WO2019030209A1

WO2019030209A1 - Katrafay essential oil composition and use as cosmetic, medicament and/or nutritional supplement

Info

Publication number: WO2019030209A1
Application number: PCT/EP2018/071353
Authority: WO
Inventors: Luisa Pecorari; Tatiana VIGNUDELLI
Original assignee: Ophera Srl
Current assignee: Ophera Srl
Priority date: 2017-08-09
Filing date: 2018-08-07
Publication date: 2019-02-14
Anticipated expiration: 2020-02-09
Also published as: IT201700092271A1

Abstract

La présente invention concerne une composition à base d'huile essentielle de Katrafay (Cedrelopsis grevei) comprenant de 20 % à 40 % en poids d'ishwarane, par rapport au poids total de la composition. La composition peut éventuellement comprendre également du limonène, dans une quantité telle que le rapport en poids d'ishwarane : limonène est compris entre 50:1 et 100:1. L'invention concerne en outre une formulation comprenant la composition précitée. L'invention concerne également un procédé de préparation de la composition, ainsi que son utilisation comme produit cosmétique, médicament ou complément nutraceutique, en particulier dans le traitement d'états inflammatoires.The present invention relates to a composition based on Katrafay essential oil ( Cedrelopsis grevei ) comprising from 20% to 40% by weight of ishwarane, relative to the total weight of the composition. The composition may optionally also include limonene, in an amount such that the weight ratio of ishwarane: limonene is between 50: 1 and 100: 1. The invention further relates to a formulation comprising the above composition. The invention also relates to a process for the preparation of the composition, as well as its use as a cosmetic product, a medicament or a nutraceutical supplement, in particular in the treatment of inflammatory conditions.

Description

KATRAFAY ESSENTIAL OIL COMPOSITION AND USE AS COSMETIC, MEDICAMENT AND/OR NUTRITIONAL SUPPLEMENT

DESCRIPTION

TECHNICAL FIELD

The present invention relates to a composition based on katrafay essential oil (Cedrelopsis grevei) and formulations containing said composition, particularly for use in the treatment of inflammation states.

BACKGROUND ART

The use in cosmetics and medicine of herbs, plants and their derivatives or extracts has very ancient origins and spread all over the world. Extracts and essential oils of vegetable origin have long been used for therapeutic purposes or to prevent the occurrence of various types of conditions. Currently there is a growing interest and a consequent increase in the use of vegetable preparations in the field of cosmetics and pharmaceuticals, accompanied by a concomitant increase in the scientific literature of information regarding the biological properties of plant species.

The katrafay essential oil (Cedrelopsis grevei), an aromatic plant native to Madagascar, is extracted from the bark and leaves and is widely used in folk medicine. The katrafay essential oil (hereafter referred to as "katrafay oil") has been shown to have antioxidant, cytotoxic (has been effective against the MCF-7 tumor cell line), antimalarial (is active against Plasmodium falciparum, one of the species that causes malaria in humans) and, above all, antimicrobial and anti-inflammatory (demonstrated in an in vitro test based on the measurement of the inhibition of 5- lipoxygenase activity, from which an IC50 value of 21,33 mg/1).

Patent application JP 2005213202 A discloses the preparation of an antiinflammatory, anti-aging, antioxidant agent which can be used for cosmetic preparations and which contains Cedrelopsis grevei extract.

WO 2008034821 Al discloses the use of active ingredients extracted from various plant species, including Cedrelopsis grevei, for the formulation of cosmetic or pharmaceutical preparations intended to protect the skin from dehydration and to relieve skin from chapping, stings, itching.

The main constituents of most essential oils, including katrafay oil, belong to the category of terpenes, biomolecules consisting of multiples of the isoprenic unit, obtained in biological systems by the reaction of several units of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). In plants, terpenes act both as a defense against parasites and as a source of reserve energy.

The katrafay essential oil contains various terpenes, from which the antimicrobial and anti-inflammatory action depend and associated with the oil itself. The main terpenes contained in the katrafay essential oil belong to the subclass of monoterpenes (e.g. limonene, alpha-pinene, beta-pinene) or to the subclass of sesquiterpenes (e.g. ishwarane, beta-elemene, alpha-copaene). The terpenes have the ability to add ozone to the double bonds of isoprenic units (ozonation reaction), forming the corresponding ozonides. Ozonides derived from terpenes (or ozonized terpenes) remain stable for long periods in an anhydrous environment and free from reducing agents, although they have the ability to act as fat-soluble oxidants and to release free oxygen.

