Bioinspired peptide and its use for the treatment of wrinkles and expression lines

Abstract

The present invention relates to a peptide (Indió SEQ ID No. 1) with botox-like action for the treatment and smoothing of wrinkles and expression lines, to a cosmetic or dermopharmaceutical composition containing this peptide as an active ingredient, and to use of this composition, developed from a bio-inspired peptide from the venom of social wasps. Wrinkles are natural characteristics of the physiological ageing of the skin, whether genetic or environmental. Currently, preventive routines and treatments are in high demand, but they can be costly or risky and may require supervision by a professional beautician. This cosmetic composition, being applied topically in a non-invasive and painless manner, can be used daily without the need for a specialist in the field, and is not as expensive as other well-known techniques. Peptide SEQ ID No. 1 acts by inhibiting the contraction of myoblasts.

Description

Bio-inspired peptide and its use in the treatment of wrinkles and expression lines. FIELD OF THE INVENTION

[0001] The present invention relates to a bioinspired compound of peptides from the venom of social wasps from the Cerrado biome, called Indió (SEQ ID No. 1), from the Tupi-Guarani language meaning "cerrado," with a Botox-like effect; a cosmetic or dermopharmaceutical composition containing this peptide as an active ingredient; and the use of this composition in the prevention and smoothing of wrinkles and expression lines. The composition acts by inhibiting the contractile capacity of muscle cells. Being a cosmetic composition, with non-invasive and painless topical application, it can be used daily without the need for a specialist in the field of aesthetics, and is not as expensive as the techniques currently known for the prevention and smoothing of wrinkles and expression lines. STATE OF THE ART

[0002] Naturally, due to age and genetic factors, human skin inevitably undergoes aging processes. These processes result from the loss of fibroblast function and a reduction in the dermal extracellular matrix, representing a reduction in collagen and elastic fibers. Consequently, there is a loss of skin elasticity and volume. These changes may be accompanied by an increase in the contractile activity of skin cells, which together cause visible changes in skin characteristics, such as the appearance of wrinkles and expression lines, mainly on the face and neck area.

[0003] Environmental stressors and harmful habits can worsen the appearance of wrinkles, such as exposure to sunlight. Without proper protection, smoking, poor diet, insufficient water intake, excessive repetitive muscle movements, insufficient sleep, inadequate skin cleansing and hydration, among other factors, can all contribute to skin cancer.

[0004] Increasingly, the consumer market is seeking rejuvenating treatments that prevent the appearance or reduce the appearance of wrinkles and expression lines. Therefore, the current trend is for such treatments to begin earlier in life, with the use of preventative creams or massages. In this way, over time, the costs of maintaining more youthful-looking skin are lower and the treatments less invasive.

[0005] Wrinkle reduction treatments vary depending on the number, location, and depth of wrinkles. For fine lines and shallow wrinkles, creams, acids, gels, and other formulations containing retinoids and retinol (a derivative of Vitamin A), Vitamin C, glycolic acid, and niacinamide as active ingredients can be used. This topical treatment is performed at home by patients themselves as part of a skincare routine recommended by a dermatologist. However, facial skin is not uniform, and therefore, in most cases, these compounds are used in conjunction with other aesthetic treatments.

[0006] Among these aesthetic treatments, the most common are microneedling, massage techniques for mobilizing facial muscles, radiofrequency, laser treatments, peels, and botulinum toxin (Botox®) injections. Although well-known, many of these aesthetic treatments do not have good adherence rates or are not accessible to most of the population, either due to post-treatment care or high costs. This leaves these treatments restricted to specific population groups.

[0007] Currently, the application of Botox®, a toxin produced by the bacterium Clostridium botulinum, is a very popular cosmetic treatment. Sought after for the reduction of wrinkles and expression lines caused by both age and repetitive movements. Botox works by blocking the production or release of acetylcholine at neuromuscular synapses and junctions, the physiological mechanism of muscle contraction. Therefore, Botox® permanently blocks the contraction of a muscle fiber from the moment it binds to its targets, leaving it permanently relaxed.

[0008] It is recommended to reapply Botox® every 3 to 6 months, as natural cell renewal restores the muscles' ability to contract naturally, requiring further applications to maintain the anti-wrinkle effect. Its application must be performed by a qualified professional in a facility designed for this type of procedure, as Botox has a therapeutic index close to 1, meaning the effective dose is very close to the lethal dose; therefore, errors in Botox application are very serious. Furthermore, the use of the toxin can have adverse effects in some patients.

