Use of a peptide from the venom of social wasps from the cerrado for the treatment of acne

Abstract

The present invention relates to the use of a cosmetic or dermopharmaceutical formulation containing the Polybia-MPII peptide, or "Intó" (SEQ ID NO: 1), for the treatment of acne. Acne is a chronic immunoinflammatory skin condition affecting the pilosebaceous unit. It is clinically characterised by comedones, papules, pustules, nodules or cysts. Although it does not pose a direct risk to health, this condition is associated with an increased likelihood of developing anxiety and depression, reducing quality of life and self-esteem. The treatments currently available on the market, though often producing satisfactory results, can be associated with unwanted adverse effects. The present invention provides an alternative to current treatments, showing no adverse effects during clinical trials and also demonstrating satisfactory efficacy in the treatment of acne. The cosmetic composition containing the peptide SEQ ID NO: 1 has been clinically proven to control acne as a result of to the peptide's antimicrobial action.

Description

[0001] Use of peptide from the venom of social wasps from the Cerrado region for the treatment of acne.

[0002] FIELD OF THE INVENTION

[0003]

[0001] The present invention relates to the application of a cosmetic or dermopharmaceutical composition with the peptide mastoparan, Polybia-MP IT (SEQ ID Nl ) or, also, "into", which in Tupi-Guarani means "face / countenance", in the treatment, control and prevention of acne. The peptide in question is found in the venom of social wasps of the Brazilian Cerrado of the genus Pseudopolybi and has antimicrobial action.

[0004] STATE OF THE ART

[0005]

[0002] According to the World Health Organization's (WHO) international classification of diseases, acne is a chronic, immune-inflammatory skin condition of the pilosebaceous unit. It is clinically characterized by comedones, papules, pustules, nodules, or cysts.

[0006]

[0003] Current data shows that 80-85% of adolescents and young adults worldwide have some type of acne, and the condition can persist into adulthood, especially in women, due to hormonal imbalances.

[0007]

[0004] Acne is a skin condition associated with several factors, such as genetic predisposition, lifestyle, diet, metabolism, hormonal imbalances, immunity, among others; and, if left untreated, acne can leave permanent scars on the skin (Kim et al. Exploring Acne Treatments: From Pathophysiological Mechanisms to Emerging Therapies. Vol 25 (10), 2024, doi: 10.3390 / ims25105302).

[0005] Although it does not represent a direct health risk, acne tends to be considered a burden on quality of life. Some studies point to an association between acne and anxiety and depression due to the negative impact of acne on self-esteem. Therefore, patients seek various treatments, spending time and resources in search of a healthier appearance.

[0008]

[0006] Currently, available treatments vary according to the causes of acne and may include the oral or topical use of antibiotics, retinoids, antiandrogens, phototherapy, chemical peels, hyaluronic acids, among others. Undesirable effects of oral antibiotics may include disruption of the body's healthy microbiota as a whole; the use of antiandrogens, such as the birth control pill, is associated with various effects such as blood clot formation and fluid retention, as well as mood changes; while the application of chemical peels can lead to hypersensitivity reactions. Therefore, the treatments currently available for acne, despite achieving satisfactory results in most cases, are associated with possible consequences that may decrease patient adherence (Auffret et al. Novel and emerging treatment options for acne vulgaris, Vol 32 (4, 2022, doi: 10.1684 / ejd.2022.4306).

[0009]

[0007] The present invention relates to a cosmetic or dermopharmaceutical composition with the peptide isolated from the venom of a social wasp (SEQ ID No. 1) with an antimicrobial effect through topical application, in order to prevent the imbalance of a healthy microbiota when considering the organism as a whole.

[0010] The use of antimicrobial peptides is advantageous because their mechanism of action hinders the formation of resistant strains and the formation of biofilms (Silva et al.). r Evaluation of the antimicrobial activity of the mastoparan Polybia-MPII isolated from venom of the social wasp Pseudopolybia vespicepstestacea (Vespidae, Hymenoptera), Vol 49, 2, 2017, d. the . I .

