Cosmetic, cosmeceutical and / or pharmaceutical composition, uses and methods

Abstract

The present invention describes new cosmetic, cosmeceutical and / or pharmaceutical compositions, as well as their uses and useful methods for alleviating conditions arising from inflammatory processes, sensitive skin, rosacea, atopic dermatitis, eczema, and / or for alleviating the sensation of discomfort and pain, modulating the CB2 receptor, and / or for improving problems relating to cutaneous hyperchromia and disorders relating to increased sebogenesis.

Description

COSMETIC, COSMECEUTICAL AND / OR PHARMACEUTICAL COMPOSITION, USES AND METHODS Technical Field of the Invention

[0001] The present invention describes new cosmetic, cosmeceutical and / or pharmaceutical compositions, as well as their uses and useful methods for alleviating conditions arising from inflammatory processes, sensitive skin, rosacea, atopic dermatitis, eczema; modulation of the CB2 receptor; and / or; for alleviating the sensation of discomfort and pain; and / or; to bring benefits to problems related to skin hyperchromia and disorders related to increased sebogenesis. Background of the Invention

[0002] Plants of the genus Cannabis have a long history of human use, with evidence that they were used more than 10,000 years ago for improving pottery and may have been one of the first known agricultural crops (TEXTILE WORLD, 2020).

[0003] Cannabis has been used for food, paper and textile production, and, more recently, as a raw material for biodegradable plastics. Among the secondary metabolites found in these plants are tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabigerol (CBG), tetrahydrocannabivarin (THCV), and other substances classified as cannabinoids. Like the secondary metabolites of other botanical species, cannabinoids have been studied to determine their bioactive properties in the human body.

[0004] The use of medicinal cannabis may have begun around 2,000 BC, although the compendium that documents it dates back to the Han dynasty (221 BC - 220 AD) (CROCQ, 2020). It should be noted that not all cannabinoids have psychoactive effects, with THC being the main psychoactive substance present in cannabis.

[0005] The hypothalamic-pituitary-adrenal (HPA) axis is a neuroendocrine system that regulates many bodily processes through the release of of hormones. The skin not only expresses proteins from this axis but also responds to its signaling, suggesting the existence of a brain-skin axis. Thus, it is understood that substances that have biological activity in the brain can eventually trigger biological processes in the skin. In fact, an imbalance in the brain-skin axis has been linked to inflammatory dermatological conditions such as psoriasis (MOON, 2013).

[0006] The endocannabinoid system (ECS) is a neuromodulatory system composed of cannabinoid receptors, endogenous cannabinoid substances (endocannabinoids), and a set of enzymes responsible for their synthesis and degradation. The ECS acts on the central nervous system (including the brain) and the peripheral nervous system. Among the cannabinoid receptors, cannabinoid receptor 1 (CB1) stands out for its abundance, although other receptors are also modulated by cannabinoid substances—such as cannabinoid receptor 2 (CB2), transient receptor potential channels (TRPs), and peroxisome proliferator-activated receptors (PPARs). There is evidence that the main cannabinoid receptors, CB1 and CB2, are also expressed in the skin, suggesting that the ECS may be part of the brain-skin axis (THE PROTEIN ATLAS).

[0007] Despite the proven effectiveness of CBD in several areas, such as the treatment of epilepsy and its great potential for treating other disorders such as migraine (GRECO, 2023) and dementia (ALEXANDRI, 2023), the possible presence of the psychoactive substance tetrahydrocannabinol (THC) in Cannabis extracts (PINO, 2023) means that products based on these plants are considered, at the very least, controversial - for example, THC intoxication has been reported through the use of Cannabis oil marketed as CBD oil (MOUSSA, 2023).

[0008] In addition to the toxicological aspect, which could be overcome by conducting chemical analyses to evaluate and control narcotic substances in CBD oil, there are other elements that hinder the complete dissemination of this ingredient in the market, such as regulatory and legal issues. and supply chain.

[0009] In Brazil, for example, although the sale of CBD-based medicines has been authorized by the National Health Surveillance Agency (Anvisa), local production of CBD (at least from cannabis) remains prohibited. Such restrictions make CBD inaccessible, both economically and practically.

[0010] In the cosmetics sector, the first consumer products containing CBD hit the market around 2018, where the substance was primarily used as an anti-inflammatory agent for the skin. In 2020, nearly 300 new CBD-containing products were launched—and since then, that number has been declining. This can be explained, in part, by the controversial nature of this ingredient.

