FR3151483A1 - 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for the treatment and/or prevention of pre-cancerous or cancerous skin lesion - Google Patents

Abstract

The present invention relates to 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine or a composition comprising it for use in the prevention and/or treatment of a pre-cancerous or cancerous skin lesion; or for use in the repair of DNA damage in skin cells and/or in the prevention and/or reduction of a deficiency in repair of DNA damage in skin cells.

Description

5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for the treatment and/or prevention of pre-cancerous or cancerous skin lesion TECHNICAL FIELD OF THE INVENTION

The present invention relates to 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine, as well as a composition comprising it, for use in the prevention and/or treatment of a pre-cancerous skin lesion such as actinic keratosis or a cancerous skin lesion; or in the repair of DNA damage in skin cells and/or in the prevention and/or reduction of a deficiency in repair of DNA damage in skin cells.

STATE OF THE ART

Excessive exposure to the sun disrupts the proper functioning of skin tissue and is responsible for more or less harmful damage depending on the time and frequency of exposure. Overexposure to ultraviolet (UV) radiation, whether solar or artificial, has major harmful effects on living systems. These biological effects are mainly linked to ultraviolet B and A (UVB and UVA). In humans, UV rays are responsible for erythema, immunosuppression, and lead in the long term to the appearance of skin wrinkles (photo-aging) and more seriously to the formation of skin cancers. The toxicity of UV rays is linked to their absorption by different molecules present in the skin. The compounds that absorb photons are called chromophores. They can be present naturally in skin tissue (DNA, melanin) or of external origin (xenobiotics, drugs).

The ability of UV to induce skin tumors is mainly explained by its genotoxic and mutagenic properties. Indeed, sunlight is a proven cause of the development of skin cancers, particularly squamous cell and basal cell carcinomas involving keratinocytes. A number of experimental evidences reveal that UVB (280-320 nm) constitute the most harmful part of the solar spectrum (Marzouki et al. 2017). Although they are minority components of solar radiation (<5% of total UV), UVB are directly absorbed by DNA. They induce photochemical reactions on the DNA double helix and modify the chemical structure of the pyrimidine bases (C and T) which are found in the form of photoproducts of two main types: cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimidone photoproducts (6-4PP) and their Dewar valence isomers. These photoproducts can form at the levels of the four possible bipyrimidine sequences (TT, TC, CT and CC) but in different proportions. These lesions induce a slight distortion of the DNA double helix causing genomic instability. During replication, the presence of these lesions can lead to the incorporation of a bad nucleotide resulting in the formation of a mutation. Mutations caused by UV are characterized by a very high frequency of transitions in dipyrimidine sequences containing cytosines. The most frequent are C→T and CC→TT transitions. These damages, if not repaired correctly, are a source of mutations that constitute the first step towards the formation of tumors. UVB are therefore capable of initiating a process of carcinogenesis. However, the mutagenic power of UVA should not be neglected because it has been shown that they are also capable of inducing the formation of CPD in DNA even if they are much less powerful than UVB. UVA are at least twenty times more intense than UVB in solar radiation but are two to three orders of magnitude less harmful in damaging DNA.

Solar radiation is one of the few environmental carcinogens that is capable of inducing the 3 phases of cancerization: initiation, promotion and progression. CPD photolesions are suspected of being the cause of the majority of DNA mutations during photocarcinogenesis and are relatively resistant to cellular repair. Sun-induced immunosuppression may also promote immune tolerance to tumor cells, and the decreased repair and regeneration capacities of the skin during photoaging may contribute to tumor development. This damage, if not repaired properly, is a source of mutations that constitute the first step towards tumor formation.

Chronic exposure to UV rays is initially responsible for the formation of benign precancerous lesions (actinic keratoses) that can degenerate into skin cancers, the number of which is increasing worldwide. Their frequency varies according to the cellular origin: epithelial carcinomas or melanomas. Epithelial cancers include basal cell tumors that result from an alteration of the keratinocytes of the basal layer of the epidermis and squamous cell carcinomas dependent on supra-basal cells (squamous cell carcinomas). Carcinomas represent 90% of skin cancers with 70% of basal cell tumors and 20% of squamous cell carcinomas.

Actinic keratoses, also called solar keratoses or pre-epitheliomatous keratoses, are pre-cancerous skin lesions resulting from the proliferation of atypical keratinocytes. The prevalence of actinic keratoses varies from 6 to 60%, depending on main factors such as age, phototype or other risk factors such as immunodeficiency or an outdoor occupation (Dréno et al. 2014). Actinic keratoses are very common and are constantly increasing due to the aging of the population.

Clinically, actinic keratoses are often more palpable than visible, giving a rough appearance under the finger, often small in size, sometimes isolated or conversely multiple, in large poorly defined fields. Lesions vary in size from 2 mm to large plaques, but most measure between 2 and 6 mm in diameter. Actinic keratoses can be light in color, darkly pigmented, erythematous or tanned. Actinic keratoses of the head and neck are flat, whereas those on the back of the hands or forearms are often thicker.

Actinic keratoses are very common in photo-exposed areas, particularly in people with fair skin. Actinic keratoses develop under the effect of UV rays, mainly UVB but also UVA. These cause damage to the DNA of keratinocytes, the cells that make up the epidermis.

The theory of somatic mutagenesis indicates that the malignant transformation of cells is intimately linked to the formation of DNA damage and the risk that these can lead to mutations that favor anarchic cell growth. Certain changes in the genetic information of somatic cells (point mutations and/or chromosomal rearrangements) are a primary cause of malignant transformation of cells. The accumulation of these alterations in important genes that play a role in maintaining cellular integrity, such as cell cycle control, repair and apoptosis, can lead to the anarchic multiplication of abnormal cells and the formation of a tumor.

Mutation spectra, particularly in the p53 tumor suppressor gene, have made it possible to identify mutations in a given type of cancer based on their nucleotide position. The most frequently mutated positions differ depending on the cancer. The p53 gene is very frequently mutated in more than 50% of human tumors and has a mutation spectrum or signature specific to UVB. Tumor analysis has revealed “hot spots” of greater sensitivity to UV rays and therefore reflect the signature of sun exposure. Finally, it has recently been shown that the genome of human keratinocytes also has “hypersensitivity zones” of CPD mutations to UVB, particularly in the promoters of genes coding for ETS family transcription factors with a sensitivity up to 550 times greater than in the average genome. Thus, when these mutations affect genes that are involved in tumor elimination and DNA repair, they lose their protective function against cancer cells. This contributes to the emergence of abnormal keratinocytes called "atypical" which proliferate, clinically resulting in actinic keratoses. Natural but also artificial UV delivered in the aesthetic cabin are responsible for their occurrence.

The main risk of actinic keratoses is their progression to an invasive lesion such as cutaneous squamous cell carcinoma. There is a continuum from actinic keratosis to carcinoma in situ (or Bowen's disease), to invasive squamous cell carcinoma. But actinic keratoses can also regress spontaneously or remain stable (Werner et al. 2013). Their potential for malignant transformation (squamous cell carcinoma) is approximately 10% of cases. Studies have clearly demonstrated that, in humans, sunlight can act as both an initiator and a promoter of non-melanoma skin cancer. The exact number of actinic keratoses that will induce carcinomas depends on additional factors that promote conversion.

Treatment options for actinic keratoses depend on their number, depth, and extent. Cryotherapy is the preferred treatment method for isolated or small lesions. The main immediate side effects are pain, swelling, and blistering. In the long term, hyperpigmented or hypochromic scars may persist. Photo-directed treatment with porphyrins and light (photodynamic therapy) may be useful in the management of larger actinic keratoses. The main limitations of photodynamic therapy are the need for a specific lamp by the dermatologist, and the pain during the procedure, which can sometimes be unbearable, requiring treatment to be stopped. Full-thickness elliptical excision for actinic keratosis is rarely performed by dermatologists, but is regularly performed by plastic surgeons, general surgeons, and general practitioners. This form of therapy is considered by most health authorities to be inappropriate and is therefore not recommended.