Likewise, also other molecules in essential oils are able to form ozonides following ozonation. For example, the unsaturated triglycerides may go towards the ozonation reaction at the level of double bonds, forming ozonized derivatives having antimicrobial and anti-inflammatory action. However, as regards the antimicrobial and anti-inflammatory action, the ozonized terpenes have two advantages compared to the unsaturated ozonized triglycerides. First of all, the ozonized terpenes have a significantly lower molecular weight (about 1/5 or 1/6 of the unsaturated ozonized triglycerides), a feature that gives them a much higher capacity to spread through biological tissues and an affinity towards water considerably more high, which results in greater reactivity in biological matrices. Moreover, the ozonized terpenes have a higher specific content of active oxygen with respect to unsaturated ozonized triglycerides, i.e. a greater content of active oxygen per mole of substance.

For these reasons, the antimicrobial and anti-inflammatory power of ozonized terpenes is much higher than that of unsaturated ozonized triglycerides. Moreover, ozonized terpenes also offer the advantage of not presenting the toxicity characterizing many of the respective terpenes.

The ozonized terpenes therefore have antimicrobial, antiviral, anti-inflammatory, and vulnerary (cicatrizant) activities and their use is known in the form of essential oils containing thereof or other formulations optimized according to application type, for the treatment of diseases such as: infections of various types, wounds, bedsores, arthritic pains, rheumatism, muscle injuries resulting from sports activities, dermatitis, psoriasis, gastroduodenal ulcer, etc..

Ishwarane is a sesquiterpenes in various plant species (including Cedrelopsis grevei, Bixa orellana, Aristolochia indica). Currently it is not known that ishwarane which is isolated from the extract has anti-inflammatory properties.

Limonene is a monoterpene that takes its name from the lemon tree {Citrus limon), but it is also in other plant species. Limonene exists naturally in two enantiomeric forms: lemon peel mainly contains L-limonene, while orange peel contains predominantly D-limonene. D-limonene is also present in many other botanical species, such as dill oil (Anethum graveolens) and cumin oil (Carum carvi), of which it is more than 30%; cumin oil also contains 48-65% of the oxygenated derivative of D-limonene ((+) - carvone), which has shown antitumor properties. D-limonene and carvone have also shown to exert significant anti-inflammatory effects.

It has also been described that D-limonene has neutralizing effects on gastric acids and is able to support normal peristalsis, characteristics that justify its use against gastroesophageal reflux disease. Furthermore, it has been described that D-limonene has chemopreventive properties for many types of tumors, particularly for breast, prostate, colon and skin cancer.

D-limonene is known for its high penetration capacity, particularly appreciable at the skin level, which is the reason why it is referred to as an "enhancer", to support the action of other molecules that alone would have little ability to cross epithelial barriers.

Among the molecular mechanisms that contribute to explaining the antiinflammatory properties of D-limonene are the inhibition of angiogenesis and the production of TNFa (Tumor Necrosis Factor a) and IL-6 (Interleukin-6), known molecular mediators of the inflammatory process.

Furthermore, D-limonene has shown to have intrinsic epithelial tissue repair properties, as its perillyl acid metabolite (POH) appears to be the key molecule in restoring homeostasis and skin integrity. In addition to its anti-inflammatory properties, D-limonene has remarkable properties to oppose to aging and promote skin rejuvenation.

The main source of D-limonene is the oil obtained from citrus peel by cold pressing, which avoids the thermal degradation of components, unlike the process of hydrodistillation generally used to obtain essential oils.

In Europe the use of limonene in cosmetic products is allowed, but being classified as an allergenic substance, its presence must be indicated in the list of ingredients, if it exceeds a certain level (0,001% for products intended to remain on the skin, as creams, ointments, deodorants, and 0,01% in products intended to be rinsed away from the skin, such as soaps or shampoos). However, the non-oxidized D-limonene is inert and well tolerated; contact dermatitis can occur instead against oxidized D- limonene, which may be allergenic and irritating. In the context of the present invention, the term limonene refers indifferently to D- limonene, L-limonene or to a mixture of D-limonene and L-limonene.

In light of the above, the possibility of developing a composition with increased antiinflammatory efficacy is therefore of interest. The primary task of the present invention is therefore to develop a composition based on katrafay essential oil (as well as a formulation which includes this oil) having superior anti-inflammatory efficacy with respect to the known compositions based on this essential oil.

Within this aim, an object of the invention is to provide a composition based on katrafay essential oil with high anti-inflammatory activity which is easy to administer, safe and non-toxic for the final consumer.

Another object of the invention is to provide a composition based on katrafay essential oil, the preparation of which is simple to make.

The aim and objects of the invention are solved by a composition as defined in the appended claims and described below, also with reference to figures 1-4.

DISCLOSURE OF INVENTION

The present invention relates to a composition based on katrafay essential oil (Cedrelopsis grevei) comprising from 20% to 40% by weight with respect to the total weight of the ishwarane composition.

