Terpene Extract for the Treatment of Hair Loss Terpenextrakt Zur Behandlung Von Haarausfall Extrait De Terpène Pour Le Traitement De La Perte Des Cheveux

(19) TZZ __T

(11) EP 2 666 461 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 31/573 (2006.01) A61K 31/575 (2006.01) 01.07.2015 Bulletin 2015/27 A61K 31/704 (2006.01) A61K 36/42 (2006.01) A61Q 5/02 (2006.01) A61K 8/63 (2006.01) (2006.01) (21) Application number: 12382191.0 A61Q 7/00

(22) Date of filing: 23.05.2012

(54) Terpene extract for the treatment of hair loss Terpenextrakt zur Behandlung von Haarausfall Extrait de terpène pour le traitement de la perte des cheveux

(84) Designated Contracting States: (56) References cited: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB WO-A1-2010/035299 US-A- 5 925 356 GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR •GREIGE-GERGES ET AL: "Cucurbitacins from Ecballium elaterium juice increase the binding of (43) Date of publication of application: bilirubin and ibuprofen to albumin in human 27.11.2013 Bulletin 2013/48 plasma", CHEMICO-BIOLOGICAL INTERACTIONS, ELSEVIER SCIENCE IRLAND, (73) Proprietor: Tergum S.L. IR, vol. 169, no. 1, 13 July 2007 (2007-07-13) , 41702 Dos Hermanas, Sevilla (ES) pages 53-62, XP022152868, ISSN: 0009-2797, DOI: 10.1016/J.CBI.2007.05.003 (72) Inventor: Rodríguez Marín, Carlos Luis • DATABASE WPI Week 200669 Thomson 41702 Sevilla (ES) Scientific, London, GB; AN 2006-662383 XP002688880, & JP 2006 206537 A (KOSE KK) 10 (74) Representative: Hoffmann Eitle August 2006 (2006-08-10) Patent- und Rechtsanwälte PartmbB Arabellastraße 30 81925 München (DE)

Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 666 461 B1

Printed by Jouve, 75001 PARIS (FR) 1 EP 2 666 461 B1 2

Description hair regrowth was achieved in 33 out of 34 sites injected with triamcinolone hexacetonide in 11 patients and in 16 Field of the Invention out of 25 sites injected with triamcinolone acetonide in 17 patients, with the affects lasting about nine months. [0001] The present invention relates to compositions 5 However, intralesional corticosteroid administration is comprisingcucurbitacins for usein thetreatment and pre- most suitable for treating patchy hair loss of limited extent vention of hair loss conditions and disorders. and for cosmetically sensitive sites such as the eyebrows, with skin atrophy at the site of injection a consist- Background of the invention ent side-effect. Systemic corticosteroid administration 10 has achieved some hair regrowth in alopecia suffers. [0002] The following discussion on the background of However, studies have demonstrated only 30-47% of pa- the invention is merely provided to aid the reader in un- tients showed significant hair regrowth and thus in most derstanding the invention and is not admitted to describe patients the response achieved is insufficient to justify or constitute prior art to the present invention. the risks associated with prolonged treatment. [0003] Hair loss is an extremely prevalent condition 15 [0007] Contact dermatitis inducers work by sensitising and it is estimated that approximately 20 to 25% of the the immune system. A low level of the drug is initially population suffer from some type of hair loss disorder. applied and adjusted until a reaction is established. Con- Whilst hair loss is more common in adult males, hair loss tact dermatitis inducers utilised for the treatment of alo- disorders also affect women and children. Although most pecia areata include dinitrochlorobenzene (DNCB), hair loss disorders are not in themselves damaging to 20 diphenylcyclopropenone (DPCP) and squaric-acid-dib- the health of an individual, many sufferers report asso- utylester (SADBE). Reviews of these treatments have ciated psychological problems including anxiety and de- concluded that the range of response rates is wide, at pression. reportedly 9 to 87% (Rokhsar et al. J Am Acad Dermatol [0004] Despite the prevalence of hair loss disorders, 1998:39:751-761). Side effects including severe derma- few efficacious treatments are available. Surgical treat- 25 titis and occipital and/or cervical lymphadenopathy are ments are available for the restoration of hair, particularly often common. to the scalp. However, these techniques are painful, ex- [0008] Specific immunosuppressants for the treatment pensive, and suffer from the inherent dangers associated of alopecia include cyclosporins and tacrolimus (FK506). with any invasive surgery. Furthermore, hair restoration These drugs act by inhibiting T-cell activation. While re- surgeries typically require at least some areas of dense 30 sults have demonstrated some efficacy for cyclosporins, hair growth to be available on the patient for transplant side-effects are a major consideration and results may or relocation to other areas of the body. Yet further, sur- often not justify the associated risks. gical treatments are rarely applicable to sufferers of alo- [0009] Alternative treatments such as essential oils pecia areata in which spontaneous regrowth of hair which and acupuncture and experimental and theoretical treat- subsequently falls out again often occurs. 35 ments such as cytokines, biologicals, desensitisation, [0005] Hair loss compositions are thus an attractive oral tolerance and gene therapy have been variously em- proposition for those suffering from hair loss conditions. ployed in the treatment of alopecia with varying degrees A range of therapies are available for which partial suc- of success. cess in the treatment of alopecia has been claimed. The [0010] Therefore, despite active research in this area various therapies available can be divided up into several 40 and a large number of proposed active ingredients there groups comprising non-specific immunosuppressants remains the need for a hair loss composition which dem- such as corticosteroids or UVA treatment, contact der- onstrates efficacy in a wide populace with minimal side matitis inducers, specific immunosuppressants such as effects. cyclosporins, non-specific irritants and other treatments such as alternative and experimental therapies. 45 Brief description of the invention [0006] Non-specific immunosuppressants such as corticosteroids function by mimicking the steroidal hor- [0011] A first aspect of the present invention refers to mones produced by the adrenal glands to suppress in- a composition (from hereinafter composition of the inven- flammation. Corticosteroids can be topically adminis- tion) comprising cucurbitacins and their glucoside deriv- tered to the affected area, through intralesional injec- 50 atives, wherein cucurbitacins D, I, L and A and their glu- tions, or systemically via injection or oral medication, de- coside derivatives are present, as a whole, in terms of pending on the severity of the condition. While topical percentage by weight, in an amount greater than 25% of corticosteroids are commonly used to treat alopecia ar- the total weight of the cucurbitacins and their glucoside eata, there is little evidence that they promote hair re- derivatives, and wherein cucurbitacins B, E and the de- growth. Intralesional administration of corticosteroids55 rivative of cucurbitacin E, ecballic acid, are each present has demonstrated more efficacy in the treatment of alo- at a concentration below 2.5% of the total weight of the pecia areata, with a study reported by Porter and Burton cucurbitacins and their glucoside derivatives. (Br. J. Dermatol. 1971:85, 272-273) demonstrating that [0012] In a preferred embodiment of the present inven-

