(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization I International Bureau (10) International Publication Number (43) International Publication Date WO 2013/050902 Al 11 April 2013 (11.04.2013) P O P C T

(51) International Patent Classification: (74) Agent: CELESTINO, Marco; Viale Giovanni Pisano, 3 1, A61K 36/185 (2006.01) A61K 36/534 (2006.01) 1-56123 Pisa (IT). A61K 36/28 (2006.01) A61P 17/02 (2006.01) (81) Designated States (unless otherwise indicated, for every A61K 36/61 (2006.01) A01N 65/00 (2009.01) kind of national protection available): AE, AG, AL, AM, A61K 36/36 (2006.01) AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (21) International Application Number: BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, PCT/IB2012/055081 DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, (22) International Filing Date: KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, 24 September 2012 (24.09.2012) ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, (25) Filing Language: Italian NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, (26) Publication Language: English TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, (30) Priority Data: ZM, ZW. PI201 1A000101 23 September 201 1 (23.09.201 1) IT (84) Designated States (unless otherwise indicated, for every (72) Inventor; and kind of regional protection available): ARIPO (BW, GH, (71) Applicant : DEL CORSO, Simone [IT/IT]; Via Sesia, 9, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, 1-56 122 Pisa (IT). UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, (72) Inventors: BIANUCCI, Anna Maria; Via Tagliamento, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, ΓΓ, LT, LU, LV, 12, 1-56122 Pisa (ΓΓ). BORGHINI, Alice; Via Vecchializ- MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, ia, 48, 1-56017 San Giuliano Terme (pi) (ΓΓ). PIETRA, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Daniele; Via Vecchializia, 48, 1-56017 San Giuliano Ter ML, MR, NE, SN, TD, TG). me (pi) (IT). Published: — with international search report (Art. 21(3))

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(54) Title: JOJOBA OIL EXTRACT AND COMPOSITIONS THEREOF, IN PARTICULAR, FOR TREATING A SKIN CONDITION

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5 - 50 125 100 - 75 - 50 25 - 0 -

-2,5 -2.0 -1.5 -1.0 0.0 Concentrazione (Log % o (57) Abstract: A method for making an extract of a of the genus Helichrysum in Jojoba oil or wax, in which a step is provided of joining the Helichrysum, preferably the upper parts of the plant, with a Jojoba wax having an acid number higher than 0.3, in par © ticular higher than 0.32, more in particular higher than 0.36. Such method makes it possible to obtain an extract with a high flavon - oid content useful for stimulating the keratinocytes of the epidermis, characterised by a content of Chrysin higher than 80 mg kg of © fatty acids having 20 carbon atoms with one unsaturation, in particular higher than 84 mg/kg, more in particular, higher than 104 mg/kg and/or makes it possible to obtain an extract with a low contained of allergens, characterised by a ratio between the amounts o of and β- Selinene lower than 0.4, in particular lower than 0.08, more in particular, lower than 0.06. The step of joining is preferably obtained by a dynamic process, for example by percolation or by centrifugation. A medical or cosmetic composition com prising such an extract for treating or preventing a disease and/or a condition that requires a stimulation of cell proliferation of kerat - o inocytes, among which wounds, burns, skin ulcers, scars, skin aging, stretch marks, wrinkles and others. An insect -repellent compos - ition comprising the extract. w o 2013/050902 A llll II II 11III III III I II 11 II 11III II I II before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h)) TITLE

JOJOBA OIL HELICHRYSUM EXTRACT AND COMPOSITIONS THEREOF,

IN PARTICULAR FOR TREATING A SKIN CONDITION

DESCRIPTION

Field of the invention The present invention relates to an extract of a plant of the genus

Helichrysum in Jojoba oil, and to a method for producing such extract. Furthermore, the invention relates to a medicament, to a medical device or to a cosmetic that contains an extract of a plant of the genus Helichrysum in

Jojoba oil, and that is suitable for treating particular skin diseases and/or conditions in a human being or in an animal. The invention relates also to a composition containing such an extract, having an insect-repellent effect.

Prior Art

In the past, many attempts have been made to develop synthetic drugs or natural remedies for treating various skin diseases. However, the extracts obtained from a variety of types of , or from different parts of a same plant, have respective different characteristics and effects.

In particular, the use is known of compositions containing derivatives of plants of the genus Helichrysum and Jojoba wax as a medicinal herb and as a cosmetic product.

Helichrysum The genus Helichrysum belongs to the family of the and comprises about six hundred species of flower plants. Various species of Helichrysum are known that contain potentially useful substances for treating skin diseases, for instance substances that have an antioxidant power. Among these species, Helichrysum italicum or angustifolium, is particularly used, which grows spontaneously in Southern Europe.

Helichrysum italicum grows in a rocky environment, up to about 800 metres of altitude. It has a bushy look, a silver-green colour, and may achieve a height of 30-40 cm. It flowers from the end of spring to the beginning of autumn. The flowers are gathered in flower-heads that have an outer wrapper consisting of dark yellow petals and have a conical central receptacle that houses flowers one by one. The flowers are light yellow and have a tubular shape. Helichrysum italicum contains many types of substances such as volatile compounds, , terpenoids, etc. Other species of the genus Helichrysum that contain substances potentially useful for treating skin diseases are, for instance, H . frigidum, H . lithoreum, H. melitense, H . montelinasanum, H . nebrodense, H . rupestre (var. errerae), H . Rupestre (var. melitense), H . rupestre (var. messerii), H . Rupestre (var. Pendulum), H. rupestre (var. rupestre), H . saxatile, H . Siculum, H . Stoechas.

Chrysin and other flavonoids The antioxidant, anti-inflammatory and antimicrobial properties of such flavonoids as Chrysin, Galangin, Pinocembrin are well known. These flavonoids are known to be present in many plants of the genus Helichrysum.

More in detail, Chrysin (5,7-Dihydroxy-2-phenyl-4H-benzo[b]pyran-4-one) can attenuate apoptosis, reduce reactive oxygen species (ROS), and also lower the expression of cyclooxygenase-2 (COX-2) that is caused by UVB and

UVA. Chrysin reverses in a predominant way the expression of Aquaporin 3

(AQP-3) caused by the UVB. It reverses in a predominant way the activation of JNK and inhibited slightly the activation of p38 caused by the UVA and by the UVB. The topical application of Chrysin is followed by an effective percutaneous absorption and does not generate irritation to the skin. Globally, the results have shown significant benefits of Chrysin on the protection of keratinocytes against injuries induced by cytotoxic factors such as UVA and UVB. See Wu, Nan-Lin et al., Chrysin Protects Epidermal Keratinocytes from UVA- and UVB-lnduced Damage, Journal of Agricultural and Food Chemistry, 201 1 59 8391-8400.

FR 2906716 describes the use of Chrysin in cosmetic compositions for treatment of the greasy skin, and its effectiveness in inhibiting the proliferation of human sebocytes.

US 2005/0049206 describes some flavonoids and extracts that contain

them, which have pharmaceutical properties useful in the therapy of fibrotic diseases, for protecting or treating or repairing fibrotic damaged tissues.

European Journal of Medicinal Chemistry, 46, 1, 201 1, 393-398 deals with Chrysin and analogues thereof as immunosuppressive and potentially useful agents against such diseases as psoriasis.

Volatile compounds of Helichrysum WO 2007/066305 describes analgesic effects of a composition containing

at least one in the human or animal body. Trans-2-decenal ((E)-2-Decenale) and Nonanal aldehydes have insect- repellent properties. See, for instance, US 2007/01 54504, JP 2006/327957, which describe their use against Dermatophagoides farinae, and US 6,524,605, which describes compositions comprising a monoterpenoid or a

sesquiterpenoid that is effective as a repellent for arthropods such as cockroaches, mosquitoes, ticks, spiders, and the like.

Furthermore, trans-2-decenal and Nonanal aldehydes are classified in the Cosing database as cosmetic ingredients, in particular as perfume agents.

Jojoba Jojoba oil or Jojoba wax are obtained from Jojoba or Simmondsia chinensis seeds, a plant that has a shrubby look and grows in some desert areas of Northern America. Jojoba oil is present in the seeds, in a weight percentage normally higher than 50%. See, for instance, D.J. Undersander, et al., Jojoba. Alternative Field Crops Manual, 1990.

Jojoba wax has a structure very similar to human sebum, with which it is completely miscible. For this reason, it is tolerated by human skin far better than other vegetable oils, and does not obstruct the pores. Since Jojoba wax does not evaporate, it provides a more long-lasting hydration than other water-

based moisturizing formulations. Furthermore, Jojoba wax is chemically very stable, does not go rancid and does not lose its antioxidant agents even after a long preservation period.