[0011] In the search for the state of the art, the following patent documents were detected. W02008014414 published on January 31, 2008 describes methods for attenuating the release of inflammatory mediators using the MANS peptide (MA-GAQFSKTAAKGEAAAERPGEAAVA - SEQ ID NO. 1) and derivatives, where "MA" is a myristoyl group attached to the N-terminal portion. Regarding the published experimental data, it is not possible to state that any terminal portion of the MANS peptide has inhibitory activity against inflammatory mediators. That is, the peptides MA-SKTAAKGEAAAERPGEAAVA (SEQ ID NO. 15) and MA-RPGEAAVA (SEQ ID NO. 153) did not show any inhibition of EPO release, and the peptide Ac-RPGEAAVA did not show any inhibition of lysozyme release. The peptide closest to the present invention tested was RPGEAAVA, with only 50% identity to the peptide tested by the present invention, but with a totally different structure due to the number of amino acids present (8 vs 4).Additionally, there is no experimental data indicating action on the levels of IL-6, IFN-gamma and KC (analog of IL-8 in murine) for peptides containing tetrapeptide sequences. The indicated tests with the MANS peptide (with only 17% identity to. tetrapeptides claimed herein) are entirely abstract, and no factual experimental results are presented. Results obtained with other sequences that presented results similar to SEQ ID NO. 153 (50% identity), as well as SEQ ID. 106, are also highlighted. None of them are identical to the peptides tested in this patent application. If any effect is observed, it would be with SEQ ID NO. 106, which does not contain "AAVA." Therefore, it is clear that, with regard to the information provided in WO2008014414, the sequence AAVA could not be inferred as responsible for the effects. Therefore, it is possible to conclude that the technical information provided in the present patent application under analysis demonstrates the surprising technical effects found.

[0012] WO2023212443, published on November 2, 2023, describes methods of treating a disease or condition that produces inflammation and / or itching of the skin. Although the document generically indicates that it uses peptide fragments of the protein titled by the document as MARCKS (GAQFSKTAAKGEAAAERPGEAAVA), the only factual / experimental test is with a peptide with a sequence entirely different from that claimed in the present patent application and sequence (Ac-GAQFSKTAAK-OH). Thus, no tetrapeptide or similar is supported or sufficiently described to be relevant to this patent application.

[0013] Therefore, as several effective therapeutic applications of cannabinoids have already been demonstrated, it is important to search for substances that act on the endocannabinoid system in a safe and effective manner as an alternative to those found naturally in plants of the Cannabis genus. Summary of the Invention

[0014] In this sense, the patent application presents new cosmetic, cosmeceutical and / or pharmaceutical compositions of peptides containing at least one peptide with the general formula:

[0015] R1 -A1 -A2-A3-A4-R2 where: A1 is an amino acid containing a non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly, norleucine, norvaline, 2-aminobutyric acid and sarcosine; A2 is an amino acid containing non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly, norleucine, norvaline, 2-aminobutyric acid and sarcosine; A3 is an amino acid containing non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly, norleucine, norvaline, 2-aminobutyric acid and sarcosine; A4 is an amino acid containing non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly-, norleucine, norvaline, 2-aminobutyric acid and sarcosine. R1 is an N-terminal chain modifier selected from the group consisting of H, substituted or unsubstituted acyclic aliphatics, substituted or unsubstituted cyclic aliphatics, substituted or unsubstituted heterocyclics, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted acyl, substituted or unsubstituted carbonyls; R2 is selected from the group consisting of -NR3R4, OR3 and -SR3, where R3 and R4 are independently selected from the group consisting of H, substituted or unsubstituted acyclic aliphatics, substituted or unsubstituted cyclic aliphatics, substituted or unsubstituted heterocyclics, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted acyl, substituted or unsubstituted carbonyls, as well as their stereoisomers, mixture and / or their cosmetically or pharmaceutically acceptable salts.

[0016] These peptides are particularly useful for acting on the endocannabinoid system (more specifically, because they have a high affinity for the CB2 receptor; and concomitant low or no affinity for CB1 receptor), thus providing the therapeutic benefits of a cannabinoid but without producing psychotropic effects. This peptide can be produced with extremely high purity, eliminating adverse effects such as those observed in some oils extracted from Cannabis.

[0017] Furthermore, in the studies carried out and which will be demonstrated throughout this patent application, it was observed that the peptide acts as an anti-inflammatory and / or antinociceptive substance, in addition to modulating melanin production, with performance equivalent to or superior to other substances used for the same purposes in the cosmetic area.