For patients with multiple lesions or in the field of cancerization, topical agents (5-fluorouracil, imiquimod, masoprocol, tretinoin, ingenol mebutate, diclofenac) are also frequently prescribed. These topical agents have been shown to effectively treat actinic keratoses but cause side effects, mainly local, such as pain, burning, erythema, edema or persistent scars. Dermabrasion, chemical peels and carbon dioxide laser resurfacing can also be used in the treatment of extensive actinic keratoses, as well as surgery, which remains the standard treatment in the event of a suspicious or treatment-resistant lesion, possibly preceded by a biopsy. Surgery also allows control of the excision margins, but does not prevent the reappearance of actinic keratosis at the edge of the scar. Treatments therefore exist but they are relatively heavy treatments, presenting multiple possible side effects.

The removal of these pre-cancerous lesions undoubtedly reduces the risk of developing squamous cell carcinomas and is therefore appropriate for optimal patient management.

There is therefore still a need for treatments for pre-cancerous or cancerous skin lesions.

The present invention therefore relates to 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for its use in the prevention and/or treatment of a pre-cancerous skin lesion, such as actinic keratosis, or of a cancerous skin lesion.

The present invention also relates to the use of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine for the prevention and/or treatment of a pre-cancerous skin lesion, such as an actinic keratosis, or of a cancerous skin lesion.

The present invention also relates to the use of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine for the preparation of a medicament or a cosmetic or pharmaceutical composition (e.g. dermatological) intended for the prevention and/or treatment of a pre-cancerous skin lesion, such as actinic keratosis, or of a cancerous skin lesion.

The present invention also relates to a method of preventing and/or treating a pre-cancerous skin lesion, such as actinic keratosis, or a cancerous skin lesion comprising administering to a person in need thereof an effective amount of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine.

The present invention also relates to a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for its use in the prevention and/or treatment of a pre-cancerous skin lesion, such as actinic keratosis, or of a cancerous skin lesion.

The present invention also relates to the use of a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for the prevention and/or treatment of a pre-cancerous skin lesion, such as an actinic keratosis, or of a cancerous skin lesion.

The present invention also relates to the use of a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for the preparation of a medicament intended for the prevention and/or treatment of a pre-cancerous skin lesion, such as actinic keratosis, or of a cancerous skin lesion.

The present invention also relates to a method of preventing and/or treating a pre-cancerous skin lesion, such as an actinic keratosis, or a cancerous skin lesion comprising administering to a person in need thereof an effective amount of a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient.

The present invention also relates to 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for its use in the repair of DNA damage in skin cells and/or in the prevention and/or reduction of a deficit in the repair of DNA damage in skin cells.

The present invention also relates to the use of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine for the repair of DNA damage in skin cells and/or for the prevention and/or reduction of a deficiency in repair of DNA damage in skin cells.

The present invention also relates to the use of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine for the preparation of a medicament or a cosmetic or pharmaceutical (e.g. dermatological) composition intended for the repair of DNA damage in skin cells and/or for the prevention and/or reduction of a deficit in the repair of DNA damage in skin cells.

The present invention also relates to a method of repairing skin cell DNA damage and/or preventing and/or reducing a skin cell DNA damage repair deficiency comprising administering to a person in need thereof an effective amount of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine.

The present invention also relates to a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for its use in the repair of DNA damage to skin cells and/or in the prevention and/or reduction of a deficit in the repair of DNA damage to skin cells.

The present invention also relates to the use of a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for the repair of DNA damage in skin cells and/or for the prevention and/or reduction of a deficit in the repair of DNA damage in skin cells.

The present invention also relates to the use of a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for the preparation of a medicament intended for the repair of DNA damage in skin cells and/or for the prevention and/or reduction of a deficit in the repair of DNA damage in skin cells.

The present invention also relates to a method for repairing skin cell DNA damage and/or preventing and/or reducing a skin cell DNA damage repair deficiency comprising administering to a person in need thereof an effective amount of a composition, preferably a cosmetic or pharmaceutical (e.g. dermatological) composition, comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient.

The present invention also relates to a method for the therapeutic treatment of skin prone to actinic keratosis consisting of the application of 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine, or of a composition, preferably a cosmetic or pharmaceutical composition (e.g. dermatological), comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient, before exposing the skin to radiation, in particular solar radiation, in order to prevent the deficit in repair of DNA damage in skin cells.

DETAILED DESCRIPTION OF THE INVENTION

5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine

5,6,5',6'-Tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine (CAS No.: 55514-22-2, trade name: Triasorb ^TM ), also called phenylene bis-diphenyltriazine, is a sunscreen, more specifically an ultra-broad spectrum sunscreen capable of filtering the harmful part of the solar spectrum, which includes UV radiation (UVB and UVA), but also visible high-energy blue light. In the remainder of the description, the names 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine and phenylene bis-diphenyltriazine are used interchangeably.

The term “ultraviolet radiation” or “UV radiation”, commonly abbreviated “UV”, refers to solar ultraviolet radiation but also to artificial ultraviolet radiation, generated by tanning lamps for example.

By "UVA radiation" or simply "UVA" is meant ultraviolet rays having a wavelength λ between 320 and 400 nm.

By "UVB radiation" or simply "UVB" is meant ultraviolet rays having a wavelength λ between 290 and 320 nm.

By “high energy visible blue light” or “HEV” is meant rays having a wavelength λ between 400 and 450 nm, in particular between 400 and 450 nm.

The term "sunscreen" refers to a substance capable of filtering solar radiation to protect a surface, typically the skin or hair, from the harmful effects of this radiation.

• les filtres stricto sensu, qui majoritairement absorbent le rayonnement, et
• les écrans, qui absorbent et réfléchissent le rayonnement.

Depending on the radiation filtration mechanism, that is to say depending on whether the radiation is absorbed and/or reflected, or even diffused by the solar filter, we distinguish, strictly speaking:

• filters in the strict sense, which mainly absorb radiation, and
• screens, which absorb and reflect radiation.

However, in the remainder of the description, the term “solar filter”, sometimes abbreviated “filter”, will be used interchangeably, whether it is a filter or a screen. The term “UV filter” can also be used instead of “solar filter”.

A sun filter is called a “UVA filter” or “UVB filter” depending on whether it mainly filters UVA or UVB rays.

A so-called "broadband" or "broad spectrum" sun filter filters both UVA and UVB. A so-called "ultra broadband" or "ultra broad spectrum" sun filter is a broadband sun filter as defined above that also filters high-energy visible blue light.

A sunscreen may be “liposoluble” or “water-soluble”, depending on whether it dissolves more easily in lipids or in water, or “non-soluble”. A non-soluble sunscreen is typically in particulate form, and may nevertheless be integrated into a sunscreen composition, in particular in a dispersed form, in particular hydrodispersed when it is dispersed in an aqueous phase.

Phenylenese bis-diphenyltriazine is a non-soluble type filter. In particular, it is a water-dispersible filter, which is typically in the form of an aqueous dispersion, comprising between 20% and 50%, in particular between 40% and 50% by weight of active material relative to the total weight of the dispersion.

Advantageously, the process for preparing the aqueous dispersion consists of grinding the insoluble organic filter in the form of particles, using a grinding apparatus and in the presence of a grinding aid. Conventionally, wet grinding processes are used (wet grinding, wet mixing) and the grinding aid allows better dispersion of the particles. These techniques are well known to those skilled in the art. The grinding apparatus may be, for example, a microbead mill, a vibrating mill or a ball mill. Depending on the grinding process, the grinding aid is chosen from the group consisting of anionic, nonionic or amphoteric surfactants, emulsifiers and dispersants such as PPG-1-PEG-9 Lauryl Glycol Ether (Eumulgin® L, marketed by BASF).

In a particular embodiment, the size D50 of the phenylene bis-diphenyltriazine particles is between 100 and 1,000 nm, more particularly between 100 and 500 nm, even more particularly between 120 and 400 nm, in particular between 120 and 250 nm, in particular between 120 and 200 nm, typically between 150 and 200 nm.

By "D50" or "median" particle size is meant the size for which the cumulative function F(D) is equal to 50%, F(D) being defined according to the following relation:

in which:

f(D) is the particle size distribution in number, and

D _i is a size class.