Products based on Helichrysum and Jojoba

An example of Jojoba wax Helichrysum composition can be found in the homeopathic product "Eczema no more". However, the type of Helichrysum that is used is not known. Furthermore, it is understood that Helichrysum essential oil is used, i.e. a product obtained by a Helichrysum stripping process.

Another example is disclosed in ITPI2001A000056, which relates to a method for making a Jojoba wax Helichrysum italicum extract, in which

Helichrysum flower-heads are soaked in a Jojoba oil, in particular at a Helichrysum/oil ratio set between 0.2 and 0.3. Soaking may be followed by a pressing step and by an addition of a camomile essential oil. However, neither the chemical composition of the final product, nor the composition of the starting material is specified. The following publications KR201 10023257, US2004258783A1 , WO2008053246A1 , GB2472379, EP2018892 relate to the presence of substances deriving from Helichrysum, and preferably from Jojoba oil in preparations for treating some skin conditions.

Insect-repellent substances Insect-repellent substances are also known. Such substances are useful in some areas, in particular of the tropical regions, where insects carry dangerous diseases such as Malaria, West Nile Fever, Dengue Fever, and the like. o o

As well known, the skin comprises a thin surface layer, the epidermis, which provides resilience and barrier properties to the skin. The epidermis comprises a wide range of cell types. Most of epidermis cells are keratinocytes, which in a human being form 75 to 80% of the epidermal cells.

In many skin diseases and/or conditions, it may be desirable to stimulate the proliferation of epidermal keratinocytes cells. ln fact, the skin has regeneration and repair mechanisms that work both physiologically and in case of particular conditions such as injuries. It is known that keratinocytes proliferation mechanisms are involved in many skin diseases and/or conditions, such as skin aging, bums, in particular sun burns, and so on.

Skin complications are particularly frequent and not negligible at dermal stoma, for instance in patients who suffer from cancer. The skin complications due to the presence of a stoma have an incidence between 55 and 61%, according to literature. These complications may be caused by a skin reaction to the glues of the aids that are used to treat the wounds, and/or by a continuous contact with faeces, and/or by a repeated skin traumatism due to a frequent change of the aid itself. The complications may be of different seriousness, and comprise skin lesions such as dermatitis, skin necrosis up to ulceration. The following documents:

Helichrysum oil, Pure Organic Essential oil, The Ananda Apothecary, 4

July 2009, in http://www.anandaapothecary.com/aromatherapy-essential- oils/helichrysum-essential-oil.html;

US 2004/234628 A ; Mastelic et al., Contribution to analysis of the Helichrysum italicum essential oil (Roth) G . Don. - Determination of Ester Bonded acid and Phenols,

Molecules, vol. 13, no. 4 , pp. 795-803;

"Italienische Strohblume", in http://de.wikipedia.org/w/index.php?title= ltalienische~Strohblume&oldid=79268296&printable=yes all refer to Helichrysum essential oils.

An essential oil is a final product obtained by a steam stripping process of a plant portion, whereas an oleolite is a product obtained by an oil-extraction of a portion of a plant. In the case of Helichrysum, the essential oil and the oleolite in Jojoba oil have completely different compositions. In particular, the oleolite contains flavonoids, which are substantially absent in the essential oil, since they cannot be extracted by steam stripping. On the contrary, essential oils contain compounds such as Nerol and Neryl acetate, which are skin-irritant substances.

In particular, the products described in Helichrysum oil, Pure Organic

Essential oil, The Ananda Apothecary, 4 July 2009, as disclosed in http:/Avww.anandaaDothecarv.com/aromatheraDv-essential-oilslHelichrvsum- essential-oil.html . and in US 2004/234628 A 1 may contain Jojoba oil as a carrier for skin application, but Jojoba oil may be replaced with another carrier.

In practice, Jojoba oil is mixed with the essential oil and no oil extraction is provided of the compounds that are present in the plant. US 2004/234628 A 1 also relates to the use of a product for treating insect bites, but not to the use as an insect-repellent substance. Mastelic shows the results of a gas chromatography and of a gas chromatography - mass spectrometry of a

Helichrysum italicum essential oil, which confirm the absence of flavonoids, and the presence of compounds that are, on the contrary, substantially absent in an oleolite, which may have a beneficial activity for the skin. LV P C ET AL: Synthesis, biological evaluation of Chrysin derivatives as potential immunosuppressive agents, European journal of Medicinal chemistry, Editions Scientifiques Elsevier, Paris, vol. 46, no. 1, pp. 393-398 describes only immunosuppressive properties of flavonoids, in particular of Chrysin. Nan-Lin Wu et al., Chrysin Protects Epidermal Keratinocytes in UVA- and UVB-lnduced Damage, Journal of Agricultural and food chemistry, vol. 59, no.

15, pp. 8391-8400 describes the effectiveness of Chrysin to protect keratinocytes from UV waves, but no regenerating properties of medical interest are described.

It is therefore felt the need of a medicament or of a cosmetic or of a medical device that is effective for treating skin diseases and/or conditions like those above indicated. In particular, the need is felt of a medicament or of a cosmetic or of a medical device that is adapted to stimulate the proliferation of keratinocytes.

In particular, it is desired to provide a medicament that is useful for treating skin diseases like those that occur as a consequence of stomata.

It is also felt the need of a medicament or of a cosmetic or of a medical device that has an insect-repellent effect.

Summary of the invention

It is therefore a feature of the invention to provide an extract of a plant of the genus Helichrysum which is extracted in Jojoba oil, for treating various skin diseases, for example skin aging, and burns of various types, in a way more effective than the products and the extract based on Helichrysum presently known.

It is also a feature of the invention to provide an extract of a plant of the genus Helichrysum in Jojoba oil for stimulating the proliferation of keratinocytes of the epidermis, for treating skin diseases.

It is a particular feature of the invention to provide a medicament useful for treating the skin diseases that occur typically in the peristomal skin of subjects affected by stoma.

It is also a feature of the invention to provide such an extract that has a lower tendency to cause skin-sensitization phenomena and/or allergic phenomena than the prior art products, for protection and the treatment of skin.

It is also a feature of the present invention to provide a Jojoba oil

Helichrysum extract that is effective as a repellent for many types of insects.

It is also a feature of the present invention to provide a method to prepare such a Helichrysum extract. These and other objects are achieved by a method for preparing an extract of a plant of the genus Helichrysum in Jojoba oil, the method comprising the steps of: prearranging a plant portion selected among the upper parts of the plant,

in particular the flowers; prearranging a Jojoba oil having an acidity higher than 0.3, expressed as acid number; joining said plant portion of a plant of the genus Helichrysum and said

Jojoba oil in an extraction condition;

recovering the extract obtained in the step of joining.

By upper parts the non-woody parts of the plant are meant, in particular the parts at up to 10-15 cm from the top of the plant, for example the flower- heads, the stems top, and the first leaves.

The acid number is defined as the amount of potassium hydroxide (KOH) in milligrams that it is required for neutralizing a gram of oil.

It has been in fact surprisingly discovered that a Jojoba oil that is considered of an inferior quality for many applications of this oil because it is more acid, has a high dissolving power for flavonoids. In other words, it enhances the extraction of flavonoids, increasing their concentration in the extracts.

Furthermore, it has been surprisingly discovered that the extracts obtained starting from an oil of acidity as above indicated are of better quality, since they have an unexpectedly low allergen concentration. These effects of the above Jojoba oil acidity on the dissolving power of the oil, with respect to flavonoids and allergens, is unknown nor can it be deduced from the prior art, comprising the documents cited at the beginning.

In particular, the acidity can be higher than 0.32, more in particular, higher than 0.36.

This method makes it possible to obtain an extract of a plant of the genus

Helichrysum in Jojoba oil that has a particularly high content of Chrysin (5,7-

Dihydroxy-2-phenyl-4H-benzo[b]pyran-4-one), in particular it has a content higher than 80 mg per kg of fatty acids that have twenty carbon atoms, and that have one unsaturation, contained in Jojoba oil. The fatty acids having 20 carbon atoms and one unsaturation are provided in a ratio from about 65% to about 80% of Jojoba oil. Preferably, the content of Chrysin is higher than 84 mg per kg of fatty acids having 20 carbon atoms and one unsaturation, more preferably is higher than 104 mg/kg of fatty acids having 20 carbon atoms and one unsaturation.