The composition can optionally also include limonene, in such amount that the ratio in weight ishwarane: limonene is between 50: 1 and 100: 1.

The invention also relates to a formulation which includes the composition as described above. Part of the invention is also a process for preparing the composition as well as its use as a cosmetic, as a medicine, or as a nutraceutical supplement, in particular for the treatment of inflammation states.

BRIEF DESCRIPTION OF THE DRAWINGS - Figure 1 shows a graph showing the effect of compositions comprising katrafay essential oil and various concentrations of ishwarane (0%, 5%, 10%, 20%>, 30%>, 40%, 50%, 60%) on expression of the inflammatory cytokine TNFa by cells of the THP-1 cell line, at basal conditions. OEK = katrafay essential oil; Ishw = ishwarane; pure ishw = composition comprising 100% ishwarane; Tnfa = TNFa; C- = negative control (untreated cells).

- Figure 2 shows a graph showing the effect of compositions comprising katrafay essential oil and various concentrations of ishwarane (0%>, 5%, 10%>, 20%>, 30%>, 40%, 50%, 60%) on expression of the inflammatory cytokine TNFa by cells of the THP-1 cell line, subjected to pre-stimulation with lipopolysaccharide (LPS). OEK = katrafay essential oil; Ishw = ishwarane; pure ishw = composition comprising 100% ishwarane; Tnfa = TNFa; C- = negative control (untreated cells, neither with OEK, nor with LPS); C + = positive control (cells treated only with LPS).

- Figure 3 shows a graph showing the effect of compositions comprising ishwarane and limonene in various weight ratios on the expression of the inflammatory cytokine TNFa by cells of the THP-1 cell line, subjected to pre-stimulation with lipopolysaccharide (LPS). Column 3 (Ishw) shows the data related to cells treated with only 0,002% ishwarane. Ishw = ishwarane; limon = limonene; OEK = katrafay essential oil; OE LAV = lavender essential oil; Tnfa = TNFa; C- = negative control (untreated cells, neither with OEK, nor with LPS); C + = positive control (cells treated only with LPS). - Figure 4 shows a graph showing the effect of compositions comprising katrafay essential oil and various concentrations of ishwarane (0%>, 5%, 10%>, 20%>, 30%>, 40%, 50%) on the expression of inflammatory cytokine TNFa by cells of the THP-1 cell line, subjected to pre-stimulation with lipopolysaccharide (LPS). OEK = katrafay essential oil; Ishw = ishwarane; pure ishw = composition comprising 100% ishwarane; Tnfa = TNFa; C- = negative control (untreated cells, neither with OEK, nor with LPS); C + = positive control (cells treated only with LPS).

DETAILED DESCRIPTION OF THE INVENTION

The composition according to the present invention comprises katrafay essential oil (Cedrelopsis grevei) and is characterized by a content of ishwarane (which may be naturally present in the katrafay essential oil or added thereto) ranging between 20% and 40% by weight with respect to the total weight of the composition. As mentioned above, the katrafay essential oil is known for its anti-inflammatory and lenitive properties, above all, but not exclusively, in the skin and mucous membranes. The inventors have surprisingly found that the presence of ishwarane terpene in the range of 20-40% by weight is able to enhance the anti-inflammatory and lenitive effect normally associated with the essential oil of katrafay. In particular, it has been observed that the anti-inflammatory and lenitive effect is synergistically enhanced, i.e. this effect is greater than the sum of the effect that would be obtained by administering both components individually.

Preferably, the composition of the invention can comprise from 25% to 35% of ishwarane by weight with respect to the total composition weight.

In one embodiment, the composition may further comprises also the limonene terpene: the weight ratio between ishwarane and limonene must be between 50: 1 and 100: 1. The presence of limonene in the composition of the invention, in the weight ratio with respect to the aforementioned ishwarane, further increases the effects of the composition itself; also in this case, the inventors have observed that the enhancement of the effects is of synergic type: it should be emphasized, however, that this synergistic strengthening occurs only when the weight ratio between ishwarane and limonene is between 50: 1 and 100: 1. The limonene can be naturally present in the katrafay essential oil or added to it. The anti-inflammatory and lenitive effect of synergistic type is manifested by inhibiting the biological response to pro-inflammatory stimuli, for example with the inhibition of the production of inflammatory mediators, typically cytokines and in particular interleukins and TNF (Tumor Necrosis Factor).

The composition of katrafay oil is closely related to its anti-inflammatory, lenitive, moisturizing properties. In vitro experimental studies conducted by the inventors of the present patent application have made it possible to highlight how the content of some components (in particular, in fact, ishwarane and limonene) is decisive in terms of effectiveness.

