WO2018211297A1

WO2018211297A1 - Improved dermatological formulations for cosmetic and pharmacological use, and cosmetic and pharmacological product prepared by means of said formulations

Info

Publication number: WO2018211297A1
Application number: PCT/IB2017/000589
Authority: WO
Inventors: Nicola PACINI
Original assignee: Pacini Nicola
Priority date: 2017-05-18
Filing date: 2017-05-18
Publication date: 2018-11-22

Abstract

Dermatological formulations for cosmetic and pharmacological use comprising, in combination with one another, 5-hydroxy-N-acetyltryptamine or N- Acetylserotonin (NAS) and melatonin or 5-methoxy-N-acetyltryptamine (MLT). The formulations of the invention induce an evident arrangement of the cytoskeletal structures, with reduction of the microfilaments or F-actin bundles, and reinforce the microtubules and the microtubular network, they considerably strengthen the effects of the MLT on its own and give the cytoskeletal structures a remodulating effect, which is not obtained with MLT alone.

Description

IMPROVED DERMATOLOGICAL FORMULATIONS FOR COSMETIC AND PHARMACOLOGICAL USE, AND COSMETIC AND PHARMACOLOGICAL PRODUCT PREPARED BY MEANS OF SAID FORMULATIONS

The present invention relates to improved dermatological formulations for cosmetic and dermatological use, and cosmetic and dermatological products obtained with these formulations.

The protective action of vitamin A and its derivatives on the epithelium regulating cell growth of the derma and epidermis and ail the epithelial tissues has been known for some time (Goldblatt H, Benischek M. 1925, Shapiro SS et al. 2013).

Several reports exist on the use of these substances in cosmetics and dermatology (Amann PM et al. 2014, Bandyopadhyay D 2009).

Vitamin A is a terpene alcohol, formed by the condensation of 4 units of isoprene to 20 atoms of carbon; it is metabolized in vivo in 2 derivatives resulting from oxidation of the alcohol function, thus distinguishing retinol or vitamin A, retinaldehyde and retinoic acid or all-trans-retinoic acid or ATRA.

These molecules perform a complex series of biological functions - from a dolichol effect similar in the processes of glycosylation of proteins, for vitamin A, to an action regulating cell growth and differentiation of the epithelium linings and mesenchymal cells - especially retinoic acid.

Many of these actions are carried out by bonding of the above-mentioned molecules with RAR and RXR type nuclear receptors for regulation of the respiratory complexes at mitochondrial level (Xun Z et al 2012, Urvalek A et al 2014, Das BC et al 2014).

Current knowledge considers vitamin A, and in particular retinoic acid or all-trans-retinoic acid (ATRA), as one of the derivatives most active in promoting and accelerating epithelial renewal and in increasing collagen synthesis, with a relaxing effect on the epithelial folds and renewal effect on the surface layer of the derma.

This substance has been widely used in the treatment of acne and other inflammatory pathologies of the derma and epidermis; it has been used as a spot removing or as an anti-melasma agent and against age spots or accumulations of lipofuscin, both alone and in combination with cortisone- and hydroquinone- based drugs; furthermore, due to its cell growth regulation properties, it has also been used to treat verrucas and many other problems in the dermatological and cosmetic field.

However, despite many positive aspects and encouraging results, said substance has some side effects which in many cases limit its use, including: a strong peeling effect which, although part of its main mechanism of action, can cause accentuated local irritation and reddening. Furthermore the use of vitamin A and ATRA on the skin can induce areas of non-keratinization, and dermatitis caused by hypersensitivity phenomena. It has also been observed that its antiwrinkle-antiage effect and stimulation of collagen synthesis is achieved only after several weeks of treatment, from 4 to 12 or more, thus entailing greater exposure to side effects and undesired effects.

Ubidecarenone or coenzyme Q10 is a benzoquinone, with a long isoprene chain formed by 10 molecules of isoprene; this highly liposoluble molecule is the main carrier of electrons through the various mitochondrial respiratory complexes and has been widely used both in formulations for oral administration, for example ubimaior or ubicol, and for topical use due to its assumed antioxidant and cytoprotective properties.