2 3 EP 2 666 461 B1 4 tion, cucurbitacins D, I, L and A and their glucoside de- b. extracting the vegetable cucurbitacins from said rivatives are present, as a whole, in terms of percentage portion by a heating process and by using a liquid by weight, in an amount greater than 30% of the total extractant capable of solubilising said compounds. weight of the cucurbitacins and their glucoside derivatives, more preferably in an amount greater than 35%, 5 [0018] In a preferred embodiment of this aspect of the 40% or 45%. invention, the plant portion pertaining to the cucurbitace- [0013] In a more preferred embodiment of the present ae family is Ecballium elaterium. invention, cucurbitacins D, I, L and A and their glucoside [0019] In another embodiment of this aspect of the in- derivatives are present, as a whole, in terms of percent- vention, the heating process is conducted at a tempera- age by weight in an amount between 25 to 50% of the 10 ture from 120 C° to 180 C°, preferably at a temperature total weight of the cucurbitacins and their glucoside de- of approximately 150 C°. rivatives. [0020] In another embodiment of this aspect of the in- [0014] In a still more preferred embodiment of the invention, the plant portion is selected from the group convention, the composition of the invention further compris- sisting of: roots, stems, leaves or fruits or any combina- es: 15 tion thereof. Preferably the plant portion is a root. [0021] In a still further embodiment of this aspect of a. Cucurbitacin O; the invention, the extractant is olive oil, wherein prefer- b. Cucurbitacin P; ably the extractant is enriched with natural or added con- c. Cucurbitacin C; tent of antioxidants. d. Cucurbitacin H; 20 [0022] A fourth aspect of the invention refers to the e. Cucurbitacin J; composition of any of the previous aspects for its use in f. Cucurbitacin F; or therapy. g. Cucurbitacin Q; [0023] A fifth aspect of the invention refers to the composition of any of the previous aspects for its use in the wherein each of these compounds is present in an25 treatment or prevention of hair loss. In a preferred em- amount greater than 2.5% of the total weight of the cu- bodiment of the invention, hair loss is caused by hormo- curbitacins and their glucoside derivatives. nal deregulations, androgenic alopecia or from alopecia [0015] In a still more preferred embodiment of the in- of seborrheic origin. vention, the composition of the invention further compris- [0024] In a preferred embodiment, the composition of es olive oil. 30 any of the previous aspects is use as a cosmetic com- [0016] A second aspect of the invention refers to a veg- position. etable extract obtained from Ecbalium Elaterium wherein [0025] A further aspect of the invention refers to a proc- said vegetable extract comprises cucurbitacins and/or ess for the preparation of a composition comprising cu- their glucoside derivatives, wherein cucurbitacins D, I, L curbitacins and/or their glucoside derivatives, wherein and A and/or their glucoside derivatives are present, as 35 cucurbitacins D, I, L and A and their glucoside derivatives a whole, in terms of percentage by weight, in an amount are present, as a whole, in terms of percentage by weight, greater than 25% of the total weight of the cucurbitacins in an amount greater than 25% of the total weight of the and their glucoside derivatives, and wherein cucur- cucurbitacins and their glucoside derivatives, and where- bitacins B, E and the derivative of cucurbitacin E, ecballic in cucurbitacins B, E and the derivative of cucurbitacin acid, are each present at a concentration below 2.5% of 40 E, ecballic acid, are each present at a concentration be- the total weight of the cucurbitacins and their glucoside low 2.5% of the total weight of the cucurbitacins and their derivatives. glucoside derivatives, which comprises the following [0017] A third aspect of the invention refers to a com- steps: position (from hereinafter composition of the third aspect of the invention) comprising cucurbitacins and/or their 45 a. Obtaining a portion of a plant pertaining to the glucoside derivatives, wherein cucurbitacins D, I, L and cucurbitaceae family and A and their glucoside derivatives are present, as a whole, b. extracting the vegetable cucurbitacins from said in terms of percentage by weight, in an amount greater portion by a heating process and by using a liquid than 25% of the total weight of the cucurbitacins and their extractant capable of solubilising said compounds. glucoside derivatives, and wherein cucurbitacins B, E 50 and the derivative of cucurbitacin E, ecballic acid, are [0026] In a preferred embodiment of this aspect of the each present at a concentration below 2.5% of the total invention, the plant portion pertaining to the cucurbitace- weight of the cucurbitacins and their glucoside deriva- ae family is Ecballium elaterium. tives, wherein this composition is obtainable by a leach- [0027] In another embodiment of this aspect of the in- ing process comprising the steps of: 55 vention, the heating process is conducted at a temperature from 120 C° to 180 C°, preferably at a temperature a. Obtaining a portion of a plant pertaining to the of approximately 150 C°. cucurbitaceae family and [0028] In another embodiment of this aspect of the in-

3 5 EP 2 666 461 B1 6 vention, the plant portion is selected from the group con- [0039] An eighth aspect of the present invention refers sisting of: roots, stems, leaves or fruits or any combina- to the shampoo of the invention or to the lotion of the tion thereof. Preferably the plant portion is a root. invention, for its use in the treatment or prevention of hair [0029] In a still further embodiment of this aspect of loss. the invention, the extractant is olive oil, wherein prefer- 5 [0040] A ninth aspect of the present invention refers to ably the extractant is enriched with natural or added con- a pharmaceutical composition (from hereinafter the phar- tent of antioxidants. maceutical composition of the invention) comprising the [0030] A sixth aspect of the present invention refers to composition as defined in any of the first, second or third a shampoo composition (from hereinafter shampoo com- aspects of the present invention for its use in therapy. position of the invention) comprising: 10 Preferably, the pharmaceutical composition of the invention is use in the treatment or prevention of hair loss. a. A cleansing base; and More preferably, the pharmaceutical composition of the b. Active ingredients, invention is use in the treatment or prevention of hair loss caused by hormonal deregulations, androgenic alopecia wherein at least one of the active ingredients comprises 15 or from alopecia of seborrheic origin. thecomposition as defined in anyof the previous aspects. [0041] A preferred embodiment of this aspect of the [0031] In a preferred embodiment of this aspect of the invention refers to the pharmaceutical composition of the invention, the cleansing base comprises surfactants of invention further comprising a pharmaceutically accept- the types: alkyl sulphates and/or an alkyl ether sulphates. able excipient. [0032] In another embodiment of this aspect of the in- 20 [0042] In a preferred embodiment of this aspect of the vention,the shampoo composition of theinvention further invention the pharmaceutical composition of the inven- comprises one or more of the following components or tion comprises a vehicle suitable for topical administra- any combination thereof: tion. [0043] In a still further embodiment of this aspect of a. Thickeners for surfactant preparation; 25 the invention the pharmaceutical composition of the in- b. Pearlescents and opacifiers; vention comprises a vehicle suitable for intradermal ad- c. Perfumes; ministration. d. Colorants; or e. Stabilizing agents. Brief description of the figures 30 [0033] In a further embodiment of this aspect of the [0044] invention, the shampoo composition of the invention is an anti-dandruff composition comprising active ingredi- Fig. 1: Figure 1 illustrates a leaching process for the ents capable of re-establishing the natural microbial flora production of the composition of the invention. on the skin surface. 35 [0034] In a further embodiment of this aspect of the Fig. 2: Figure 2 illustrates the i) initial state, ii) evo- invention, the shampoo composition of the invention is a lution after treatment for 6 six months and iii) final shampoo for greasy hair comprising a cleansing base results obtained after one year of treatment, for the capable of eliminating excess sebum. treatment of Example 4. [0035] In a further embodiment of this aspect of the 40 invention, the shampoo composition of the invention is Fig. 3: Figure 3 illustrates the results before treat- for sensitive scalp and comprises mild anionic sur- ment and after treatment for a period of 6 months, phactants and amphoterics and/or non-ionic surfactants. for the treatment of Example 5. [0036] In a still further embodiment of this aspect of the invention, the shampoo composition of the invention 45 Fig. 4: Figure 4 illustrates the results before and after is for dry hair and comprises: treatment for the treatment of Example 6.

