WO2013020719A2

WO2013020719A2 - New use of compositions to delay the onset of the catagen phase of the hair

Info

Publication number: WO2013020719A2
Application number: PCT/EP2012/053929
Authority: WO
Inventors: Saad Harti
Original assignee: Legacy Healthcare Holding Ltd
Priority date: 2011-08-09
Filing date: 2012-03-07
Publication date: 2013-02-14
Also published as: WO2013020719A3; AR087488A1

Abstract

A new use of compositions administered by topical route, containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma, to increase to its normal level the intracellular level of the anti-apoptotic protein Bcl-2 so as to delay the onset of the catagen phase of the hair cycle. A method of intervention to increase to its normal level the intracellular level of the anti-apoptotic protein Bcl-2 so as to delay the onset of the catagen phase of the hair cycle comprising the administration by topical route of compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma.

Description

NEW USE OF COMPOSITIONS TO DELAY THE ONSET OF THE CATAGEN PHASE

OF THE HAIR

BACKGROUND AND SUMMARY

The invention concerns a new use of compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species, to delay the onset of the catagen phase of the hair and methods thereof. More particularly the invention concerns a new use of compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species, to increase to its normal level the intracellular level of Bcl-2 anti-apoptotic protein in the hair follicle in order to delay the onset of the catagen phase of the hair cycle, resulting in a prolongation of the growth phase, or anagen phase of the hair.

From patent application WO 2008/1 13912 it is known that, compositions containing as active ingredient an extract of Allium species, an extract of Citrus species and

- either an extract of Paullinia species and an extract of Theobroma species

- or an extract of Salix species and zinc sulphate increase the hair growth

Hair loss disorders, such as Androgenic Alopecia, Telogen Effluvium and Alopecia Areata originate from the premature on-set of the catagen phase of the hair.

The catagen phase consists in the detachment of the hair from the dermal papilla, due of the entry into apoptosis (death process) of the cells linking the hair to the dermal papilla. It has been shown that application of compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species enables the increase of BCL2 (an anti-apoptotic protein) to its normal level, hence enabling the cells linking the hair to the dermal papilla not to enter into premature apoptosis, provoked by androgenic alopecia, telogen effluvium and alopecia areata.

The delay of the premature onset of the catagen phase enables the hair remain into its anagen phase longer, and therefore grow longer and thicker.

Hair growth is a complex and successive cyclical process involving DNA, RNA and protein synthesis , hair growth, regression of the hair follicle (HF), detachment of hair from the dermal papilla and finally shedding of the hair shaft. The hair cycle is thus represented by the anagen, followed by catagen, telogen, exogen (also called teloptosis) and kenogen phases. During anagen phase, the hair actively grows, with a highly increased cell proliferation rate in all epithelial compartments of hair follicle. It is subdivided into 6 successive substages. The first 5 anagen phases (I to V) correspond to the hair follicle regeneration and the anagen VI phase represents the continuous visible hair growth. Predetermined genetically, such an Anagen growth phase can last from a few weeks to several years. At this stage, dermal papilla cells are fibroblastic in shape, widely interspersed by large amounts of extracellular matrix, and are supported by blood vessels. Following this growth period, the hair follicle stops its proliferative activity and enters the degenerative catagen phase characterized with massive cell apoptosis in hair follicle, starting from the melanogenic area (late anagen), propagating to the hair matrix (early catagen) and then to the proximal/central outer and inner root sheaths and hair shaft (mid- and late catagen), leading to regression of the hair follicle and the hair gets gradually detached from the dermal papilla. However, apoptotic cells are never seen in the dermal papilla of catagen HFs (Lindner et al., 1997; Matsuo et al., 1998; Soma et al., 1998) although the extracellular matrix of the dermal papilla is markedly diminished, tightly packed with greatly reduced intercellular space, and cells in lower portion of the hair follicle attach to the hair shaft. The next telogen phase is the ultimate period of the hair cycle during which the hair is fully keratinized and the hair follicle has diminished to half of its previous size. The dermal papilla now has much reduced extracellular matrix and is no longer enveloped by epithelial cells. Dermal papilla cells become quiescent as DNA, RNA and protein synthesis are insignificant, only with few exceptions such as keratin 14, to which telogen hair fiber anchors. At this stage, the hair is on its way to detach before shedding.

The exogen phase is the actual shedding of the hair. However, this is a highly controlled and active process contrary to the quiescent telogen phase. It is therefore implausible that the newly formed hair pushes the resting shaft outward to achieve shedding. Kenogen phase is the period before a new anagen hair emerges. Nevertheless, the hair follicle is still empty. The duration of "hair vacancy" is reported to be extended in androgenetic alopecia patients.