However, more recent studies have extensively modified these concepts since it has been observed that the molecule can not only mediate reducing reactions but, as an oxidation-reduction system, it can also trigger cascades of free radicals or ROS (Reactive Oxygen Species) which, via the regulation of complex mechanisms, can induce important biological effects, hydrogen peroxide being a by-product of cell respiration, a potential second messenger (de Luca C et al 2007, Kumar A et al 2015).

Hydroquinone is a very different molecule from ubidecarenone, and although it possesses the same benzoquinone nucleus, it has effects on melanogenesis and on melasma since it has a different pharmacokinetic and pharmacodynamic profile from ubidecarenone (Puizina-lvic N et al 2010). Vitamin E

From the chemical point of view, the term "vitamin E" refers to a group of eight homologues derived from 6-chromanol, which in position 2 have a methyl group and a lateral chain with 16 atoms of carbon. This lateral chain can be saturated or unsaturated, and therefore two groups are distinguished: the tocopherols (with saturated lateral chain) and the tocotrienols (with three double bonds); for each of the two groups there can be four homologues (α, β, γ, δ) according to the number and position of the methyl groups on the aromatic ring.

This molecule, like all its homologues, to a more or less varying extent, has marked reducing properties, interferes with various mitochondrial respiratory processes, and is known for its eutrophic effects on the derma and on the epidermis and for its antimelanogenic action (Jiang Q 2014).

Melatonin or 5-methoxy-N-acetyltryptamine (MLT) is an indolamine produced by the pineal gland or epiphysis, and by the gastrointestinal tract, by the cells of the retina and by many other histotypes. This substance, in vitro and in vivo, has shown considerable and pronounced reducing and scavenging activities of free radicals. However, more recent studies document a much wider and heterogeneous spectrum of activity and biochemical-functional effects than a generic antioxidant or reducing activity. In fact, it performs a series of important actions: on regulation of the respiratory complexes, at the level of the electron mitochondrial transport chain, on regulation of the acto-myosin filaments and actin components of the cytoskeleton; it also mediates many effects via GPCR (G protein-coupled receptors) and nuclear membrane receptors (Reiter RJ et al 1999, Slominski RM et al 2012, Cardinal! DP et al 2013, Pacini Borziani 20 1).

This substance was initially isolated in 1958 by the dermatologist Aaron

Bunsen Lerner from the bovine pineal gland, since, as had been known since 1917, pineal gland extracts contain a factor that regulates pigmentation in amphibians (lightening effect). Said factor was then identified as MLT.

Although successive studies have not provided conclusive and definitive results, and the literature in this regard continues to be controversial, it has been repeatedly demonstrated in amphibians and other models that MLT counters the dispersing action of the melanosomes by the peptide a-MSH or melanocyte stimulating hormone. From the data in the literature it can be seen that this functional antagonism by the MLT towards the a-MSH, with overall lightening effect, could be mediated by the microtubules, i.e. the MLT could prevent the gel- sol transition performed by the a-MSH on the cutaneous melanophores.

The mechanism of this effect is neither clear nor known today, furthermore even the actual existence of this mechanism in mammals and its extent is currently widely debated.

However, the action of MLT as a reducing substance or powerful and effective antioxidant with the capacity to penetrate the lipophilic membranes, penetrate the nuclear membrane and have a stabilizing and cytoprotective effect has meant that this substance has become very popular and widely studied, and there are several patent applications for oral or parenteral formulations that involve MLT.

For example, a patent by Murray N. Waldman, Jerry Vriend (CA 2230389 A1 ) 1999 describes a new system for use of the substance in cream bonded with albumin, in order to reduce percutaneous absorption and facilitate its conservation. As a cream formulation, another patent exists for a formulation comprising melatonin, ginko biloba and biotin by Hans W. Schmid (US 8062648 B2). Then there is a patent by Michael Cohen dated 1989 (WO 1992011855 A1) for the use of melatonin and its derivatives in association with estrogens/progestins in tablets, capsules, pessaries and vaginal suppositories in order to alleviate the symptoms of the menopause.

From studies performed on globular osmotic resistance of erythrocytes of rats, men and rabbits, it emerged that MLT increases erythrocyte deformability and interferes with polymerization of the F-actin filaments, structures also called microfilaments which, together with the microtubules and the intermediate filaments, constitute the cellular cytoskeletal system, responsible for the processes of exo/endocytosis, chemotaxis, diapedesis, migration and all contractile phenomena.