a. A fat absorber; and/or Description of the invention b. An abrasive to dislodge soil; and/or c. An alkalizer 50 Definitions

[0037] A seventh aspect of the present invention refers [0045] For the following defined terms, these defini- to a hair lotion (from hereinafter lotion of the invention) tions shall be applied, unless a different definition is given comprising the composition as defined in any of the first, in the claims or elsewhere in this specification. second or third aspects of the present invention. 55 [0046] The terms "cucurbitacin", "cucurbitacin com- [0038] In a preferred embodiment of this aspect of the pound", and "cucurbitacin derivative" are used inter- invention, the lotion is in the form of liquid, gel, cream, changeably in the specification to mean a group of trit- an emulsion, oil or serum. erpenoid substances which are structurally character-

4 7 EP 2 666 461 B1 8 ized by the tetracyclic cucurbitane nucleus skeleton, fication to mean compounds having the following chem- namely, 19-(10->9beta)-abeo-10alpha-lanost-5-ene (al- ical formula: so known as 9beta-methyl-19-norlanosta-5-ene), with a variety of substitutions at different positions of the struc- ture such as H, O, hydroxyl, alkyl, alkenyl, alkynyl, halo- 5 gen, alkoxy, aryl and heteroaryl. [0047] The term "cucurbitacin D" is used in the specification to mean compounds having the following chemical formula: 10

[0051] The term "cucurbitacin O" is used in the specification to mean compounds having the following chemical formula: 20

[0048] The term "cucurbitacin I" is used in the specification to mean compounds having the following chemical 25 formula:

[0052] The term "cucurbitacin P" is used in the speci- 35 fication to mean compounds having the following chemical formula:

40 [0049] The term "cucurbitacin L" is used in the specification to mean compounds having the following chemical formula:

50 [0053] The term "cucurbitacin C" is used in the specification to mean compounds having the following chemical formula:

[0050] The term "cucurbitacin D" is used in the speci-

5 9 EP 2 666 461 B1 10

[0054] The term "cucurbitacin H" is used in the speci- [0057] The term "cucurbitacin Q" is used in the specification to mean compounds having the following chem- 15 fication to mean compounds having the following chemical formula: ical formula:

30 [0055] The term "cucurbitacin J" is used in the speci- [0058] The term "ecballic acid" is used in the specifi- fication to mean compounds having the following chem- cation to mean compounds having the following chemical ical formula: formula:

[0056] The term "cucurbitacin F" is used in the speci- [0059] The term "cucurbitacin glucoside" is used in the fication to mean compounds having the following chem- specification to mean cucurbitacin derivatives formed by ical formula: 50 condensation of cucurbitacins with glucose units at the hydroxyl positions. The formation of glucoside derivatives is common in plant metabolisms as a pathway des- tined to store secondary metabolites involved in defense mechanisms in tissues or as a strategy to increase sol- 55 ubility of non-polar metabolites. [0060] The term "total weight of the cucurbitacins and their glucoside derivatives" is used in the specification to mean the total concentration of cucurbitacins and gluco-

6 11 EP 2 666 461 B1 12 side derivatives measured by a chromatographic method and pentangular. Tendrils are present at 90° to the leaf such as that reported by Greige-Gerges et al. (Chemico- petioles at nodes. Leaves are exstipulate alternate sim- Biological Interactions 169 (2007) 53-62). This method ple palmately lobed or palmately compound. The flowers is based on the analysis of the extract by liquid chroma- are unisexual, with male and female flowers on different tography with mass spectrometry detection (LC-MS) pro- 5 plants (dioecious) or on the same plant (monoecious). viding a chromatographic peak signal for each cucur- The female flowers have inferior ovaries. The fruit is often bitacin and glucoside derivative (limit of detection 0.5-1.0 a kind of modified berry called a pepo. mg/L). The peak area of the signal associated to each [0064] The term "Ecballium elaterium" (L.) ("squirting cucurbitacin or derivative is a function of its concentration cucumber" or "donkey’s green") as used herein is taken in the extract. Quantitation of each cucurbitacin or deriv- 10 to be a perennial plant from the family Cucurbitaceae, a ative is carried out by preparation of a calibration model Mediterranean medicinal plant that has been investigat- using the same analytical protocol with standard solu- ed for several of its pharmacological properties. Ecbal- tions of cucurbitacin E at different concentrations. This lium elaterium has a large fleshy root, from which several model enables to obtain a calibration curve by plotting round, thick, rough stems rise, branching and trailing like peak area of cucurbitacin E chromatographic signals ver- 15 the common cucumber, but without tendrils. The leaves sus cucurbitacin E concentration in the solutions used to are petiolate, large, rough, irregularly cordate, and of a prepare the calibration model. The interpolation in the grayish-green color. The flowers are yellow and axillary. calibration curve of the peak area for each cucurbitacin The fruit has the shape of a small oval cucumber, about or derivative provided by analysis of the extract enables an inch and a half long and one inch thick. It has a green- to estimate the concentration of each cucurbitacin or de- 20 ish or grayish color, and is covered with stiff hairs or prick- rivative expressed relatively as cucurbitacin E. The total les. When fully ripe, it separates from the peduncle, and concentration of cucurbitacins and derivatives is calcu- throws out its juice and seeds with considerable force lated by summing up the concentrations of individual cu- through an opening at the base, where it was attached curbitacins and derivatives. The total concentration of to the footstalk. The name of squirting cucumber was cucurbitacins and derivatives is expressed as percent- 25 derived from this circumstance, and the scientific and age concentration (total weight of the cucurbitacins and official title (elaterium) is supposed to have had a similar their glucoside derivatives) referred to the total weight of origin, though some authors maintain that the term elat- plant material used for extraction. As exposed above, erium was applied to the drug rather from the mode of cucurbitacins D, I, L and A and their glucoside derivatives its operation upon the bowels than from the projectile should be present in the extract as percentage by weight, 30 property of the fruit. The word elaterium was used by in an amount greater than 25% of the total weight of the Hippocrates to signify any active purge. cucurbitacins and their glucoside derivatives measured [0065] The term "leaching process for the extraction of by the test described in this paragraph. On the other terpenes" as used herein is taken to be a solid-liquid ex- hand, cucurbitacins B, E and the derivative of cucur- traction from the raw material (i.e. a plant portion of bitacin E, ecballic acid, are each present at a concentra- 35 ecbalium elaterium) using as extractant a non-polar liquid tion below 2.5% of the total weight of the cucurbitacins phase capable of solubilising terpenes (compounds of and their glucoside derivatives. non-polar nature). [0061] As used herein, "pharmaceutically acceptable carrier" means any material which, when combined with Detailed description the composition of the invention, allows the compound 40 to retain biological activity. [0066] The applicant has now surprisingly discovered [0062] The term "alopecia" as used herein is taken to that a composition comprising cucurbitacins and/or their mean a disease in which the hair falls out and includes, glucoside derivatives, wherein cucurbitacins D, I, L and but is not limited to alopecia areata, male and female A and their glucoside derivatives are present, as a whole, pattern baldness, alopecia androgenetica, alopecia tota- 45 in terms of percentage by weight, in an amount greater lis, alopecia universalis, alopecia partialis, alopecia bar- than 25% of the total weight of the cucurbitacins and their bae, alopecia cicatricial, alopecia areata ophiasis, alo- glucoside derivatives, and wherein cucurbitacins B, E pecia areata diffusa, reticular alopecia areata, sisaipho and the derivative of cucurbitacin E, ecballic acid, are alopecia areata, congenital alopecia, drug-induced alo- each present at a concentration below 2.5% of the total pecia, alopecia mucinosa, alopecia marginalis, androge- 50 weight of the cucurbitacins and their glucoside deriva- netic alopecia, traction alopecia and syphilitic alopecia. tives, is highly effective in compositions for the treatment [0063] The term "cucurbitaceae family" as used herein and prevention of hair loss. is taken to consist of approximately 125 genera and 960 [0067] In order to demonstrate the above, the applicant species, mainly in tropical and subtropical regions. All has prepared a series of vegetable extracts by using 5 g species are sensitive to frost. Most of the plants in this 55 of the root of Ecbalium elaterium as raw material for the family are annual vines, but there are also woody lianas, leaching process detailed in example 1. These vegetable thorny shrubs, and trees (Dendrosicyos). Many species extracts contained a total content of cucurbitacins and have large, yellow or white flowers. The stems are hairy their glucoside derivatives, as detailed in example 2, in