[0018] The peptides presented in this invention are safe and effective substances in reducing inflammation, as proven through studies in in vitro, ex vivo and in vivo models. Description of Figures

[0019] Figure 1: Effect of the peptide on the synthesis of IL-1 a in human keratinocytes, after 6 hours of treatment in comparative differences between the groups tested. ## P<0.01 compared to the basal control group. **P<0.01 compared to PMA / LPS (Dunnett's multiple comparisons test).

[0020] Figure 2: Effect of the peptide on the synthesis of IL-6 in human keratinocytes, after 48 hours of treatment in comparative differences between the groups tested. ## P<0.01 compared to the basal control group. **P<0.01 compared to PMA / LPS (Dunnett's multiple comparisons test).

[0021] Figure 3: Semi-quantification of melanin levels. Data obtained by recording images of histological sections of human skin fragments previously treated with a formulation without active ingredient (Placebo) and treated with the same formulation containing the peptide SEQ ID No. 1 (Succinyl-L-alanyl-L-alanyl-L-valyl-L-alanine) (at 200 ppm or 100 ppm). ##P<0.01 compared to the basal control group. **p<0.01 compared to the placebo group. Detailed Description of the Invention

[0022] The present detailed description of the invention establishes some non-limiting definitions of the main terminologies and technical characteristics employed throughout this patent application, as well as provides examples of some of the embodiments of the present invention so that it can be reproduced by a person skilled in the art.

[0023] The present invention describes the use of tetrapeptides capable of alleviating conditions arising from inflammatory processes, such as, but not limited to, sensitive skin, rosacea, atopic dermatitis, eczema, among others. Simultaneously, the present invention is intended for applications to alleviate the sensation of discomfort and pain. Additionally, the invention provides benefits for problems related to skin hyperchromia and disorders related to increased sebogenesis.

[0024] In a preferred embodiment of the present invention, R1 and R2 described in the general formula defining the peptide group are not proteinogenic amino acids.

[0025] For the description of amino acids in this document, 1- or 3-letter abbreviations may be used, as widely adopted by the scientific community and the International Nucleotide Sequence Database.

[0026] This invention further includes any and all conservative substitutions of the sequence SEQ ID No. 1. Conservative substitution means the replacement of one amino acid by another with similar hydrophobicity, polarity and / or side chain, so that there is little or no three-dimensional or functional change in the peptide.

[0027] Examples of these peptides include Succinyl-Ala-Ala-Val-Ala-OH (SEQ ID No. 1), Pal-Ala-Ala-Val-Ala-OH (SEQ ID No. 2), Ac-Ala-Ala-Val-Ala-OH (SEQ ID No. 3), Succinyl-Gly-Ala-Val-Ala-OH (SEQ ID No. 4), and Ac-Gly-Ala-Val-Ala-OH (SEQ ID No. 5), with the prefix Pal- identifying an N-palmitoylated peptide and the prefix Ac- identifying an N-acetylated peptide.

[0028] Such peptides can be obtained through various methods, preferably by chemical synthesis in solid phase or in homogeneous systems, without excluding other possibilities, such as biotechnology, hydrolysis and / or protein purification and other methods that may exist or may come to exist.

[0029] These peptides can be used in a concentration range between 0.001% (w / w) to 10.00% (w / w), preferably close to 0.01% (w / w), applied in a cosmetic or pharmaceutical formulation suitable for human or veterinary use.

[0030] A cosmetic or pharmaceutical formulation suitable for human or veterinary use is understood to include any and all types of solvents, dispersion media, encapsulations, permeation enhancing agents, surfactants, preservatives and other ingredients, alone or in combination, that are physiologically compatible.

[0031] Example 1: In vitro evaluation of inhibition of IL-1 alpha production.

[0032] Human keratinocytes were seeded in 75 cm flasks 2 cultivated and expanded in a humidified greenhouse at 37 SC in the presence of 5% CO2, using a specific culture medium. Upon reaching 70–80% confluence, the cells were trypsinized and seeded in 6-well plates for subsequent incubation with the test product and evaluation of the proposed parameter. The cells were incubated with two non-cytotoxic concentrations of the peptide (120 ppm and 60 ppm). The cells were then incubated under the same culture conditions mentioned above for 6 hours for subsequent collection of the cell lysate. Subsequently, the pro-inflammatory mediator IL-1 alpha was quantified using the ELISA technique.

[0033] The results expressed in Figure 1 demonstrate the anti-inflammatory capacity of the peptide Succinyl-Ala-Ala-Val-Ala-OH (SEQ ID No. 1). At a concentration of 120 ppm, the peptide reduces IL-1α levels by 31%, compared to the group stimulated with PMA / LPS.