[0009] Among the complications that can be mentioned are eyelid ptosis, resulting from very high concentrations of the toxin, injections too close to the orbital rim, massages or intense manipulation of the area after application, causing greater diffusion of the toxin preparations; difficulty in closing the eyelids (lagophthalmos) in the treatment of periorbital wrinkles; eyebrow ptosis and a significant decrease in the expressiveness of the upper third of the face; lateral eyebrow ptosis; the appearance of asymmetrical points on the face; upper lip ptosis; in addition to difficulty in moving the lower lip, causing impairment to biting and speech.

[0010] Due to the risks and the need to visit a specialist clinic for reapplication, less invasive products that can be used at home have emerged on the market promising to produce an effect similar to that of botulinum toxin. These are the so-called Botox-like (BL) products. Thus, BL products have... The objective is to promote muscle relaxation in a non-invasive way, reducing wrinkles and expression lines.

[0011] Therefore, the present invention consists of a treatment option for wrinkles and expression lines through a simple cosmetic or dermocosmetic composition, for non-invasive and painless topical application, which can be used daily by the patient, being effective in reducing skin wrinkles, acting non-invasively in the repair of superficial wrinkles and fine lines, promoting skin rejuvenation and youthfulness. The invention concerns a bioinspired peptide from the venom of social wasps of the Cerrado, called Indió (SEQ ID No. 1), a dermocosmetic composition containing the peptide, and its use in the treatment and prevention of wrinkles and expression lines, for simple and daily use, ensuring greater practicality and accessibility to wrinkle treatment.

[0012] In searches conducted in databases, documents describing compositions with a Botox-like effect, wrinkle-reducing and anti-aging properties, comprising peptides, were found, such as document BR1120220149441A2, which deals with a composition with peptides composed of 6 to 17 amino acids derived from amino acid sequences of human SNAP25, Syntaxin la and VAMP2; document BR1120200040886A2 reports compositions formed by a peptide with skin wrinkle-improving activity, such peptide consists of four amino acid sequences: KWGGGRY, ILGRWCG, GPVH, EDEFKPPAAGR; document BR1120200173975A2 provides two synthetic tetrapeptides and compositions with them to inhibit muscle cell contraction and reduce the appearance of wrinkles; Document BR1120150150217B1 describes cosmetic compositions with polypeptides derived from fish egg hatching fluids to reduce or prevent the appearance of wrinkles or fine lines;Documents W02020050427A1 and KR101967630B1 refer to adiponectin-derived peptides and fragments thereof; active ingredients for cosmetic compositions that improve skin elasticity and wrinkles; document EP2420510A1 deals with a cosmetic composition with neuropeptides linked to nicotinic acid with anti-inflammatory and anti-wrinkle action, good collagen regeneration, low cytotoxicity, water solubility and good stability; documents SG11202100812U and WO2020138674A1 describe the KFLIK peptide sequence with muscle relaxation effect, improvement of skin wrinkles and improvement of acne; US patent document US20090280078 presents a cosmetic composition with several peptides, including Matrixyl™ 3000, two anticholinergic peptides (SYN®-AKE and acetyl hexapeptide-3), hexapeptide-11 isolated from yeast, as well as other compounds intended to prevent and treat skin wrinkles caused by sun or age;Document EP4230638 describes peptides derived from Taki fugu rubripes (Japanese pufferfish, tiger pufferfish) with anti-wrinkle activity and promoting muscle relaxation; documents PI 1105822-6A2, US 8987212B2, AU2011265587 and WO2012091743 describe a pentapeptide (Lysine-Threonine-Threonine-Lysine-Serine) linked to oleanolic acid used in a cosmetic composition to reduce signs of aging, skin wrinkling, fine lines, dark circles, hyperpigmentation. BRIEF DESCRIPTION OF THE FIGURES

[0013] Figure 1 illustrates the percentage of cell viability after treatment with the sample containing Indiol (SEQ ID No. 1). Figure 2 shows the percentage reduction in the volume of the three-dimensional matrix containing myocytes after treatment with the sample containing Indiol (SEQ ID No. 1). Figure 3 shows the three-dimensional matrices after 72 hours of incubation with the medium according to the experimental group. Figure 4 illustrates the subjective clinical efficacy results of the placebo group in clinical trials. Figure 5 illustrates the subjective clinical efficacy results of the group treated with the Indió strain (SEQ ID No. 1) in clinical trials. Figure 6 demonstrates the evaluation of cosmetic appeal in clinical trials of the placebo group after 30 + / - 2 days of daily use. Figure 7 presents the evaluation of cosmetic appeal in clinical tests of the treatment group with the Indió sample (SEQ ID No. 1) after 30 + / - 2 days of daily use. DETAILED DESCRIPTION OF THE INVENTION

[0014] The present invention relates to a bioinspired peptide (Indio - SEQ ID No. 1), a cosmetic or dermopharmaceutical composition containing the bioinspired peptide (Indio - SEQ ID No. 1) and its use in reducing and preventing wrinkles and expression lines.

