Use of cordia verbenacea essential oil, stable dermocosmetic composition, product for treating acne vulgar and method for treating acne vulgar

Abstract

The present invention relates to a stable dermocosmetic composition, for topical application, effective in the treatment of acne vulgaris, with multifactorial efficacy, which comprises an active ingredient based on Cordia verbenacea, in a silicone base, which has a dry touch and is suitable for acne-prone skin, as well as a cosmetic product, uses and method for treating acne vulgaris related thereto.

Description

USE OFCORDIA VERBENACEAESSENTIAL OIL, STABLE DERMOCOSMETIC COMPOSITION, PRODUCT FOR TREATING ACNE VULGAR AND METHOD FOR TREATING ACNE VULGARField of the Invention

[0001] The present invention relates to a stable dermocosmetic composition, for topical application, effective in the treatment of acne vulgaris, with multifactorial efficacy, which comprises an active ingredient based onCordiaverbenacea, in a silicone base, which has a dry touch and is suitable for acne-prone skin, as well as a cosmetic product, related uses and method for treating acne vulgaris.Background of the Invention

[0002] Acne vulgaris can be considered the most common non-contagious dermatological condition of multifactorial origin in the general population. It is characterized by the formation of comedones, papules, pustules, nodules and / or cysts as a result of obstruction and inflammation of the pilosebaceous unit of the skin (hair follicles and their accessory sebaceous glands).

[0003] In general, acne develops on the face and upper torso and, as it is a multifactorial condition, it can affect different age groups.

[0004] From a pathophysiological point of view, acne vulgaris occurs through the interaction of 4 main factors or pillars: (1) colonization of follicles byCutibacteriumacnesorC. acnes (formerlyPropionibacterium acnes), a gram-positive anaerobic bacteriumnormally present on human skin; (2) follicular closure with sebum and keratinocytes or hyperkeratosis; (3) excess sebum production or hyperseborrhea; and (4) release of multiple inflammatory mediators or inflammation.

[0005] Diagnosis is made by clinical examination and current treatment is based on the severity of the condition and may involve a variety of topical and systemic agents aimed at reducing sebum production, infection, comedone formation, inflammation, bacterial counts and normalizing keratinization.

[0006] Among the agents used in the treatment, it is found:a. Topical retinoids that act through gene modulation, influencing the expression of different proteins that contribute to improving the differentiation and formation of the epithelial barrier, helping to unclog pores, control sebum and, consequently, reduce inflammation stimulated by C. acnes.b. Peroxides that release free radicals, which kill C. acnes bacteria present on the skin or in follicles, reducing colonization, in addition to having a slight sebum-regulating effect.c. Topical or systemic antimicrobials.d. Hormonal regulators that provide gene regulation, causing cell renewal, a sebum-regulating effect and a reduction in hyperkeratosis.

[0007] These agents do not act in a multifactorial manner and any anti-inflammatory effect eventually observed should be considered secondary, as a consequence of the primary effect of retinoids on cell differentiation, the reduction of C. acnes and less obstruction of pores due to the sebum-regulating effect, among others.

[0008] For this reason, the use of multiple agents in the treatment is known, for example, by the combination of anti-inflammatory (e.g. clindamycin, erythromycin, tetracycline, tretinoin, isotretinoin), antibacterial (e.g. benzoyl peroxide, clindamycin, erythromycin, tetracycline, doxycycline, minocycline, azithromycin), anti-sebum (e.g. spironolactone, flutamide, adapalene, tretinoin, isotretinoin) and keratolytic (e.g. salicylic, adapalene, tretinoin, isotretinoin).

[0009] There is no evidence that traditional anti-inflammatory drugs are effective in treating acne or that they may have a multifactorial effect in treating acne, according to data from international associations such as the American Academy of Dermatology, Journal of the American Academy of Dermatology – Expert Board, Dermatological Society of Singapore, Canadian Medical Association, Cochrane Reviews or NHS. For this reason, they have not been recommended for the treatment of acne.