[0011] 10.1016 / j . ij antimicag .2016.11.013)

[0012]

[0008] In searches conducted in patent databases, documents were found that describe the use of compositions containing antimicrobial peptides or peptides useful in the treatment of acne, or even compositions containing ingredients of natural origin for the treatment of acne, such as document BR 112020026825-9 A2, which describes a composition containing a peptide with various physiological actions, including oil control, which helps control acne; document CN108495646B describes 24 peptides composed of 6 amino acids following a basic structure that can be used in skin care compositions that can be applied to the treatment of acne; while document CN102985091B describes a composition including a chelated peptide with antimicrobial action for the treatment of skin conditions; and document BR 11 2024 000048 6 A2 describes bacterial strains and their use in the treatment of acne;Document BR 10 2022 011004 2 A2 describes a biodegradable film composition comprising a yeast for the treatment of acne; document BR 11 2021 025797 7 A2 describes a cream for the treatment of melasma and acne containing extracts of the roots, stems and leaves of medicinal plants, not including peptides; finally, document BR 11 2021 004823 5 A2 describes conjugates of montelukast and peptides that aid in the treatment of acute inflammation, being useful in the treatment of acne.

[0013] BRIEF DESCRIPTION OF THE FIGURES

[0014]

[0009] Figure 1 illustrates the percentage of cell viability after treatment with the Into sample (SEQ ID No. 1) considering concentrations relative to the recommended concentration for use (0.0001%).

[0010] Figure 2 illustrates the subjective clinical efficacy results of the placebo group in clinical trials.

[0015]

[0011] Figure 3 illustrates the subjective clinical efficacy results of the group treated with the Into sample (SEQ ID No. 1) in clinical trials.

[0016]

[0012] Figure 4 shows the first part of the cosmetic appreciability assessment in clinical trials of the placebo group after 30 + / - 2 days of daily use.

[0017]

[0013] Figure 5 shows the second part of the cosmetic appreciability assessment in clinical trials of the placebo group after 30 + / - 2 days of daily use.

[0018]

[0014] Figure 6 presents the first part of the cosmetic appreciability assessment in clinical tests of the treatment group with the Into sample (SEQ ID No. 1) after 30 + / - 2 days of daily use.

[0019]

[0015] Figure 7 presents the second part of the cosmetic appreciability assessment in clinical trials of the treatment group with the Into sample (SEQ ID No. 1) after 30 + / - 2 days of daily use.

[0020] DETAILED DESCRIPTION OF THE INVENTION

[0021]

[0016] The present invention relates to the use of a peptide (Intó SEQ ID N° 1) extracted from the venom of wasps of the genus Pseudopolybí a in a cosmetic or dermopharmaceutical composition for the treatment and reduction of acne.

[0022]

[0017] The peptide of the present invention (SEQ ID No. 1) has a theoretical mass of 1612.90 Da (Purity >95%) and has the following amino acid sequence: Ile-Asn-Trp-Leu-Lys-Leu-Gly-Lys-Met-Val-Ile-Asp-Ala-Leu-NH2 (INWLKLGKMVIDAL-NH2). It was identified in the venom of the social wasp Pseudopolybia vespiceps testacea.

[0023]

[0018] Furthermore, for the purposes of the present invention, sequences in which the amino acid Isoleucine is replaced by the amino acid with the same chemical properties Leucine and vice versa may be considered.

[0024]

[0019] The formulation containing the peptide consists of water, Into peptide (SEQ ID No. 1) from 0.01% to 0.0001% and / or peptides obtained by replacing the amino acid Isoleucine with Leucine in the structure of SEQ ID No. 1.

[0025]

[0020] In vitro cytotoxicity studies were conducted using eukaryotic dermatological cell culture systems with the peptide SEQ ID No. 1. Regarding clinical trials, trials were conducted with volunteer individuals (n=30, CEP / Plataforma Brasil number 6.679.704) with the objective of evaluating the safety profile (dermatological acceptability) and subjective clinical efficacy of Into (SEQ ID No. 1). In addition, a cosmetic appreciability analysis of Into (SEQ ID No. 1) was also performed in the clinical trials.

[0026]

[0021] Technology can be better understood from the following examples, which are not exhaustive.