In fact, the inventors have observed that a Jojoba oil Helichrysum extract, in the conditions according to the invention, is particularly rich in the following flavonoids: Chrysin, Galangin, Pinocembrin, which increases the antioxidant, antiinflammatory, antimicrobial effects of the composition. Furthermore, the inventors have surprisingly found that a Jojoba oil

Helichrysum extract that is particularly rich in such flavonoids as Chrysin, Galangin, Pinocembrin, has particular features stimulating the proliferation of keratinocytes, independently from the action of such cytotoxic factors as UV-waves. Such technical effect of obtaining a high concentration of such flavonoids as

Chrysin, Galangin, Pinocembrin, is capable of assisting the proliferation of keratinocytes, and is not described in literature, nor can it be deduced between the documents cited in the introductory part. More in detail, as shown in the examples below, a Jojoba oil Helichrysum extract having a Chrysin concentration higher than 80 mg/Kg, as obtainable using for the extraction a Jojoba oil with acid number higher than 0.30, has a capacity to promote the proliferation of keratinocytes. Such capacity increases with the concentration in the extract. On the contrary, an extract with a Chrysin concentration of 75 mg/Kg, produced with a low acidity oil, does not show any significant biological activity in this sense. In particular, the Helichrysum essential oil has the opposite effect, i.e. at a certain concentration it causes the death of keratinocytes. For characterising the extract versus the content of flavonoids the sole

Chrysin is here considered, instead of the total content of flavonoids, since

Chrysin is the most abundant . The abundance of Galangin and

Pinocembrin is, respectively, lower than the abundance of Chrysin. In any case, the amount of these three flavonoids are correlated to each other. In other words, there is a substantial proportionality between the concentration of Galangin and of Pinocembrin with respect to the Chrysin concentration. Furthermore, the analytic procedure for determining the concentration of the sole Chrysin causes more easily reproducible results, and then more reliable, than the method for determining all the flavonoids.

Such method allows also to obtain a Jojoba oil, i.e. a Jojoba wax extract of a plant of the genus Helichrysum, where the weight ratio of Limonene (S-(-)- 4-isopropenil-1 -methyl-1 -cycloesene; R-(+)-4-isopropenil-1 -methyl-1 - cycloesene) with respect to β-Selinene ((3S, 4aR, 8aS)-8a-methyl-5- methylidene-3-prop-1 -en-2-il-1 ,2,3,4,4a,6,7,8-ottahydronaphtalene) is lower than 0.4.

Also in this case, in order to characterise the presence of Limonene, β-

Selinene is selected as a reference, since β-Selinene is the component with highest concentration in the volatile fraction of the extract. Furthermore, the procedure for determining the amount of Limonene provides more reliable results.

The ratio Limonene : β-Selinene can be determined as the ratio between the areas of the respective peaks of a graph obtained by a chromatographic analysis.

In particular, the inventors have observed that the volatile part of a Jojoba wax Helichrysum extract is particularly rich in: β-Selinene; trans-2-decenal; o Selinene; AR-Curcumene; Nonanal, whose analgesic effects are known, as indicated above. On the contrary, surprisingly, this extract is particularly poor in

Limonene, which is a well-known allergenic agent. Unlike the essential oil, the inventors have also surprisingly found that the Jojoba Helichrysum extract according to the invention contains much less Limonene than Helichrysum essential oil. Therefore, the extract according to the invention has a lower tendency to cause skin-sensitization phenomena and/or allergic phenomena than the known products. Furthermore, the extract according to the invention provides formulations for treating particular skin conditions that take advantage from an action that stimulates the proliferation of keratinocytes, and that is also hypoallergenic and analgesic. As described, some of the substances of the volatile part, for example trans-2-decenal, AR-Curcumene, Nonanal and Octanal are also insect- repellent substances. Preferably, the weight ratio Limonene^-Selinene is lower than 0.08 and even more preferably this weight ratio is lower than 0.06. In particular, the Helichrysum plant belongs to the species Helichrysum italicum, in particular Helichrysum italicum subspecies italicum; Helichrysum italicum subspecies microphyllum; Helichrysum italicum subspecies serotinum; Helichrysum italicum subspecies siculum; Helichrysum italicum variant pseudolitoreum; Helichrysum italicum variant typicum; Helichrysum italicum variant picardii; Helichrysum italicum variant HyperCreso; Helichrysum italicum subspecies italicum variant corsa.

In particular, the plant of the genus Helichrysum belongs to the species Helichrysum stoechas.

In particular, the plant of the genus Helichrysum belongs to the species .

The enrichment in the above mentioned compounds may be obtained by properly selecting the Jojoba oil, in particular by selecting an oil that has an acidity higher than 0.3.

In addition, or alternatively, the enrichment in the compounds above indicated may be obtained by carrying out the extraction, i.e. the contact between the plant of the genus Helichrysum and Jojoba oil, in a dynamic step, where Jojoba oil is in movement with respect to a mass of Helichrysum, for example a step of percolation and/or a step of centrifugation.

As is described hereinafter, the plant portion that is chosen for the extraction, i.e. the choice of the upper parts, more in particular, the flowers, enables to obtain a concentration of compounds such as flavonoids, terpenoids and other, higher than the known Helichrysum extracts and the known products based on Helichrysum and Jojoba oil. The method may comprise a step of adding pharmaceutically and/or cosmetically acceptable components that are adapted to increase the solubility of Chrysin and of other flavonoids, these components selected, in particular, among propylene glycol, glycerine and cyclodextrins. A particularly high content of useful active principles such as flavonoids, shown above, may be obtained by selecting the extraction method. The step of joining the part of Helichrysum italicum and Jojoba oil may be carried out by any known technique of extraction. Advantageously, the step of joining may be dynamic, i.e. the Jojoba oil is in movement with respect to a mass of Helichrysum.

Preferably, the step of joining is carried out by a percolation technique. In fact, by a percolation technique an extract is obtained that has at the same time a higher flavonoid content and a lower Limonene content than the extract that can be obtained from the same plant and by using the same oil, and by carrying out the further steps of the method in the same operating conditions.

Advantageously, the step of joining is carried out with an amount of

Helichrysum set between 70 and 300 g per 1000 ml of Jojoba oil, in particular between 100 and 200 g per 000 ml of Jojoba oil, more in particular between

110 and 150 g per 000 ml of Jojoba oil. Alternatively, the step of joining may comprise a step of centrifuging the mass of Helichrysum in Jojoba oil. The percolation and centrifugation methods are two possible dynamic contact methods between the mass of Helichrysum and Jojoba oil.

The step of joining may also comprise a step of soaking. In this case, the step of joining is carried out with an amount of Helichrysum set between 150 and 350 g per 1000 ml of Jojoba oil, in particular between 200 and 320 g per

1000 ml of Jojoba oil, more in particular, between 220 and 300 g per 1000 ml of Jojoba oil. Advantageously, the step of joining is carried out at a contact temperature set between the room temperature, in particular 20°C, and an extraction temperature lower than a maximum extraction temperature, wherein a maximum extraction temperature is 70°C, in particular a maximum extraction temperature is 60°C, more in particular, a maximum extraction temperature is

50°C, even more in particular, a maximum extraction temperature is 45°C. This way, it is possible to further enhance the dissolving power of Jojoba oil with respect to Helichrysum flavonoids, without affecting their features. In particular, this way, it is possible to obtain a flavonoid concentration higher than the prior art products, even if the step of joining is carried out in static conditions, for example in static soaking conditions. Advantageously, the step of prearranging said plant portion for the extraction comprises a step of drying said plant portion. Another advantage of the use of dried upper parts consists of forming an extract that is substantially free from moisture. This makes it possible to exploit the antioxidant character of some Jojoba oil components, which do not require the use of preservatives such as synthetic antioxidants. Furthermore, the removal of thermolabile substances that takes place during the drying step allows using a higher process temperature without any risk of forming degradation products, which would derive from the residual thermolabile substances. Advantageously, the step of prearranging said plant portion for applications in extraction comprises a step of preparing an infusion-type cut product, i.e. of a step of crushing.