[0010] Recent studies indicate the lack of clinical efficacy of traditional anti-inflammatory drugs in the treatment of acne. In a comparative study between the effect of treatment with benzoyl peroxide and the effect of treatment with benzoyl peroxide + diclofenac (COX inhibitor, used in musculoskeletal inflammation), it was found that the addition of diclofenac to the treatment did not bring superiority in improving acne, showing that “traditional” anti-inflammatory drugs do not contribute to the treatment of the pathology (Tantipunpipat et al.,Clinical comparison between topical benzoyl peroxide 2.5% and diclofenac gel 1% versus benzoyl peroxide 2.5% and placebo in the treatment of mild to moderate facial acne vulgaris, Surg Cosmet Dermatol., 2020).

[0011] Natural active ingredients are also used to combat acne, however, the use of many of them comes from popular knowledge and without proven clinical efficacy.

[0012] Ideally, the selection of candidates that are effective in acne should take into account the treatment pillars discussed herein. However, there are few possibilities for multifactorial ingredients.

[0013] Cordiaverbenacea, which has the scientific synonymCordiacurassavica, also known as erva baleeira or maria-milagrosa, is a plant with known healing and anti-inflammatory properties and, for this reason, researches have been done on it, especially its essential oil.

[0014] In the search for the state of the art in scientific and patent literature, the following documents related to the subject matter of the present invention were found:

[0015] PI 0300600-0 which discloses a composition associated with the chromatographic isolation process of alpha-humulene from the essential oil ofCordiacurassavica, being delivered in a pharmaceutical form for topical use, chosen from cream, ointment, gel, gel / cream, creamy lotion, emulsion, transdermal, mouthwash, vaginal cream or ointment, vaginal ovule and vaginal soft capsule, and comprising between 10 and 60 mg of alpha-humulene obtained from a specific process. Data on the efficacy of alpha-humulene, in isolation, in different models of musculoskeletal inflammation are presented.

[0016] BR 122016029094-8 which discloses an anti-inflammatory pharmaceutical composition, aimed at the treatment of inflammations of musculoskeletal origin, which comprises alpha-humulene as the sole active ingredient, in which the composition is an ointment comprising a concentration greater than 0.025 and less than 0.05% by weight, in relation to the total weight of the ointment, of alpha-humulene and pharmaceutically acceptable adjuvants and excipients.

[0017] Even with a known anti-inflammatory effect for inflammation of musculoskeletal origin,Cordiaverbenacea, particularly its essential oil, as well as other traditional anti-inflammatories, has not been researched for the treatment of acne both due to the lack of evidence of clinical efficacy of anti-inflammatories in the treatment of acne, and due to the probable failure in a multifactorial action.

[0018] In the case ofCordiaverbenaceaessential oil, there is still an obstacle, since it is an oily active ingredient, that is, a characteristic that the person skilled in the art expects not to favor the treatment of acne and which faces difficulties in stabilizing for this type of application.

[0019] Furthermore, products based onCordiaverbenaceaessential oilintended for topical application on the face and nearby areas would require the use of fragrances due to the characteristic odor of said oil. In this sense, there is also a notable instability ofCordiaverbenaceaessential oilin emulsions, especially with the use of fragrances, which would make its topical use unfeasible.

[0020] Thus, there remains a need and search for new active ingredients and dermocosmetic compositions, capable of acting effectively and multifactorially in the treatment of acne vulgaris, maintaining appropriate stability and odor.Summary of the Invention

[0021] Therefore, the present invention aims to solve the constant problem of the state of the art related to the lack of active ingredients that act in a multifactorial manner, as well as to provide a stable pharmaceutical composition for topical application, particularly in the presence of fragrances.

[0022] In a first aspect, the present invention relates to the use ofCordiaverbenaceaessential oilin the preparation of a stable dermocosmetic composition for the treatment of acne vulgaris.

[0023] In a second aspect, the present invention relates to a stable dermocosmetic composition, for topical application, effective in the treatment or prevention of acne vulgaris, with multifactorial efficacy, which comprisesCordiaverbenaceaessential oil, in a silicone base, which has a dry touch and is suitable for acne-prone skin.

