FR3063641A1 - PREVENTION OF UV-CUTANEOUS LESIONS AND MELANOMAS USING A SELECTIVE INHIBITOR OF THE PRODUCTION OF REACTIVE SPECIES OF MITOCHONDRIAL OXYGEN - Google Patents

Abstract

The present invention relates to the prevention and / or treatment of diseases in which the reactive oxygen species (or ROS) of mitochondrial origin are involved. It relates more particularly to the use of an inhibitor of the production of ROS of mitochondrial origin to prevent cutaneous lesions due to UV, in particular the signs of aging, as well as the occurrence of melanomas. This invention can be implemented using a topical composition comprising anethole trithione; this composition may further contain 4-OH-anethole trithione and UV filters.

Description

Holder (s): PETITJEAN OLIVIER, PETITJEAN GUILLAUME, PETITJEAN ELODIE, PETITJEAN GREGOIRE, CHILDS MARC, SAUZIERES JACQUES.

Extension request (s)

Agent (s): IP TRUST.

PREVENTION OF SKIN LESIONS DUE TO UV AND MELANOMA USING A SELECTIVE INHIBITOR FOR THE PRODUCTION OF REACTIVE SPECIES OF OXYGEN OF MITOCHONDRIAL ORIGIN.

FR 3 063 641 - A1 (bq) The present invention relates to the prevention and / or treatment of diseases in which the reactive oxygen species (or ROS) involved in mitochondrial origin are involved. It relates more particularly to the use of an inhibitor of ROS production of mitochondrial origin to prevent skin lesions due to UV rays, in particular the signs of aging, as well as the occurrence of melanomas. This invention can be practiced using a topical composition comprising anethole trithione; this composition may also contain 4-OHanethole trithione and UV filters.

PREVENTION OF SKIN LESIONS DUE TO UV AND MELANOMA USING A SELECTIVE INHIBITOR FOR THE PRODUCTION OF REACTIVE SPECIES OF MITOCHONDRIAL OXYGEN

The present invention relates to the prevention and / or treatment of diseases in which reactive oxygen species (or ROS) are involved of mitochondrial origin. It relates more particularly to the use of an inhibitor of the production of mitochondrial ROS, in particular anethole trithione, to prevent skin lesions due to UV rays as well as the occurrence of melanomas.

PRIOR ART

The mitochondria participate in the pathogenesis of almost all diseases associated with aging, including cardiovascular diseases, neurodegenerative diseases (Parkinson's disease, Alzheimer's disease, etc.), diabetes, as well as tissue dysfunctions of ischemic origin. . It is widely accepted that it plays a central role in the "theory of free radicals in aging". This theory states that the accumulation of damage caused by reactive oxygen species (ROS) affects many cellular functions, in particular the mitochondrial functions, which are essential for energy supply and proper functioning. cellular. The mitochondria therefore appear to be the primary targets of ROS since optimal cell functioning is crucial to supply the energy that a cell needs to repair itself.

While mitochondria are the main source of ROS, they are also particularly sensitive to the damage caused by these ROS. Consequently, the mitochondria themselves generate ROS, which causes oxidative damage which they undergo and which contribute to their dysfunction and, subsequently, to cell death.

Numerous studies have been carried out to assess the ability of antioxidants to counteract the effect of ROS. Several antioxidant molecules have given satisfaction in preclinical studies, but their effectiveness has only been partially confirmed in most clinical trials (Orr et al., Free Radie Biol. Med, 2013, 65: 1047-59).

In addition, recent studies have shown that an excessive reduction in ROS has a deleterious effect on cells, suggesting that a balanced production of ROS contributes to good cellular functioning (Goodman et al., J. Natl. Cancer Inst. , 2004, 96: 1743-50; Bjelakovic G et al., JAMA. 2007, 297: 842-57).

TECHNICAL PROBLEM AND SOLUTION PROVIDED BY THE INVENTION

Studies relating to the role of oxidative stress in many pathologies have highlighted the advantage of having a selective inhibitor of ROS production of mitochondrial origin. The antioxidants available to date do not have such a specificity with, as a consequence, the risk of occurrence of side effects when the production of cytosolic ROS is affected, especially in case of prolonged treatment; these side effects are well described.