It is important to underline that the synergistic action in terms of anti-inflammatory and lenitive effect is closely correlated to the proportions between the components. In particular, as mentioned above, the synergistic action is achieved only when the content of ishwarane in the composition is between 20% and 40% by weight with respect to the total weight of the composition; if limonene is also present in the composition, a further synergistic effect is observed when the ratio in weight ishwarane: limonene is between 50: 1 and 100: 1. Outside the indicated ranges, the anti-inflammatory and lenitive effect of katrafay oil due to the presence of ishwarane (and possibly limonene) is not significantly enhanced.

The relationship between the increase of the anti-inflammatory activity of the katrafay oil and the particular concentration range of ishwarane, and possibly of the limonene, was determined by the inventors of the composition described here by experimental analyzes. These analyzes consist of assays performed on cells of the THP-1 cell line, also stimulated with lipopolysaccharide (LPS). The THP-1 cell line is an immortalized line of human monocytes derived from a patient with acute monocytic leukemia; this cell line represents an experimental model for the in vitro study of the inflammation, which is accepted and commonly used by the scientific community. Lipopolysaccharide is a component of the cell wall of Gram-negative bacteria; it is a macromolecule consisting of a lipidic portion and a polysaccharide portion capable of causing strong immune responses in animals, including humans. The stimulation of a cell culture with lipopolysaccharide, therefore, induces the inflammatory response and the consequent production of the inflammatory cytokine TNFa by monocytes, similarly to what happens in vivo in the presence of an inflammatory stimulus. In the tests performed to evaluate the effectiveness of the composition of the invention, the THP-1 cells were pre-stimulated (or not) with LPS and treated (or not treated) with the composition of the invention to evaluate their ability to inhibit the inflammatory response by verifying the production of TNFa, as shown in examples.

The effected tests have shown that the composition according to the invention, based on katrafay essential oil and comprising ishwarane in concentrations ranging from 20% to 40%, is able to reduce the production of TNFa by THP-1 cells in a more marked way (as a result of the synergistic effect) with respect to the reduction obtained by treating the same cells, under the same experimental conditions, with only katrafay oil or with compositions based on katrafay oil comprising concentrations of ishwarane outside the abovementioned concentration range.

Similarly, the conducted tests shown that the addition of limonene to the composition of the invention further increases the anti-inflammatory action with a synergistic effect, when the weight ratio between the ishwarane and limonene components is between 50: 1 and 100: 1.

As mentioned earlier, katrafay essential oil may naturally contain a certain amount of ishwarane and limonene; katrafay oil is commonly marketed without titration in ishwarane and limonene, since the concentration of these active substances is subjected to natural variability depending on the plant part from which the oil is extracted, from the harvesting season, from the geographical plant origin and from its growth climate, as well as from the extraction method of the essential oil.

To achieve the concentrations provided for the composition of the invention it may therefore be necessary to add to the katrafay essential oil further ishwarane (previously purified) in addition to that naturally present in the oil itself. In the embodiment that also provides the presence of limonene in the expected weight ratio, it is possible that even a further amount of this component should be added to the katrafay essential oil.

In one embodiment, the ishwarane added to the katrafay essential oil may be of natural origin, and preferably may be isolated from botanical species; in another embodiment, ishwarane may be of synthetic origin, i.e. produced in the laboratory by chemical synthesis. Similarly, in one embodiment the limonene added to the katrafay essential oil may be of natural origin, and preferably may be isolated from botanical species; in another embodiment, the limonene may be of synthetic origin, i.e. produced in laboratory by chemical synthesis.

The limonene added to the katrafay essential oil may be D-limonene, L-limonene or a mixture of D-limonene and L-limonene.

The composition of the invention may be in ozonized or non-ozonized form. In a preferred embodiment, the composition may be in ozonized form. In the context of the present invention, and as is immediately clear to the person skilled in the art, by "ozonized form composition" (or more simply "ozonized composition") is meant the composition according to the invention, based on katrafay essential oil comprising 20 -40% by weight of ishwarane and, optionally, limonene in the weight ratio with the expected ishwarane, which was effected by an ozonization process. In the ozonization process, ozone is added at the double bond level of one or more of the isoprenic units present in the terpenes contained in the composition. As shown previously, the ozonized terpenes have a number of favorable biological characteristics, including lower toxicity compared to the corresponding terpenes and the ability to diffuse in biological matrices: for this reason the ozonized form composition represents a preferred embodiment of the invention.

In this embodiment where the katrafay essential oil is in ozonized form, the ozonization process is preferably carried out after the possible addition of ishwarane and/or limonene and/or after the possible mixing of the composition with any other vegetable oils; in this way all the present terpenes, both those naturally contained in katrafay oil and those added to obtain the expected quantities, as well as any unsaturated triglycerides present in vegetable oils, are ozonized.