The value of the D50 size can be determined by laser diffraction, in particular using a Mastersizer 3000 laser diffraction granulometer, more particularly in liquid form.

Composition

The composition according to the invention comprises 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine, as defined above, and at least one cosmetically and/or pharmaceutically acceptable excipient.

In the present invention, "cosmetically and/or pharmaceutically acceptable" is intended to mean that which is useful in the preparation of a composition which may have a cosmetic and/or pharmaceutical application, and which is generally safe, non-toxic and neither biologically nor otherwise undesirable and which is acceptable for human cosmetic or pharmaceutical use.

By “cosmetically and/or pharmaceutically acceptable excipient” is meant any cosmetically and/or pharmaceutically acceptable adjuvant allowing the manufacture, preservation or administration of the composition.

In a particular embodiment, the composition for use according to the invention is in the form of an emulsion, preferably oil-in-water.

For the purposes of the present invention, the term "emulsion" means any type of emulsion obtained by dispersing a discontinuous internal phase in a continuous external phase, one of these phases being an aqueous phase and the other being an oily phase. This is, for example, an oil-in-water or water-in-oil or multiple emulsion, preferably an oil-in-water emulsion.

The emulsion can be more or less fluid and can be presented in particular in the form of a white or colored cream, an ointment, a milk, a lotion, a serum, a paste, or possibly an aerosol, a mousse or a spray, and can also be water-resistant.

The composition for use according to the invention is intended more particularly for topical application to the skin.

In a particular embodiment, the phenylene bis-diphenyltriazine represents between 1% and 5%, in particular between 2% and 5%, in particular between 2% and 4%, typically between 3% and 4% by weight relative to the total weight of the composition.

It is understood that these mass percentages, as well as all those indicated in the description, are expressed as a percentage of mass of active material relative to the total mass of the composition.

In the composition according to the present invention, phenylene bis-diphenyltriazine can be combined with one or more other sunscreens, to constitute a photoprotective system which allows, after application to a surface (skin, hair, etc.), by mechanisms of absorption and/or reflection and/or diffusion of UVA and/or UVB radiation, to prevent, or at least limit, the contact of said radiation with said surface.

The photoprotective system constituting a composition according to the invention preferably comprises one or more liposoluble sun filters chosen from the following filters (i) to (v):

(i) 2-[4-(diethylamino)-2-hydroxybenzoyl]hexyl benzoate (CAS No.: 302776-68-7), also known as diethylamino hydroxybenzoyl hexyl benzoate or DHHB, marketed by BASF under the name Uvinul A Plus®.
It is a liposoluble UVA filter, with a maximum absorption wavelength λmax equal to 354 nm.
In a particular embodiment, the DHHB represents between 1% and 10%, for example between 3% and 10%, in particular between 4% and 9%, typically between 5% and 8%, in particular between 5% and 7%, in particular between 6% and 7% or advantageously between 5% and 6% by weight, relative to the total weight of the composition.

(ii) 2,4-bis[4-(2-ethylhexyloxy)-2-hydroxyphenyl]-6-(4-methoxyphenyl)-1,3,5-triazine (CAS No.: 187393-00-6), also known as bis-ethylhexyloxyphenol methoxyphenyl triazine or BEMT, marketed by BASF under the name Tinosorb S®.
It is a liposoluble broadband filter, having two absorption peaks at 310 nm and 340 nm.
In a particular embodiment, the BEMT represents between 1% and 4%, in particular between 2% and 4%, in particular between 2% and 3% by weight relative to the total weight of the composition.

(iii) 4-tert-butyl-4-methoxydibenzoylmethane or butyl methoxydibenzoylmethane (chemical name: 1,4-(1,1-dimethylethyl)phenyl-3-(4-methoxyphenyl)-1,3-propanedione; CAS No.: 70356-09-1), also called avobenzone or BMDBM, marketed in particular by DSM under the name Parsol 1789®.
It is a liposoluble UVA filter, with a maximum absorption wavelength λmax equal to 357 nm.
In a particular embodiment, the BMDBM represents between 1% and 10%, for example between 1% and 8%, in particular between 1% and 5%, typically between 2% and 4% by weight, relative to the total weight of the composition.

(iv) ethylhexyl triazone or EHT (chemical name: 4-[[4,6-bis[[4-(2-ethylhexoxy-oxomethyl)phenyl]amino]-1,3,5-triazin-2-yl]amino]benzoic acid, 2-ethylhexyl ester; CAS No.: 88122-99-00), marketed by BASF under the name Uvinul T 150®.
It is a liposoluble UVB filter, with a maximum absorption wavelength λmax equal to 314 nm.
In a particular embodiment, the EHT represents between 1% and 6%, typically between 2% and 5%, in particular between 3% and 5%, in particular between 3.5% and 4.5% by weight relative to the total weight of the composition.

(v) diethylhexyl butamido triazone or DBT (chemical name: 4,4'-[[6-[[4-[[(1,1-dimethylethyl)amino]carbonyl]phenyl]amino]-1,3,5-triazine-2,4-diyl]diimino]bis-, bis(2-ethylhexyl)benzoate; CAS No.: 154702-15-5), marketed by 3V Sigma under the name Uvasorb HEB®.
It is a liposoluble UVB filter, with a maximum absorption wavelength λmax equal to 310 nm.
In a particular embodiment, the DBT represents between 1% and 6%, typically between 2% and 5%, in particular between 3% and 5%, in particular between 3.5% and 4.5% by weight relative to the total weight of the composition.

The photoprotective system constituting a composition according to the invention may further comprise one or more additional particulate organic UV filters, in addition to phenylene bis-diphenyltriazine, chosen from the following filters (vi) and (vii):

(vi) methylene bis-benzotriazolyl tetramethylbutylphenol (chemical name: 2,2'-methylene-bis-(6-(2H-benzotriazol-2-yl)-4-(1,1,3,3-tetramethylbutyl)phenol; CAS No.: 103597-45-1) also known as MBBT, marketed by BASF under the name Tinosorb M®.
It is a broadband filter, having two absorption peaks at 306 nm and 348 nm, of non-soluble type.
MBBT typically comes in the form of an aqueous dispersion, comprising between 40% and 60%, typically 50% by weight of active material relative to the total weight of the dispersion.
In a particular embodiment, the MBBT represents between 1% and 7%, typically between 2% and 6%, in particular between 2% and 5%, in particular between 3% and 4% by weight relative to the total weight of the composition.
In a particular embodiment, the size D50 of the MBBT particles is between 50 nm and 250 nm, in particular between 60 nm and 150 nm.

(vii) 2,4,6-tris(biphenyl-4-yl)-1,3,5-triazine (CAS No.: 31274-51-8) also called trisbiphenyl triazine or TBPT, marketed by BASF under the name Tinosorb A2B®.
It is a UVB filter, with a maximum absorption wavelength λmax equal to 310 nm, of non-soluble type.
TBPT is typically in the form of an aqueous dispersion, comprising between 40% and 60%, typically 50% by weight of active material relative to the total weight of the dispersion.
In a particular embodiment, the TBPT represents between 1% and 7%, in particular between 2% and 7%, in particular between 2% and 6% by weight relative to the total weight of the composition.
In a particular embodiment, the D50 size of the TBPT particles is between 50 nm and 250 nm, in particular between 80 nm and 150 nm.

In a particular embodiment, the photoprotective system constituting a composition according to the invention may further comprise one or more particulate inorganic filters, such as titanium dioxide (TiO2) or zinc oxide (ZnO).

The particulate inorganic filter(s) optionally present in the composition according to the invention typically have a particle size D50 of between 15 nm and 150 nm.

In a particular embodiment, the composition according to the invention does not comprise ethylhexyl salicylate, octocrylene, ethylhexyl methoxycinnamate, isoamyl methoxycinnamate, homosalate, paraaminobenzoic acid (PABA), octyl dimethyl PABA, 3-methylbenzylidene camphor, 4-methylbenzylidene camphor, benzophenone-3 and/or benzophenone-4.