Recurrent premature termination of the Anagen phase and earlier onset of catagen of the follicular cycle results in Androgenetic Alopecia (AGA), subsequently leads to miniaturization of follicles and finally follicular loss associated with mild inflammation.

Although the pathogenetic mechanisms in AGA are not yet fully revealed, evidences indicate that androgens, especially 5-dihydrotestosterone (5-DHT), inhibit hair follicle activity leading to shortening of the anagen phase and induce earlier onset of catagen phase.

It was previously discovered that apoptosis may have an effect on alopecia (Cowper et al. 2002, Botchkarev et al. 2003, Winiarska et al. 2006, El-Domyati et al. 2010). In particular, testosterone (T) and 5-DHT induce apoptosis in the mesenchyme- derived Dermal Papilla in AGA by decreasing Bcl-2 expression at the mRNA and protein level. This was correlated with the increase of the pro-apoptotic Bax protein level (Wrobel A et al. 2008).

The Bcl-2 family members consist of pro-apoptotic (e.g., Bax and Bid) and anti- apoptotic (e.g., Bcl-2 and Bcl-XL) proteins, acting upstream of caspase activation. They form homodimers and heterodimers exercising essential roles in the regulation of apoptosis of the mitochondrial apoptotic pathway. Bcl-2 (B-cell lymphoma 2), an anti- apoptotic protein, is the founding member of the Bcl-2 family that localizes to the mitochondria (Hockenbery et al., 1990; Krajewski et al., 1993) and promotes cell survival by preventing cytochrome c release (Kim et al., 1997; Kluck et al., 1997; Yang et al., 1997; Shimizu et al., 1998; Newmeyer et al., 2000), contrary to the pro-apoptotic Bcl-2 family member Bax that promotes cytochrome c release (Jurgensmeier et al., 1998). In consequence, the cellular fate is therefore determined by the ratio of pro- apoptotic to anti-apoptotic Bcl-2 family members within the cell (Oltvai et al., 1993). Bcl-2 overexpression has been shown to prevent apoptosis of neuronal cells during normal development and experimental settings (Martinou et al., 1994, Mostafapour et al., 2002). Likewise, over-expression of Bcl-2 could also significantly increased hair cell survival in an adult mouse model (Lisa L. et al., 2003). Following application of the composition described in this invention, a statistically significant improvement in the anagen/telogen ratio on alopecia patients has been detected via Phototrichogram. Besides, such observation correlates to an increase of the anti-apoptotic protein BCL2 (+89%, almost doubling) to its normal level in the skin after application of the composition.

Documented scientific data strongly suggest that a protection against pre-mature cell apoptosis in the follicular cycle, in particular anti-apoptosis of the cells in dermal papilla and the surrounding matrix cells, as well as the connection cells between the dermal papilla in the follicle and the hair shaft, may be the compelling strategy for therapeutic intervention on hair loss.

As indicated above, there are mainly three types of hair loss summarised as follows.

Androqenetic Alopecia: commonly called male pattern hair loss, which is the result of hormone binding (testosterone and 5a-dihydrotestosterone) to the androgen receptor of the matrix cell in the dermal papilla. Such a hormonal action not only leads to the miniaturisation of anagen hairs, but also decreases the duration of anagen, inhibits hair follicle activity with early induction of the catagen and prolongation of telogen phase.

Interestingly, treatment of cultured dermal papilla cells with testosterone and 5a - dihydrotestosterone induces apoptosis in a dose-dependent and time-related manner. This is essentially due to the decreased level of Bcl-2 protein, hence an increase of the bax/bcl-2 ratio. These data suggest that local pharmacological intervention to normalize Bcl-2 level may serve as an elective treatment in androgenetic alopecia. Androgenic alopecia in women is also known as female pattern alopecia or baldness (Female Pattern Hair Loss). This is seen as hair thinning predominantly over the top and sides of the head. It affects approximately one-third of all susceptible women, but is most commonly seen after menopause, although it may begin as early as puberty.

Teloqen Effluvium: Telogen effluvium is a, generally female, form of non-scarring alopecia characterized by a large percentage of scalp hairs shifting into "shedding" phase, often with an acute onset. There exists a chronic form with a more insidious onset and a longer duration (Whiting DA et al., 1996, Sinclair R. et al., 2005). Telogen effluvium is a reactive process caused by panoply of stimuli resulting in anagen arrest and early entry of hairs into the telogen phase. The shedding may be seasonal. But it may be metabolic, hormonal, nutritional, drug-associated or stress-associated. Examples of known factors are: childbirth, severe blood loss, high fevers, major illness, severe weight loss, nutritional iron or zinc deficiency, thyroid disease and chronic renal disease.