In particular, the extract of the invention may be obtained only from dry Helichrysum flowers, as well as from other parts of the plant. For example, the Helichrysum italicum flowers come preferably from natural Italian plant and are harvested during the balsamic period. An advantage of the use of upper parts and/or of a Jojoba Oil that has a strong solubilising power for flavonoids is that it the extract obtained has a volatile fraction comprising: a high amount of such flavonoids as Chrysin, Galangin, Pinocembrin, thereby obtaining an extract whose characteristic stimulating the growth

of keratinocytes have been mentioned above, and in particular: a high amount of Chrysin; a high amount of Galangin and of Pinocembrin; a high amount of β-Selinene and/or of a-Selinene and/or of trans-2- decenal and/or of AR-Curcumene and/or of Nonanal and/or of Octanal, which are known as insect-repellent substances;

a low amount of Limonene, which is an allergen. The step of recovering the extract may be carried out with any known method in the arte, in particular by a technique selected from the group consisting of: pressing, hydraulic pressing, filtering, centrifugation, or a combination of the above techniques. The extract can be used as obtained from the above steps of the procedure, or the method may also comprise a step of post-treating the extract during the sep of recovering the extract. In particular, a step of post-treatment may be:

a step of dilution, for example in order to restore the volume of used Jojoba oil; a step of sterilization; a step of further extraction; a separation step of a component. Regardless of these post-treatment steps, the method may also comprise a step of adding substances of various type. For example, a step may be provided of adding further active principles. The method may also provide a step of packaging the Helichrysum extract in a shape arranged to the distribution. Some components of the Helichrysum italicum extract of the invention can then work as active principles of a medicament or of a cosmetic, and can also be used in a medical device.

Even if the Helichrysum italicum extract of the invention is used as such, it may also be incorporated in a medicament or in a cosmetic by adding any pharmaceutically or cosmetically acceptable excipients, for example a filler, a thickener, or other excipients known in the pharmaceutical technique or in the art of preparing cosmetic products.

Accordingly, it falls within the scope of the invention also a medicament or a cosmetic product or a medical device that contains the above described extract, obtained from a plant of the genus Helichrysum, in Jojoba oil, i.e. an extract obtained with the above described method, for treating diseases and/or conditions that take advantage from a stimulative action for the proliferation of keratinocytes.

In particular, as test on patients show, this medicament or cosmetic product or medical device is well suited for treating:

skin complications in stoma-bearing patients, in particular complications caused by the reaction of the skin to glues of aids used to treat the wounds, or caused by the continuous contact with the faeces, by the repeated skin traumatism due to a frequent change of said aids; wounds, including cuts, excoriations, abrasions, insect bites, wounds by cutting arm, wounds by firearm, surgical wounds, etc.; burns, including UV burns, sun burns, radiotherapy burns, fire burns, hot contact burns, acid burns, alkali burns, etc.; skin ulcers, including decubitus ulcers, diabetic ulcers, etc.; scars, including surgical scars, scars owing to excessive growth of connective tissue after a tissue damage, atrophic scars, acne scars, etc.; skin aging, including age skin aging, UV skin aging, aging caused by environmental pollution, aging due to atmospheric factors, hormonal aging, etc.; stretch marks, including pregnancy stretch marks, stretch marks owing to important variation of body weight, hormonal stretch marks, etc.; psoriasis injuries; wrinkles, including expression wrinkles, etc.

Due to the properties of flavonoids and of the compounds present in the volatile fraction of the extract, the medicament or cosmetic product or medical device can show beneficial effects such as antimicrobial, antibacterial, protective effects against UV waves, anti-inflammatory, antioxidant, analgesic sebum normalizing effects, i.e. in treating Seborrhoeic dermatitis, for preventing the formation of fibrotic damaged tissues, for preventing psoriasis injuries.

The medicament according to the invention may be formulated in whichever form suitable for any administration procedure. The preferred administration procedure is any topical administration procedure . To preparation of the medicament or of the cosmetic product may be carried out by procedures known in the art of preparing pharmaceutical or cosmetic products. These techniques are described, for instance, in Remington's Pharmaceutical Science, 18a edition, by Gennaro et al., 1980, Mack Publishing Co., Easton, PA.

Advantageously, the composition comprises an essential oil. In particular, this essential oil is selected from the group consisting of: roman camomile essential oil; matricaria camomile essential oil; carnation essential oil; tea tree essential oil; lavender essential oil; wintergreen essential oil; origanum essential oil, or mixtures thereof. In particular, the roman camomile and/or the wintergreen essential oil, besides imparting a more agreeable flavour to the composition, enhances with a synergistic effect the anti-inflammatory power of the Helichrysum extract. It falls within the scope of the invention also the use of the above described extract in the preparation of such a medicament or cosmetic product or medical device. It falls within the scope of the invention also an insect-repellent composition that contains the above described extract of a plant of the genus Helichrysum.

It falls within the scope of the invention also the use of the above described extract in the preparation of an insect or acari-repellent composition.

Brief description of the drawings The invention will be now shown with the following description of its exemplary embodiments, exemplifying but not limitative, with reference to the attached drawings, in which: Fig. 1 shows a chromatographic profile of an essential oil used to obtain the extract according to the invention; - Fig. 2 is a micrograph of three slides of a thin layer chromatographic analysis of samples of respective Jojoba oil Helichrysum italicum extracts;

Fig. 3 shows the trend of the proliferation ratio of keratinocytes in a

culture medium in the presence of various concentration of three extracts of Helichrysum italicum in Jojoba oil, and in the presence of a

Helichrysum italicum essential oil; Fig. 4 shows the positive correlations between the amount of some

compounds found in Helichrysum italicum extract in Jojoba oil and the proliferation ratio of keratinocytes treated with the extract at the

concentration of 1% ; Fig. 5 diagrammatically shows a lesion of the skin of a stoma-bearing patient.

EXAMPLES

1. Chemical analysis of the Jojoba oils

The Jojoba oils or waxes to be used are characterized by the parameters chemical-physical analysis parameter shown in table 1. - Table 1 -

SPECIFICATION METHOD

Aspect clear liquid Colour (Gardner) 15 max. Odour slight

Specific gravity. 20°C. g u 0.860 - 0.870 ISO 6883 Retractive index, 20°C 1.4640 - 1.4680 ISO 6320

Acid valu 2 max. ISO 660 Iodine value 80 - 90 ISO 3961 Saponification value 85 - 98 ISO 3657 Peroxide value, meq g 5 max. ISO 3960 Unsaponifiable, % 45 - 55 ISO 3596-1

Fatty acid composition GC ( total fatty acids) CI6:0 3 max. C16:l 1 max. C 8:1 5 - 15 C20:l 65 - 80 C22:l 10 - 20 C24:l 1 - 3 C26:l 0 - 1

Store away from heat and light. ln the examples of the present patent application, according to the invention, a Jojoba oil was used whose characteristic are shown in detail in table 2 .