In a particular embodiment, the composition according to the invention does not comprise any water-soluble filters. The absence of water-soluble filters is particularly advantageous, in that water-soluble filters are, generally speaking, more easily assimilated by marine organisms than liposoluble or non-soluble filters. Consequently, the Applicant's choice of using only non-soluble (more particularly particulate, and in particular water-dispersed) or liposoluble filters makes it possible to minimize the impact of the composition according to the invention on the marine environment.

In another particular embodiment, the composition for use according to the invention does not comprise any particulate inorganic filter.

In a particular embodiment, the total number of UV filters present in the composition is less than or equal to 5, preferably less than or equal to 4, and/or all of the UV filters present in the composition represent between 4% and 25% by weight, in particular between 6% and 20% by weight, in particular between 7% and 18% by weight, typically between 8% and 17% by weight, relative to the total weight of the composition.

1. 5,6,5’,6’-tétraphényl-3,3’-(1,4-phénylène)-bis[1,2,4]triazine,
2. 2,4-bis[4-(2-éthylhexyloxy)-2-hydroxyphényl]-6-(4-méthoxyphényl)-1,3,5-triazine (BEMT),
3. 2-[4-(diéthylamino)-2-hydroxybenzoyl]benzoate d’hexyle (DHHB), et
4. éthylhexyl triazone (EHT) ou diéthylhexyl butamido triazone (DBT).

In an advantageous embodiment, the photoprotective system of the composition according to the invention consists of:

1. 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine,
2. 2,4-bis[4-(2-ethylhexyloxy)-2-hydroxyphenyl]-6-(4-methoxyphenyl)-1,3,5-triazine (BEMT),
3. Hexyl 2-[4-(diethylamino)-2-hydroxybenzoyl]benzoate (DHHB), and
4. ethylhexyl triazone (EHT) or diethylhexyl butamido triazone (DBT).

In particular, (d) corresponds to ethylhexyl triazone.

In this particular embodiment, the phenylene bis-diphenyltriazine represents between 1% and 5%, in particular between 2% and 5%, in particular between 2% and 4%, typically between 3% and 4% by weight relative to the total weight of the composition; the DHHB represents between 1% and 10%, for example between 3% and 10%, in particular between 4% and 9%, typically between 5% and 8%, for example between 5% and 7%, in particular between 6% and 7% or advantageously between 5% and 6% by weight, relative to the total weight of the composition; the BEMT represents between 1% and 4%, in particular between 2% and 4%, in particular between 2% and 3% by weight of the composition relative to the total weight of the composition; and the EHT or DBT represents between 1% and 6%, typically between 2% and 5%, in particular between 3% and 5%, in particular between 3.5% and 4.5% by weight relative to the total weight of the composition.

The compositions according to the invention may thus comprise cosmetically and/or pharmaceutically (e.g. dermatologically) acceptable excipients conventionally used in the art, in particular chosen from liquid lipophilic compounds, water-miscible polyols, emulsifiers, consistency agents, water-retentive agents, texturizing agents, opacifiers, coloring pigments, anti-foaming agents, perfumes, preservatives, polymers, fillers, sequestrants, bactericides, odor absorbers, alkalizing or acidifying agents, surfactants, free radical scavengers, vitamins, alpha-hydroxy acids, active agents (in particular moisturizing agents, antioxidants) or any other ingredient usually used for the manufacture of anti-sun compositions.

The composition may thus comprise a liquid lipophilic compound or a combination of several liquid lipophilic compounds, typically a liquid lipophilic emollient or a combination of liquid lipophilic emollients, in particular a liquid fatty substance or a combination of liquid fatty substances.

By “liquid lipophilic compound” is meant a compound which has an affinity for apolar substances such as lipids, and is in liquid form, particularly at a temperature between 5°C and room temperature (between 20°C and 25°C). It typically comprises one or more polar chemical function(s) chosen from ester (-CO ₂ -), alcohol (-OH), ether (-O-), carbonate (-OCO ₂ -) and carboxylic acid (-CO ₂ H) functions, and one or more lipophilic group(s), i.e. one or more aliphatic and/or aromatic hydrocarbon group(s), in particular one or more saturated or unsaturated, linear or branched hydrocarbon chain(s), comprising between 6 and 20 carbon atoms, for example between 7 and 18 carbon atoms, in particular between 8 and 16 carbon atoms. It may be chosen in particular from C12-C15 alkyl benzoate, caprylic/capric triglycerides, dicaprylyl carbonate, cococaprylate, diisopropyl sebacate, diethylhexyl succinate, isopropyl palmitate, isopropyl myristate, propylheptyl caprylate, phenethyl benzoate, dibutyl adipate, diisopropyl adipate, glyceryl tri-2-ethylhexanoate, pentaerythrityl tetra-2-ethylhexanoate, cetyl 2-ethylhexanoate, isononyl isonanoate, butylene glycol dicaprylate/dicaprate, octyldodecanol, propylene glycol dicaprylate/dicaprate and dicaprylyl ether, in particular from caprylic/capric triglycerides, dicaprylyl carbonate, isopropyl palmitate, propylheptyl caprylate, phenethyl benzoate, diisopropyl adipate, glyceryl tri-2-ethylhexanoate, pentaerythrityl tetra-2-ethylhexanoate, cetyl 2-ethylhexanoate, isononyl isononoate, butylene glycol dicaprylate/dicaprate, octyldodecanol, propylene glycol dicaprylate/dicaprate, dicaprylyl ether and combinations thereof.

The liquid lipophilic emollient(s) are chosen in particular from C ₁₂ -C ₁₅ alkyl benzoate, caprylic/capric triglycerides and dicaprylyl carbonate.

In a particular embodiment, the composition according to the invention does not comprise diisopropyl sebacate, diethylhexyl succinate, isopropyl myristate and/or dibutyl adipate. Typically, it does not include dicaprylyl carbonate, coco-caprylate, diisopropyl sebacate, diethylhexyl succinate, isopropyl palmitate, isopropyl myristate, propylheptyl caprylate, phenethyl benzoate, dibutyl adipate, diisopropyl adipate, glyceryl tri-2-ethylhexanoate, pentaerythrityl tetra-2-ethylhexanoate, cetyl 2-ethylhexanoate, isononyl isononoate, butylene glycol, dicaprylate/dicaprate, octyldodecanol, propylene glycol dicaprylate/dicaprate and/or dicaprylyl ether.

The composition may further comprise a polyol miscible with water at room temperature (approximately 25°C), in particular chosen from polyols having from 2 to 20 carbon atoms, preferably having from 2 to 10 carbon atoms, and preferentially having from 2 to 6 carbon atoms, such as glycerin; glycol derivatives such as propylene glycol, butylene glycol, pentylene glycol, hexylene glycol, dipropylene glycol, diethylene glycol; glycol ethers such as C1-C4 alkyl ethers of mono-, di- or tri-propylene glycol, C1-C4 alkyl ethers of mono-, di- or tri-ethylene glycol and mixtures thereof.

The composition may also comprise one or more anionic, cationic and/or nonionic emulsifiers selected from glucose derivatives such as cetearyl glucoside, arachidyl glucoside, lauryl glucoside, polyglyceryl-3 methylglucose distearate and methyl glucose sesquistearate; sucrose derivatives such as sucrose polystearate and sucrose palmitate; fatty acid glycerides such as glyceryl stearate, glyceryl oleate and glyceryl stearate citrate; glutamic acid derivatives such as sodium stearoyl glutamate; sulfosuccinic acid derivatives such as disodium cetearyl sulfosuccinate and sodium dioctyl sulfosuccinate; phosphoric acid derivatives such as potassium cetyl phosphate; polyglyceryl fatty acid esters such as polyglyceryl-3-diisostearate and polyglyceryl-2-dipolyhydroxystearate; sorbitol derivatives such as polysorbate, PEG-10 sorbitan laurate and sorbitan esters; fatty alcohol and fatty acid polyglycol ethers such as ceteareth-20, beheneth-25, steareth-2, trideceth-9, PEG-5 ethylhexanoate and PEG-100 stearate; sodium lauryl sulfate, sodium stearate, organomodified silicone/polysiloxane/polyalkyl/polyether oxyalkenyl copolymers and derivatives, and mixtures thereof.