[0015] The peptide of the present invention (SEQ ID No. 1) has a theoretical mass of 1937.41 Da (Purity >95%) and possesses the following amino acid sequence: H-Ile-Leu-Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg-Ile-Leu-Gly-Lys-Ile-Ile-NH2 (ILRPPGFTPFRILGKI I-NH2). It was bioinspired and developed from peptides identified in the venom of the social wasp Polybía occidentalis.

[0016] The formulation containing the peptide consists of water, 1,3-butylene glycol, and Indiol peptide (SEQ ID No. 1) from 0.01% to 0.0001%.

[0017] In vitro cytotoxicity studies were conducted using eukaryotic dermatological cell culture systems, as well as assays to inhibit the contractile action of fibroblasts in a collagen matrix using the SEQ ID No. 1 peptide. Regarding clinical trials, tests were performed with volunteer individuals (n=30, CEP / Plataforma Brasil number 6.850.780) to evaluate the safety profile. (dermatological acceptability) and subjective clinical efficacy of Indió (SEQ ID No. 1). In addition, a cosmetic appreciability analysis of Indió (SEQ ID No. 1) was also performed after the clinical trials.

[0018] Technology can be better understood from the following examples, which are not exhaustive. EXAMPLES OF IMPROVEMENT OF THE INVENTION

[0019] EXAMPLE 1 - IN VITRO BIOLOGICAL ASSAYS USING EUKARYOTIC DERMAL CELL SYSTEMS IN CELL CULTURE

[0020] To evaluate the in vitro cytotoxic potential of the Indio peptide (SEQ ID No. 1) in Balb / c 3T3 clone 31 cell cultures maintained in 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.

[0021] Sample preparation Indió (SEQ ID No. 1): Sample preparation conditions in the starting solution: sample was diluted to 10% in culture medium;

[0022] Preparation of concentrations: serial dilution in 5 decreasing concentrations on a base of 10;

[0023] Appearance of the solubilized sample: completely solubilized.

[0024] Preparation of control groups:

[0025] Control group: supplemented cell culture medium;

[0026] Positive control group: sodium lauryl sulfate diluted in supplemented cell culture medium (100 pg / ml)

[0027] 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 Incubation was carried out 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.

[0028] 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.

[0029] Figure 1 indicates the percentage of cell viability found after treatment with the sample compared to the control with serial dilutions between 10% and 0.001%. For Indio (SEQ ID No. 1), no cytotoxicity was observed at any of the concentrations tested. The positive control group demonstrated cell viability of 22.4%.

[0030] EXAMPLE 2 - IN VITRO BIOLOGICAL ASSAYS USING EUKARYOTIC CONTRACTILE CELL SYSTEMS IN CELL CULTURE TO STUDY CELL CONTRACTION INHIBITION CAPACITY

[0031] In this study, myoblast cell cultures were used, maintained in culture with DMEM (Dulbecco's Modified Eagle's Medium) with the addition of supplements, in an incubator at 37°C and 5% CO2, and manipulated inside a laminar flow hood. Initially, cytotoxicity analysis was performed to determine the concentration evaluated as having the potential to inhibit cell contraction.

[0032] To stimulate cell contraction, the cells were immersed in a three-dimensional collagen matrix. After polymerization of the matrix containing the myoblasts, the culture medium was added according to the experimental groups.

[0033] Sample preparation: sample preparation conditions in the starting solution: non-cytotoxic concentration evaluated: 5.0%; appearance of the solubilized sample: completely solubilized;

[0034] Preparation of control groups: the basal control group was in cell culture medium and the negative control group was the absence of contractile cells in the matrix.

[0035] After adding the culture medium according to each experimental group, the plates were kept in an incubator under the conditions described above for 72 hours, and at the end of this period, each matrix was measured for final volume calculation.

[0036] The results were evaluated using GraphPad Prism software. One-way ANOVA and Bonferroni tests were applied, and statistical significance was considered when p<0.05. The final volume of the negative control was normalized to 100%, and the other groups were compared to it. The baseline control showed an 85.6% reduction in matrix volume, and the group that received the test product showed a 49.3% reduction in the volume of the negative control. Statistical analyses resulted in p<0.001, representing statistical significance, as illustrated in Figures 2 and 3.