This datum is further confirmed since it increases the action of various toxins that interfere with the actin filaments and with their polymerization, including latrunculin, cytochalasins B and D, whereas it has little effect on, and partly favours, the action of colchicine, a metabolic toxin that interferes with the microtubules; these studies were always conducted in vitro in relation to globular osmotic resistance.

Similar studies were conducted in vitro (acellular system) where polymerization and depolymerization of the actin filaments was monitored in spectrophotometry; important results were obtained also in vitro on neoplastic cells of various lines, where the capacity of this indolamine to interfere with the processes of growth, apoptosis and cell death, but also and in particular on cell migration, emerged.

If there is an effect on these structures, there should also be an effect on leukocyte chemotaxis and diapedesis, since the MLT is totally atoxic. For this purpose the inventor performed a study on 25 healthy human volunteers, where the administration of 1 mg/kg at 9 p.m., followed by sampling and haemocytometer examination, literally modified the leukocyte formula, inducing a drastic reduction in the level of leukocytes in circulation well beyond the possible circadian variations, as shown in the following Table 1 :

Table 1 - Leukocyte formula variation on 35 persons, sampling at 9 p.m.

and 9 a.m.

±

The variation in the number of white globules is 22% (p<0.0001). In addition to the mean variation, it is interesting to note that not only the latter varies, but also the general variation in the distribution of the values in this group of persons if the mean variation in the number of leukocytes circulating after the administration of MLT has dropped by 22%. Equally significant is the fact that of the 35 persons examined, 10 show a reduction greater than 30%, two show a drop of over 50%, while one person shows a variation of 66%, with a modest leukopenia.

Indicating a direct effect on the processes of leukocyte chemotaxis and diapedesis, all phenomena largely cytoskeleton-dependent and especially actin- mediated, these and many other elements have led to the conclusion that MLT exercises a direct action on polymerization of the F-actin, reduces the intensity and distribution of said filaments and in particular directly modifies the assembly and disassembly kinetics of the G-actin in the F-actin polymers. The above- mentioned effects were evident only at pharmacological doses between 100μΜ and 1mM. On the basis of this, the substance has a strong stabilizing effect on the dynamics of the microtubules, as already evident in literature, which are stabilized and maintained functional.

5-hydroxy-N-acetyltryptamine or N-Acetylserotonin (NAS) is the derivative acetylated on the amine nitrogen of the serotonin, which leads to the synthesis of an amide. This compound is also an intermediate in the biosynthesis of melatonin, from which it differs due to the methoxylation in position 5'. Like MLT, the amide NAS has shown marked reducing or antioxidant properties and a spectrum of activity which in many aspects recalls that of MLT, but does not coincide perfectly as the ratios of power and effectiveness on the various receptors differ. In fact, compared to the MLT receptors, NAS is less effective and powerful in strictly pharmacological terms, and in some cases can function as a partial agonist and not a full agonist.

The compounds considered in the present invention, their molecular formula and the relative names are given in the following Table 2:

Table 2

The pigmentation of the skin in individuals producing eumelanin is regulated by fine molecular mechanisms and by the activity of the melanocytes, which form from the melanoblasts that derive, by migration during embryonal development, from the neuroectoderm; during the maturation phase these cells migrate to the level of the epidermis passing the junction between the connective and the epithelial tissue and settle between the keratinocytes of the basal layer. The melanocyte has a dendritic form, i.e. it has a cell body from which a certain number of branches depart which infiltrate the spaces in the intercellular labyrinth. Due to the three-dimensional development of its branches, a melanocyte can be in contact with thirty or so keratinocytes with which it constitututes the melanic unit.

As is widely known, within these cells, synthesis of the dark pigment melanin takes place, the precise structure of which is still debated; the key enzyme is tyrosinase which converts tyrosine into dopamine and from there, via various reactions, synthesis of the pigment is obtained.

Each melanocyte is in contact with various keratinocytes in the melanic unit, and the melanin synthesized in the cytoplasm is accumulated in structures called melanosomes. In basal conditions, the melanosome is distributed in the area of the perikaryon around the nucleus, and under stimuli like UV radiations it is decentralized from the periphery to the dendritic extensions, which are in contact with the keratinocytes and reabsorbed by them by cytocrinia.