The oxidative stress caused by exposure to UV, that is to say to high energy radiation, is responsible for an overproduction of ROS by melanocytes, especially in subjects with a high phaeomelanin: eumelanin ratio. i.e. in subjects most at risk of developing melanoma (Liu-Smith et al., Exp Dermatol, 2015, 24: 171-174). To date, the source of this overproduction of ROS is still under discussion. According to the literature, it should be attributed to melanosomes, the hypothesis so far the most studied, with the participation of NOX (NADPH Oxydase phagolysosomes, enzyme system precursor of ROS) and that of neuronal NOS (nNOS or NO synthase type I cytosolic) (Venza et a!., Oxid Med Cell Longev, 2015, ID 481782). On the other hand, the involvement of mitochondrial production (complexes I and III of the respiratory chain) does not seem to have been looked at, perhaps for reasons of technical difficulties. However, unlike the overproduction of ROS of iatrogenic origin which most often targets in a very specific way a particular structure - lysosomes in the case of aminoglycosides or mitochondria in the case of anthracyclines, for example we are entitled to think that in the case of a production linked to exposure to radiation, it is the whole of the cellular structures producing ROS which will be simultaneously stimulated by the global inflammatory reaction thus induced which includes mitochondrial production.

Furthermore, UV rays, by activation of a transcription factor, USF-1, stimulate the proliferation of melanocytes and their activity, which contributes mechanically to the increase in ROS production.

The cellular antioxidant systems are then quickly overwhelmed and excess ROS will thus be able to induce the synthesis of a-Melanocyte Stimuloting Hormone (α-MSH or melanotropic hormone) (Huang et al., Int J Mol Sci, 2014, 15: 16665 -16679) and thereby the production of melanin. However, if melanin plays a protective role, therefore beneficial, with regard to UV exposure, an excess of production can have an opposite effect and become deleterious for the cell (Chaiprasongsuk et al., Redox Biol, 2016, 8: 79-90). Indeed, the binding of the α-MSH to its MC1R receptor (type 1 melanocortin receptor) induces an increase in the intracellular concentration of cyclic AMP and in PKA (protein kinase A) activity, which causes all both a stimulation of melanogenesis as well as a mitogenic effect on melanocytes, a source of cellular dysplasia.

At the same time, the overproduction of ROS, from a certain threshold, is ultimately responsible for a destructuring of the melanosomes with leakage of its various components, in particular ROS (in large quantities in the case of phaeomelanosomes), RNS (Reactivated nitrogen species , products of the action of NOS), melanin and amyloid substance; however, the latter potentiates the inducing effect of ROS towards the melonomo form in the same way as DNA breaks caused directly by UV (Liu-Smith, 2015).

Reduce the production of ROS by the cell and contain it at a level which allows the clearance of cells with damaged DNA (preventive effect of a malignant transformation) and which promotes, moreover, a fair increase in melanin synthesis (protective effect with regard to UV rays, melanin coming into contact with the keratinocyte nucleus protecting the cell's DNA) should constitute a good preventive approach to melanoma.

To date, there is no preventive treatment for melanomas, other than sun creams, the benefits of which are limited.

In the present invention, the inventors propose to reduce the production of ROS at the mitochondrial level in order to benefit from the preventive effect for the cell without undergoing the deleterious side effects of non-selective antioxidants. Thus, the inventors have established the advantage of administering a specific inhibitor of the production of mitochondrial ROS, in particular anethole trithione (ATT), to prevent skin damage linked to UV exposure as well as in the occurrence of melanomas.

The ATT already benefits from a marketing authorization under the trade name of Sulfarlem® to increase the secretion of bile and saliva. It is used to treat difficult digestion and dry mouth. In the 70 years that this product has been marketed in France, no side effects have been reported to date relating to long-term use of this molecule.

Indeed, the mechanism of action of ATT can be deduced from the results of another experiment not published to date: the inventors have successfully treated a cat victim of acute poisoning with organochlorines. This result demonstrates that the target of ΙΆΤΤ is the complex I (or III) or, possibly, the mitochondrial l-lll supercomplex and this in connection with the results described in 2012 by Tebourbi (Tebourbi et ai., Molecular mechanism of pesticide toxicity. In: STOYTCHEVA M. Ed. Pesticides in the modem world. Pests control and pesticides exposure and toxicity assessment. InTech publisher, 2012, chapter 15.). While knowing that as soon as ROS are overexpressed by the mitochondrion, this induces a simultaneous overproduction of cytosolic ROS, whose production source is NADPH oxidase (or NOX, in general NOX4 and NOX2), some authors arrive at the conclusion that NOX are the first responsible when it is actually a mitochondrial production (Gosh R. et al., J. Clin. Diagn. Res., 2017, 11: BC09- BC12; Mangum LC et al ., Chem. Res. Toxicol, 2015, 28: 570-584).