[0027] EXAMPLES OF IMPROVEMENT OF THE INVENTION

[0028]

[0022] EXAMPLE 1 - IN VITRO BIOLOGICAL ASSAYS USING EUKARYOTIC DERMAL CELL SYSTEMS IN CELL CULTURE

[0029]

[0023] To evaluate the in vitro cytotoxic potential of the Intó peptide (SEQ ID No. 1) in Balb / c 3T3 clone 31 cell cultures maintained in culture with DMEM (Dulbecco's Modified Eagle's Medium) with added supplements, in an incubator at 37 °C and 5% CO2 and manipulated within a laminar flow hood. For the study, the cells were distributed in 96-well plates and maintained in culture under the same conditions described.

[0030]

[0024] Sample preparation Into (SEQ ID No. 1): Sample preparation conditions in the starting solution: sample was diluted to 10% in culture medium;

[0031]

[0025] Preparation of concentrations: serial dilution in 5 decreasing concentrations on a 10 basis;

[0032]

[0026] Appearance of the solubilized sample: completely solubilized.

[0033]

[0027] Preparation of control groups:

[0034]

[0028] Control group: supplemented cell culture medium;

[0035]

[0029] Positive control group: sodium lauryl sulfate diluted in supplemented cell culture medium (100 pg / ml)

[0036]

[0030] Incubation and cell viability assessment: solutions containing the control, positive control and sample groups were applied to the cell culture, 100 pL per well in quadruplicate for each group, followed by incubation in an oven at 37 °C and 5% CO2 for 24 hours. After washing, cell viability was analyzed with neutral red indicator, 100 pL per well. After incubation, the reaction was revealed and absorbance was determined at 540 nm.

[0037]

[0031] The results were evaluated using Microsoft Excel software. The control group was normalized to 100%, and the percentage of cell viability in the sample and in the positive control was calculated in relation to the control. Samples are considered cytotoxic when they show a 30% or greater reduction in cell viability.

[0038]

[0032] Figure 1 indicates the percentage of cell viability found after treatment with the sample compared to the control with serial dilutions at concentrations relative to the recommended use concentration (0.0001%) between 100x and 0.001x. For Into (SEQ ID No. 1), cytotoxic activity was observed only when the concentration was equal to 100x the recommended use concentration. No cytotoxicity was observed at any of the other concentrations tested. The positive control group showed cell viability of 36.4%.

[0039]

[0033] EXAMPLE 2 - EVALUATION OF DERMATOLOGICAL ACCEPTABILITY

[0034] In order to evaluate the safety and efficacy of Intó (SEQ ID No. 1) in humans, clinical trials were conducted with 30 volunteers of both sexes, aged 18 to 35 years, who used the product containing the peptide for 30 days.

[0040]

[0035] An initial medical assessment was carried out at the time of participant enrollment to verify the absence of initial clinical signs incompatible with participant enrollment.

[0041]

[0036] After 30 ± 2 days of using the products, the participants returned to the Institution for final medical evaluation of the clinical signs presented and questioning about the sensations of discomfort felt.

[0042]

[0037] The data from the medical assessment were recorded in the investigation notebook. The physician was available throughout the study to assess possible adverse events.

[0043]

[0038] The results were evaluated as follows: Sensations of discomfort: participants were questioned about the sensations of discomfort felt, in parallel with the clinical examination. The sensations of discomfort reported were described in relation to their nature (example: burning, tingling, itching, pulling, cooling, heating, etc.); they were classified according to intensity as: mild, moderate or intense; according to location; and according to duration; and the attributability to the test product was verified.

[0044]

[0039] Clinical signs: were classified according to the presence of clinical signs such as edema, papules, blisters, vesicles, erythema and nothing to declare and the causal link of the reactions to the products was investigated.

[0040] No participant reported feelings of discomfort and no clinical signs were detected after using the product. Therefore, it supports the claim "Dermatologically tested".

[0045]

[0041] EXAMPLE 3 - EVALUATION OF SUBJECTIVE CLINICAL EFFICACY

[0042] For the evaluation of the subjective clinical efficacy of these products, clinical trials were conducted with 30 volunteers of both sexes, aged 18 to 35 years, who used the product containing the peptide for 30 days.

[0046]

[0043] The volunteers were evaluated at baseline (DO) and after 30 ± 2 days of using the products at home (D30) with the parameters described in Table 1.