The emulsifier(s) are chosen in particular from potassium cetyl phosphate, lauryl glucoside and glyceryl stearate citrate.

In a particular embodiment, the composition according to the invention does not comprise an ethoxylated emulsifier and/or comprising a PEG chain. In particular, the composition does not comprise sorbitol derivatives such as a polysorbate and PEG-10 sorbitan laurate; polyglycol ethers of fatty alcohol and fatty acid such as ceteareth-20, beheneth-25, steareth-2 and PEG-100 stearate; and organomodified oxyalkenyl silicone/polysiloxane/polyalkyl/polyether copolymers and derivatives.

The composition may also comprise one or more solid consistency agents selected from fatty alcohols such as cetyl alcohol, cetearyl alcohol and stearyl alcohol; fatty acids such as stearic acid; fatty acid esters such as myristyl stearate; polysaccharides or derivatives such as xanthan gum, guar gum, agar gum, alginates, gellan gum and carrageenan; cellulose derivatives such as hydroxypropylcellulose; waxes such as beeswax, carnauba wax, microcrystalline wax, ceresin, and ozocerite; crosslinked polyacrylates or homopolymers of acrylic acids or polyacrylamides such as carbomers, acrylate copolymers, crosslinked acrylate/(C10-C30)alkylacrylate copolymers, acrylate/behenethyl-25 methacrylate copolymers; silicate derivatives such as magnesium silicates and mixtures thereof.

In a particular embodiment, the composition according to the invention comprises one or more consistency agents chosen from fatty alcohols such as cetyl alcohol, cetearyl alcohol and stearyl alcohol; fatty acids such as stearic acid; fatty acid esters such as myristyl stearate; polysaccharides or derivatives such as xanthan gum, guar gum, agar gum, alginates, gellan gum and carrageenan; cellulose derivatives such as hydroxypropylcellulose, and mixtures thereof.

In a particular embodiment, the composition does not comprise a consistency agent chosen from acrylates and their derivatives.

The composition may also comprise one or more water-retentive agents selected from a copolymer of N-vinylpyrrolidone (VP) and eicosene (INCI: VP/eicosen copolymer), a crosslinked polymer of trimethylpentanediol, adipic acid and glycerol (INCI: trimethylpentanediol/adipic acid/glycerin crosspolymer), polyamide 3 (INCI), a copolymer of acrylate and octylacrylamide (INCI: acrylates/octylacrylamide copolymer), and triacontanyl PVP (INCI).

Preferably, the composition for use according to the invention comprises VP/eicosene copolymer as a water-retentive agent.

The composition may also comprise one or more texturizing agents in the form of powders, in particular chosen from ingredients from the starch family, such as rice, corn and tapioca starches; talcs; cellulose derivatives such as microcrystalline cellulose, and mixtures thereof.

In a particular embodiment, the composition according to the invention does not comprise a texturing agent chosen from nylon derivatives and polymethyl methacrylate (PMMA).

The composition may also comprise coloring pigments, in particular chosen from CI77891, CI77491, CI77492 and CI77499 (INCI designations).

In a particular embodiment, the composition according to the invention comprises an amount of C1-C4 alcohols, such as ethanol and isopropanol, less than or equal to 4% by weight, preferably less than or equal to 2% by weight, in particular less than or equal to 1% by weight, in particular less than or equal to 0.5% by weight, relative to the total weight of the composition.

In another particular embodiment, the composition is free of ethanol and isopropanol.

In a particular embodiment, the composition according to the invention comprises an amount of volatile ingredients less than or equal to 5% by weight, preferably less than or equal to 2% by weight, in particular less than or equal to 1% by weight, in particular less than or equal to 0.5% by weight, relative to the total weight of the composition.

For the purposes of the present invention, the term “volatile ingredients” means non-alcoholic volatile organic or inorganic compounds such as esters, silicones or volatile alkanes, characterized by a flash point below 60°C.

For the purposes of the present invention, the term “flash point” (denoted PE) means the minimum temperature at which a combustible substance emits vapours in sufficient concentration to form a gas mixture with the ambient air which ignites on contact with a flame or a hot point, but insufficient for combustion to propagate by itself in the absence of the so-called pilot flame. According to Regulation (EC) No 1272/2008 on the classification, labelling and packaging of substances and mixtures, a substance having a PE such that 23°C ≤ PE < 60°C belongs to category 3 “low flammability”.

Volatile esters include isohexyl neopentanoate. Volatile silicones include cyclopentasiloxane. Volatile alkanes include isohexadecane.

In another particular embodiment, the composition for use according to the invention is free of volatile ingredients within the meaning of the present invention.

The composition may also comprise alkalizing or acidifying agents, and more particularly acids and bases for adjusting the pH range of said composition. The bases may be mineral (soda, potash, ammonia, etc.) or organic, such as mono-, di- or triethanolamine, an aminomethylpropanediol, N-methylglucamine, basic amino acids such as arginine and lysine, and mixtures thereof. The acids may be mineral (hydrochloric acid, etc.) or organic, such as lactic acid and citric acid.

The composition according to the invention may further comprise additional active agents chosen in particular from moisturizing agents, desquamating agents, agents improving the barrier function, depigmenting agents, antioxidant agents, dermocontracting agents, anti-glycation agents, agents stimulating the synthesis of dermal and/or epidermal macromolecules and/or preventing their degradation, agents stimulating the proliferation of fibroblasts or keratinocytes and/or the differentiation of keratinocytes, NO synthase inhibitors, agents increasing the activity of the sebaceous gland, tightening agents, liporestructuring agents, slimming agents, agents promoting cutaneous microcirculation, soothing and/or irritating agents, sebum-regulating or antiseborrheic agents, astringent agents, healing agents, anti-inflammatory agents, anti-acne agents, and mixtures thereof.

In a particular embodiment, the composition according to the invention is free of silicone.

In a preferred embodiment, the composition according to the invention comprises one or more antioxidant agents, typically chosen from vitamin E (i.e. tocopherols and tocotrienols) and its derivatives.

By "tocopherols" or "tocotrienols" is meant the compounds α-, β-, γ- and δ-tocopherol or tocotrienol. The term "tocopherol" usually refers to α-tocopherol. Among the derivatives of tocopherol(s), we can notably cite ester derivatives, such as tocopheryl acetate, and glycosylated derivatives, such as tocopheryl glucoside (INCI: tocopheryl glucoside).

In a preferred embodiment, the composition for use according to the invention comprises one or more antioxidants chosen from tocopherol and its derivatives, in particular from tocopherol, tocopherol glycoside and their mixtures, in an amount typically between 0.01% and 1% by weight relative to the total weight of the composition.

Applications

The subject of the present invention is 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine or a composition comprising it as defined above for its use in the prevention and/or treatment of a pre-cancerous or cancerous, preferably pre-cancerous, skin lesion.

The pre-cancerous skin lesion will more specifically be an actinic keratosis.

Preferably, the cancerous skin lesion is not a melanoma. The cancerous skin lesion may be a cancerous skin lesion due to a deficiency in repair of DNA damage in skin cells, the deficiency in repair of DNA damage being in particular due to exposure of the skin to high-energy visible blue light, in particular from the sun, and therefore in particular may be due to exposure to sunlight. Preferably, the cancerous skin lesion is a carcinoma, for example a squamous cell carcinoma.

5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine or the composition according to the invention may be administered more particularly to a patient with sensitive or hypersensitive skin and/or skin that is reactive to the sun and/or subject to actinic keratosis.

5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine or the composition according to the invention may be administered topically, more particularly by application to the skin, the skin being more particularly sensitive or hypersensitive skin and/or reactive to the sun and/or prone to actinic keratosis.

The treatment based on 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine or a composition comprising it as defined above can be combined, preferably sequentially, with photodynamic therapy (PDT), in particular in the case of a pre-cancerous skin lesion, and more particularly of an actinic keratosis.