- Table 2 - 3 October 2010 ANALYSIS FILE Botanic Name: HELICRISUM ITALICUM Common Name: HELICHRYSUM ITALICUM Batch number: 02-AZEO 2010 Origin: SARDINIA AZ.AGR.ERBORISTERIA OFFICINALE Plant portion:— TOP FLOWERS Analysis features: CPG 5890 / MS 5970 - Column: polar HP INNOWAX: 60 m x 0.25 mm x 0.5 pm, CPG 5890 FID - Column: polar HP INNOWAX: 60 m x 0.25 mm x 0.5 pm. Temperature program: 6 min at 50°C - 2°C/ min - 250 C - 20min at 250°C. Carrier gas He: 30 psis/FID, 23 psis/MS. Injector: split. Sample: 1 µ Ι of 5% hexane solution. Mass range: 30 to 350. The compounds of the essential oils are identified by a combined search of mass spectra (NIST library 75 KL and proprietary library) and of retention times. The % are calculated starting from the areas of the peaks obtained by GC/FID without using any correction factor. Chromatographic profile 20.00 40.00 60.00 80.00 100.00 0 500Ό 00 1Ό 00Ό 00 l'500'OOO 2Ό 00Ό 00 2'500 Ό 00 3Ό 00Ό 00 3'500 Ό 00 4Ό 00Ό 00 4'500 Ό 00 Time- Abundance TIC: HIL10086.D Table of the results 1: HELICHRYSUM ITALICUM Peaks of the time for retention - Components % 1 7.6 3-METHYL-CYCLOHEXANENE 0.04 2 15.4 a-PINENE 0.21 3 17.8 a-FENCHENE 0.15 4 18.4 CANFENE 0.03 5 19.0 4-METHYL-3-ESANENE 0.02 6 2 1.0 β-ΡΙΝΕΝΕ 0.06 7 22.1 SABINENE 0.01 8 25.1 β-MYRCENE 0.01 9 25.9 ISO BUTYL-2-METHYL BUTYRATE 0.02 10 27.1 ISOAMYL ISOBUTYRATE 0.05 11 28.2 LIMONENE 1.57 12 29.0 1,8 CINEOLE 0.99 13 3 1.9 Y-TERPINENE 0.10 14 32.1 Trans-p-CIMENE 0.09 15 33.7 p-CIMENE 0.09 16 34.0 2-METHYL BUTYL 2-METHYLBUTYRATE 0.05 17 34.6 TERPINOLENE 0.04 18 34.8 ISOBUTYL ANGELATE 0.22 19 35.2 3-METHYL BUTYL 3-METHYLBUTYRATE 0.08 20 42.5 2-EPTANONE 0.06 2 1 42.8 ISOAMIL ANGELATE 0.76 22 47.6 ANGELIC ESTER 0.05 23 48.3 SESQUITERPENE 0.07 24 49.6 YLANGENE 0.07 25 50.1 SESQUITERPENE 0.03 26 50.3 a-COPAENE 1.22 27 50.9 ISOITALICENE 1.96 28 53.1 a-GURJUNENE 0.13 29 53.3 LINALOOL 2.45 30 5 1.7 ITALICENE 6.71 3 1 54.9 a-CEDRENE 0.05 32 55.4 ITALICENE ISOMERO 0.20 33 55.5 a-cis-BERGAMOTENE 2.25 34 56.6 a-trans-BERGAMOTENE 1.26 35 57.2 β-ELEMENE 0.13 36 57.3 2-UNDECANONE 0.04 37 57.5 3,5-DIMETHYL OPTAN-4.6-DIONE Mol wt =170 0.04 38 57.8 TERPINEN-4-OL 0.18 39 573 β- CARYOPHYLLENE 1.14 40 58.3 β-GUAIADIENE 0.14 4 1 58.6 AROMADENDRENE 0.33 42 59.4 GURJUNENE ISOMER 0.14 43 60.1 β-SANTALENE 0.03 44 60.4 SESQUITERPENE 0.03 45 6 1.2 ALLO-AROMADENDRENE 2.73 46 6 1.6 Ε-β-FARNESENE 0.22 47 62.0 β-CADINENE 0.26 48 62.2 SESQUITERPENE 0.05 49 62.8 a-HUMULENE 0.16 50 83.0 ISOACORADIENE 0.88 5 1 63.3 CURCUMENE ISOMER 0.76 52 63.5 Ζ -β-FARNESENE 0.24 53 63.8 Y-CURCUMENE +Y-MUUROLENE 19.05 54 64.1 ANGELIC ESTER 0.14 55 64.2 SESQUITERPENE 0.27 56 64.3 LEDENE 0.27 57 65.3 GERMACRENE D 0.36 58 65.6 NERYL ACETATE 1.69 59 65.7 CADINENE ISOMER 0.12 60 65.9 a-BISABOLENE 0.38 6 1 66.1 a-MUUROLENE 1.85 62 66.2 β-SELINENE 0.85 63 66.4 a-SELINENE 0.84 64 66.8 β-CURCUMENE 0.53 65 66.9 DIENE ALCOHOL 0.88 66 67.1 OXYGENATED SESQUITERPENE OXYGENATED 0.02 67 67.9 a-FARNESENE ISOMER 0.20 68 68.3 δ -CADINENE 5.49 69 68.6 Y-CADINENE 4.04 70 68.9 a-BISABOLENE 0.1 5 7 1 69.0 a-CURCUMENE 9.97 72 69.4 NERYL PROPIONATE 0.91 73 69.9 CADINA-1 ,4-DIENE 0.42 74 70.0 NEROL 0.43 75 70.5 a-AMORPHENE 0.58 76 72.6 TRANS-CARVEOL 0.03 77 73.1 CALAMENENE 0.47 78 73.5 OXYGENATED COMPOUND OXYGENATED 0.10 79 73.6 NERYL BUTYRATE 0.03 80 74.8 ISONERYL BUTYRATE 0.41 8 1 J5.3 AROMATIC COMPOUND 0.10 82 75.7 ITALIDIONE I Mol wt.=21 0 0.06 83 75.9 ITALIDIONE II Mol wt.=224 0.04 84 76.7 AROMATIC COMPOUND 0.04 85 78.1 CALACORENE 0.17 86 78.8 PALUSTROL 0.1 3 87 79.0 AROMATIC COMPOUND 0.05 88 80.4 ITALIDIONE III 0.04 89 80.6 NERYL VALERATE 0.03 §0 8 1.8 AROMATIC ESTER 0.1 3 9 1 82.1 AROMATIC ESTER 0.03 92 83.1 DIENE DERIVATIVE 0.03 93 83.6 NEROLIDOL 0.23 94 84.1 SESQUITERPENOL Mol wt =222 0.10 95 84.4 LEDOL 0.52 96 85.8 EPI-CUBENOLO 0.20 97 86.2 CUBENOL 0.27 98 86.3 SESQUITERPENE EPOXIDE Mol wt.=220 0.07 99 86.7 GLOBULOL 0.06 100 87.1 GUAIOL 1.62 10 1 88.0 AROMATIC COMPOUND 0.14 02 88.4 TRIMETHYL-PENTADECANONE 0.13 103 89.3 EUDESMA-5-ENE-1 1-a-OL 10.31 161 90.4 AROMATIC COMPOUND 0.14 105 90.6 SESQUITERPENOL Mol wt.=220 0.45 106 9 1.2 Y-EUDESMOL 0.90 107 9 1.6 EUDESMOL ISOMER 1.00 108 92.1 SESQUITERPENOL Mol wt.=222 9.08 109 92.4 a-MUUROLOLO 0.24 110 92.6 SESQUITERPENOL Mol wt. 222 0.16 111 92.8 CADINOL ISOMER 0.37 112 93.2 EUDESMOL ISOMER 0.45 113 93.6 BULNESOL ISOMER 0.48 114 94.4 a-EUDESMOL 0.28 115 94.7 a-CADINOL 0.37 116 94.9 β-EUDESMOL 0.49 117 95.2 DITERPENE Mol wt.=272 0.05 118 95.7 ALIPHATIC DIONE + CAPRIC ACID 0.19 119 96.0 SESQUITERPENOL Mol wt =222 0.10 120 96.5 ALIPHATIC DIONE 0.09 121 97.9 SESQUITERPENOL 0.08 122 10 1.1 ALIPHATIC DIONE 0.16 123 10 1.0 AROMATIC COMPOUND 0.06 124 102.0 AROMATIC COMPOUND ISOMER 0.06 125 104.3 ALIPHATIC DIONE 0.05 126 104.2 ALIPHATIC DIONE 0.04 127 112.9 ALIPHATIC DIONE 0.18 128 113.0 ALIPHATIC DIONE 0.23 TOTAL 100.00

2 . Materials and conditions of the extraction step.

Example 1

Extraction by soaking . The following operating conditions were observed: temperature: room; - overall soaking time: 28 days; Jojoba oil acidity = 0.38; amount of oil used = 1000 ml; type of plant used: Dried Helichrysum italicum, infusion cut; amount of plant used = 250 g ; - Helichrysum : oil ratio = 0.25 g/ml; Jojoba oil acidity = 0.38; Helichrysum : oil ratio = 0.25.

Example 2

Extraction by percolation , in a 1 litre micro extractor without double piston, therefore with percolation by simple oil drop through a Helichrysum bed, and with no external oil recirculation. The following operating conditions were observed: temperature: room; pressurization time = 3 minutes; - depressurization time = 6 minutes; number of cycles = 88; overall cycle time: 792 minutes; Jojoba oil acidity = 0.38; amount of oil used = 1100 ml; - type of plant used: Dried Helichrysum italicum, infusion cut; amount of plant used = 275 g ; Helichrysum : oil ratio = 0.25 g/ml.

Example 3

Extraction by forced percolation in a quick extractor Timatic 5p model, equipped with a double piston. The oil is withdrawn from and recirculated into the extraction chamber. The following operating conditions were observed: temperature: room; number of forced percolations: 5 , with external active recirculation; pressurization time = 3 minutes; - depressurization time = 6 minutes; operating pressure = 8.0 bar; extraction overall time = 891 minutes; number of cycles = 99; Jojoba oil acidity = 0.2; - Amount of oil used = 6000 ml; type of plant used: dried Helichrysum italicum, infusion-type cut; amount of plant used = 800 g ; Helichrysum : oil ratio = 0.1 3 g/ml.

3 . Analysis of flavonoids in the extracts

Flavonoids were analysed by HPLC at Chelab company (Resana, TV, Italia) under constraint of confidentiality. The detailed description of the analytic procedure is available at this firm. The results of the analysis are given in the Tables 3a, 3b, 3c for the extract of examples 1, 2 , 3 , respectively. - Table 3a - Analysis of the flavonoids of the extract of Example 1

FLAVONOIDS (Met.: MP0679 rev. 4 , 201 1) Rutin

- Table 3b - Analysis of the flavonoids of the extract of Example 1 FLAVONOIDS (Met.: MP0679 rev. 4, 201 1) Rutin

FLAVONOIDS (Met.: MP0679 rev. 4 , 201 1) Rutin

4 . Gas chromatographic analysis of the volatile fraction of the extracts

The volatile fraction was analyzed by gas-chromatography at Chelab (Resana, TV, Italia).