7 13 EP 2 666 461 B1 14 the range from 200 mg/L to 1 g/L; wherein the most con- f. Cucurbitacin F; or centrated cucurbitacins detected in the extracts were the g. Cucurbitacin Q; D, I, L and A forms, which constituted approximately, as a whole expressed as percentage by weight, an amount wherein each of these compounds is present in an ranging from 25 to 50% of the total weight of the cucur- 5 amount greater than 2.5% of the total weight of the cu- bitacins and their glucoside derivatives. Other cucur- curbitacins and their glucoside derivatives. bitacins such as cucurbitacin O, P, C, H, J, F and Q were [0074] Additionally, the applicant has discovered the also present at lower concentrations (each of these cu- presence of phenolic compounds such as hydroxytyro- curbitacins were present in an amount greater than 2.5 sol, tyrosol, flavonoids (luteolin, apigenin), simple phe- weight percent of the total weight of the cucurbitacins 10 nols (vanillic acid, vanillin), secoiridoids (3,4-DHPEA- and their glucoside derivatives). EDA, 3,4-DHPEA-EA, p-HPEA-EDA, p-HPEA-EDA) and [0068] However, surprisingly cucurbitacins B, E and other triterpenic compounds characteristic of olive oil the derivative of cucurbitacin E, ecballic acid, which are (oleanolic acid and maslinic acid) in the vegetable ex- the most concentrated cucurbitacins or derivatives found tracts obtained in example 1. The antioxidant capacity of in Ecballium elaterium, were detected at trace levels15 olive phenols enhances the biological activity of cucur- (concentration below 2.5% of the total weight of the cu- bitacins and glucoside derivatives improving their thera- curbitacins and their glucoside derivatives) or not detect- peutic effect. ed at all. [0075] Thus, in a further embodiment, the composition [0069] These vegetable extracts were used by the ap- of the invention further comprises phenols or phenolic plicant to prepare the shampoo composition illustrated 20 compounds, preferably phenols or phenolic compounds in example 3. This shampoo composition was tested, obtained from olive oil. alone or in combination with other treatments, on patients [0076] A second aspect of the present invention refers suffering from different types of alopecia as illustrated in to a vegetable extract obtained from Ecbalium Elaterium examples 4 to 8. The results show that these types of wherein said vegetable extract comprises cucurbitacins compositions are highly effective for the treatment and 25 and/or their glucoside derivatives, wherein cucurbitacins prevention of hair loss. D, I, L and A and/or their glucoside derivatives are [0070] Accordingly, the first aspect of the present in- present, as a whole, in terms of percentage by weight, vention provides a composition (composition of the in- in an amount greater than 25% of the total weight of the vention) comprising cucurbitacins and their glucoside de- cucurbitacins and their glucoside derivatives, and where- rivatives, wherein cucurbitacins D, I, L and A and their 30 in cucurbitacins B, E and the derivative of cucurbitacin glucoside derivatives are present, as a whole, in terms E, ecballic acid, are each present at a concentration be- of percentage by weight in an amount greater than 25% low 2.5% of the total weight of the cucurbitacins and their of the total weight of the cucurbitacins and their glucoside glucoside derivatives. derivatives, and wherein cucurbitacins B, E and the de- [0077] In a preferred embodiment of this aspect of the rivative of cucurbitacin E, ecballic acid, are each present 35 invention, the vegetable extract contains, before dilution, at a concentration below 2.5% of the total weight of the a total content of cucurbitacins and their glucoside deriv- cucurbitacins and their glucoside derivatives. atives in the range from 200 mg/L to 1 g/L. In a more [0071] The composition of the invention preferably preferred embodiment of the invention, the vegetable ex- comprises cucurbitacins D, I, L and A and their glucoside tract further comprises phenols or phenolic compounds, derivatives, as a whole, as percentage by weight, in an 40 preferably phenols or phenolic compounds obtained from amount greater than 30% of the total weight of the cu- olive oil. More preferably, the vegetable extract compris- curbitacins and their glucoside derivatives, more prefer- es olive oil. ably in an amount greater than 35%, 40% or 45%. [0078] A third aspect of the invention refers to a com- [0072] In a more preferred embodiment of the present position (from hereinafter composition of the third aspect invention, cucurbitacins D, I, L and A and their glucoside 45 of the invention) comprising cucurbitacins and/or their derivatives are present, as a whole, as percentage by glucoside derivatives, wherein cucurbitacins D, I, L and weight in an amount between 25 and 50% of the total A and their glucoside derivatives are present, as a whole, weight of the cucurbitacins and their glucoside deriva- in terms of percentage by weight, in an amount greater tives. than 25% of the total weight of the cucurbitacins and their [0073] In a still more preferred embodiment of the in- 50 glucoside derivatives, and wherein cucurbitacins B, E vention, the composition of the invention further compris- and the derivative of cucurbitacin E, ecballic acid, are es: each present at a concentration below 2.5% of the total weight of the cucurbitacins and their glucoside deriva- a. Cucurbitacin O; tives, wherein this composition is obtainable by a leach- b. Cucurbitacin P; 55 ing process comprising the steps of: c. Cucurbitacin C; d. Cucurbitacin H; c. Obtaining a plant portion of a plant pertaining to e. Cucurbitacin J; the cucurbitaceae family and