By "photodynamic therapy", also called "photodynamic therapy" or "PDT" for short, is meant, within the meaning of the present invention, a technique for treating a pre-cancerous or cancerous skin lesion using a photosensitizing chemical substance administered, preferably by topical application to the lesion, in combination with exposure to light. The photosensitizing chemical substance is then activated by the light to become cytotoxic. Said treatment technique therefore has the advantage of being local and non-invasive. Preferably, the photosensitizing chemical substance is methyl aminolevulinate (MAL).

The present invention also relates to 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine or a composition comprising it as defined above for its use in the repair of DNA in skin cells and/or in the prevention and/or reduction of a deficit in the repair of DNA damage in skin cells.

For example, DNA may have damage to repair following exposure to radiation, i.e. photoinduced damage, the radiation having caused said damage. The radiation may be UV radiation, in particular UVB and/or UVA, in particular from the sun. This UV radiation may have been associated with high-energy visible blue light, in particular from the sun, which prevented the DNA from repairing itself. Thus, DNA may have damage to repair following exposure to sunlight.

In particular, DNA may present lesions to be repaired at the level of bipyrimidine sequences TT, TC, CT and CC, in particular in the form of cyclobutane dimers of pyrimidines (CPD) or even pyrimidines 6-4,pyrimidone (6-4PP) and their Dewar valence isomers.

The deficit in repair of DNA damage in skin cells may in particular be due to exposure of skin cells, and therefore of the skin, to high-energy visible blue light, particularly from the sun, and therefore may in particular be due to exposure to solar radiation.

The skin may be particularly sensitive or hypersensitive and/or reactive to the sun and/or prone to actinic keratosis.

5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine or the composition according to the invention may be administered topically, more particularly by application to the skin, the skin being more particularly sensitive or hypersensitive skin and/or reactive to the sun and/or prone to actinic keratosis.

EXAMPLES

Example 1: Effects of HEV blue light on DNA damage

Visible light, which is radiation with wavelengths ranging from 400 to 700 nm and accounts for approximately 50% of all solar radiation reaching the Earth, has recently been highlighted as a contributing factor to skin photodamage, in addition to UV radiation. Specifically, high-energy visible (HEV) light (400–450 nm) belonging to the blue light waveband (400–500 nm) can penetrate from the epidermis to the upper dermis of the skin, where it can have a major effect on skin cells. Blue light has been shown to induce pigmentary disorders and, although clinical data are scarce, may also contribute to signs of photoaging.

Activation of biological pathways can affect cellular macromolecules (DNA, proteins, and lipids), leading to structural damage and impaired biological function. Although conflicting results have been obtained due to differences in light sources and exposure between studies, HEV/blue light radiation has been shown to cause increased melanin production and transfer, promoting cell differentiation, inhibiting cell growth, and even inducing cell death by apoptosis. These cellular effects can then manifest on the skin as erythema, pigmentary changes, wrinkles, and in some cases, photodermatosis.

Although there is evidence that blue light has harmful effects on the skin, it may also have beneficial effects such as anti-proliferative and anti-inflammatory properties. Blue light phototherapy has been studied as a potential treatment for psoriasis and Alzheimer's disease. The overall physiological effect of blue light on the skin therefore depends on the underlying skin condition, as well as the duration, dose, and irradiance of exposure.

Therefore, the aim of this study is to evaluate the effect of HEV radiation on the DNA damage repair process after UVB exposure. Secondly, the protective effect of a formulation comprising phenylene bis-diphenyltriazine is evaluated.

In this study, the model used is a human epidermal follicular model reconstructed in vitro.

Epidermis were reconstructed from keratinocytes from the outer epithelial sheath of human hair follicles obtained from healthy volunteers who gave informed consent. The 3D reconstruction method described by Guiraud et al. 2014 was used. Human hair follicle outer epithelial sheath cells are a readily available and non-invasive source of keratinocytes for epidermal reconstruction.

Briefly, cells (keratinocytes) were isolated from the outer epithelial sheath of hair follicles, then cultured before being seeded on culture inserts immersed in culture medium, then the culture inserts were placed at the air/liquid interface in an incubator at 37°C in a humidified atmosphere with 5% _CO2 , to induce epidermal differentiation and form the stratum corneum . At 16 days (D16), a reconstructed epidermis with a surface area of 1.12cm² was formed. The culture medium was renewed every 24 hours.

Method :

Three different formulations are studied in this model. Each is applied at a dose of 2 mg/cm² on the surface of a polymethyl methacrylate (PMMA) plate. This plastic is naturally transparent and has a higher light transmission than glass. The PMMA plate is then placed on the reconstructed epidermis model obtained according to Guiraud et al. 2014 just before exposure to HEV blue light stress (60J/cm²). After HEV blue light stress, the PMMA plate is removed and the reconstructed epidermis are exposed to UVB at a dose of (50mJ/cm²). DNA damage is then analyzed immediately at the end of UVB irradiation (T0) or after 48h of re-culture (T48h).

• Formulation A comprenant de la 5,6,5’,6’-tetraphényl-3,3’-(1,4-phénylène)-bis[1,2,4]triazine comme seul filtre solaire et dont la composition est donnée dans le tableau 1 ;
• Formulation B comprenant du BEMT, du DHHB et de la BEMT comme filtres solaires, en plus de la 5,6,5’,6’-tetraphényl-3,3’-(1,4-phénylène)-bis[1,2,4]triazine (système photoprotecteur à 4 filtres), et dont la composition est donnée dans le tableau 2 ;
• Formulation C correspondant à une composition solaire commerciale ne comprenant pas de 5,6,5’,6’-tetraphényl-3,3’-(1,4-phénylène)-bis[1,2,4]triazine comme filtre solaire et dont la liste des ingrédients (noms INCI) est donnée ci-après dans l’ordre de prépondérance, les filtres solaires étant identifiés par un astérisque :
  AQUA • DIISOPROPYL SEBACATE • ALCOHOL DENAT. • ETHYLHEXYL SALICYLATE* • ETHYLHEXYL TRIAZONE* • BIS-ETHYLHEXYLOXYPHENOL METHOXYPHENYL TRIAZINE* • DIISOPROPYL ADIPATE • PEG-8 • DIPROPYLENE GLYCOL • C12-22 ALKYL ACRYLATE/ HYDROXYETHYLACRYLATE COPOLYMER • GLYCERIN • DROMETRIZOLE TRISILOXANE* • ALUMINUM STARCH OCTENYLSUCCINATE • BUTYL METHOXYDIBENZOYLMETHANE* • METHOXYPROPYLAMINO CYCLOHEXENYLIDENE ETHOXYETHYLCYANOACETATE* • SILICA • TOCOPHEROL • DIETHYLAMINO HYDROXYBENZOYL HEXYL BENZOATE* • ACRYLATES COPOLYMER • ACRYLATES/C10-30 ALKYL ACRYLATE CROSSPOLYMER • CAPRYLYL GLYCOL • HYDROXYACETOPHENONE • PENTAERYTHRITYL TETRA-DI-T-BUTYL HYDROXYHYDROCINNAMATE • SODIUM POLYACRYLATE • TEREPHTHALYLIDENE DICAMPHOR SULFONIC ACID* • TRIETHANOLAMINE • TRISODIUM ETHYLENEDIAMINE DISUCCINATE.