[0034] Example 2: In vitro evaluation of inhibition of IL-6 production.

[0035] Human keratinocytes were seeded in 75 cm flasks 2 cultivated and expanded in a humidified greenhouse at 37 S C in the presence of 5% CO2, using a specific culture medium. Upon reaching 70-80% confluence, the Cells were trypsinized and seeded in 6-well plates for subsequent incubation with the test product and evaluation of the proposed parameter. The cells were incubated with two non-cytotoxic concentrations of the peptide (120 ppm and 60 ppm). The cells were then incubated under the same culture conditions mentioned above for 48 hours for subsequent collection of the cell supernatant. Subsequently, the pro-inflammatory mediator IL-6 was quantified using the ELISA technique.

[0036] The results expressed in Figure 2 demonstrate the anti-inflammatory capacity of the peptide Succinyl-Ala-Ala-Val-Ala-OH (SEQ ID No. 1). At a concentration of 60 ppm, the peptide reduces IL-6 levels by 33%, compared to the group stimulated with PMA / LPS.

[0037] Example 3: Evaluation of inhibition of melanin synthesis in human skin fragments.

[0038] Skin explants, obtained from a surgical procedure performed by an ophthalmological plastic surgery clinic, were treated for 72 hours with a formulation containing the peptide Succinyl-Ala-Ala-Val-Ala-OH (SEQ ID No. 1) at 200 ppm or 100 ppm and with the same formulation without the addition of the peptide (placebo).

[0039] After incubation with the test products as described above, the skin fragments were fixed in 4% paraformaldehyde (pH 7.4) for 24 hours. After fixation, the material was transferred to a 30% sucrose solution for cryopreservation and then embedded in tissue freezing medium. Serial sections of approximately 10 μm thickness were cut using a cryostat and collected directly onto silanized glass slides. The skin sections were then stained using the Fontana-Masson technique. Subsequently, the slides were analyzed and recorded using an optical microscope coupled to a camera. The results can be seen in Figure 3.

[0040] Example 4: Preparation of a cosmetic composition comprising peptide SEQ IP No. 1.

[0041] A cosmetic composition was prepared whose ingredients are shown in Table 1 below. In a suitable container, the Phase B ingredient was added and heated to 75-80°C. Once this temperature was reached, the Phase A ingredient was added, maintaining the temperature for 10 minutes. Then, cooling was initiated to 40-45°C with stirring. Subsequently, the Phase C ingredients were added, one by one, with stirring and homogenized for another 10 minutes, obtaining a cosmetic composition with the proportions shown in Table 1. The composition is a cream suitable for topical use.

[0042] Table 1. Cosmetic composition comprising peptide SEQ ID No. 1.

[0043] Example 5: In vivo study with the composition of Example 4, testing the efficacy of the peptide SEQ IP No. 1 for anti-erythema action induced in volunteers.

[0044] This study included 15 volunteers of both sexes, phototypes II or III, between 20 and 55 years old. In the back region, each volunteer had two 2 cm quadrants 2 demarcated, and both exposed to UV-B radiation with a solar simulator (ULTRAVITALUX LAMP - OSRAM), at a dose of 4J / cm 2 . Immediately after, a formulation (placebo cream - without active ingredient - cream containing the peptide at 100 ppm) was applied to each quadrant. Colorimetric measurements to assess the reduction in erythema (using the a* parameter); in addition to subjective measurement of the sensation of heat (guided by an opinion survey questionnaire with psychometric responses to identify the level of affirmation of the observed efficacy) were performed immediately (T0), 30 minutes (T30), 1 h (T60) and 2h (T120) after application of the formulations. The results for reduction of induced erythema and relief of the sensation of heat are shown in Tables 2 and 3.

[0045] Product efficacy is assessed by evaluating the whitening parameters (L, ITA, Melanin Index): L (Luminosity); ITA (Individual Typology Angle); Melanin Index. Table 2 shows the percentage variations in relation to the initial time of the parameters related to pigmentation.

[0046] Table 2. Variation of the parameter a*.

[0048] The products resulting from the use of the knowledge of the present invention can be used as cosmetics, cosmeceuticals, or pharmaceuticals, for human or veterinary use, for topical use, capable of alleviating conditions resulting from inflammatory processes, such as, but not limited to, sensitive skin, rosacea, atopic dermatitis, eczema, among others. Simultaneously, the present invention is intended for applications to alleviate the sensation of discomfort and pain. Additionally, the invention provides benefits for problems related to skin hyperchromia and disorders related to increased sebogenesis. This list of applications includes cosmetics, creams, lotions, serums, ointments, and rinse-off products.