The pigmentation of individuals of various races differs not so much due to the synthesis of melanin, which is approximately the same, but to its arrangement (sol-gel transition), the captation capacity on the part of the keratinocytes via the processes of cytocrinia and the capacity to decentralize the melanosomes from the perikaryon towards the dendritic extremities. These mechanisms are under the control of the acto-myosin networks and of the microtubules, with the microfilaments contributing to dispersion of the melanosomes and to drastically increasing the cutaneous pigmentation. The microtubules, on the other hand, make an essential contribution to condensation of the melanosomes around the perikaryon. These mechanisms are performed by the MLT.

The inventor has now surprisingly found that, for the described effects of the MLT to be even more evident and pronounced, the co-presence of NAS is necessary. The effects of NAS, MLT and NAS+MLT are illustrated in the electron micrographs of figures 1 to 3 respectively, where figure 1 is the control.

According to the invention, the co-administration of the two substances NAS and MLT: a) induces an evident arrangement of the cytoskeletal structures, with reduction of the microfilaments or F-actin bundles and reinforces the microtubules and the microtubular network;

b) this association considerably strengthens the effects of the MLT compared to the use thereof on its own;

c) gives the cytoskeletal structures a remodulating effect, which is not obtained with MLT alone.

Although some effects of MLT are known both on the cytoskeleton and on the melanosomes of amphibians, the effect of MLT on the microfilaments, or on the actin component of the cytoskeleton, is not currently known. Above all, the effect of the NAS+MLT association on the cytoskeleton is not known. Previous studies document a diametrically opposite effect produced by serotonin, a precursor of NAS, on the cytoskeleton and on the melanosomes, and on the aggregation of the melanosomes (Miller LJ et al 1989, Olivereau M 1978).

The inventors have surprisingly discovered that the NAS hugely strengthens the above-mentioned mechanisms and that said effects are maximum and easily seen with a NAS/MLT concentration ratio from 1/10 to 10/1 , preferably 1/10. In particular in the formulations of the invention, 1.5 to 0.01 % by weight of NAS and 1.5 to 0.1% weight of MLT are present, preferably but not exclusively in a NAS/MLT ratio = 0.1.

The formulations of the invention are distinguished in particular by being a topical drug containing: MLT 0.2% NAS 0.02% ATRA 0.1% as sodium salt, coenzyme Q10 0.2 % w/w.

The effects of the cited association of substances on the mechanisms described above is documented by the results shown in the following Table 3: Table 3 - Leukocyte formula variation on 35 persons, sampling at 9 p.m.

and 9 a.m.

The variation in the number of white globules is 29% (p<0.0001). Also in this case it is interesting to note not only the overall mean, and its statistical variation, but also the distribution, in fact if the mean variation in the number of leukocytes circulating after administration of MLT dropped by 29%. Equally significant is the fact that of the 35 persons examined, 12 show a reduction greater than 36% and 4 show a reduction greater than 50%

The attached figures from 1 to 3 show the electron myographs on a control sample, represented by untreated mammary cancer cells MCF-7, on a sample treated with MLT and on a sample treated with the NAS+MLT system in a ratio of 1 : 10 in a concentration of 0.00001 μΜ and 0.0001 μΜ respectively.

Thus, in short, the NAS+MLT system according to the invention, via a mediated cytoskeletal mechanism, influences the distribution of the melanosomes and their aggregation, partly countering the action of the a-MSH, and of various factors which induce the dispersion of the melanosomes and cytocrinia, with an overall action of redistribution of the cutaneous pigmentation.

Again according to the invention, this formulation combines stimulation of the collagen synthesis with acceleration of the epithelial renewal, of the Vitamin A or ATRA. For their part, the beneficial effects of MLT and of ubidecarenone and the homologue derivatives of Vitamin E on the respiratory complexes and on the electron transport chain ETC at mitochondrial level induce a stabilization of the generation of ROS and reduce an important stimulus for the induction of melanogenesis.

Furthermore, as per the tests performed, the synergic effect of these

NAS+MLT molecules conserves the benefits and known effects of the ATRA but reduces the unpleasant side effects, clearly improving the pharmacological profile of the ATRA which does not coincide with the ATRA alone.