[0047] Table 1: Parameters used in the evaluation of Into and placebo.

[0048] Oiliness 0 = absent

[0049] Eaters

[0050] 1 = light

[0051] 2 = moderate

[0052] 3 = intense

[0053] Papules: Quantity count in the experimental area. Pustules.

[0054]

[0044] After 30 ± 2 days of using the placebo product, 40% of participants showed improvement in skin oiliness, 47% showed improvement in the quantification of papules, 47% showed improvement in the quantification of pustules, and 40% of participants showed improvement in the classification of comedones (Figure 2).

[0055]

[0045] After 30 ± 2 days of using the test product, as illustrated in Figure 3, 67% of participants showed improvement in skin oiliness, 67% showed improvement in the quantification of papules, 60% showed improvement in the quantification of pustules, and 67% of participants showed improvement in the classification of comedones (Figure 3).

[0046] EXAMPLE 4 - EVALUATION OF COSMETIC APPRECIABILITY (OPINION OF RESEARCH PARTICIPANTS)

[0056]

[0047] Participants were instructed to answer a questionnaire containing the questions and possible answers listed below, after 30 ± 2 days of using the investigational products.

[0057]

[0048] A cosmetic appreciability questionnaire was applied. The questionnaire assessed some of the volunteers' perceptions, based on "Yes" or "No" answers. The questions evaluated in relation to the questions are listed below.

[0058] Did the product help control oiliness?

[0059] Did it reduce acne (pimples and blackheads)?

[0060] Does this product dry out acne?

[0061] - Did it soften the appearance of blemishes caused by acne? - Did you notice that the product prevented and treated acne? - Did it reduce the appearance of pores?

[0062] Have you noticed an improvement in your skin texture?

[0063] Did the product leave your skin moisturized?

[0064] Did the product make your skin healthier?

[0065] - Have you noticed an improvement in the appearance and overall quality of your skin? - Is the product producing good results?

[0066] Would you buy this product?

[0067]

[0049] Figures 4 and 5 show the results after 30 ± 2 days of using the placebo product, namely: 33% felt that the product controlled oiliness; 40% reported that the product reduced acne; 33% reported that the product acted as a drying agent; 27% reported that the product softened the appearance of blemishes caused by acne; 33% noticed that the product prevented and treated pimples; 33% noticed an improvement in the appearance of pores; 40% noticed an improvement in skin texture; 33% noticed an improvement in skin hydration; 40% reported that the product left their skin healthier; 40% noticed an improvement in the overall appearance and quality of their skin; 33% reported that the product has good results; and 33% would buy this product.

[0050] Figures 6 and 7 show the results after 30 ± 2 days of using the test product, namely: 87% felt that the product controls oiliness; 87% reported that the product reduced acne; 87% reported that the product acts in a drying way; 93% reported that the product softened the appearance of blemishes caused by acne; 87% noticed that the product prevented and treated pimples; 87% noticed an improvement in the appearance of pores; 87% noticed an improvement in skin texture; 93% noticed an improvement in skin hydration; 93% reported that the product left their skin healthier; 87% noticed an improvement in the overall appearance and quality of their skin; 87% reported that the product has good results; and 87% would buy this product.

Claims

CLAIMS 1- COSMETIC OR DERMO-PHARMACEUTICAL COMPOSITION, characterized by consisting of water, peptide SEQ ID No. 1 from 0.01% to 0.0001%, 1,3-butylene glycol. 2- COSMETIC OR DERMO-PHARMACEUTICAL COMPOSITION, characterized by consisting of peptide SEQ ID No. 1 from 0.01% to 0.0001%; ​​1,3-butylene glycol and serum base. 3- COSMETIC OR DERMO-PHARMACEUTICAL COMPOSITION, according to claim 1, characterized in that the peptide of the composition may result from substitutions of the amino acid Ie, of SEQ ID No. 1, by the amino acid Leu and vice versa. 4- USE OF THE COSMETIC OR DERMO-PHARMACEUTICAL COMPOSITION, as described in claims 1, 2 and 3, characterized by being applied to the skin for the treatment of acne, acting as an antimicrobial agent.

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