As can be understood in the light of the foregoing, ozonation improves the antiinflammatory, lenitive and moisturizing effect of katrafay oil regardless of the specific composition of the invention, since the terpenes in ozonized form, as we said, are more effective in reaching the biological action site and then in carrying out their action. However, when ishwarane and possibly limonene are present in the indicated percentage ratios, and therefore the synergic effect is obtained, ozonation allows to obtain an even more intense anti-inflammatory, lenitive and moisturizing action.

The experimental studies conducted by the inventors have shown that the treatment, with ozonized compositions based on katrafay essential oil, determines an inhibition of the production of TNFa by the THP-1 cells much higher than the inhibition of the production of TNFa obtained with the treatment of cells with the corresponding compositions based on non-ozonized katrafay essential oil.

Another aspect of the invention relates to a formulation comprising the composition based on katrafay essential oil comprising from 20% to 40% by weight of ishwarane with respect to the total weight of the composition itself and one or more acceptable excipients from pharmaceutical, and/or cosmetic and/or nutritional point of view. In one embodiment, the formulation of the invention may further comprise one or more carriers. In consideration of the chemical nature of the composition, the excipients and the carriers can be preferably lipophilic or alcoholic, in particular they can be vegetable oils and/or mixtures thereof. The excipients and carriers are chosen appropriately according to the formulation in which the composition will be incorporated, as it is known to a person skilled in the pharmaceutical, cosmetic and/or nutraceutical formulation field. In another embodiment, the formulation of the invention can further comprise one or more additional vegetable oils, preferably essential vegetable oils, in particular the lavender essential oil. Some studies have suggested that lavender essential oil, the use of which is known for various applications, would be able to enhance the effects of other compositions based on essential oils, including katrafay, probably thanks to an interaction between active components of various oils.

In another embodiment, the formulation of the invention can further comprise one or more anti-inflammatory and/or lenitive components.

The formulation of the invention can preferably comprise the composition as previously described in a percentage by weight typically included between 0,001% and 40%, preferably between 0,1 % and 10%>. These percentage ranges are applied particularly, but not exclusively, to cosmetic formulations, preferably compositions for topical applications on skin and mucous membranes. With regard to food formulations (for example, supplements or nutraceuticals) they may preferably comprise the composition of the invention in an amount such that the daily administration of the composition is between 30 and 50 mg per kg of weight of the subject.

It is also possible to use the composition of the invention in pure form (i.e. without adding any other excipient, carrier or other ingredient), for example to make "touches" on small areas of the skin and mucous membranes.

The formulation comprising the composition according to the invention can take different physical forms, depending on the type of administration to which it is intended. By way of example but not limitative, the formulation can be a cream, a gel, a solution, an ointment, a suspension, a formulation in liposomes, a powder, an aerosol, a capsule, a tablet, a pessary, a paste, a spray, a suppository.

In one embodiment, the formulation can preferably be selected from a cream, a gel and a solution. A solution can for example be represented by a mouthwash, a lavender, ear drops, eye drops. In another embodiment, the formulation can preferably be selected from a powder, a capsule and a tablet. Furthermore, the formulation can for example be incorporated into transdermal patches, inhalers, impregnated scaffolds, ointments.

In another embodiment, the formulation comprising the composition of the invention may be a functional food and/or a supplement and/or a nutraceutical. In a preferred embodiment, for example, such functional food, integrator, and/or nutraceutical may be specific for the treatment of inflammation of the locomotor and/or gastrointestinal, and/or nervous apparatus,.

Another aspect of the invention relates to a process for preparing the composition according to the invention, which comprises the steps of:

(a) obtaining an essential katrafay oil;

(b) verify the ishwarane title of the essential oil obtained in step (a) and, if necessary, add ishwarane until it reaches a percentage by weight with respect to the total weight of the composition between 20% and 40%;

(c) mixing the components together so as to obtain a homogeneous composition; where (b) and (c) steps are carried out at a temperature not exceeding 80°C and avoiding direct exposure to light.

In one embodiment, in step (b) it is also possible to verify the limonene title of the essential oil obtained in step (a) and, if necessary, add limonene in an amount such that the ratio by weight between ishwarane and limonene in the composition it is between 50: 1 and 100:1.

In another embodiment, the process described herein may comprise a further step (d) of ozonization of the homogeneous composition obtained in step (c): this further step leads to obtaining a composition based on katrafay essential oil in which the contained terpenes are ozonized. Another aspect of the invention relates to the use of the composition described herein (or to formulations comprising the composition) as cosmetic, and/or as a medicament, and/or as a nutraceutical/supp lenient. The use as a medicament may preferably be in the prevention or treatment of inflammation states. In one embodiment, the states of inflammation may be acute; in another embodiment, the states of inflammation may be chronic.