Alopecia Areata: Alopecia Areata is an immunological disease under genetic control, which results in circular patches of complete baldness to appear on the scalp or elsewhere on the body. There are three types of Alopecia Areata; Alopecia areata, alopecia areata totalis and alopecia areata universalis. In all forms of alopecia areata, the hair follicles remain alive and are ready to resume normal hair production whenever they receive the appropriate signal or the inflammation ceases. The inflammation around the hair bulb is thought to be autoimmune in nature, causes premature anagen arrest and hair shedding. Follicle miniaturization is also one consequence.

It has been discovered that the administration by topical route of a composition containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species has an effect to delay the onset of the catagen phase. This means that hair will remain connected with the dermal papilla longer and continue its growth. As a result, this is an invention that could alter the natural annual biological process of hair shedding that takes place in all mammals, including humans.

Such a composition can be applied in order to successfully treat all types of alopecia in men and women. This composition can also be used in combination with hair transplant as well as for other prevention and treatment of alopecia.

Hence the present invention concerns a new use of topical compositions according to the invention to normalize the hair cycle. Precisely, to delay the onset of the catagen phase of the hair by increasing to its normal level the intracellular anti- apoptotic protein Bcl-2 in the hair follicle, and therefore extending the anagen phase. As the intracellular level of Bcl-2 in alopecia patients' hair follicle is abnormally low (hence unbalanced ratio of pro-apoptotic and anti-apoptotic proteins), application of this composition can normalize the ratio of pro-apoptotic / anti-apoptotic proteins, in particular, by raising the intracellular level of the anti-apoptotic protein Bcl-2. As a result, it renders the cells, and stem cells, in dermal papilla and the surrounding matrix cells, as well as the connection cells between the dermal papilla in the follicle and the hair shaft, more resistant against pre-mature cell apoptosis during the follicular cycle.

Thus the present invention concerns a new use of compositions administered by topical route, containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species, to normalize the hair cycle of hair loss patients by delaying the onset of the catagen phase of hair, thanks to an increase to its normal level the intracellular Bcl-2 anti-apoptotic protein in the hair follicle, and therefore lengthening anagen phase. The present invention concerns a new use of compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species in the Androgenic Alopecia, Telogen Effluvium and Alopecia Areata, Cicatricial Alopecia and extending the life of hair implants.

Another object of the invention is also a new use of topical compositions containing an aqueous-alcoholic extract of Allium species, an aqueous-alcoholic extract of Citrus species, an aqueous alcoholic extract (atomised or not) of Paullinia species and an aqueous-alcoholic extract (atomised or not) of Theobroma species, to increase to its normal level the intracellular level of Bcl-2 anti-apoptotic protein.

Among the new use of topical compositions according to the invention to delay the onset of the catagen phase of hair, those which are of more particular interest are the compositions in which the preferred topical composition contains from 65% to 93% of an aqueous-alcoholic extract of Allium species, from 5% to 33% of an aqueous- alcoholic extract of Citrus species, from 0.25% to 2.5% of an aqueous alcoholic extract (atomised or not) of Paullinia species and from 0.25% to 2.5% of an aqueous-alcoholic extract (atomised or not) of Theobroma species. Among the new use of compositions of the invention to delay the onset of the catagen phase of hair, according to the invention, those which are the more preferred interest are the uses in which the compositions are containing from 65% to 93% of an aqueous-alcoholic extract of Allium species, from 5% to 33% of an aqueous-alcoholic extract of Citrus species, from 0.25% to 2.5% of an aqueous-alcoholic extract (atomised or not) of Paullinia species and from 0.25% to 2.5% of an aqueous-alcoholic extract (atomised or not) of Theobroma species and especially those containing from 65% to 93% of an aqueous-alcoholic extract of Allium cepa, from 5% to 33% of an aqueous-alcoholic extract of Citrus lemon, from 0.25% to 2.5% of an aqueous-alcoholic extract (atomised or not) of Paullinia species and from 0.25% to 2.5% of an aqueous-alcoholic extract (atomised or not) of Theobroma species.