[0024] In a third aspect, the present invention relates to a product for treating acne vulgaris comprising the stable dermocosmetic composition according to the present invention.

[0025] In a fourth aspect, the present invention relates to methods of treating acne vulgaris, comprising a step of applying, to the skin, essential oil ofCordiaverbenaceaor a layer of a product as defined in the third aspect.

[0026] Furthermore, the inventive concept common to all the protection contexts described and claimed is that they all refer to the essential oil ofCordiaverbenaceaand the stable dermocosmetic composition comprising it.

[0027] These and other objects of the invention will be immediately appreciated by those skilled in the art and by companies with interests in the segment, and will be described in sufficient detail for their reproduction in the following description.Brief Description of Figures

[0028] In order to better define and clarify the content of the present invention, the following figures are presented:

[0029] shows the amount of non-inflammatory lesions after 28 days of treatment with the composition comprisingCordiaverbenaceaessential oilaccording to the present invention.

[0030] shows the amount of inflammatory lesions after 28 hours of treatment with the composition comprisingCordiaverbenaceaessential oilaccording to the present invention.

[0031] shows the oiliness index after 28 days of treatment with the composition comprisingCordiaverbenaceaessential oilaccording to the present invention.

[0032] shows the reduction of C. acnes after 28 days of treatment with the composition comprising the essential oil ofCordiaverbenaceaaccording to the present invention.Detailed Description of the Invention

[0033] Cordiaverbenaceaessential oil has been found to have multifactorial activity against acne vulgaris.

[0034] According to the present invention, “multifactorial” means obtaining simultaneous antibacterial, anti-inflammatory, anti-oiliness and keratolytic effects.

[0035] Initially, with regard to the inflammatory pillar, studies have shown that C. acnes activates the receptor on polysebaceous cells, leading to the production of multiple pro-inflammatory cytokines, including interleukin 6 and 8, and that the levels and metabolic pathways of several lipid mediators, such as prostaglandin E2, are abnormal in acne lesions, suggesting an important role for the COX-2 signaling pathway in the polysebaceous unit.

[0036] TheCordiaverbenaceaessential oil according to the present invention acts via the TLR-2 and MCH-II pathways and not via the arachidonic acid pathway.

[0037] With regard to the keratolytic pillar, it is known that the proliferation of keratinocytes increases the content of keratinous material on the skin, which leads to the obstruction of the polysebaceous follicle, causing the formation of microcomedones that are precursors of acne lesions.

[0038] TheCordiaverbenaceaessential oilaccording to the present invention was able to promote a keratolytic effect in cell culture, which allows cell renewal, prevents the accumulation of lipids in the pilosebaceous follicle, reducing oiliness and acne.

[0039] Furthermore, theCordiaverbenaceaessential oilaccording to the present invention, surprisingly, acted directly on the antibacterial activity, inhibiting the bacterial concentration and preventing the initial formation of the bacterial biofilm.

[0040] Thus, unlike what is taught in the state of the art, theCordiaverbenaceaessential oilaccording to the present invention presents multifactorial activity against acne vulgaris, that is, simultaneously, anti-bacterial, anti-inflammatory, anti-sebum and keratolytic effects.

[0041] In one aspect, the present invention relates to the use ofCordiaverbenaceaessential oilin the preparation of a dermocosmetic composition for multifactorial treatment of acne vulgaris, i.e., with simultaneous antibacterial, anti-inflammatory, anti-oiliness and keratolytic effects, which eliminates the need to use complementary products for the treatment.

[0042] According to the present invention, the term “essential oil” can be understood as oil derived fromCordiaverbenacea, particularly obtained from the leaves. The essential oil according to the present invention can be obtained through any process known in the state of the art, preferably those that maintain the quality and amount so that it can perform its cosmetic functions.

[0043] By “derivatives” it is meant that the biological components or mixtures thereof, obtained fromCordiaverbenacea, can be isolated or enriched, in order to include those that have some components in amounts not naturally achievable, without human intervention.