8 15 EP 2 666 461 B1 16

d. extracting the vegetable cucurbitacins from said demonstrated utility in the treatment of hair loss and ac- portion by a heating process and by using as an ex- cordingly these compounds may be added to the com- tractant a non-polar liquid phase capable of solubi- position of any of the previous aspects. lising said compounds. [0088] Anti-fungal materials have also demonstrated 5 utility in the treatment of hair loss. Accordingly, the com- [0079] In a preferred embodiment of this aspect of the position of any of the previous aspects may comprise an invention, the plant portion pertaining to the cucurbitace- anti-fungal material. Suitable antifungal materials include ae family is Ecballium elaterium. azole antifungal agents such as ketoconazole, micona- [0080] In another embodiment of this aspect of the in- zole, clotrimazole, econazole, bifonazole, butoconazole, vention, the heating process is conducted at a tempera- 10 fenticonazole, isoconazole, oxiconazole, sertaconazole, ture from 120 °C to 180 °C, preferably at a temperature sulconazole, tioconazole, fluconazole, itraconazole, isa- of approximately 150 °C. vuconazole, ravuconazole, posaconazole, voriconazole [0081] In another embodiment of this aspect of the in- and terconazolepolyene; polyene antifungal agents such vention, the plant portion is selected from the group con- as natamycin, rimocidin, filipin, nystatin, amphotericin B sisting of: roots, stems, leaves or fruits or any combina- 15 and candicin; allylamines such as terbinafine, amorolf- tion thereof. Preferably the plant portion is a root. ine, naftifine and butenafine; echinocandin antifungal [0082] In a still further embodiment of this aspect of agents such as anidulafungin, caspofungin and micafun- the invention, the extractant comprises phenols or phe- gin, and other known antifungal agents such as benzoic nolic acids, preferably the extractant is olive oil, more acid in combination with a keratolytic agent, ciclopirox, preferably the extractant is enriched with natural or added 20 flucytosine, griseofulvin, gentian violet haloprogintolnaf- content of antioxidants. tate, undecylenic acid and tea tree oil. The azole antifun- [0083] A fourth aspect of the invention refers to the gal agents are of particular utility since they also inhibit composition of any of the previous aspects for its use in the synthesis of testosterone, which function can also be therapy. useful in the treatment or prevention of hair loss condi- [0084] A fifth aspect of the invention refers to the com- 25 tions and disorders as described above. position of any of the previous aspects for its use in the [0089] Anti-inflammatory agents are effective at reduc- treatment or prevention of hair loss. Disorders and con- ing inflammation and pain and may have a secondary ditions that can result in hair loss include hereditary and effect on hair loss disorders. Anti-inflammatory agents congenital alopecia, hypotrichosis, trichotillomania, tinea suitable for use in the composition of any of the previous capitis, telogen effluvium, anagen effluvium, monilethrix, 30 aspects include, but are not limited to, 2-arylpropionic trichorrhexis nodosa (trichonodosis), infections, which in- acids (profens), salicylates, arylalkanoic acids, N-arylan- volve the hair or scalp such as mycotic infection, infes- thranilic acids (fenamic acids), pyrazolidine derivatives, tations involving the hair or scalp, endocrine conditions oxicams, COX-2 inhibitors, sulphonanilides and other such as hyperthyroidism, hypothyroidism and diabetes known anti-inflammatory agents such as licofelone and mellitus, lupus, hormone imbalance, for example as a 35 omega-3 fatty acids. Most preferably ibuprofen is com- result of polycystic ovary syndrome, and sebaceous prised as the anti-inflammatory agent. cysts. [0090] Antipruritic agents or constituents that provide [0085] Preferably, the composition is for use in the a mild analgesic may be added to the composition to treatment or prevention of alopecia or hypotrichosis. reduce associated side effects such as irritation or itch- [0086] It has been reported that reduced production of 40 ing. Such compositions include, but are not limited to zinc androgens, estrogens and epidermal growth factor oxide (calamine), methol, phenol and camphor. (EDG) can be useful in the treatment and prevention of [0091] Preferably, the composition is suitable for topi- hair loss. Accordingly, steroids which block or inhibit a cal administration to a patient. The compositions con- hormone or factor associated with hair loss can optionally templated by this invention include compositions adopt- be added to the composition of any of the previous as- 45 ed for topical application to the human scalp and/or skin. pects. As an example, 5[alpha]-reductase inhibitors Conventional composition forms for this purpose include block the conversion of testosterone to the active 5[alpha] ointments, lotions, pastes, jellies, gels, mousses, sprays, -dihydrotesterone (DHT) form and have been used in the foams, aerosols, powders and similar known composi- treatment of hair loss disorders. Suitable steroids for the tion forms. reduced production and inhibition of these compounds 50 [0092] Therefore, the composition may also comprise and factors include testolactone, pregnenolone, dehy- a suitable carrier or diluent. Suitable carriers or diluents droepiandrosterone (DHEA) diosgenin, spironolactone, may be aqueous and/or alcoholic and may include a vis- finasteride and tamoxifen. cous base to retain in situ the composition in use. Suitable [0087] Indole-based compounds such as indole, me- diluents for use as carriers to form a lotion include water latonin, N-[2-(5-methoxy-1H-indol-3-yl)ethyl]acetamide, 55 and lower alcohols or polyols, such as methanol, ethanol, skatole and indole-3-carbinol; certain bioflavenoids such isopropanol, glycerol or propylene glycol. To form a as quercetin methyl chalcone, and anti-inflammatory fat- cream or ointment a paraffinic fraction and an emulsion ty acids such as TES TRIOLATE, or PX-13 have also base may be used. The carrier may also comprise other