[0049] The examples disclosed here are intended to merely exemplify some of the numerous ways of realizing and using the present invention, without limiting the interpretation of its scope and breadth, as well as, on possible alternative forms and configurational variations thereof that will be defined based on the claims presented here.

Claims

Claims 1. Cosmetic, cosmeceutical and / or pharmaceutical composition characterized by comprising at least one compound comprising the general formula: R1 -A1 -A2-A3-A4-R2 in which: A1 is an amino acid containing a non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly, norleucine, norvaline, 2-aminobutyric acid and sarcosine; A2 is an amino acid containing non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly, norleucine, norvaline, 2-aminobutyric acid and sarcosine; A3 is an amino acid containing non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly, norleucine, norvaline, 2-aminobutyric acid and sarcosine; A4 is an amino acid containing non-polar side chain and selected from the group Ala, He, Leu, Val, Gly, Pro, He, Gly-, norleucine, norvaline, 2-aminobutyric acid and sarcosine; wherein; R1 is an N-terminal chain modifier selected from the group consisting of H, substituted or unsubstituted acyclic aliphatics, substituted or unsubstituted cyclic aliphatics, substituted or unsubstituted heterocyclics, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted acyl, substituted or unsubstituted carbonyls; R2 is selected from the group consisting of -NR3R4, OR3, and -SR3, where R3 and R4 are independently selected from the group consisting of H, substituted or unsubstituted acyclic aliphatics, substituted or unsubstituted cyclic aliphatics, substituted or unsubstituted heterocyclics, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted acyl, substituted or unsubstituted carbonyls; their stereoisomers, mixtures and / or their cosmetically or pharmaceutically acceptable salts.

2. Composition according to claim 1, characterized in that the compound is at least one peptide selected from Succinyl-Ala-Ala-Val-Ala-OH (SEQ ID No. 1), Pal-Ala-Ala-Val-Ala-OH (SEQ ID No. 2), Ac-Ala-Ala-Val-Ala-OH (SEQ ID No. 3), Succinyl-Gly-Ala-Val-Ala-OH (SEQ ID No. 4) and Ac-Gly-Ala-Val-Ala-OH (SEQ ID No. 5), where the prefix Pal- identifies an N-palmitoylated peptide and the prefix Ac- identifies an N-acetylated peptide.

3. Composition, according to claim 1 or 2, characterized by being for the inhibition of the production of IL-1 alpha, inhibition of the production of IL-6, inhibition of the synthesis of melanin in fragments of human skin and anti-erythema action; and / or; to alleviate conditions resulting from inflammatory processes, sensitive skin, rosacea, atopic dermatitis, eczema; and / or; to alleviate the sensation of discomfort and pain; and / or; to bring benefits to problems related to skin hyperchromia and disorders related to increased sebogenesis.

4. Composition according to claim 1, 2 or 3, characterized in that it is for topical use.

5. Composition according to any one of the preceding claims, characterized in that it is in the form of a cosmetic, cream, lotion, serum, ointment or rinse-off product.

6. Composition according to any one of the previous claims, characterized in that the peptides are in a concentration range between 0.001% (w / w) and 10.00% (w / w).

7. Composition according to any one of the preceding claims, characterized in that the peptides are in a concentration range between 0.001% (w / w) and 0.1% (w / w).

8. Use of a composition as defined in any one of claims 1 to 7, characterized in that it is for producing a composition cosmetic, cosmeceutical and / or pharmaceutical for the inhibition of IL-1 alpha production, inhibition of IL-6 production, inhibition of melanin synthesis in human skin fragments and anti-erythema action; and / or; for topical use to alleviate conditions resulting from inflammatory processes, sensitive skin, rosacea, atopic dermatitis, eczema; and / or; to alleviate the sensation of discomfort and pain; modulation of the CB2 receptor and / or; to bring benefits to problems related to skin hyperchromia and disorders related to increased sebogenesis in a human or animal.

9. Method for inhibiting the production of IL-1 alpha, inhibiting the production of IL-6, inhibiting the synthesis of melanin in fragments of human skin and anti-erythema action; and / or; to alleviate conditions arising from inflammatory processes, sensitive skin, rosacea, atopic dermatitis, eczema; and / or; to alleviate the sensation of discomfort and pain; modulation of the CB2 receptor and / or; to bring benefits to problems related to skin hyperchromia and disorders related to increased sebogenesis in humans or animals, characterized by comprising the application to a human or animal of a cosmetically, cosmeceutically and / or pharmaceutically effective amount of a composition as defined in any one of claims 1 to 7.

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