By way of non-limiting example, the following creams were prepared with the formulations of the invention:

Example 1: topical drug NAS+MLT

Ratio (% weight) CONCENTRATION

5-methoxy-N- acetyltryptamine or Melatonin. MLT 0.2

5-hydroxy-N-acetyltryptamine or N-acetylserotonin NAS 0.02

All-trans-retinoic acid or tretinoin 0.1

Ubidecarenone or coenzyme Q10 0.2

a-tocopherol acetate enantiomer D, or D-tocopherol or

Minimum 4% max 8% γ-tocotrienol

EDTA 0.01 %

Oil/water emulsion

% 100 g of finished product Example 2: cosmetic NAS+MLT

These two formulations, the first of which is a topical drug containing (ATRA) and the n°2 which is cosmetic containing retinyi palmitate, or retinol or retinaldehyde, were used for various preparations also with different concentrations but always maintaining a concentration ratio MLT/NAS=10 MLT/ATRA=2 or in the case of the cosmetic MLT/Retinol or derivative = 2 and MLT/Coenzyme Q10=1.

Some examples are given below by way of example. If not specified otherwise, the contents of the above examples 1 and 2 always apply.

Example 3: topical drug NAS+MLT higher concentration

Ratio (% weight) CONCENTRATION (W W)

5-methoxy-N-acetyltryptamine or Melatonin. MLT 1

5-hydroxy-N-acetyltryptamine or N-acetylserotonin NAS 0.1

All-trans-retinoic acid or tretinoin 0.5

Ubidecarenone or coenzyme Q10 1

a-tocopherol acetate 8

EDTA 0.01%

Oil/water emulsion According to EU regulations

% 100 g of finished product Example 4.1: topical drug NAS+MLT

Concerning the importance of the ratios indicated in the previous tables, it can be hypothesised (UV and IR spectrophotometric data, in aqueous and ethanol solution, and in the solid state) that their essential importance in the stability of the finished product derives from the establishment of strong TT-stacking interactions between the aromatic nuclei of the compounds MLT and NAS and the diene groups of the ATRA or Retinol. Another important role is played by the Van der Waals forces (London forces) and by the hydrophobic bond.

EFFECTS AND USES

It has been observed and studied that both the creams of the previous examples have a strong peeling effect, accelerate epithelial renewal, have a marked remodelling and modulating action vis-a-vis cutaneous melanophores, induce a considerable improvement in age spots, and are useful in a series of dermatological diseases and aesthetic alterations.

The cream of the topical drug is used medically, in particular for acne, pathologies connected with alteration of the pigmentation, seborrheic dermatitis, cutaneous manifestations of scleroderma, 4th stage melanoma, as per Test no. 4 below, for palliative compassionate purposes, ulcerative lesions of breast cancer, as a topical palliative, verrucous affections and in cosmetic medicine as effective medical peeling and collagen synthesis accelerant, with consequent medical tensor and anti-wrinkle effect.

The cosmetic cream is used mainly as a cosmetic, non-medical peeling cream, as a skin tensor, as a broad spectrum anti-wrinkle and anti-ageing product.

The data described above refer to observational tests on 35 healthy persons with average age 45±8, 28 female and 7 male, after detailed informed consent. None of the above-mentioned persons manifested toxic or allergic effects or effects of other types. The cream of example 2 (topical drug) was applied in a random manner on 16 persons, while the cream of example 4 (cosmetic product) was tested on 19 persons.

At the end of a 30-day study each person was asked to fill in a questionnaire from which it emerged that: no-one manifested toxic effects, allergies or local hypersensitization.

In 4 persons there was a very slight reddening, due to excessive use of the compound.

MANUFACTURING PROCESS

The main components are mixed, dispersing the MLT and the NAS in absolute alcohol, then the ATRA is added or the retinol and derivatives, mixing in an environment with infrared lights or in a mixer not exposed to the light, since the components are photosensitive. The oil in water emulsion is prepared by means of a turboemulsifier (e.g. cetomacrogol with 4% glycerol and lipophilic phase with beeswax, previously emulsified containing tocopherol and derivatives in the percentages described above and the EDTA in addition to any stabilizers). Having reached a temperature of 35-40°C, the mixture is incorporated with the components in the emulsifier and the emulsification begins. Emulsion times of approximately 30'-40' guarantee almost total evaporation of traces of alcohol.

The following tests describe and report the results of application of the products of the invention.