The three formulations tested are as follows:

• Formulation A comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine as the sole sunscreen and the composition of which is given in Table 1;
• Formulation B comprising BEMT, DHHB and BEMT as sunscreens, in addition to 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine (4-filter photoprotective system), and the composition of which is given in Table 2;
• Formulation C corresponding to a commercial sunscreen composition not comprising 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine as a sunscreen and whose list of ingredients (INCI names) is given below in order of predominance, the sunscreens being identified by an asterisk:
  AQUA • DIISOPROPYL SEBACATE • ALCOHOL DENAT. • ETHYLHEXYL SALICYLATE* • ETHYLHEXYL TRIAZONE* • BIS-ETHYLHEXYLOXYPHENOL METHOXYPHENYL TRIAZINE* • DIISOPROPYL ADIPATE • PEG-8 • DIPROPYLENE GLYCOL • C12-22 ALKYL ACRYLATE/ HYDROXYETHYLACRYLATE COPOLYMER • GLYCERIN • DROMETRIZOLE TRISILOXANE* • ALUMINUM STARCH OCTENYLSUCCINATE • BUTYL METHOXYDIBENZOYLMETHANE* • METHOXYPROPYLAMINO CYCLOHEXENYLIDENE ETHOXYETHYLCYANOACETATE* • SILICA • TOCOPHEROL • DIETHYLAMINO HYDROXYBENZOYL HEXYL BENZOATE* • ACRYLATES COPOLYMER • ACRYLATES/C10-30 ALKYL ACRYLATE CROSSPOLYMER • CAPRYLYL GLYCOL • HYDROXYACETOPHENONE • PENTAERYTHRITYL TETRA-DI-T-BUTYL HYDROXYHYDROCINNAMATE • SODIUM POLYACRYLATE • TEREPHTHALYLIDENE DICAMPHOR SULFONIC ACID* • TRIETHANOLAMINE • TRISODIUM ETHYLENEDIAMINE DISUCCINATE.

Ingredients of formulation A % (w/w) 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine 5 Glycerin 5 Demineralized water Qsp 100 Xanthan gum 0.2 C12-C15 alkyl benzoate 12.0 Dicaprylyl carbonate 9.0 Caprylic/capric triglycerides 9.0 Conservatives Qs Stearyl alcohol 1.0 Glyceryl monostearate 0.5 Glyceryl behenate 0.4 PEG-100 stearate 0.5 VP/eicosene copolymer 1.0 Polyacrylate 0.2 Potassium cetyl phosphate 2.0

Ingredients of formulation B % (m/m) 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine 3-3.5 Bis-Ethylhexyloxyphenol Methoxyphenyl Triazine 2.5-3 Diethylamino hydroxybenzoyl hexyl benzoate 5-6 Ethyl Hexyl Triazone 3.5-4 Glycerin 5.0 Demineralized water Qsp 100 Xanthan gum 0.2 C12-C15 alkyl benzoate 12.0 Dicaprylyl carbonate 9.0 Caprylic/capric triglycerides 9.0 Conservatives Qs Stearyl alcohol 1.0 Glyceryl monostearate 0.5 Glyceryl behenate 0.4 PEG-100 stearate 0.5 VP/eicosene copolymer 1.0 Polyacrylate 0.2 Potassium cetyl phosphate 2.0

The reconstructed epidermis on which the PMMA plates coated with the formulations to be tested are applied are irradiated by a simulated HEV stress using a blue light LED lamp (Kessil PR160). The wavelength is 427±30 nm (HEV spectrum 400-450 nm) and a dose of 60J/cm² was applied to the reconstructed epidermis for an average duration of 39 minutes. The power of the LED lamp is approximately 176 mW/cm². After this HEV blue light stress, the PMMA plate is then removed.

The reconstructed epidermis are then exposed to a specific UVB stress using a Biosun RMX irradiator (Vilber Lourmat, Germany) equipped with a monochromatic illumination lamp at 312 nm. The lamp power is approximately 3 mW/cm ² . The skin models are exposed to a single dose of 50 mJ/cm².

The reconstructed epidermis are then harvested just after UVB stress for half of them (T0) and 48 hours after this UVB stress for the second half (T48h), and all are frozen at -80°C for LC/MS analysis (liquid chromatography coupled with mass spectrometry).

All experimental conditions were performed with n=3 technical replicates and 1 donor (N=1, n=3), except for the UVB controls (UVB stress alone) and HEV + UVB (HEV blue light stress then UVB stress) performed with 3 donors (N=3, n=3).

DNA is extracted by an approach combining tissue grinding in a Qiagen “TissueLyser” system based on the use of metal beads in a buffer, followed by destructuring of the epidermis by suitable buffers and proteinase K according to the supplier’s recommendations. DNA is then isolated on a column using the DNAeasy Blood and Tissue kit (Qiagen). DNA is obtained in solution in water and hydrolyzed into monomers by successive incubation with two enzymatic cocktails (endonucleases, exonucleases, phosphodiesterases).

The hydrolyzed samples are then injected into a reverse-phase HPLC (high-performance liquid chromatography) system. At the column outlet, the eluent passes through a UV detector that quantifies the normal bases of DNA. The flow is then directed to the inlet of an electrospray mass spectrometer with triple quadrupole analyzer (MS/MS). Detection is done in multiple reaction monitoring (MRM) mode, which sensitively and specifically follows the production of fragments characteristic of targeted analytes. In the DNA of the reconstructed epidermis, the pyrimidine dimers produced by UV will be sought, namely the cyclobutane dimers of pyrimidines (CPD), the pyrimidine (6-4) pyrimidone photoproducts (6-4PP) and their Dewar valence isomers (Dewar), and this for each of the bipyrimidine dinucleotides (TT, TC, CT). The precise amount of each analyte detected in MS/MS is determined by isotopic dilution using TpT photoproducts [M+12]. The amount of DNA analyzed is determined simultaneously by measuring the normal nucleosides 2’-deoxyguanosine and thymidine by a UV detector and quantified by an external calibration.

The raw data will then be processed to express the results in number of photoproducts per million normal bases. This approach makes it possible to overcome the inevitable variations in the quantities of DNA extracted from one sample to another.

Results

The results for the photoproduct levels observed for the UFV and HEV + UVB controls are presented in Table 3 below. The photoproduct level is expressed in dimer per million normal bases), Data are means ± sem (standard error of the mean).

Quantified photoproducts UVB control HEV + UVB control T0 T48h T0 T48h TT CPD 76.9 ± 4.9 46.5 ± 3.6 79.2 ± 3.6 69.7 ± 3.5 TC CPD 38.7 ± 2.7 23.4 ± 1.6 41.9 ± 2.2 34.9 ± 2.1 CT CPD 12.8 ± 1.1 4.4 ± 0.4 13.8 ± 1.6 9.0 ± 1.2

Table 4 presents a one-way statistical analysis (ANOVA) followed by the Tuckey multiple comparison test.

Quantified photoproducts Impact of HEV blue light Evidence of DNA repair At T0 T0 compared to T48h UVB control
vs HEV + UVB Control UVB control HEV + UVB control TT CPD NS P<0.001 NS TC CPD NS P<0.001 NS CT CPD NS P<0.001 NS

Exposure to UVB alone induces a significant and important increase in DNA photoproducts just after irradiation. The rate of TT CPD lesions is higher than that of TC CPD and CT CPD. After 48 hours, the inventors observe a significant decrease (***p<0.001) in photoproducts, which is explained by the fact that there has been a repair of DNA lesions.

When HEV blue light irradiation is performed before UVB exposure, the induction of DNA photoproducts is not significantly different from UVB exposure only at T0. Exposure to HEV blue light does not cause DNA damage. However, after 48 hours, photoproducts are found in quantities equivalent to T0. Under these conditions, there was no DNA repair. These results demonstrate the negative impact of HEV blue light exposure on the DNA repair process.

Similar results are found when analyzing TT CPD, TC CPD or CT CPD photoproducts.

The results of the tests carried out with formulations A, B or C on the efficiency of DNA repair are summarized in Table 5 below. The efficiency of DNA repair is calculated under the different conditions. The statistical analysis is performed by comparing the data of each condition at T48h with the UVB control conditions at T0.

Photoproducts tested TT CPD TC CPD CT CPD UVB control 39.5% ± 4.6 39.7% ± 4.1 65.9% ± 3.5 HEV + UVB control 14.8% ± 5.5 16.6% ± 5.0 35.0% ± 8.6 Formulation A 30.0% ± 5.3* 34.1% ± 4.9* 58.8% ± 4.4* Formulation B 38.4% ± 5.4*** 35.0% ± 7.9* 45.9% ± 7.9* Formulation C -1.0% ± 9.6 1.8% ± 8.03 -0.5% ± 14.5

*p<0.05; ***p<0.001

Formulation A containing 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine at 5% effectively protects against the effects of HEV blue light, since a 30% DNA repair is observed for the TT CPD photoproduct.