[0037] EXAMPLE 3 - EVALUATION OF DERMATOLOGICAL ACCEPTABILITY

[0038] With the aim of evaluating the safety and efficacy of Indió (SEQ ID No. 1) in humans, clinical trials were conducted with 30 volunteers of both sexes, aged 35 to 59 years, who used the product containing the peptide for 30 days.

[0039] An initial medical assessment was conducted at the time of participant enrollment to verify the absence of initial clinical signs inconsistent with participant inclusion.

[0040] After 30 ± 2 days of using the products, the participants returned to the Institution for a final medical evaluation of the clinical signs presented and questioning about any discomfort felt.

[0041] The data from the medical evaluation were recorded in the investigation notebook. The physician remained available throughout the study to assess any potential adverse events.

[0042] The results were evaluated as follows:

[0043] Sensations of discomfort: participants were asked about the sensations of discomfort they experienced, in parallel with the clinical examination. The reported sensations of discomfort were described in relation to their nature (e.g., 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 attribution to the test product was verified.

[0044] 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.

[0045] No participants reported any discomfort, and no clinical signs were detected after using the product. Therefore, it supports the claim "Dermatologically tested".

[0046] EXAMPLE 4 - EVALUATION OF SUBJECTIVE CLINICAL EFFICACY

[0047] To evaluate the subjective clinical efficacy of these products, clinical trials were conducted with 30 volunteers of both sexes, aged 35 to 59, who used the product containing the peptide for 30 days.

[0048] The volunteers were evaluated at baseline (D0) and after 30 ± 2 days of using the products at home (D30) with the following parameters:

[0049]

[050] After 30 ± 2 days of using the placebo product, 40% of participants showed improvement in crow's feet wrinkles; 33% of participants showed improvement in nasolabial folds; 47% had improvement in forehead wrinkles; 33% had improvement in skin elasticity; 33% had improvement in skin firmness; and 40% had improvement in skin hydration (Figure 4).

[0051] After 30 ± 2 days of using the test product, as illustrated in Figure 5, 67% of participants showed improvement in crow's feet wrinkles; 60% of participants showed improvement in nasolabial folds; 67% had improvement in forehead wrinkles; 60% had improvement in skin elasticity; 67% had improvement in skin firmness; and 73% had improvement in skin hydration.

[0052] EXAMPLE 5 - EVALUATION OF COSMETIC APPRECIABILITY (OPINION OF RESEARCH PARTICIPANTS)

[0053] Participants were instructed to complete a questionnaire containing the questions and possible answers listed below, after 30 ± 2 days of using the investigational products.

[0054] A cosmetic appreciation questionnaire was administered. The questionnaire assessed several of the volunteers' perceptions, based on "Yes" or "No" answers. The questions evaluated are listed below.

[0055] Does the product have a "botox-like" effect?

[0056] Did the product leave your skin looking more radiant?

[0057] Did the product leave your skin more hydrated?

[0058] Did the product soften the appearance of fine lines on the skin?

[0059] Have you noticed an improvement in your skin texture?

[0060] Does the product work well?

[0061] Would you buy this product?

[0062] Figure 6 shows the results after 30 ± 2 days of using the placebo product, namely: 33% of participants reported feeling a "botox-like" effect with the use of the product; 60% noticed more radiance in the skin; 67% noticed more hydrated skin; 27% reported smoothing of expression lines; 27% reported improvement in skin texture; 27% consider that the product had a good result; and 27% of participants would buy the product.

[0063] Figure 7 shows the results after 30 ± 2 days of using Indió (SEQ ID No. 1). 80% of participants reported feeling a "botox-like" effect with the use of the product; 87% noticed more radiance in their skin; 93% noticed more hydrated skin; 80% reported a smoothing of expression lines; 73% reported an improvement in skin texture; 80% considered the product to have had a good result; and 80% of participants would buy the product.

Claims

MODIFIED CLAIMS Received by the International Secretariat on January 21, 2026 (21.01.2026) 1- PEPTIDE with the exact amino acid sequence described in SEQ ID No.

1. 2- PEPTIDE according to claim 1, characterized by being used in the preparation of cosmetic or dermopharmaceutical compositions for the treatment of wrinkles and facial expression lines. 3- 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. 4- 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. 5- USE OF THE COSMETIC OR DERMO-PHARMACEUTICAL COMPOSITION, as described in claims 3 and 4, characterized by being applied to the skin for the treatment of wrinkles and expression lines with a botox-like function. MODIFIED SHEET (ARTICLE 19)

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