Test no. 1: application of the topical drug of Example 1

The test was conducted on an elderly female patient (85 years old) affected by infiltrating ulcerated basal cell carcinoma (basalioma) in the region of the nose. In view of the general state of health compromised due to age-related pathologies and the simultaneous presence of suspect myeloma and blindness due to thrombosis of the retina, given the depth of the lesion which was infiltrative and locally extensive, and following the advice of expert dermatologists, the possibility of surgical removal of the lesion was ruled out in favour of possible subsequent cryotherapy with nitrogen after palliative treatment.

Instead of surgery, a palliative application of the retinoic acid-based cream of Example 2 was carried out for 40 days, every evening. 1 night-time application of 8 hours in the evening until washing in the morning at approximately 8 a.m. After 1 year the result is stable.

4 months after the treatment the patient is stable without recurrence and the product is applied 2 or 3 times a week without surgery or cryotherapy.

Test no. 2: application of the topical drug of Example 1

Trophic ulcer of the right ankle in a 72 year-old female patient affected by chronic venous insufficiency. The patient presented a severe form of chronic venous insufficiency, which had induced a state of edema in the legs and mainly (but not only) in the right leg, accompanied by the formation of an ulcerated lesion on the right ankle, persistent and recurring despite previous treatments. The patient underwent a course of elastic compression bandaging with the usual vasotonic products, a treatment that had already been used previously, but without a clear improvement in the small trophic ulcer. In particular every 48 hours elastic compression bandaging was performed after medication with disinfecting saline solution and application of the product based on tretinoin and/or salts thereof.

After 8 days of treatment with the cream of example 2 of the invention, a scab formed on the early-stage trophic ulcer with the initiation of a healing process which, as widely known, is difficult to obtain for trophic ulcers.

Test no. 3: application of the topical drug of Example 4.1

This test reports the effects of the product of Example 2 at 24, 48 and 72 hours in a male patient affected by SLE (Systemic Lupus Erythematosus) with eosinophilia in progress and high phlogosis indexes (PCR 9.8 Eosinophilia 8.6%) on an exudative eczematous lesion.

The benefit identified in terms of symptomatology was almost immediate.

Each application entailed use of the product in the evening with occlusive bandaging for a minimum of 8-10 hours. It was also interesting to note, in addition to the evident clinical improvement, the benefit in terms of lightening and tensor effect on the skin.

Test no. 4: application of the topical drug of Example 1

The test concerns the treatment of a basal cell carcinoma (epithelioma) in a 45 year-old male patient affected by cystic fibrosis who had undergone a lung transplant following everolimus-based immunosuppressive treatment. The patient presented over the years various recurrences of basal cell carcinoma (basalioma) (4 in 6 years) in the lateral region of the ears and right jaw.

After 30 days of treatment with the topical drug of example 2 of the invention, the lesion appeared reduced and almost resolved, to the extent that the dermatologist in charge opted not to remove it.

Test no. 5: application of the topical drug of Example 2

48 year-old female patient affected for approximately 25 years by a recurrent form of psoriasis, located mainly on the decollete and legs. The patient used the product of the invention every evening for 40 days and subsequently 3-4 times a week before going to bed. Almost total disappearance of the rash was observed.

Test no. 6: Application of the product of example 2 preceded by application of a product containing only MLT and not NAS

In a 48 year-old female patient, with histological diagnosis of malignant melanoma free from BRAF mutation (B-Raf proto-oncogene, serine/threonine kinase). With cerebral, pulmonary and hepatic metastasis and several evident skin lesions.

In view of the gravity and extent of the disease, the patient was declared untreatable.

Solely for compassionate palliative purposes and exclusively for the aspects concerning the skin lesion, after informed consent, a formulation containing exclusively retinoic acid 1% and MLT 2% NAS 0.2% W/W formulation example 3, as documented by photographic material, was applied in the evenings with bandaging for 8 hours for a total of 25 applications, after which almost total regression of the lesion is documented. Subsequently a formulation was applied containing only ATRA 1% MLT 2%, in addition to the other components excluding the NAS, in an extremely brief interval over a period of 15 days, after which total resumption of the melanogenesis, documented by a vast skin lesion, occurred. The photos in figures 4 and 5 show the initial situation, the photo in figure 6 the situation after 6 days of treatment, with the formulation 3, the photos in figure 7 after 10 days, in figure 8 after 12 days, in figure 9 after 15 days while the photos in figures 10 and 11 show the situation after 20 and 25 days of treatment respectively with the formulation 3 containing NAS.