The term extract of Allium species or aqueous-alcoholic extract of Allium species refers particularly to aqueous-alcoholic extracts and native extracts obtained from all species of the genus Allium (family Liliaceae) and especially Allium cepa. The term extract of Citrus species or aqueous-alcoholic extract of Citrus species refers particularly to aqueous-alcoholic extracts and native extracts obtained from all species of the genus Citrus (family Rutaceae) and especially Citrus lemon. The term extract (atomised or not) of Paullinia species or aqueous-alcoholic extract (atomised or not) of Paullinia species refers particularly to aqueous-alcoholic extracts and native extracts obtained from all species of the genus Paullinia (family Sapindaceae) and especially Paullinia cupana. The term extract (atomised or not) of Theobroma species or aqueous-alcoholic extract (atomised or not) of Theobroma species refers particularly to aqueous-alcoholic extracts and native extracts obtained from all species of the genus Theobroma (family Malvaceae) and especially Theobroma cacao.

Finally, the invention relates a method of intervention to increase to its normal level of the intracellular anti-apoptotic protein Bcl-2 and to delay therefore the onset of the catagen phase of the hair cycle comprising the administration by topical route of a composition containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species, more particularly in the treatment of Androgenic Alopecia, Telogen Effluvium, Alopecia Areata, Cicatricial Alopecia and extending the life of hair implants.

The compositions of the invention can be used as a drug in the treatment of: male androgenetic alopecia (male balding, male pattern balding, male pattern hair loss), Female pattern hair loss (androgenetic alopecia, androgenic alopecia, common baldness in women, diffuse alopecia in women, female pattern baldness, diffuse hormonal alopecia )

Primary cicatricial alopecia (primary scarring alopecia)

Secondary cicatricial alopecia (secondary scarring alopecia, secondary fibrosing alopecia)

Alopecia areata (all forms of alopecia aerata, pelade).

The compositions of the invention can also be used as cosmetics to extend the life of hair implants.

The most preferred compositions used according to the invention are: those containing approximately 87% of an aqueous-alcoholic extract of Allium cepa, approximately 12% of an aqueous-alcoholic extract of Citrus lemon, approximately 0.5% of an aqueous-alcoholic extract (atomised or not) of Paullinia cupana and approximately 0.5% of an aqueous-alcoholic extract (atomised or not) of Theobroma cacao,

These compositions are prepared as indicated in patent application WO 2008/1 13912. These pharmaceutical compositions are prepared by conventional methods, in which pharmaceutically inert, organic or inorganic excipients are added to the compositions obtained according to the invention. These compositions may be administered by local topical application to the skin. They may be solids or liquids, and may be presented in any of the pharmaceutical or cosmetic forms used in human medicine or cosmetics in the form of, but not limited to, ointments, solutions, sprays, lotions, films, suspensions, creams, serums or aerosols.

According to the invention the composition is administered daily during a period of several months with a composition containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species.

In order to obtain an impressive effect on the onset of the catagen phase of the hair it is necessary to perform the administration of the compositions during at least 3 months. When using the compositions obtained according to the invention, doses may vary within relatively wide limits and must be set according to the person being treated and the condition concerned. Pharmaceutical compositions normally contain from 0.2 to 500 mg, preferably from 1 to 200 mg, of active ingredients as defined above, in the form of dry extract. EXAMPLE OF TREATMENT

During a period of 4 months, the patients have received, every day on the skull a lotion containing :

- an aqueous-alcoholic extract of Allium cepa: 87.04 %

- an aqueous-alcoholic extract of Citrus lemon: 1 1 .96 %

- an atomised aqueous-alcoholic extract of Paullinia cupana: 0.50 %

- an atomised aqueous-alcoholic extract of Theobroma cacao: 0.50 %

(hereafter composition A).

This lotion has been prepared as indicated in example 1 of patent application WO 2008/1 13912.

Composition A which is a mixture of four natural ingredients has been reported to beneficially delay the onset of the catagen phase, meaning that hair will remain connected with the dermal papilla longer and continue its growth. In addition, people using the product occasionally accelerated re-growth of hair. This has suggested that composition A is capable of affecting positively the hair cycle after the shedding (telogen) process, in order to provoke a quicker restart next hair growth (anagen) phase, as such shortening the resting phase.

Examples of Compositions

As a drug

Hair Lotion

Active ingredient: Composition A ..................... 40 g per 100 ml

Excipients: aqueous ethanol at 55 °

Tretment : Application 1 -2 times daily friction

A cosmetics 1 °) Hair Lotion

Active ingredient: Composition A........................21 g per 100 ml

Ingredients: aqua, alcohol, betaine, glycerin, polysorbate 20, maltodextrin, silica.

Treatment : Application 1 to 2 times daily friction.

2°) Hair Gel

Active ingredient: Composition A..... .21 %

Excipients: 96% ethanol (10%), betaine 2%, 5% glycerol, Ultragel 300 (1 %), Sterile demineralized water qs 100%.

Treatment : Application 1 to 2 times daily massage.