[0044] In particular, theCordiaverbenaceaessential oilaccording to the present invention comprises alpha-pinene, beta-pinene, 1,8-cineole, trans-caryophyllene, alpha-humulene, beta-farnesene, allo-aromadendrene, germancrene d, bicyclogermancrene, beta-bisabolol, or mixtures thereof.

[0045] Preferably, theCordia verbenaceaessential oilis obtained through a steam distillation process described in patent PI 0203067-5, by Aché Laboratórios Farmacêuticos SA, the content of which is incorporated herein as a reference.

[0046] In a particular embodiment, theCordiaverbenaceaessential oilof the present invention is in oily form, and its delivery is expected in an emulsion base (oil in water, for example). However, clinical adherence to the treatment of acne vulgaris requires providing compositions appropriate for application to oily skin, so that the delivery of the active ingredient according to the present invention would be impaired for the intended application.

[0047] In order to overcome the challenge of the state of the art, the present invention provides a silicone-based composition that is suitable for sensitive skin, such as oily and acne-prone skin.

[0048] Thus, in another aspect, the present invention relates to a stable dermocosmetic composition, for topical application, effective in the treatment of acne vulgaris, with multifactorial efficacy, which comprisesCordiaverbenaceaessential oil as an active ingredient, in a dermocosmetically acceptable silicone base, which has a dry touch and is suitable for acne-prone skin.

[0049] The stability of the composition of the present invention is directly related to its quality and integrity, as well as its useful life, that is, without phase separation.

[0050] This result was only achieved using the specific composition according to the present invention, which combines particular amounts of the active ingredient according to the present invention with a silicone base developed from specific excipients.

[0051] For the purposes of the present invention, unless otherwise stated, percentage values refer to weight / weight percentage (%w / w). Thus, percentages mean the weight of one or more ingredients in relation to the total weight of the composition.

[0052] In one embodiment of the present invention, Cordiaverbenaceaessential oil is used in a concentration of about 0.05 to about 0.5% w / w.

[0053] It was also observed that the use ofCordiaverbenaceaessential oil according to the present invention in amounts above about 0.5% presents a very evident and undesirable characteristic odor for the intended application. On the other hand, the use of fragrances in concentrations above about 0.5% was found to be unfeasible, if outside the scope of the present invention, since phase separation is observed. This is because high concentrations of fragrance tend to destabilize silicone-based compositions.

[0054] It is known that clinical adherence is also related to the touch of the final composition and its odor, which would make the use ofCordiaverbenaceaessential oilunfeasiblebecause it has a characteristic odor in certain concentrations. On the contrary, the composition according to the present invention has a dry touch that is highly desirable for its intended application and allows the use of fragrances in amounts above 0.5% of active ingredient, in a stable manner, that is, without phase separation.

[0055] Therefore, it is also an object of the present invention, in a particular embodiment, to use theCordiaverbenaceaessential oilaccording to the present invention in amounts of about 0.5% to about 3%, particularly about 2.5% w / w with fragrances. In one embodiment of the present invention, the fragrance is present in an amount of about 0.05% to 0.2% w / w.

[0056] The silicone base according to the present invention is made of the following ingredients: cyclopentasiloxane, dimethicone, dimethiconol, dimethicone crosspolymer, peg / ppg-18 / 18-dimethicone, or mixtures thereof. It is also an object of the present invention, in a particular embodiment, that the silicone base comprises from about 2 to about 8% silicone, in particular about 5.5% silicone.

[0057] The other excipients may be those known in the art, for example, selected from the classes of solvents, anionic polyelectrolytes, glycol ethers, fat-soluble vitamins, high-density glycols, surface-active polymers, salts, non-ionic surfactants, or mixtures thereof.

[0058] Regarding clinical efficacy, the specifically balanced composition according to the present invention provides a non-comedogenic effect, in addition to contributing to the non-aggravation or prevention of acne lesions, reducing skin inflammation and oiliness.

[0059] In another aspect, the present invention relates to a product for treating acne vulgaris comprising the stable dermocosmetic composition according to the present invention, intended for topical application.