Preferably, the composition is applied in dermatological field, for the prevention of irritation or inflammation states affecting the skin or mucous membranes. With regard to skin irritation or inflammation, the composition can be used for example to prevent or treat dermatitis of various types (such as pruriginous and/or seborrhoeic dermatitis), sunburn, insect bites, acne. With regard to irritation or inflammation of the mucous membranes, the composition can be used, for example, to prevent or treat gingivitis, inflammation of the oral or nasal mucosa, asthma, vaginitis. The composition was also evaluated for use on body sensitive areas and subject to irritation such as the genitals and it was effective and well tolerated.

The use as a medicament may preferably be in the treatment of erectile dysfunction, thanks to the vasodilatory properties of the active components of the composition itself.

The composition according to the invention (and the formulations comprising thereof) can be administered, for example, topically or systemically. Topical administration is performed by applying the composition or formulation at the interested site, in an appropriate physical form. An example of topical administration may be the application of the composition or formulation (preferably in the form of a cream, gel, solution) on the skin. Another example of topical administration may be vaginally, preferably where the formulation is in the pessary form. Yet another example of topical administration can be nasally, preferably where the formulation is in the form of a solution or aerosol. Systemic administration can typically occur by mouth and involves formulations comprising the composition; the composition of katrafay oil in pure form is in fact not appropriate for systemic administration. In this case the formulation may therefore preferably have the shape of a capsule, a tablet or a powder. Furthermore, a formulation comprising the composition of the invention can be used as a nutraceutical supplement, incorporating it to a food or a beverage, thus obtaining a functional food.

The composition according to the present invention, the process for its preparation, the formulations which comprise it and the uses to which it is intended, conceived as described herein, are susceptible of numerous modifications and variations, all of which are within the scope of the inventive concept; furthermore, all the details may be replaced by other equivalent elements whose correspondence is known to the person skilled in the art.

Furthermore, it must be understood that the characteristics of the embodiments described with reference to an aspect of the invention are to be considered valid also with regard to the other aspects of the invention, even if not explicitly repeated. EXEMPLES

EXAMPLE 1 : ANTIINFIAMMATORY ACTIVITY OF KATRAFAY AND ISHWARANE ESSENTIAL OIL COMPOSITIONS ON CELLS OF THP-1 CELLULAR LINE AT BASIC CONDITIONS.

The expression of the inflammatory cytokine TNFa, an indicator of inflammatory conditions, was evaluated by ELISA tests carried out in vitro using cells of the THP- 1 cell line, under basal conditions. Cells were treated in vitro with katrafay essential oil compositions (0,02 μΐ/ml of cell culture medium), added with ishwarane at different concentrations. The cells were placed for 24 hours in an incubator at a constant temperature of 37°C, with humidity between 90% and 95% and 5% carbon dioxide. After this incubation time, ELISA tests were performed to quantify the production of TNFa by the cells. The ELISA assays were conducted in 96-sink plates according to the indirect sandwich mode: at the bottom of the plate sinks, the capture antibodies are immobilized to bind the TNFa present in the samples when they are deposited in the respective wells; the capture phase is then followed by a series of washings and then by the detection phase which takes place by adding a marked second anti-TNFa antibody with alkaline phosphatase, an enzyme capable of developing the colorimetric reaction that is detected and quantified by spectrophotometric reading. Incubation times and conditions, antibodies and reagents amounts, and spectrophotometric reading mode are those indicated in the ELISA assay kit (Human TNF-a ELISA Kit, Thermo Fisher) used in this experiment.

With reference to figure 1, it is noted that the compositions comprising katrafay essential oil and ishwarane, the latter in a concentration between 20% and 40% by weight with respect to the total weight of the composition of essential oil + ishwarane, are found to have maximum anti-inflammatory activity (indicated by the lower production of TNFa). These results show that, within the concentration range of 20%) to 40%) by weight of ishwarane, a synergistic effect is established between this compound and the katrafay oil, which allows to obtain an anti-inflammatory activity which cannot be achieved with the use of the two components in a disjointed manner and greater than the sum of the anti-inflammatory effects mediated by the two components separately (consider that the treatment of cells with ishwarane alone is not associated with any anti-inflammatory activity in these experimental conditions, as shown by the last right column of the graph in figure 1).

EXAMPLE 2: ANTIINPHIAMMATORY ACTIVITY OF COMPOSITIONS INCLUDING KTRAFAY AND ISHWARANE ESSENTIAL OIL ON THP-1 CELLULAR LINE CELLS AFTER STIMULATION WITH LIPOPOLYSACCHARIDE.