3°) Shampoo

Active ingredient: Composition A 10 %

Excipients: sodium dodecyl sulfate at 25% (10%), sodium chloride, 3% citric acid 0.1 %, 0.5% imidazolydinylureum, demineralized water sterile qs 100%

Treatment : 1 shampoo every day.

4°) After Shampoo

Active ingredient: Composition A 20 %

Excipients: Montanov 68 (5%), DM Amonyl 2%, 3% dimethicone, imidazolydinyluree 0.5%, sterile demineralized water qs 100%.

Treatment : 1 shampoo every day.

Clinical Experiment

Study population

Twenty volunteers presenting with clinically diagnosed AGA were recruited by the Dermatology Ambulatory Hospital /SaoPaulo/ Brazil. The study was carried out in accordance with the Declaration of Helsinki, while laboratory analyses complied with the principles set out by the Best Laboratory Practice guidelines.

Participants had to meet the following inclusion criteria: men, ages 20-40 years, Fitzpatrick skin phototypes II and III, presenting with AGA, classified as type l-VII according to the modified Norwood-Hamilton scale, absence of treatment with minoxidil or finasteride for at least 5 months prior to recruitement, and absence of modifications in habits and hair styling during the study period.

Methods

Participants had to attend two clinical examinations for data collection and scalp biopsies (the first time before the start of treatment (W0 phase) and the second 12 weeks after treatment (W12 phase) with composition A. Side effects and cosmetic tolerance were evaluated by the investigator using a scale of 1 -3 for intensity and 1 -8 for the dermatological examination.

Biopsies from vertex scalp were obtained.

Treatment

The investigator supplied each Volunteer with a specific treatment. Specifically the solution contained 210 mg/ml of active ingredient (Composition A). The treatment began after the SO phase, with volunteers applying approximately 1 ml of the topical solution on either dry or wet scalp twice a day (total daily : 2ml) at 12 h intervals over a period of 12 weeks.

Results

Twenty participants of an average of 32.5 years (range 23-40) were recruited to take part of the study. Five participants refused to undergo scalp biopsy in the W12 phase, while one participant did not answer the questionnaire about efficacy. Evaluation by volunteers showed good effectiveness and cosmetic acceptance given that 90% of participants observed new hair growth, 63-73% observed faster hair growth, 84%observed more hair, 68 % experienced a pleasant sensation following the application and 65 % classified the product as good or very good.

Claims

1. A new use of compositions administered by topical route, containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma, to normalize the hair cycle of hair loss patients by delaying the onset of the catagen phase of hair, thanks to increase to its normal level of the intracellular anti-apoptotic protein Bcl-2 and to delay therefore the onset of the catagen phase of the hair cycle.

2. A new use of topical compositions containing an aqueous-alcoholic extract of Allium species, an aqueous-alcoholic extract of Citrus species, an aqueous alcoholic extract (atomised or not) of Paullinia species and an aqueous-alcoholic extract (atomised or not) of Theobroma species, according to claims 1 , to increase to its normal level of the intracellular anti-apoptotic protein Bcl-2.

3. A new use of topical compositions containing from 65% to 93% of an aqueous-alcoholic extract of Allium species, from 5% to 33% of an aqueous-alcoholic extract of Citrus species, from 0.25% to 2.5% of an aqueous alcoholic extract (atomised or not) of Paullinia species and from 0.25% to 2.5% of an aqueous-alcoholic extract (atomised or not) of Theobroma species, according to claims 1 and 2, to increase to its normal level of the intracellular anti-apoptotic protein Bcl-2.

4. A new use of topical compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species, to increase to its normal level of the intracellular anti-apoptotic protein Bcl-2, according to claims 1 to 3, wherein the composition is administered daily during a period of several months.

5. A new use of topical compositions containing as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma species, according to claims 1 to 4, in the Androgenic Alopecia, Telogen Effluvium, Alopecia Areata, Cicatricial Alopecia and extending the life of hair implants.

6. A new use according to claims 1 to 5, wherein the compositions contain from 0.2 to 500 mg of active ingredients in the form of dry extract.

7. A new use according to claim 6, wherein the compositions contain from 1 to 200 mg of active ingredients in the form of dry extract.

8. A method of intervention to increase to its normal level, the intracellular level of the anti-apoptotic protein Bcl-2 so as to delay the onset of the catagen phase of the hair cycle comprising the administration by topical route of a composition containg as active ingredient an extract of Allium species, an extract of Citrus species, an extract of Paullinia species and an extract of Theobroma.

9. A method according to claim 8, in the treatment of Androgenic Alopecia, Telogen Effluvium, Alopecia Areata, Cicatricial Alopecia and extending the life of hair implants.