In addition, ΓΑΤΤ is, unlike the majority of other antioxidants, a fat-soluble molecule which gives it excellent tissue and cell penetration (the Vd of ADT is, in rats, of the order of 2L / kg , or almost 10 times the volume of extracellular fluids [Yu HZ et al. J Pharm Sci, 2011, 100: 5048-5058]), explaining its access to the mitochondria. Thus, ΓΑΤΤ acts at concentrations of the order of a micromolar, 10 μΜ in the work of Pouzaud et al. (2004), concentration which is easy to reach at the usual doses; and this molecule seems to have persistent effects over time, certainly for several hours which should facilitate the method of administration of the product.

Finally, after administration in animals as in humans, ΓΑΤΤ rapidly undergoes demethylation forming a derivative itself very likely to be active, 4-OHanetholtrithione (or ATX), carrying an alcohol function on the aromatic cycle; this function is esterifiable and can thus give access to water-soluble esters even though ΓΑΤΤ and its ATX metabolite are insoluble in water.

The inventors have thus established that ΓΑΤΤ, unlike conventional antioxidants, acts directly and selectively on the production of mitochondrial ROS, mainly at the level of complex I (or complex III) of the mitochondrial respiratory chain, sites which are both the main sites of ROS production and the main sites of mitochondrial dysfunctions (Tebourbi O., 2012, idem above).

ATT is therefore the first drug for human use authorized by the FDA and ΓΕΜΑ which prevents the mitochondria from producing ROS at the level of complexes I or III of the respiratory chain.

The present invention proposes, in a completely surprising and innovative way, to use this drug in a new therapeutic indication, namely to prevent skin damage linked to UV exposure as well as to prevent the occurrence of melanomas.

DETAILED DESCRIPTION OF THE INVENTION

A first object of the present invention relates to the use of a specific inhibitor of ROS production of mitochondrial orignin to prevent skin damage due to exposure to UV and melanomas.

The term “specific inhibitor of ROS production of mitochondrial origin” means any compound capable of specifically inhibiting the production of ROS in the mitochondrial respiratory chain, without affecting the production of cellular ROS in the cytosol; this specificity is essential because it prevents the side effects associated with a defect in ROS in the cytosol, as can be observed in the event of excessive inhibition of ROS production by a non-selective antioxidant. In a preferred embodiment, this inhibitor is capable of inducing a specific inhibition of the production of ROS at the level of the complex I (or III) of the mitochondrial respiratory chain.

Such inhibitors are, for example, ΙΆΤΤ, ΓΑΤΧ or NC-POBS, but any other compound having the same specificity of inhibition is suitable.

Within the meaning of the invention, the use of "an inhibitor" means the use of at least one specific inhibitor of the production of mitochondrial ROS; it can therefore be an inhibitor or a combination of several inhibitors, as described below.

ATT, for anethole trithione, is a 5- (4-methoxyphenyl) -3H-1,2-dithiole-3-thione. It is also known as ADT. Its formula is as follows:

ATX corresponds to the phenolic form of ΓΑΤΤ as it is metabolized by the liver, both in humans and in animals. This 4-OH-anethole trithione form has been described previously (Li et al., J Pharm Biomed Anal, 2008, 47: 612-617). The structure of ATT being conserved during this metabolization, there is every reason to believe that the antiROS activity carried by ATT is found in ATX, all the more so since after oral administration which is currently the marketed form, essential of the circulating product found is ΓΑΤΧ (Yu, 2011). In addition, ΙΆΤΧ carries a phenol group in para which allows the formation of esters. In a particular embodiment, ΙΆΤΧ is used in its esterified form, for example in the form of an ester: of phosphate, of ethylidenephosphate, of sulfate, of hemisuccinate, of acetate, of propionate, of isobutyrate, of hexanoate, pivalate, ethoxycarbonate, nicotinate, or an ester of amino acids such as glycine, diethylglycine or valine ester, and the list is not exhaustive.

NC-POBS corresponds to N-cyclohexyl-4- (4-nitrophenoxy) benzenesulfonamide, the only molecule described in the literature to date as a specific inhibitor of the production of mitochondrial ROS at site _Q of the respiratory chain ( Orr et al., 2013, Free Radie. Biol. Med., 65: 1047-1059).

In a preferred embodiment, the specific inhibitor is chosen ΙΆΤΤ, ΙΆΤΧ or an ATX ester. In a particular embodiment, the inhibition of damage linked to UV is obtained thanks to the association of at least two molecules among ΓΑΤΤ, ΙΆΤΧ and an ester of ATX.