Formulation B containing a 4-filter photoprotective system including 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine also effectively protects against the effects of HEV blue light, a 39% DNA repair is observed for the TT CPD photoproduct.

In contrast, Formulation C not containing 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine has no significant effect on the deleterious effects of HEV blue light.

Similar results are observed for TC CPD and CT CPD photoproducts.

The inventors therefore demonstrate that only formulations containing 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine protect against damage caused by HEV blue light and provide protection for DNA damage repair enzymes.

It is important to remember that these DNA lesions are responsible for the transformation of keratinocytes in an “atypical” mode, leading to a proliferation of keratinocytes which results in actinic keratosis.

The inventors therefore demonstrate that formulations containing 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine are capable of preventing and limiting the formation of pre-cancerous skin lesions of the actinic keratosis type.

Example 2: Clinical study on healthy volunteers

2.1. Study without application of composition according to the invention

• Une zone A exposée seulement à un rayonnement d’UVB selon les conditions du stress à un rayonnement UVB du protocole ci-dessous ;
• Une zone B exposée à une lumière bleue HEV et à un rayonnement UVB selon le protocole ci-dessous.

This study is carried out on 3 healthy volunteers. Each volunteer is exposed locally on 2 distinct areas at the back, namely:

• A zone A exposed only to UVB radiation according to the UVB radiation stress conditions of the protocol below;
• A zone B exposed to HEV blue light and UVB radiation according to the protocol below.

Protocol

Patients are exposed to stress sequences with HEV blue light (60J/ ^cm2 for each sequence), 2 times per day (the sequences being spaced approximately 4 hours apart) and for 3 successive days, in order to mimic real exposure conditions. Approximately 30 minutes after the ^6th sequence, stress with UVB radiation is performed (150mJ/cm2).

Biopsies with a diameter of approximately 4 mm are taken from zones A and B. They are collected only after 48 hours to assess the possible effect of HEV blue light on DNA repair. To limit biopsies, no samples were taken at T0.

Results

The rates of photoproducts (expressed as photoproduct per million normal bases) in the biopsies of the 3 donors are reported in Table 6 below.

Samples TT CPD TC CPD CT CPD Patient 1, zone A 8.16 1.30 0.65 Patient 1, zone B 11.33 5.74 0.77 Patient 2, zone A 6.49 4.02 0.32 Patient 2, zone B 7.69 3.06 0.12 Patient 3, zone A 4.04 2.37 0.00 Patient 3, zone B 5.34 2.53 0.00

The analyses show that a satisfactory quantity of DNA was obtained, on average 5µg per biopsy.

Table 7 below shows the average values for the 3 patients in TT CPD following UVB stress, preceded or not by HEV blue light stress.

Condition / Parameter TT CPD UVB stress 6.23 ± 1.2 per million normal bases (B) HEV Stress + UVB Stress 8.12 ± 1.7 per million normal bases (HB) Ratio (HB-B)/Bx100 30%

The averaged values above clearly highlight a higher rate of residual TT CPD when the skin is previously exposed to HEV blue light (zone B) before exposure to UVB radiation. This difference in the CPD rate observed at 48 hours, between the two zones, reveals that there was a higher repair of DNA lesions of the order of 30% in the absence of exposure to HEV blue light.

2.2. Study with application of composition according to the invention

• Une zone A exposée seulement à une lumière bleue HEV ;
• Une zone B exposée seulement à un rayonnement d’UVB selon les conditions du stress à un rayonnement UVB du protocole de la section 2.1 ci-dessus ;
• Une zone C exposée à une lumière bleue HEV et à un rayonnement UVB selon protocole de la section 2.1 ci-dessus ;
• Une zone D exposée à une lumière bleue HEV et à un rayonnement UVB selon protocole de la section 2.1 ci-dessus, l’exposition à la lumière bleue HEV étant réalisée après application sur la zone C de la formulation B (cf. tableau 2).

This study is carried out on healthy volunteers with skin phototype III or IV. Each volunteer is exposed locally on 4 distinct areas on the back, namely:

• A zone A exposed only to HEV blue light;
• A zone B exposed only to UVB radiation according to the UVB radiation stress conditions of the protocol of section 2.1 above;
• A zone C exposed to HEV blue light and UVB radiation according to the protocol in section 2.1 above;
• A zone D exposed to HEV blue light and UVB radiation according to the protocol in section 2.1 above, exposure to HEV blue light being carried out after application to zone C of formulation B (see table 2).

Pigmentation and erythema measurements are taken at 20h and 48h after the end of exposure to UVB radiation.

Biopsies with a diameter of approximately 4 mm are taken from zones A and B. They are collected only 48 hours after the end of exposure to UVB radiation to assess the possible effect of HEV blue light on DNA repair. To limit biopsies, no samples were taken at T0.

Bibliographic references

• Dréno et al. 2014, J Eur Acad Dermatol Venereol, 28, 1141-1149.
• Guiraud et al. 2014, Int J Cosmet Sci, 36, 485-493.
• Marzouki et al. 2017, Life Sci, 188, 10-16.
• Werner et al. 2013, Br J Dermatol, 169, 502-518
  

Claims (14)

5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for use in the prevention and/or treatment of a pre-cancerous skin lesion or a cancerous skin lesion. 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for its use according to claim 1, characterized in that the pre-cancerous skin lesion is an actinic keratosis and the pre-cancerous skin lesion is a carcinoma, in particular a squamous cell carcinoma. Composition comprising 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for its use in the prevention and/or treatment of a pre-cancerous skin lesion or a cancerous skin lesion. Composition for its use according to claim 3, characterized in that the pre-cancerous skin lesion is an actinic keratosis and the pre-cancerous skin lesion is a carcinoma, in particular an epidermoid carcinoma. Composition for its use according to claim 3 or 4, characterized in that it comprises a photoprotective system consisting of:
(a) 5,6,5',6'-tetraphenyl-3,3'-(1,4-phenylene)-bis[1,2,4]triazine,
(b) 2,4-bis[4-(2-ethylhexyloxy)-2-hydroxyphenyl]-6-(4-methoxyphenyl)-1,3,5-triazine,
(c) hexyl 2-[4-(diethylamino)-2-hydroxybenzoyl]benzoate, and
(d) ethylhexyl triazone or diethylhexyl butamido triazone,
the photoprotective system representing between 4% and 25% by weight relative to the total weight of the composition,
said composition not comprising any sunscreen other than those constituting said photoprotective system. Composition for use according to any one of claims 3 to 5, characterized in that the 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine represents between 1% and 5% by weight relative to the total weight of the composition. Composition for use according to any one of claims 3 to 6.
characterized in that 2,4-bis[4-(2-ethylhexyloxy)-2-hydroxyphenyl]-6-(4-methoxyphenyl)-1,3,5-triazine represents between 1% and 4% by weight relative to the total weight of the composition. Composition for use according to any one of claims 3 to 7, characterized in that the hexyl 2-[4-(diethylamino)-2-hydroxybenzoyl]benzoate represents between 1% and 10% by weight relative to the total weight of the composition. Composition for use according to any one of claims 3 to 8, characterized in that the sunscreen (d) represents between 1% and 6% by weight relative to the total weight of the composition. Composition for use according to any one of claims 3 to 9, characterized in that the sunscreen (d) is ethylhexyl triazone. 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for use in the repair of skin cell DNA damage and/or in the prevention and/or reduction of a skin cell DNA damage repair deficiency. Composition comprising 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine and at least one cosmetically and/or pharmaceutically acceptable excipient for use in repairing DNA damage in skin cells and/or in preventing and/or reducing a deficit in repairing DNA damage in skin cells. 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for its use according to claim 11 or composition for its use according to claim 12, characterized in that the DNA lesions are photoinduced, in particular by UV radiation. 5,6,5’,6’-tetraphenyl-3,3’-(1,4-phenylene)-bis[1,2,4]triazine for its use according to claim 11 or 13 or composition for its use according to claim 12 or 13, characterized in that the DNA damage repair deficit is due to exposure of skin cells to visible high-energy blue light.