9 17 EP 2 666 461 B1 18 conventional carriers or diluents, for example, glucose, tion include, but are not limited to, parabens such as me- lactose, corn starch, starch paste, gum acacia, gelatin, thyl and propyl paraben, sorbic acid, potassium sorbate, mannitol, magnesium trisilicate, potato starch, urea, ker- quaternium-15, methylchloroisothiazolinone, and iodo- atin or colloidal silica. propynyl butylcarbamate (IPBC) and natural preserva- [0093] The term "ointment" includes creams having 5 tives such as citrus oils. oleaginous water soluble emulsion bases, for example [0104] The composition may additionally comprise a lanolin, petrolatum, glycols, glycerin and similar. skin absorption enhancer to facilitate absorption of the [0094] The compounds may be liposomal preparations composition through the skin of the patient. Suitable skin or liquid emulsions or be dissolved in conventional sol- absorption enhancers for this purpose include pentane vents such as acetonitrile, dimethylformamide (DMF), 10 1,5-diol, N-dodecyl-2-pyrrolidone and its acetate ana- dimethylacetamide (DMA), or alcohols such as propanol logue, fatty acids such as oleic acid, terpenes, esters and similar. such as isopropyl myristate, khellin and khellin ana- [0095] Optionally, the composition may be adapted for logues, methyl nicotinate, MSM-decylmethylsulfoxide, oral administration to a patient. Conventional composi- diethylene glycol, citric acid, pyruvic acid, phenoxyetha- tion forms for this purpose include tablets, coated tablets, 15 nol, transcutol, phosphatidyl choline, a medium chain caplets, troches, lozenges, dispersions, suspensions triglyceride oil (MCT oil) and water. and capsules and similar known composition forms. [0105] Administration of the composition to the affect- [0096] Alternatively, the composition may be adapted ed area of a patient may be carried out in any manner for parenteral administration. Conventional routes of pa- which will result in delivery of an effective amount of the rental administration include intravenous, intramuscular, 20 composition to the affected area of the patient. Prefera- intralesional, intraarterial, subcutaneous, intradermal, bly, the composition is applied topically to the affected transdermal, transmucosal and inhalational and similar area of the patient, which may be the scalp or another known routes. Preferably, the composition is adminis- body surface. Preferably, the composition is adminis- tered by intramuscular or intralesional routes. tered topically to the affected area or areas between one [0097] The composition may further comprise at least 25 and three times daily, and most preferably twice daily. one component selected from the group consisting of a [0106] Preferably, the composition in the form of a mineral supplement, a vitamin supplement, essential shampoo may be used daily for treatment of the condition oils, fragrances, colouring agents, preservatives and a either alone, or in combination with the topical adminis- skin absorption enhancer. tration. [0098] Suitable mineral supplements for use in the30 [0107] Thus, a sixth aspect of the present invention composition include, but are not limited to, iron, zinc, cop- refers to a shampoo composition (from hereinafter sham- per, magnesium and calcium and combinations thereof. poo composition of the invention) comprising: [0099] Suitable vitamin supplements for use in the composition include, but are not limited to, vitamin A (reti- c. A cleansing base; and nol), B group vitamins such as vitamin B1 (thiamine), 35 d. Active ingredients, vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine pyridoxal or wherein at least one of the active ingredients comprises pyridoxamine), B7 (Biotin), B9 (folic acid) and B12 (co- the composition as defined in any the previous aspects. balamin), vitamin C (ascorbic acid), vitamin D (calciferol), [0108] The cleansing base is constituted by sur- vitamin E (tocopherol) and vitamin K (phyllochinone) and 40 phactants preferably of the types alkyl sulphates and/or combinations thereof. an alkyl ether sulphates. [0100] Suitable essential oils for use in the composition [0109] The term "active ingredients" are the condition- include, but are not limited to, jasmine oil, tea tree oil and ing agents whose roles are to provide care for and beau- citrus oils. tification of the hair, and the specific active ingredients [0101] Suitable fragrances for use in the composition 45 to mitigate and correct an imbalance affecting the scalp include menthol, benzyl alcohol, eugenol, phenoxyetha- such as the composition of the invention. nol, isopropyl palmitate, isopropyl myristate, benzyl sal- [0110] In a preferred embodiment, the shampoo com- icylate, phenylethyl salicylate, thymol, isoamyl salicylate, position of the invention further comprises any of the fol- triton X-100 surfactant, benzoic acid, benzyl benzoate, lowing components or any combination thereof: methyl salicylate, phenol, oleic acid, caproic acid or car- 50 baryl. f. Thickeners for surfactant preparation; [0102] Conventional colouring agents suitable for use g. Pearlescents and opacifiers; in the composition include, but are not limited to, tartra- h. Perfumes; zine, quinoline yellow, sunset yellow, amaranth, ponceau i. Colorants; or 4R, erythrosine, red 2G, allura red AC, patent blue V, 55 j. Stabilizing agents. indigo carmine, brilliant blue FCF, fast green FCF, green S and iron oxides. [0111] In a further embodiment, the shampoo compo- [0103] Suitable preservatives for use in the composi- sition of the invention is an anti-dandruff composition

10 19 EP 2 666 461 B1 20 comprising active ingredients capable of re-establishing rized with 10 bar by opening the inlet valve V1 and closing the natural microbial flora on the skin surface. the restrictor valve V2 to assure the liquid state of the [0112] In another embodiment, the shampoo compo- extractant. Subsequently, the temperature in the oven sition of the invention is a shampoo for greasy hair com- was increased to the working temperature (150 °C) and 5 prising a cleansing base capable of eliminating excess kept under these conditions for 15 min. Then, valve V1 sebum. was switched to allow N2 to push the extract, which was [0113] In yet another embodiment, the shampoo com- cooled in coil c and finally collected in erp. position of the invention is for sensitive scalp and comprises mild anionic surphactants and amphoterics and/or Example 2: Analysis of different extracts obtained as il- non-ionic surphactants. 10 lustrated in example 1 [0114] In a still further embodiment, the shampoo composition of the invention is for dry hair and comprises: [0120] The total content of cucurbitacins and their glucoside derivatives in the erp extracts, as measured by d. A fat absorber; and/or liquid chromatography-mass spectrometry using a high e. An abrasive to dislodge soil; and/or 15 accuracy time of flight mass analyzer (limit of detection f. An alkalizer of the method: 0.5-1.0 mg/L), was determined to be in the range from 200 mg/L to 1 g/L. [0115] The frequency of the application as well as the [0121] On average the most concentrated cucur- application time may be varied according to the individual bitacins in the extracts were the D, I, L and A forms, which objective. For example, the hair loss composition of the 20 constituted approximately, as a whole, from 25 to 50% invention may be applied as a shampoo which is imme- of the total weight of the cucurbitacins and their glucoside diately rinsed out or may be applied as a lotion, shampoo, derivatives. Other cucurbitacins such as cucurbitacin O, ointment or gel which is applied to the affected area and P, C, H, J, F and Q were also present at lower concen- left to penetrate into the skin for a period of time. The trations (each of these cucurbitacins were present in an ointment formulations are particularly suitable for use on 25 amount greater than 2.5% of the total weight of the cu- facial areas such as the eyebrows or beard, whereas the curbitacins and their glucoside derivatives). However, it spray formulations are preferred for diffuse types of hair is worth pointing out that cucurbitacins B, E and the de- loss such as male or female pattern hair loss or telogen rivative of cucurbitacin E, ecballic acid, which are the effluvium. Gel and lotion formulations are suitable for all most concentrated cucurbitacins or derivatives in vegetal types of hair loss. Liposome formulations may be used 30 material obtainable from Ecballium elaterium, were de- for all types of hair loss, but are particularly suitable in tected at trace levels (concentration below 2.5% of the cases where the patient is devoid of any existing hair, for total weight of the cucurbitacins and their glucoside de- examplesevere cases ofalopecia areata, alopecia totalis rivatives, in this specific example below 5 mg/L of the and alopecia universalis. total volume of the extract) or not detected in the extracts [0116] In each case, it is recommended that the hair 35 with the method used here due to hydrolysis during the treatment composition is used on a regular basis and for leaching process. a period of time to meet the specified objective of prevention of hair loss and/or stimulation of regrowth. Example 3: Preparation of 250 mL of the shampoo com- [0117] An eighth aspect of the present invention refers position of the invention. to the shampoo of the invention or to the lotion of the 40 invention, for its use in the treatment or prevention of hair [0122] The ingredients used for the preparation of 250 loss. mL of the shampoo composition of the invention were: [0118] The following examples serve to illustrate the presentinvention; these examples arein no way intended Example 3.a. Neutral shampoo to limit the scope of the invention. 45 [0123] Examples 7% Ether lauryl sodium sulphate Example 1: Preparation of the composition of the inven- 5% Cocamidopropyl betaine tion. 50 1.5% Sulfosuccinic acid 0.15% polyquatemium-10# [0119] The procedure is based on the extraction sys- 0.4% Extract obtained in example 1. tem shown in Fig. 1 in which 5 g of the root of Ecbalium Elaterium was placed in the extraction cell ec. The cell Example 3.b. Antiseborrheic shampoo was assembled, as the scheme shows, and located in 55 the oven o. The cell was filled with extra virgin olive oil [0124] (EVOO) as extraction solvent with the high-pressure pump (EVOO volume 20 mL). The system was pressu- 7.5% Ether lauryl sodium sulphate

11 21 EP 2 666 461 B1 22

5.5% Cocamidopropyl betaine and after treatment for a period of 6 months. 1.25% Sulfosuccinic acid [0132] As shown, the patient experienced substantial 0.15% polyquatemium-10# hair recovery after treatment. 0.5% Zinc pyrithione 0.4% Extract obtained in example 1. 5 Example 6: Treatment of alopecia areata.