After this period of 25 days the treatment is suspended and a formulation is applied containing all the active ingredients of the type 3 formulation, but without the presence of NAS. Over a period of 15 days resumption of the melanogenesis is documented with the formation of a vast lesional area occupying the entire umbilical region. Photos in figures 12 and 13.

Extensive photographic documentation documents blocking of the melanogenesis and substantial remission of the skin lesion. It should be pointed out that, in clinical terms, other untreated lesions showed practically no modification, furthermore the patient in question did not undergo any other medical therapy, apart from supporting treatments.

The result in question is indicative of an antiproliferative action of the composition concerned and synergy of the indols MLT and NAS in redistribution of the cutaneous melanophores and blocking of the melanogenesis.

Although metastatic melanoma remains an incurable disease, this example demonstrates that: A) The product is particularly active on the melanogenesis and therefore valid also for aesthetic depigmentation purposes. B) The simultaneous presence of MLT + ATRA is not sufficient in an extreme case of high melanogenesis, like the present one, to obtain blocking of the melanogenesis and/or redistribution of the cutaneous melanophores. C) In fact, the prompt resumption of the melanogenesis after suspension of the type 3 formulation and the use of a product without NAS indicates that there is not a remission from necrotic lesions but there is an effective and prompt anti- melanogenetic effect associated with inhibition of the proliferation.

Test no. 7: application of the cosmetic product of Example 4

The cosmetic formulation of Example 4 of the invention was given to 36 healthy persons aged 48±3.8, 24 of whom were female and 12 male. Out of the 36 persons, only 4 (8.33%) complained of a slight irritating effect (peeling), resulting from an arbitrarily abnormal use of the product. 31 out of 36 persons subject to the treatment (86.11 %) said they were satisfied with the quality of the product, which was also judged to be better than others previously used for cosmetic purposes.

To the invention as described above modifications can be made to obtain variations that however fall within the protective scope of the following claims. In particular, without prejudice to the decisive co-presence of the substances NAS and MLT, the formulations of the invention can contain even only one or more of the remaining substances indicated in examples 2 and 4. References:

Goldblatt H, Benischek M. (1927) VITAMIN A DEFICIENCY AND METAPLASIA. J Exp Med. 1927 Oct 31 ; 46(5):699-707

Shapiro SS, Seiberg M, Cole CA. Vitamin A and its derivatives in experimental photocarcinogenesis: preventive effects and relevance to humans. J Drugs Dermatol. 2013 Apr; 12(4): 458-63. Review

Everts HB. Endogenous retinoids in the hair follicle and sebaceous gland. Biochim Biophys Acta. 2012 Jan; 1821(1): 222-9. doi: 10.1016/j.bbalip.2011.08.017. Epub 2011 Sep 3. Review. Amann PM, Merk HF, Baron JM. (2014) Retinoids in dermatopharmacology Hautarzt. 2014 Feb; 65(2): 98-105. doi: 10.1007/s00105-013-2649-3. Review.

Bandyopadhyay D. Topical treatment of melasma. Indian J. Dermatol. 2009, 54(4):303-9. doi: 10.4103/0019-5154.57602.

Urvaiek A, Laursen KB, Gudas LJ. (2014) The roles of retinoic acid and retinoic acid receptors in inducing epigenetic changes. Subcell Biochem. 2014; 70:129- 49. doi: 10. 007/978-94-017-9050-5_7. Review.

Xun Z, Lee DY, Lim J, Canaria CA, Barnebey A, Yanonne SM, McMurray CT. (2012) Retinoic acid-induced differentiation increases the rate of oxygen consumption and enhances the spare respiratory capacity of mitochondria in SH- SY5Y cells. Mech Ageing Dev. 2012 Apr; 133(4): 176-85. doi: 10:1016/j.mad.2012.01.008. Epub 2012 Feb 8

Das BC, Thapa P, Karki R, Das S, Mahapatra S, Liu TC, Torregroza I, Wallace DP, Kambhampati S, Van Veldhuizen P, Verma A, Ray SK, Evans T. (2014) Retinoic acid signaling pathways in development and diseases. Bioorg Med Chem. 2014 Jan 15; 22(2):673-83. doi: 10.1016/j.bmc.2013.11.025. Epub 2013 Nov 22. Review.