In particular, the study of the volatile fraction composition, i.e. the study of the "active" flavouring fraction, was made by GC/MS and GC/FID analysis of the extract obtained. Different extraction techniques were used, but complementary to one another other with respect to the kind of information obtained. This had the object of assessing which is the best analysis protocol for studying the aromatic profile.

The protocol comprising an SDE analysis of the extract obtained turned out to be the best suited for characterising and comparing the samples, since it provides the best reproducibility and the best sensitivity with respect to some important compounds. The composition of the volatile fractions of the extract of examples ,

2 , 3 and 4 is given in table 4 , with reference to the compounds that are considered to be the most important. The relative distribution values of the listed compounds, evaluated as percentage areas in the gas chromatography diagram, were normalized with respect to the total of considered volatile compounds only.

For comparison purpose, table 1 and Fig. 1 show the analysis of the Helichrysum essential oil used as a reference, purchased from Daniele

Patrizia Erboristeria Balaina, Lu Capruleddu, Luogosanto, Olbia NU, Sardinia, Italy.

Moreover, as in Parasitol. Res. (2010) 107, 1455-146, a chromatographic study of some Helichrysum essential oil samples has shown that the main components are: Neryl acetate 25.3%; a-Pinene 14.5%; Limonene 12.3%; γ -Curcumene 8.7%; - Neryl propionate 6.4%; Nerol 5.2%, according to the data of table 1 and Fig. 1.

From the data of tables 1 and 4 and of Fig. 1, it is observed that the composition of the essential oil is extremely different from the composition of the Jojoba wax extract of examples 1, 2 and 3 . - Table 4 - Composition of the volatile fraction of the extracts

DESCRIPTION AND SAMPLE Example 1 Example 2 Example 3

COMPOUNDS % % % beta-Selinene 15,94 18,36 10.59 trans-2-Decenal 11, 17 16,61 17,53 alpha-Selinene .04 12,98 7,57 AR-Curcumene 9,53 12.97 7,21 Nonanal 8,31 5,62 5,00 Limonene 6,40 1,19 0,92 Rosifoliol 3,70 6,05 3,16 Optanal 3,60 2,65 2,31 2-Etilesilhexanol 3,56 0,24 2,25 beta-Cariofiillene 2,62 2,69 1,55 trans-Anetol + 2-Dodecanone 2,44 1,67 1,02 beta-Himachalene 2,33 1,33 1,20 alpha-Pinene 1,97 0,39 0,35 trans-2-Undecenal 1,74 1,51 3,41 Heptanal 1,49 0,40 1,38 Nerol 1,45 1,57 0,91 Optanol 1,20 1,74 1,03 delta-Cadinene 0,95 1,26 0,42: Elemol 0,91 0,95 0,55 trans-2-Nonenal 0,67 0,33 1,32 alpha-Terpineol 0,62 1,23 0 ,30' Tridecene 0,62 1,33 0,66 Linalylacetate 0,59 0,64 0 ,71 Carvone 0,57 0,46 0,45 4-terpineol 0,56 0,60 0,36 trans-2-Heptenal 0.55 0,23 1,38 Dodecane 0,53 0,20 0,24 Menthol 0,53 0,61 0,3 Camphor 0,42 0,58 0,41 Decanal 0,37 0,31 0,18 gamma-Terpinene 0,37 0,12 0,12 trans-2-Optenal 0,37 0. 13 0,91 trans-alpha-Bergamotene 0,37 0,37 0,25 2-Optanone 0,35 0,21 0,23 neryl Acetate 0,32 0,44 0,36 Borneol 0,31 0,27 0,38 p-Cimene 0,31 0,13 0,12 2-Dodecanone 0,26 0,27 0,24 unidentified alcohol 1 <0,1 0 <0,10 4,33 Tetradecane 0,25 0,53 0,60 Isomenthol 0, 7 0,21 0,24 2-Heptanone 0,16 0,12 0,08 unidentified alcohol 1 < , <0,10 17,42 Results expressed as % GC area 5 . Thin layer chromatography analysis of the extract (Examples 1 and 2)

A qualitative thin-layer chromatography (TLC) analysis of samples of the

Helichrysum italicum extract in Jojoba oil of examples 1 and 2 was carried out, as well as in other oils, for checking possible differences.

Analysed Samples

I - Example 1 (soaking);

II - Example 2 (percolation by gravity);

III - Jojoba wax alone.

In Fig. 2 , concerning the TLC profile, sample I (Example 1) is indicated as

"D", sample II (Example 2) as Έ " , and the sample III as "G". The samples were analysed after treating 2 droplets of extract with 5 droplets of methanol, by maintaining a vortex stirring for 10 seconds and then allowing the two phases to stratify. The methanol phase was sent to TLC, as shown hereinafter: - stationary phase: TLC Silica Gel F254 plates, by Merck, Code 1.05554.0001 ;

mobile phase: 1) Dichloromethane 100%; 2) dichloromethane/n-esane 50%/50%;

deposition of the samples: in an amount of 2 µ Ι or of 3 µ Ι by microcapillary pipettes; spot display: by UV irradiation of 254 nm and 365 nm wavelength, and

then spraying the plate with a concentred solution of 5% Vanilline in sulphuric acid.

As shown in Fig. 2 , a comparison was carried out between samples I, II and III for detecting the differences between percolate extract, soaked extract and Jojoba wax as such. The presence of a spot 30, shown in the box only for sample II, concerning the percolate extract, indicates that the percolate extract contains one compound in a far higher concentration than the soaked extract.

This compound is not present in Jojoba oil as such (sample III), therefore it is considered that it comes from Helichrysum. This spot 30 is the only one which had a slight fluorescence at 254 nm.

6 . Formulation examples

Qleolite formulation

Helichrysum italicum extract; Simmondsia chinensis seeds oil; Anthemis Nobilis oil.

Shampoo/shower Formulation

Water, Sodium cocoamphoacetate, Cocamidopropyl Betaine, Acrylates C/10-

C30, crossipolimer acrylate, Simmondsia chinensis seeds oil, Helichrysum italicum extract, Anthemis nobilis oil, Methylisothiazolinone, Methylchloroisothiazolinone, Lactic acid, Sodium hydroxide.

Solid soap Formulation

Cocos Nucifera, water, Elaeis guinensis, Olea europea, sodium hydroxide,

Simmondsia chinensis seeds oil, Anthemis nobilis oil, Helichrysum italicum extract, Tricticum vulgare face and body Cream Formulation

Water, Butyrospermum parkii (butter of Galam), Stearic acid, Glycerine, of Simmondsia chinensis oil, Helichrysum italicum extract, Methylglucose dioleate, Caprylic/capric triglyceride, hydrogenated beaver oil, phenoxyethanol, Methyl paraben, Butyl paraben, Propyl paraben, Isopropyl paraben,

Ethanolamide, Tocopheryl acetate, Anthemis nobilis oil.

7 . Effect of the extracts on cell proliferation of dermal keratinocytes

The substances examined were evaluated in a range from 1% v/v to 0.0001% v/v. Human keratinocytes (HEK, Human Epidermis Keratinocytes, ECACC) were grown in the Complete Keratinocyte Growth Medium (ECACC) according to the supplier's instructions, and were used for the intermediate test between the first passage and the sixth passage.

The test was carried out using cell proliferation Reagent WST-1 kit, provided by Roche Diagnostic. The test was carried out according to the supplier's instructions.

More in detail, the cells were sown in a 96-well plate for absorbance reading. The cell density ranged from 2.000 to 8.000 cells per well in a volume of 100 microlitres per well. The substance examined had a concentration set within the above defined range. In one control, no product had been added to the cells.

The tests were made in quadruplicate. Furthermore, a blank was associated characterised by the medium without the product to be analysed (control) and with the product to be analysed, at the concentration as defined above, in single.

A first step of incubating the cells lasted 8 hours. Then, the reactive was added ( 0 Ι for each well) and the incubation was carried on during another 2 hours, or during another 4 hours. At the end of the incubation step, an absorbance reading was carried out during 1 s at the wavelength of 450 n in a Victor 2 Wallac microplate reader by Perkin Elmer.

The results are given in the graphs of Figs. 3 and 4 , where cell proliferation is expressed as a proliferation ratio (%), defined as it follows:

Proliferation % = [(absorbance of the treated cells - absorbance of the blank) / (absorbance of the control - absorbance of the blank)] x 100.