[0125] In both cases, the ingredients were mixed by [0133] The affected zone from a man suffering from stirring in order to obtained a homogenous solution, alopecia areata was treated two times per week for a which was thus ready for application. period of two months. The treatment was as follows: 10 Example 4: Treatment of alopecia caused by hormone • Cleaning the head skin with ozone vapour for 5 min. deregulations. • Washing with the shampoo of example 3a. • Stimulation of blood circulation in the affected zone [0126] A woman with alopecia attributed to a post-par- by acupuncture (needle hits). tum hormone imbalance was treated with the shampoo 15 • Absorption favoured by application of a low-frequen- composition of example 3a. cy current. [0127] In this case the treatment was carried out three • Washing with the shampoo of example 3a. times per week for a period of one year, with daily use of the recommended shampoo. Each treatment session [0134] Figure 4 illustrates the results before and after involved: 20 treatment. [0135] As shown, hair regeneration was achieved after a. Cleaning the head skin with ozone vapour for 5 treatment. min. b. Washing with the shampoo of example 3a. Example 7: Treatment of seborrheic dermatitis andro- c. Stimulation of blood circulation in the affected zone 25 genic alopecia by acupuncture (needle hits). d. Absorption favoured by application of a low-fre- [0136] In this example a man suffering from seborrheic quency current. dermatitis androgenic alopecia having significant hair e. Washing with the shampoo of example 3a. loss (> 100 hairs / day) was treated with the shampoo 30 composition of example 3b. [0128] Figure 2 shows the initial state, evolution after [0137] During the first week of treatment involved treatment for 6 months, and final results after one year washing the hair with the shampoo composition of ex- treatment. ample 3b twice daily, leaving the shampoo for 5 minutes [0129] The results illustrate that after one-year of treat- before rinsing. During the second week of treatment, the ment the affected zone was completely recovered. 35 used of the shampoo composition was limited to once per day. Example 5: Treatment of androgenetic alopecia or alo- [0138] After the second week of treatment we ob- pecia of seborrheic origin. serveda significant reduction in hairloss (< 80 hairs /day). Treatment was maintained for 6 months, observing 25% [0130] Androgeneticalopecia oralopecia of seborrheic 40 hair regeneration. origin is the most common type of alopecia affecting the masculin gender. In this case, the patient suffered from Example 8: Treatment of pityriasis rosea seborrhea with deterioration of the capilar ramifications. Thetreatment inthis case consistedin a chemical peeling [0139] In this example, a man suffering from pityriasis by using ethanol containing 1% iodine and cristallized 45 rosea was treated with the shampoo composition of ex- mint in ozone vapour. The objective of this first step was ample 3a. As with example 7 the treatment involved to removed epidemial layers and renew them with higher washing the hair with the shampoo composition of ex- quality layers. After this step, the treatment was similar ample 3a twice daily, leaving the shampoo for 5 minutes to the one exemplified in the previous example, namely: before rinsing. During the second week of treatment, the 50 used of the shampoo composition was limited to once • Washing with the shampoo of example 3b. per day. The treatment was maintained for 6 months, • Stimulation of blood circulation in the affected zone observing 50% hair regeneration. by acupuncture (needle hits). • Absorption favoured by application of a low-frequency current. 55 Claims • Washing with the shampoo of example 3b. 1. A composition comprising cucurbitacins and their [0131] Figure 3 illustrates the results before treatment glucoside derivatives, wherein cucurbitacins D, I, L

12 23 EP 2 666 461 B1 24

and A and their glucoside derivatives are present, 9. The composition of any one of claims 7 or 8, wherein as a whole, in terms of percentage by weight in an the heating process is conducted at a temperature amount greater than 25% of the total weight of the from 120 °C to 180 °C. cucurbitacins and their glucoside derivatives, and wherein cucurbitacins B, E and the derivative of cu- 5 10. The composition of any one of claims 7 to 9, wherein curbitacin E, ecballic acid, are each present at a con- the plant portion is a root. centration below 2.5% of the total weight of the cucurbitacins and their glucoside derivatives. 11. The composition according to any one of claims 7 to 10, wherein the extractant is olive oil. 2. The composition of claim 1, wherein cucurbitacins 10 D, I, L and A and their glucoside derivatives, as a 12. A shampoo composition which comprises: whole, are present in amounts greater than 35% of the total weight of the cucurbitacins and their gluco- a. A cleansing base; and side derivatives. b. Active ingredients, 15 3. The composition of claim 1, wherein cucurbitacins wherein at least one of the active ingredients com- D, I, L and A and their glucoside derivatives, as a prises the composition as defined in any of claims 1 whole, are present in amounts from 25 to 50% of the to 11. total weight of the cucurbitacins and their glucoside derivatives. 20 13. A hair lotion comprising the composition as defined in any of claims 1 to 11. 4. The composition of any one of claims 1 to 3, wherein the composition further comprises: 14. The composition of any one of claims 1 to 11, for its use in therapy. a. Cucurbitacin O; 25 b. Cucurbitacin P; 15. The composition of any one of claims 1 to 11 or the c. Cucurbitacin C; shampoo composition of claim 12 or the hair lotion d. Cucurbitacin H; of claim 13, for its use in the treatment or prevention e. Cucurbitacin J; of hair loss. f. Cucurbitacin F; and 30 g. Cucurbitacin Q; Patentansprüche wherein each of these compounds is present in an amount greater than 2.5% of the total weight of the 1. Zusammensetzung, die Cucurbitacine und deren cucurbitacins and their glucoside derivatives. 35 Glykosid-Derivate umfasst, wobei Cucurbitacine D, I, L und A und deren Glykosid-Derivate insgesamt 5. The composition of anyone of claims 1 to 4, wherein in Gewichtsprozent in einer Menge größer als 25% the composition further comprises olive oil. des Gesamtgewichtes der Cucurbitacine und deren Glykosid-Derivate vorliegen, und wobei Cucurbita- 6. AnEcbalium Elateriumvegetable extract which com- 40 cine B, E und das Derivat von Cucurbitacin E, Ec- prisesa composition as defined in any of theprevious ballinsäure, jeweils in einer Konzentration von unter claims. 2,5% des Gesamtgewichtes der Cucurbitacine und deren Glykosid-Derivate vorliegen. 7. The composition of anyone of claims 1 to 4, wherein the composition is obtainable by a leaching process 45 2. Zusammensetzung nach Anspruch 1, wobei Cucur- comprising the steps of: bitacine D, I, L und A und deren Glykosid-Derivate insgesamt in Mengen größer als 35% des Gesamt- h. Obtaining a plant portion of a plant pertaining gewichtes der Cucurbitacine und deren Glykosid- to the cucurbitaceae family and Derivate vorliegen. i. extracting the vegetable cucurbitacins from 50 said portion by a heating process and by using 3. Zusammensetzung nach Anspruch 1, wobei Cucur- as an extractant a non-polar liquid phase capa- bitacine D, I, L und A und deren Glykosid-Derivate ble of solubilising said compounds. insgesamt in Mengen von 25 bis 50% des Gesamt- gewichtes der Cucurbitacine und deren Glykosid- 8. The composition of claim 7, wherein the plant portion 55 Derivate vorliegen. pertaining to the cucurbitaceae family is Ecballium elaterium. 4. Zusammensetzung nach einem der Ansprüche 1 bis 3, wobei die Zusammensetzung ferner:

13 25 EP 2 666 461 B1 26

a. Cucurbitacin O; 14. Zusammensetzung nach einem der Ansprüche 1 bis b. Cucurbitacin P; 11, für dessen Verwendung in der Therapie. c. Cucurbitacin C; d. Cucurbitacin H; 15. Zusammensetzung nach einem der Ansprüche 1 bis e. Cucurbitacin J; 5 11 oder Haarwaschmittelzusammensetzung nach f. Cucurbitacin F; und Anspruch 12 oder Haarlotion nach Anspruch 13, für g. Cucurbitacin Q; dessen Verwendung bei der Behandlung oder der Vorbeugung gegen Haarausfall. umfasst, wobei jede dieser Verbindungen in einer Menge größer als 2,5% des Gesamtgewichtes der 10 Cucurbitacine und deren Glykosid-Derivate vorliegt. Revendications

5. Zusammensetzung nach einem der Ansprüche 1 bis 1. Compositioncomprenant des cucurbitacineset leurs 4, wobei die Zusammensetzung ferner Olivenöl um- dérivés glucosidiques, dans laquelle les cucurbita- fasst. 15 cines D, I, L et A et leurs dérivés glucosidiques sont présents, dans leur ensemble, en termes de pour- 6. Ecballium elaterium - Pflanzenextrakt, der eine Zu- centage en poids dans une quantité supérieure à 25 sammensetzung nach einem der vorhergehenden % du poids total des cucurbitacines et de leurs dé- Ansprüche umfasst. rivés glucosidiques, et dans laquelle les cucurbita- 20 cines B, E et le dérivé de cucurbitacine E, l’acide 7. Zusammensetzung nach einem der Ansprüche 1 bis ecballique, sont chacun présents à une concentra- 4, wobei die Zusammensetzung durch ein Auslau- tion inférieure à 2,5 % du poids total des cucurbita- gungsverfahren erhältlich ist, das folgende Schritte cines et de leurs dérivés glucosidiques. umfasst: 25 2. Composition de la revendication 1, dans laquelle des h. Erhalten eines Pflanzenteils einer der Cucur- cucurbitacines D, I, L et A et leurs dérivés glucosi- bitaceae-Familie angehörenden Pflanze, und diques, dans leur ensemble, sont présents dans des i. Extrahieren der pflanzlichen Cucurbitacine quantités supérieures à 35 % du poids total des cu- dieses Teils mittels eines Erwärmungsverfah- curbitacines et de leurs dérivés glucosidiques. rens und indem als Extrationsmittel eine nicht- 30 polare flüssige Phase verwendet wird, die in der 3. Composition de la revendication 1, dans laquelle des Lage ist, diese Verbindungen aufzulösen. cucurbitacines D, I, L et A et leurs dérivés glucosidiques, dans leur ensemble, sont présents dans des 8. Zusammensetzung nach Anspruch 7, wobei der der quantités comprises entre 25 et 50 % du poids total Cucurbitaceae-Familie angehörende Pflanzenteil 35 descucurbitacines et de leursdérivés glucosidiques. Ecballium elaterium ist. 4. Composition de l’une quelconque des revendica- 9. Zusammensetzung nach einem der Ansprüche 7 tions 1 à 3, dans laquelle la composition comprend oder 8, wobei das Erwärmungsverfahren bei einer en outre : Temperatur von 120 °C bis 180 °C durchgeführt wird. 40 a. Cucurbitacine O ; 10. Zusammensetzung nach einem der Ansprüche 7 bis b. Cucurbitacine P ; 9, wobei der Pflanzenteil eine Wurzel ist. c. Cucurbitacine C ; d. Cucurbitacine H ; 11. Zusammensetzung nach einem der Ansprüche 7 bis 45 e. Cucurbitacine J ; 10, wobei das Extraktionsmittel Olivenöl ist. f. Cucurbitacine F ; et g. Cucurbitacine Q ; 12. Haarwaschmittelzusammensetzung, umfassend: dans laquelle chacun de ces composés est présent a. eine Reinigungsgrundlage; und 50 dans une quantité supérieure à 2,5 % du poids total b. Wirkstoffe, descucurbitacines et de leursdérivés glucosidiques.

wobei mindestens einer der Wirkstoffe die Zusam- 5. Composition de l’une quelconque des revendica- mensetzung nach einem der Ansprüche 1 bis 11 um- tions 1 à 4, dans laquelle la composition comprend fasst. 55 en outre de l’huile d’olive.

13. Haarlotion, welche die Zusammensetzung nach ei- 6. Extrait végétal d’Ecballium elaterium qui comprend nem der Ansprüche 1 bis 11 umfasst. une composition telle que définie dans l’une quel-

14 27 EP 2 666 461 B1 28

conque des revendications précédentes.

7. Composition de l’une quelconque des revendications 1 à 4, dans laquelle la composition est obtena- ble par un procédé de lixiviation comprenant les éta- 5 pes de :

h.Obtention d’une portionde plante d’uneplante appartenante à la famille des Cucurbitaceae et i. extraction des cucurbitacines végétales de la- 10 dite portion par un procédé de chauffage et en utilisant, comme agent d’extraction, une phase liquide non polaire capable de solubiliser lesdits composés. 15 8. Composition de la revendication 7, dans laquelle la portion de plante appartenante à la famille des Cu- curbitaceae est l’Ecballium elaterium.

9. Composition de l’une quelconque des revendica- 20 tions 7 ou 8, dans laquelle le procédé de chauffage est réalisé à une température comprise entre 120 °C et 180 °C.

10. Composition de l’une quelconque des revendica- 25 tions 7 à 9, dans laquelle la portion de plante est une racine.

11. Composition selon l’une quelconque des revendications 7 à 10, dans laquelle l’agent d’extraction est de 30 l’huile d’olive.

12. Composition de shampoing qui comprend :

a. Une base nettoyante ; et 35 b. Des ingrédients actifs,

dans laquelle au moins l’un des ingrédients actifs comprend la composition telle que définie dans l’une quelconque des revendications 1 à 11. 40

13. Lotion capillaire comprenant la composition telle que définie dans l’une quelconque des revendications 1 à 11. 45 14. Composition de l’une quelconque des revendications 1 à 11, pour son utilisation en thérapie.

15. Composition de l’une quelconque des revendications 1 à 11 ou la composition de shampoing de la 50 revendication 12 ou la lotion capillaire de la revendication 13, pour son utilisation dans le traitement ou la prévention de la perte des cheveux.

15 EP 2 666 461 B1

16 EP 2 666 461 B1

17 EP 2 666 461 B1

REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

Non-patent literature cited in the description

•PORTER; BURTON. Br. J. Dermatol., 1971, vol. 85, • GREIGE-GERGES et al. Chemico-Biological Inter- 272-273 [0006] actions, 2007, vol. 169, 53-62 [0060] • ROKHSAR et al. J Am Acad Dermatol, 1998, vol. 39, 751-761 [0007]