, wherein said (d) rutin is present as an extract of Fagopyrum esculentum.

E3. A composition comprising the combination or mixture (a)-(d) according to any one of E1-E2, and food or pharmacologically acceptable additives and/or excipients.

E4. The composition according E3, wherein diosmin (a) is present in an amount comprised from 1 mg to 5000 mg, preferably in an amount comprised from 10 mg to 3000 mg, more preferably in an amount comprised from 20 mg to 2000 mg, even more preferably in an amount comprised from 200 mg to 700 mg.

E5. The composition according to any one of E3-E4, wherein the extract of Betula pendula (b) is present in an amount comprised from 0.1 mg to 2000 mg, preferably in an amount comprised from 1 mg to 1000 mg, more preferably in an amount comprised from 5 mg to 800 mg, even more preferably in an amount comprised from 6 mg to 100 mg.

E6. The composition according to any one of E3-E5, wherein the extract of Hamamelis virginiana (c) is present in an amount comprised from 1 mg to 2500 mg, preferably in an amount comprised from 5 mg to 1500 mg, more preferably in an amount comprised from 10 mg to 1000 mg, even more preferably in an amount comprised from 50 mg to 200 mg.

E7. The composition according to any one of E3-E6, wherein rutin (d) is present in an amount comprised from 1 mg to 3000 mg, preferably in an amount comprised from 5 mg to 2000 mg, more preferably in an amount comprised from 10 mg to 1200 mg, even more preferably in an amount comprised from 15 mg to 300 mg. E8. The composition according to any one of E3-E7, wherein said composition is in the form of tablet, granulate, or oral solution, wherein:

- said tablet comprises:

(c) an extract of Hamamelis virginiana : from 11 mg to 150 mg, more preferably from 12 mg to 70 mg, even more preferably 20 mg;

(d) rutin: from 20 mg to 200 mg, more preferably from 25 mg to 150 mg, even more preferably 50 mg; or

- said granulate, preferably a granulate dispersible in water, comprises:

(a) diosmin: from 200 mg to 500 mg, more preferably from 210 mg to 500 mg, even more preferably 300 mg;

(b) an extract of Betula pendula : from 6 mg to 80 mg, more preferably from 7 mg to 50 mg, even more preferably 20 mg;

(c) an extract of Hamamelis virginiana'. from 11 mg to 180 mg, more preferably from 12 mg to 120 mg, even more preferably 40 mg;

(d) rutin: from 40 mg to 250 mg, more preferably from 50 mg to 200 mg, even more preferably 100 mg; or

- said oral solution comprises:

E9. The composition according to any one of E3-E8, wherein said composition is a pharmaceutical composition, or a medical device composition, a dietary supplement composition, or food for special medical purposes composition. E10. The composition according to any one of E3-E9, wherein said composition is for use in a method for the preventive or curative treatment of a disorder, or of an ailment, or of a symptom, or of a disease of the circulatory system in a subject, preferably in a mammal, more preferably in a human (human being) or animal.

E11. The composition for use according to E10, wherein said disorder, or ailment, or disease is venous insufficiency, preferably chronic venous insufficiency, or wherein said symptom is associated with said venous insufficiency, preferably chronic venous insufficiency. E12. The composition for use according to E11, wherein venous insufficiency, preferably chronic venous insufficiency, is functional venous insufficiency.

E13. The composition for use according to E11, wherein venous insufficiency, preferably chronic venous insufficiency, is organic venous insufficiency.

Claims

1. A combination or mixture comprising or, alternatively, consisting of:

(a) diosmin;

(b) an extract of Betula pendula;

(c) an extract of Hamamelis virginiana; e

(d) rutin.

2. The combination or mixture according to claim 1, wherein said (d) rutin is present as an extract of Fagopyrum esculentum.

3. A composition comprising the combination or mixture (a)-(d) according to any one of the preceding claims, and food or pharmacologically acceptable additives and/or excipients.

4. The composition according to claim 3, wherein diosmin (a) is present in an amount comprised from 1 mg to 5000 mg, preferably in an amount comprised from 10 mg to 3000 mg, more preferably in an amount comprised from 20 mg to 2000 mg, even more preferably in an amount comprised from 200 mg to 700 mg.

5. The composition according to any one of claims 3-4, wherein the extract of Betula pendula (b) is present in an amount comprised from 0.1 mg to 2000 mg, preferably in an amount comprised from 1 mg to 1000 mg, more preferably in an amount comprised from 5 mg to 800 mg, even more preferably in an amount comprised from 6 mg to 100 mg.

6. The composition according to any one of claims 3-5, wherein the extract of Hamamelis virginiana (c) is present in an amount comprised from 1 mg to 2500 mg, preferably in an amount comprised from 5 mg to 1500 mg, more preferably in an amount comprised from 10 mg to 1000 mg, even more preferably in an amount comprised from 50 mg to 200 mg.

7. The composition according to any one of claims 3-6, wherein rutin (d) is present in an amount comprised from 1 mg to 3000 mg, preferably in an amount comprised from 5 mg to 2000 mg, more preferably in an amount comprised from 10 mg to 1200 mg, even more preferably in an amount comprised from 15 mg to 300 mg.

8. The composition according to any one of claims 3-7, wherein said composition is in the form of tablet, granulate, or oral solution, wherein: - said tablet comprises:

- said granulate, preferably a granulate dispersible in water, comprises:

- said oral solution comprises:

9. The composition according to any one of claims 3-8, wherein said composition is a pharmaceutical composition, or a medical device composition, a dietary supplement composition, or food for special medical purposes composition.

10. The composition according to any one of claims 3-9, wherein said composition is for use in a method for the preventive or curative treatment of a disorder, or of an ailment, or of a symptom, or of a disease of the circulatory system in a subject, preferably in a mammal, more preferably in a human (human being) or animal.

11. The composition for use according to claim 10, wherein said disorder, or ailment, or disease is venous insufficiency, preferably chronic venous insufficiency, or wherein said symptom is associated with said venous insufficiency, preferably chronic venous insufficiency.

12. The composition for use according to claim 11, wherein venous insufficiency, preferably chronic venous insufficiency, is functional venous insufficiency.

13. The composition for use according to claim 11, wherein venous insufficiency, preferably chronic venous insufficiency, is organic venous insufficiency.