Reiter RJ, Tan DX, Cabrera J, D'Arpa D. Melatonin and tryptophan derivatives as free radical scavengers and antioxidants. Adv Exp Med Biol. 1999; 467: 379-87. Slominski RM, Reiter RJ, Schlabritz-Loutsevitch N, Ostrom RS, Slominski AT. Melatonin membrane receptors in peripheral tissues: distribution and functions. Mol Cell Endocrinol. 2012;351(2): 152-66.

Cardinal! DP, Pagano ES, Scacchi Bernasconi PA, Reynoso R, Scacchi P. Melatonin and mitochondrial dysfunction in the central nervous system. Horm Behav. 2013;63(2): 322-30.

Jiang Q.(2014) Natural forms of vitamin E: metabolism, antioxidant, and antiinflammatory activities and their role in disease prevention and therapy. Free Radic Biol Med. 2014 Jul; 72:76-90. doi: 10.1016/j.freeradbiomed.2014.03.035. Epub 2014 Apr 3. Review.

Puizina-lvic N, Miric L, Carija A, Karlica D, Marasovic D.(2010) Modern approach to topical treatment of aging skin. Coll Antropol. 2010 Sep; 34(3): 1145-53. Review. de Luca C, Deeva I, Mikhal'Chik E, Korkina L.(2007) Beneficial effects of pro- /antioxidant-based nutraceuticals in the skin rejuvenation techniqueCell Mol Biol (Noisy-le-grand). 2007 Apr 15;53(1 ):94-101.

Kumar A, Singh A.(2015) A review on mitochondrial restorative mechanism of antioxidants in Alzheimer's disease and other neurological conditions. Front Pharmacol. 2015 Sep 24;6:206. doi: 10.3389/fphar.2015.00206. eCollection 2015. Review.

Miller LJ.( 989) Serotoninergic activity stimulates melanin dispersion within dermal melanophores of newts. Life Sci. 1989;44(5):355-9.

Olivereau M.(1978) Serotonin and MSH secretion: effect of parachlorophenylalanine on the pituitary cytology of the eel. Cell Tissue Res. 1978 Jul 13;191(1):83-92.

Claims

1. Dermatological formulations for cosmetic and pharmacological use, characterized in that they comprise, in combination with one another, 5-hydroxy- N-acetyltryptamine or N-Acetylserotonin (NAS) and melatonin or 5-methoxy-N- acetyltryptamine (MLT).

2. Formulations according to claim 1 , characterized in that they have a NAS/MLT concentration ratio from 1/10 to 10/1.

3. Formulations according to claim 2, characterized in that they comprise 1.5 to 0.01 % by weight of NAS and 1.5 to 0.1 % weight of MLT.

4. Formulations according to claim 1 , characterized in that they are a topical drug containing: MLT 0.2% NAS 0.02% ATRA 0.1 % such as sodium salt, coenzyme Q10 0.2% w/w.

5. Formulations according to claim 1 , characterized in that they are a cosmetic for topical use.

6. Formulations according to claim 1 , characterized in that they are a topical drug with high anti-melanogenic and depigmenting activity having the formula:

7. Formulations according to claim 1 , characterized in that they are a cosmetic product having the formula: Ratio (% weight) CONCENTRATION (W/W)

5-methoxy-N-acetyltryptamine or Melatonin. MLT 0.2

5-hydroxy-N-acetyltryptamine or N-acetylserotonin NAS 0.02

Retinol 0.1

Ubidecarenone or coenzyme Q10 0.2

a-tocopherol acetate enantiomer D, or D-tocopherol or

4

γ-tocotrienol

EDTA 0.01

Oil/water emulsion

% 100 g of finished product

8. Topical drug for dermatological use, characterized in that it is a cream containing the formulations according to one of the preceding claims.

9. Cosmetic product for dermatological use, characterized in that it is a cream containing the formulations according to one of the preceding claims.

10. Use of the topical drug formulations according to one or more of the preceding claims for the treatment of dermatological diseases, in particular basal cell carcinoma (basalioma and epithelioma), trophic ulcers, SLE, psoriasis and metastatic melanoma.