With reference to Figs. 3 and 4, the extract of example , which provided a good cell proliferation stimulation on keratinocytes, has a content of Chrysin of 123 mg/kg oil. Therefore, a threshold could be 123 x 0.65 = 80 mg/kg oil. Similarly, for the extract of example 2 a threshold preferred value could be 130 x 0.65 = 84 mg/kg oil and a still more preferred value could be 130 x 0.80 = 104 mg/kg oil.

From the diagram of fig. 3 , it can be observed that the extract of example 3 does not cause any significant stimulation of keratinocytes proliferation at all the tested concentrations. On the contrary, the extracts of examples 1 and 2, in particular the latter, stimulate keratinocytes proliferation significantly more than the control, at least at the highest two test concentrations. At lower concentrations, i.e. with a more diluted extract, the proliferation values are similar to the control ones.

A 100% proliferation value is considered as a control value. The different behaviour of example 3 with respect to examples 1 and 2 can be explained with reference to the different chemical compositions, in particular to the different content in flavonoids Chrysin, Galangin and

Pinocembrin, as shown in Tables 3a, 3b, 3c. The content of these flavonoids in example 3 is clearly lower than in examples 1, 2 and 4 .

In independent experiments, not shown, Jojoba wax has not produced any significant effects on keratinocytes cell proliferation. Therefore, the effect on keratinocytes depends upon Helichrysum and does not depend upon Jojoba.

In the graph of Fig. 4 all the positive correlations are shown between the amounts of the compounds that were found in the extract samples of examples 1, 2, 3 and 4 , and the proliferation ratio with respect to HEK cells treated by an extract at the concentration of 1%. These results are also given in table 5 .

- Table 5 -

It can be observed that these positive correlations occur for the three flavonoids Galangin, Pinocembrin and Chrysin, for a-Selinene and β-Selinene terpenes, and for Curcumene. On the contrary, no positive correlation is found for the aldehydes. 8 . Effectiveness of the extract in treating skin injuries associated with stomata.

A preparation based on a Jojoba oil Helichrysum extract according to the invention was used for treating skin complications of stomata in 10 patients. The injuries were evaluated according to the gravity, and a score was given to the injuries according to four levels, from "L1" to "L4", indicating with these symbols a proliferative lesion. The peristomal skin was subdivided into 4 quadrants from T 1 to T4, and a circumferential lesion on the whole peristomal skin was indicated as T5, as shown in Fig. 5 . The therapeutic protocol consisted of the exclusive use of a preparation based on the Helichrysum oil extract as medic aid. The preparation was spread on the injuries, at a maximum amount of 0.5 cc to cover a skin extension of about 5-1 0 cm. Thereafter, the 10 patients were monitored by photography and by measuring the size of the injured skin area at the beginning of the therapy and after 1, 2 and 3 weeks of treatment. The results of these checks are given in table 6 , along with a description of the starting situation of the patients.

- Table 6 -

In summary, at the beginning of the treatment L2 and L3 injuries were observed, which extended over different peristomal quadrants;

at the second week of the treatment, an improvement was observed in

seven patients, whereas there was no injury reduction in three patients;

at the third week of the treatment, a recovery had occurred in five

patients, whereas an injury stabilization was observed in two patients,

which suggested to continue the treatment; a partial response is also

observed in the patients who had appeared as "non-responding patients" at the first week of treatment.

From the analysis of preliminary data, a reduction is observed of the injuries after treatment with Helichrysum oil in all the patients (100%) after three weeks of treatment, and a complete recovery is observed in five patients (50%). No cases of allergic reaction were observed, or of intolerance to the product based on the Jojoba oil Helichrysum extract. Therefore, Helichrysum oil is a safe and effective aid in the treatment of skin injuries in stoma-bearing patients.

9 . Effectiveness of the extract in the treatment of other skin diseases and conditions.

Tests were carried out on patients, which show the effectiveness of medicaments or medical devices in the treatment of skin diseases and conditions such as:

skin complications in stoma-bearing patients, in particular complications caused by the reaction of the skin to the glues of the aids used to treat the wounds, by a continuous contact with the faeces, by the repeated skin traumatism due to the frequent change of the same; wounds, including cuts, excoriations, abrasions, insect bites, wounds by cutting arm, wounds by firearm, surgical wounds, etc.; burns, including UV burns, sun burns, radiotherapy burns, fire burns, hot contact burns, acid burns, alkali burns, etc.; skin ulcers, including decubitus ulcers, diabetic ulcers, etc.; scars, including surgical scars, scars owing to excessive growth of connective tissue after a tissue damage, atrophic scars, acne scars, etc.; skin aging, including age skin aging, UV skin aging, aging caused by environmental pollution, aging due to atmospheric factors, hormonal aging, etc.; stretch marks, including pregnancy stretch marks, stretch marks caused by an important body weight change, hormonal stretch marks, etc.; psoriasis injuries; wrinkles, including expression wrinkles, etc. The foregoing description exemplary embodiments of the method and of the product according to the invention, will so fully reveal the invention according to the conceptual point of view, so that others, by applying current knowledge, will be able to modify and/or adapt for various applications such embodiment without further research and without parting from the invention, and, accordingly, it is therefore to be understood that such adaptations and modifications will have to be considered as equivalent to the specific embodiments. The means and the materials to realise the different functions described herein could have a different nature without, for this reason, departing from the field of the invention. It is to be understood that the phraseology or terminology that is employed herein is for the purpose of description and not of limitation. CLAIMS

1. A method for preparing an extract of an Helichrysum in Jojoba oil, said method comprising the steps of: prearranging a Helichrysum plant portion selected among the upper

parts of the plant, in particular the flowers; prearranging a Jojoba oil having an acid number higher than 0.3;

joining said Helichrysum plant portion and said Jojoba oil in an extraction condition; recovering said extract obtained during said step of joining.

2. A method according to claim 1, wherein said extract has a content of Chrysin, per kg of fatty acids having 20 carbon atoms and one unsaturation, higher than 80 mg.

3. A method according to claim 1, wherein said extract comprises Limonene and β-Selinene having a weight ratio of Limonene with respect to said β- Selinene lower than 0.4.

4. A method according to claim 1, wherein said plant of the genus Helichrysum belongs to a species selected from the group consisting of:

Helichrysum italicum, in particular Helichrysum italicum subspecies italicum and/or Helichrysum italicum subspecies microphyllum; Helichrysum stoechas; Helichrysum arenarium.

5. A method according to claim 1, wherein said Jojoba oil has an acid number higher than 0.32.

6. A method according to claim 1, wherein said Jojoba oil has an acid number higher than 0.36.

7. A method according to claim 1, wherein said step of joining comprises a

step of dynamic joining, wherein said Jojoba oil is in movement with respect to said plant portion. 8. A method according to claim 1, wherein said step of joining comprises a step of percolating.

9. A method according to claim 1, wherein said step of joining comprises a

step of centrifuging said plant portion in said Jojoba oil.

10. A method according to claim 1, wherein said step of joining comprises a step of soaking.

1 . A method according to claim 1, wherein said step of joining is carried out by maintaining said Jojoba oil at a contact temperature set between the

room temperature, in particular 20°C, and an extraction temperature lower than a maximum extraction temperature, wherein said maximum

extraction temperature is 70°C, in particular 60°C, more in particular,

50°C, even more in particular 45°C.

12. A method according to claim 1, wherein said step of prearranging a plant portion comprises a step of drying said plant portion.

13. An extract in Jojoba oil obtained by a plant of the genus Helichrysum, said Jojoba oil comprising fatty acids having 20 carbon atoms and one unsaturation, said extract comprising a predetermined content of Chrysin,

characterised in that said content of Chrysin, per kg of said fatty acids having 20 carbon atoms

and one unsaturation, is higher than 80 mg,

in particular, for use in the treatment of a disease and/or of a condition for

which a stimulation of cell proliferation of keratinocytes is required.

14. An extract according to claim 13, wherein said content of Chrysin is

higher than 84 mg per kg of said fatty acids having 20 carbon atoms and

one unsaturation, in particular said content of Chrysin is higher than 104 mg/kg of said fatty acids having 20 carbon atoms and one unsaturation.

15. An extract in Jojoba oil obtained by a plant of the genus Helichrysum in Jojoba oil, said extract comprising Limonene and β-Selinene, characterised in that said Limonene has a weight ratio with respect to said β-Selinene lower than 0.4.

16. An extract according to claim 15, wherein said weight ratio is lower than

0.08, in particular said weight ratio is lower than 0.06.