Cells from the THP-1 human monocyte cell line were pre-stimulated with ^g/ml of lipopolysaccharide; 12 hours after pre-stimulation with LPS, THP-1 monocytes were treated with katrafay essential oil compositions (0,02 μΐ/ml of cell culture medium), added with ishwarane at different concentrations. The cells were placed for 24 hours in an incubator at a constant temperature of 37°C, with humidity ranging between 90% and 95% and 5% carbon dioxide. Once this incubation time has elapsed, ELISA assays have been performed to quantify the production of TNFa by the cells, as described with reference to Example 1.

With reference to figure 2, it is observed that, even under the experimental conditions of Example 2, which simulate an ongoing acute inflammatory process, the compositions comprising katrafay essential oil (Cedrelopsis grevei) and ishwarane, the latter in a concentration between 20% and 40% by weight with respect to the total weight of the essential oil + ishwarane composition, have the highest antiinflammatory activity (indicated by the lower production of TNFa). Also from these results it is evident that, within the concentration range from 20% to 40% by weight of ishwarane, a synergistic effect is established between this compound and the katrafay oil, which allows to obtain an unreachable anti-inflammatory activity, in these experimental conditions, with the two single components in a disjointed way and greater than the sum of the anti-inflammatory effects mediated by the two components separately, considering that the only treatment of ishwarane cells is not associated with any anti-inflammatory activity even in these experimental conditions (last column on the right of the graph in figure 2).

Experiments carried out with katrafay extracts obtained from the bark, from other parts of the plant (for example, leaves), and also mixing extracts obtained from different parts of the plant, are always arrived to similar results.

EXAMPLE 3: ANTIINFIAMMATORY ACTIVITY OF ISHWARANE AND LEMONENE COMPOSITIONS ON CELLULAR LINE THP-1 AFTER STIMULATION WITH LIPOPOLYSACCHARIDE.

Cells from the THP-1 human monocyte cell line were pre-stimulated with ^g/ml of lipopolysaccharide; 12 hours after pre-stimulation with LPS, the THP-1 monocytes were treated with compositions comprising ishwarane and limonene, whose weight ratio is between 10: 1 and 200: 1. The total amount of the two components was kept equal to 0,002% by weight on the total weight of the cell culture medium in order to compare the result with the reference sample, treated with 0,002% by weight with respect to the total weight of the culture medium of the only ishwarane.

The cells were placed for 24 hours in an incubator at a constant temperature of 37°C, with humidity between 90% and 95% and 5% carbon dioxide. Once this incubation time has elapsed, ELISA assays have been performed to quantify the production of TNFa by the cells, as described with reference to Example 1.

With reference to figure 3, it is observed that the compositions comprising ishwarane and limonene exert high anti-inflammatory activity (low production of TNFa) when the weight ratio between the two components is between 50: 1 and 100: 1. Outside this range the anti-inflammatory activity decreases significantly even when the ratio shifts in favor of limonene (ishwarane: limonene 10: 1, 20: 1, 35: 1), for which antiinflammatory properties are known and described. The column of the graph marked as "OEK + total ishw:limon 50: 1" represents the composition of the invention, comprising katrafay essential oil, ishwarane and limonene, with an ishwarane weight ratio: limonene 50: 1. The fact that this column is particularly low therefore reflects the synergistic effect of the composition of the invention, which is able to reduce TNFa production far more than ishwarane alone or in combination with limonene.

The column of the graph marked as "OEK + total ishw:limon 50: 1 + OE LAV", corresponding to the composition of the invention, further added with lavender essential oil, shows how the presence of this oil further increases the anti- inflammatory efficacy of the composition.

EXAMPLE 4: ANTIINPHIAMMATORY ACTIVITY OF COMPOSITIONS INCLUDING KETRAFAY OIL OF KATRAFAY OZONIZED OR NOT OZONIZED ON CELLS OF THE THP-1 CELLULAR LINE AFTER STIMULATION WITH LIPOPOLYSACCHARIDE. Cells from the THP-1 human monocyte cell line were pre-stimulated with ^g/ml of lipopolysaccharide; 12 hours after pre-stimulation with LPS, THP-1 monocytes were treated with essential katrafay oil compositions (0,02 μΐ/ml of cell culture medium), added with ishwarane at different concentrations; for each composition the non- ozonized form and the ozonized form were tested.

With reference to figure 4, it is observed that the compositions comprising katrafay essential oil and ishwarane have a greater anti-inflammatory activity (indicated by the lower production of TNFa) when the katrafay essential oil is in ozonized form, compared to when it is in non-ozonized form, with equal percentage content of ishwarane. Similarly to what is reported in example 2, it is observed that the maximum anti-inflammatory activity occurs in the concentration range of ishwarane ranging from 20% to 40% by weight on the total weight of the composition.

Overlapping results were obtained both by performing the ozonation of the katrafay essential oil before mixing it with ishwarane, or by making the ozonation directly on the mixture.