To ensure optimal protection against UV, it is possible to combine ATT and ATX with UVA and UVB filters and, possibly, with ROS scavengers (ROS trappers) other if the level of cutaneous ROS reduction obtained with the couple ATT / ATX does not seem sufficient. As such, the choice of edaravone, marketed by the Mitsubishi Tanabe Pharma Corporation laboratories under the name of Radicut®, seems particularly interesting to us as a partner of the ATT / ATX pair, insofar as this molecule is a powerful specific inhibitor of nNOS (neuronal NOS or NOS type I) and iNOS (inducible NOS or NOS type II cytosolic) which also respects eNOS activity (or NOS type III endothelial) [Yoshida et al., CNS Drug Rev, 2006 , 12: 9-20],

Thus, in a particular embodiment, the specific inhibitor of mitochondrial ROS production is ΙΆΤΤ or a combination of ATT and ATX and is associated with UVA and / or UVB filters. It can also be associated with ROS scavengers, such as edaravone.

By "prevention" is meant in the sense of the invention the fact of inhibiting the deleterious effects of UV at the cellular and molecular level (DNA damage). Prevention also means reducing and slowing down the accumulation of damage linked to UV exposure in the skin. The prevention of UV damage extends to any pathology linked to the accumulation of DNA damage such as melanomas. The subject to be treated is preferably a human being.

“Prevention of skin damage due to UV exposure” also means prevention of skin aging, physiologically and from a cosmetic point of view.

Thus, inhibitors of mitochondrial ROS production can be used both in therapeutic applications, for example in subjects at risk of developing melanomas, and in cosmetic applications, in order to combat skin aging.

In a particular embodiment, the present invention applies to the prevention of melanomas in subjects with a strong phaeomelanin: eumelanin ratio, in fair-skinned subjects working in the sun (farmers, construction workers) and in people with fair skin residing in a country with a lot of sunshine (for example in Australia).

A second object of the present invention relates to a composition for topical use comprising ΙΆΤΤ (or one of its derivatives) and suitable excipients.

Such a composition can also comprise ΓΑΤΧ or an ATX ester, UVA and / or UVB filters and ROS trappers.

In a preferred embodiment, a composition according to the invention comprises ΙΆΤΤ and ΓΑΤΧ or a derivative.

In another preferred embodiment, a composition according to the invention comprises ATT, ΙΆΤΧ or a derivative and UVA and UVB filters. This composition can also comprise ROS trappers.

Such a composition can be used to prevent melanoma or to prevent damage linked to UV exposure, in particular aging.

Such a composition can be in the form of a sunscreen or a protective day cream against damage caused by solar UV rays.

The composition according to the invention is preferably formulated so as to be suitable for topical use. Knowing that, here, it is a question of protecting the skin from the effect of solar radiation, the forms with local application which appear appropriate are for example the emulsions Oil in Water (O / W) or Water in Oil (O / H), the latter having, as we know, the advantage of having a high absorption coefficient (between 40 and 80), and therefore potentially very good protective power. Thanks to its two phases, the emulsion makes it possible to incorporate both hydrophilic molecules and lipophilic molecules, an advantage which is used to combine different UV filters and thus increase the level of protection of sunscreen preparations. Here, ATT is strictly fat-soluble; however, in vivo, ATT is metabolized in animals as in humans to a phenolic compound by the liver, ΓΑΤΧ or 4-OH-anethole trithione (Li et al., J Pharm Biomed Anal, 2008, 47: 612- 617), which authorizes the synthesis of esters, a potential access route to water-soluble derivatives (examples: sulfate, hemisuccinate, phosphate, ethylidenephosphate, amino acid esters, etc.).

In the prevention of UV-related damage, in particular of melanoma, oral administration of the inhibitor according to the invention could be proposed in addition to the topical application of a cream containing it. This approach would, at least partially, protect from the problems of compliance with local treatments, while strengthening their action.

Claims (7)

1. Specific inhibitor of ROS production of mitochondrial origin chosen from anethole trithione (ATT), 4-OH-anethole trithione (ATX) or an ATX ester or a combination of at least two of these molecules , for its use in the prevention of skin damage due to UV exposure and melanoma. 2. Inhibitor according to claim 1, for its use in the prevention of skin aging linked to UV exposure. 3. Composition for topical use comprising ΙΆΤΤ and ATX or an ATX ester or a combination of at least two of these molecules and excipients. 4. Composition according to claim 3, further comprising UVA and / or UVB filters and / or ROS trappers. 5. Composition according to one of claims 3 or 4, intended for cosmetic use. 6. Composition according to one of claims 3 or 4, for its use in the prevention of skin damage due to exposure to UV and melanomas. 7. Composition according to claim 6, for its use in the prevention of skin aging.