17. A medical or cosmetic composition comprising an extract according to

any of claims from 13 to 16, or an extract obtained with the method

according to any of claims from 1 to 12, for treating or preventing a condition for which a stimulation of the cell proliferation of keratinocytes is required, or for treating or preventing a disease and/or a condition selected from the group consisting of:

skin complications in stoma-bearing patients, in particular complications caused by the reaction of the skin to glues of aids used to treat the wounds, by continuous contact with the faeces, by a repeated skin traumatism due to a frequent change of said aids;

wounds, in particular cuts, excoriations, abrasions, insect bites, wounds by cutting arm, wounds by firearm, surgical wounds;

burns, in particular UV burns, sun burns, radiotherapy burns, fire burns, hot contact burns, acid burns, alkali burns;

skin ulcers, in particular decubitus ulcers, diabetic ulcers;

scars, in particular surgical scars, scars owing to excessive growth of connective tissue after a tissue damage, atrophic scars, acne scars;

skin aging, in particular age skin aging, UV skin aging, aging caused by environmental pollution, aging due to atmospheric factors, hormonal aging;

stretch marks, in particular pregnancy stretch marks, stretch marks owing to important variation of body weight, hormonal stretch marks; psoriasis injuries;

wrinkles, in particular expression wrinkles. 18. A composition according to claim 17, comprising an essential oil, in particular said essential oil being selected from the group consisting of: roman camomile essential oil; matricaria camomile essential oil; carnation

essential oil; tea tree essential oil; lavender essential oil; wintergreen essential oil; origanum essential oil, or mixtures thereof.

19. An insect-repellent composition comprising an extract as by any of claims

from 13 to 16, and/or an extract obtained with a method according to any

of claims from 1 to 12.

A . CLASSIFICATION O F SUBJECT MATTER INV. A61K36/185 A61K36/28 A61K36/61 A61K36/36 A61K36/534 A61P17/02 A01N65/00 ADD. According to International Patent Classification (IPC) or to both national classification and IPC

B . FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) A61K A61P A01N

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched

Electronic data base consulted during the international search (name of data base and, where practicable, search terms used)

EPO-Internal , BIOSIS, EMBASE, WPI Data

C . DOCUMENTS CONSIDERED TO B E RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

I T PI20 010 056 Al ( ERBORI STERI A DEL C0RS0 1-19 DI SIMO [IT] ) 2 November 2001 (2001-11-02) ci ted i n the appl i cati on abstract pages 2-3 -/-

X| Further documents are listed in the continuation of Box C . See patent family annex.

* Special categories of cited documents : "T" later document published after the international filing date or priority date and not in conflict with the application but cited to understand "A" document defining the general state of the art which is not considered the principle or theory underlying the invention to be of particular relevance "E" earlier application or patent but published o n or after the international "X" document of particular relevance; the claimed invention cannot be filing date considered novel or cannot be considered to involve an inventive "L" documentwhich may throw doubts on priority claim(s) orwhich is step when the document is taken alone cited to establish the publication date of another citation or other "Y" document of particular relevance; the claimed invention cannot be special reason (as specified) considered to involve an inventive step when the document is "O" document referring to an oral disclosure, use, exhibition or other combined with one o r more other such documents, such combination means being obvious to a person skilled in the art "P" document published prior to the international filing date but later than the priority date claimed "&" document member of the same patent family

Date of the actual completion of the international search Date of mailing of the international search report

19 March 2013 26/03/2013

Name and mailing address of the ISA/ Authorized officer European Patent Office, P.B. 5818 Patentlaan 2 NL - 2280 HV Rijswijk Tel. (+31-70) 340-2040, Fax: (+31-70) 340-3016 Cami 1l eri , Al ai n C(Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

Anonymous : " El i cri so Ol eol i t o 1-19 Composto-Al bero del e Rose del l a Dott. ssa Cecchi ni " , Simone del Corso

11 June 2011 (2011-06-11) , XP002674396, Retri eved from the Internet: URL: http: //www. al berodel l erose. i t / i ndex. h p?page=shop.product_detai 1s&f lypage=garden _f 1ypage . tpl &product_i d=1950&category_i d=3 29&opti on=com_vi rtuemart&I temi d=134&l ang=i t [retri eved on 2012-04-16] the whol e document

Admi ni strator: " El i cri so (Hel i chrysum 17 , 18 i tal i cum) " , Di z i onari o di Fi toterapi a e Pi ante Medi c i nal i di Enri ca Campani ni Ed Tecni che Nuove

11 June 2011 (2011-06-11) , XP002674397 , Retri eved from the Internet: URL: http://www.al berodel l erose. i t / i ndex. ph p?vi ew=arti c l e&cati d=54%3Api ante-medi c i nal i &i d=424%3Ael i cri so-hel i chrysum-i tal i cum&t mpl =component&pri nt=l&l ayout=def aul t&page= &opti on=com_content&I temi d=147&l ang=i t [retri eved on 2012-04-13] the whol e document

"Hel i chrysum Oi l , Pure Organi c Essenti al 1-19 Oi l " , The Ananda Apothecary

4 July 2009 (2009-07-04) , XP002674398, Retri eved from the Internet: URL: http://www.anandaapothecary. com/aromat herapy-essenti al -oi 1s/hel i chrysum-essenti a 1-oi 1. html [retri eved on 2012-04-13] the whol e document

US 2004/234628 Al (KEARNS MARCY L [US] ET 17-19 AL) 25 November 2004 (2004-11-25) page 1, r i ght-hand col umn , paragraph 14-15 page 2 , r i ght-hand col umn , paragraph 27 tabl e 1 c l aims 1, 3 ,4,5 , 14, 15 ,23 ,27 -I C(Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

Y MASTELIC JOSI P, POLITEO OLIVERA, JERKOVIC 1-19 IGOR: "Contri buti on t o the Analysi s of the Essenti al Oi l of Hel i chrysum i tal i cum (Roth) G. Don . - Determi nati on of Ester Bonded Aci ds and Phenol s " , MOLECULES, vol . 13 , no. 4 , 7 Apri l 2008 (2008-04-07) , pages 795-803 , XP002674399 , Basel , Swi tzerl and D0I : 10.3390/mol ecul esl3040795 abstract tabl es 1-2 page 796, paragraph 1

Y Anonymous : " Ital i eni sche Strohbl ume" , 1-19 i ki pedi a , der frei en Enzykl opadi e

18 September 2010 (2010-09-18) , XP002674400, Retri eved from the Internet: URL: http://de.wi ki pedi a.org/w/i ndex.php?ti t l e=I tal i eni sche_Strohbl ume&ol di d=79268296 &pri ntabl e=yes [retri eved on 2012-04-20] the whol e document

Y B0HLMANN F ET AL: "Neue prenyl f l avanone 1-19 aus Hel i chrysum hypocephal um" , PHYTOCHEMISTRY, vol . 18, no. 11 , 1 January 1979 (1979-01-01) , pages 1851-1853 , XP026609283 , PERGAM0N PRESS, GB ISSN : 0031-9422 , D0I : 10. 1016/0031-9422 (79)83068-X [retri eved on 1979-01-01] abstract page 1851 , l eft-hand col umn , paragraph 1

Y LV P C ET AL: "Synthesi s , bi ol ogi cal 1-19 eval uati on of chrysi n deri vati ves as potenti al immunosuppressi ve agents" , EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, vol . 46, no. 1, 1 January 2011 (2011-01-01) , pages 393-398, XP027565727 , EDITIONS SCI ENTI FIQUE ELSEVI ER, PARIS, FR ISSN : 0223-5234 [retri eved on 2010-12-18] c i ted i n the appl i cati on abstract page 393 , l eft-hand col umn , paragraph 2

-/-- C(Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

Y NAN-LIN U ET AL: "Chrysi n Protects 1-19 Epi dermal Kerati nocytes from UVA- and UVB-Induced Damage" , JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, vol . 59 , no. 15 , 10 August 2011 (2011-08-10) , pages 8391-8400, XP055025258, ISSN : 0021-8561 , D0I : 10. 1021/j f 200931t c i ted i n the appl i cati on abstract page 8392 , l eft-hand col umn , paragraph 1-2 page 8393 , r i ght-hand col umn , paragraph 2 - page 8396, l eft-hand col umn , paragraph 1 Patent document Publication Patent family Publication cited in search report date member(s) date

IT PI20010056 A l 02- 11 -2001 NONE us 2004234628 A l 25- 11 -2004 CA 2524375 A l 09 -12 -2004 EP 1635853 A2 22 -03 -2006 US 2004234628 A l 25 -11 -2004 O 2004105689 A2 09 -12 -2004