Pure ishwarane (composition containing 100% ishwarane) also slightly increases its anti-inflammatory property as a result of ozonation, but this effect is to be considered limited and non-specific.

EXAMPLE 5: FORMULATION IN THE FORM OF CREAM FOR DERMATOLOGICAL USE INCLUDING KATRAFAY ESSENTIAL OIL

Ingredients of the formulation indicated according to the INCI nomenclature (the percentages refer to the quantities by weight with respect to the total weight of the formulation):

- Cedrelopsis grevei Bark extract, titrated in ishwarane to 33,5% e in limonene to

0,44%: 1,00%

- Glyceryl stearate: 2,00%

- Polysorbate 60 (or Tween): 1,00%

- Stearic Acid: 1,40%

- Glycerin: 0,2%

- Glycyrrhetinic acid: 2,00%

- Triethanolamine: 0,50%

- Carbomer: 0,40%

- Butyrospermum Parkii Butter (Shea olein) 12,00%

- Helianthus Annuus seed oil: 8,00%

- Prunus Amygdalus Dulcis oil: 5,00%

- Tocopherol: 0,05%

- Parfum: 0,5%

- Phenoxyethanol and Ethylhexylglycerin: 0,30%

- Water: complement to 100%

The formulation here described, in cream form, can be used in the topical treatment of skin irritation states.

EXAMPLE 6: FORMULATION IN THE FORM OF CREAM FOR DERMATOLOGICAL USE (ESPECIALLY DERMATOLOGICAL-GENITAL) INCLUDING KATRAFAY ESSENTIAL OIL.

Oryza sativa bran oil: 50%

sesamum indicum seed oil: 12%

argania spinosa seed oil: 3% - rosa moschata seed oil: 5%

- Coco-caprylate: 26%

- Cedrelopsis Grevei Bark oil, titrated at ishwarane to 34,7%

and at limonene to 0,45%>: 1,5%

- Cananga odorata flower oil: 1%

- Calophyllum inophyllum seed oil: 1%

- Tocopheryl acetate: 0,5%

The formulation described here, in the form of a cream, can be used in the topical treatment of pruriginous dermatitis, particularly in the penis. This formulation is well tolerated at the genital level and compatible with sexual activity, also thanks to a vasodilatory effect, able to contrast problems of erectile dysfunction.

Claims

Composition based on katrafay {Cedrelopsis grevei) essential oil comprising from 20% to 40% by weight of ishwarane relative to the total weight of the composition, preferably comprising from 25% to 35% by weight of ishwarane relative to the total weight of the composition.

Composition according to claim 1 further comprising limonene, wherein the weight ratio ishwarane: limonene is between 50: 1 and 100:1.

Composition according to claim 1 or claim 2, wherein the composition is in an ozonized form.

Composition according to anyone of claims 1-3 to be used as a medicine or as a nutraceutical supplement, preferably where the above-mentioned use is in the treatment of inflammation and/or erectile dysfunction, more preferably for the treatment of pruriginous and/or seborrhoeic dermatitis, particularly those affecting the penis.

Process for preparing the composition according to anyone of the claims 1-3, comprising the steps of:

a. obtaining a katrafay essential oil;

b. verifying the concentration of ishwarane in the essential oil obtained in step (a) and, if necessary, adding ishwarane to obtain a percentage by weight of ishwarane relative to the total weight of the composition comprised between 20% and 40%, and optionally adding limonene in such quantity that the weight ratio between ishwarane and limonene is between 50: 1 and 100: 1;

c. mixing the components together to obtain a homogeneous composition;

where steps (b) and (c) are performed at a temperature not exceeding 80°C and avoiding direct exposure to light.

Process according to claim 5 further comprising the ozonization step (d) of the homogeneous composition obtained in step (c).

Formulation comprising the composition according to anyone of claims 1-3 in a percentage ranging from 0,001% to 40%> by weight, preferably comprised between 0,1 % and 10%> by weight, and:

- one or more acceptable cosmetic and/or pharmaceutical and/or nutritional excipients; and/or

- one or more carriers; and/or

- one or more anti-inflammatory and lenitive components.

Formulation according to claim 7, wherein said formulation is selected from the group consisting of: cream, gel, solution, ointment, suspension, liposome formulation, powder, aerosol, capsule, tablet, pessary, paste, spray, suppository, or is incorporated into transdermal patches, inhalers, impregnated scaffolds, pomades.

Formulation according to claim 7 or claim 8 to be used as a medicine or as a nutraceutical supplement, preferably where the above-mentioned use is in the treatment of inflammation and/or erectile dysfunction, more preferably for the treatment of pruriginous and/or seborrhoeic dermatitis, particularly those affecting the penis.

Formulation to be used according to claim 9 wherein said use is by the administration of the formulation by topical way or systemic way.