[0060] The present invention also relates to methods of treating acne vulgaris, comprising a step of applying, to the skin,Cordiaverbenaceaessential oilor a layer of a cosmetic composition or product as defined in the present invention, particularly twice a day.Examples

[0061] The examples shown herein are intended only to exemplify one of the numerous ways of carrying out the invention, without limiting its scope. Their purpose is to assist in proving the technical effect, in a direct and comparative manner.Example 1 – Composition of the present invention.

[0062] The composition according to the present invention was prepared according to the following table.

[0063] [Table 1] Composition according to the present invention.IngredientConcentrationClassFunctionWater85.96%SolventVehicleGlycerin4.98%HumectantCyclopentasiloxane3.35%Volatile silicone polymerEmollientSodium polyacrylate1.50%Anionic polyelectrolyteStabilizerDimethicone1.33%Fluid siliconeEmollientPhenoxyethanol0.52%Glycol etherPreservativeCordia verbenaceaessential oil0.50%Essential oilActive ingredientTocopheryl acetate0.50%fat soluble vitaminAntioxidantDimethiconol0.31%Silicone gumEmollientCaprylyl glycol0.30%High density glycolHumectantDimethicone crosspolymer0.18%Silicone polymerStabilizerPEG / PPG-18 / 18-dimethicone0.18%PolymersEmulsifierTrideceth-60.18%Surfactant PolymerEmulsifierFragrance0.10%FragranceFragranceLaureth-230.05%Surfactant PolymerEmulsifierPotassium sorbate0.01%SaltPreservativeLaureth-40.01%Non-ionic surfactantEmulsifierExample 2 - Efficacy tests

[0064] TheCordiaverbenaceaessential oiland the composition (example 1) according to the present invention were tested and the results are discussed below.

[0065] Bactericidal and bacteriostatic effect ofCordiaverbenaceaoil

[0066] In a study that sought to evaluate the bactericidal potential ofCordiaverbenaceaoil, C. acnes strain ATCC 11827 bacteria were incubated with different concentrations ofCordiaverbenaceaessential oil. First, a 5000 μg / mL solution ofCordiaverbenaceaoil was produced and serial dilutions were performed to a concentration of 19.5 μg / mL. Subsequently, the bacterial cultures were grown in culture media and 500 μL of the different concentrations ofCordiaverbenaceaoil were added. The culture media were incubated for 7 days and after this period, the number of cultures was counted and the concentrations capable of promoting inhibition of bacterial growth (MIC) and elimination (MBC) of the bacteria were identified. At the end of the study, it was possible to observe thatCordiaverbenaceaoil promoted an inhibition of bacterial growth at a concentration of 625 µg / mL (MIC) and an elimination of the bacteria at a concentration of 2,500 µg / mL.

[0067] Effect ofCordiaverbenaceaoil on biofilm formation

[0068] Biofilms are highly organized biological communities where bacteria form structured, coordinated and functional communities. These biological communities are embedded in polymeric matrices produced by themselves. The formation of biofilms is associated with a mechanism of resistance and protection of microorganisms, therefore, the elimination of these structures contributes to a more efficient antimicrobial effect.

[0069] A study was conducted to evaluate the potential ofCordiaverbenaceaessential oilto inhibit the formation and break down bacterial biofilm. For this purpose, bacteria of the C. acnes ATCC 11827 strain were cultivated in 96-well plates containing culture medium for 12 hours in the presence of different concentrations ofCordiaverbenaceaoil. After this period, the supernatant was removed from the wells and the amount of biofilm formed was evaluated. It was observed thatCordiaverbenaceaessential oil was able to prevent biofilm formation from a concentration of 15.6 mg / mL, a result that demonstrates the bactericidal effect ofCordiaverbenaceaoilalso by inhibiting biofilm formation.

[0070] Effect ofCordiaverbenaceaoil on the specific inflammation present in acne condition

[0071] In a study that aimed to evaluate the effects ofCordiaverbenaceaessential oilon inflammatory parameters present in acne, human fibroblasts were added to a culture medium containing DMEM supplement, L-glutamine, penicillin and streptomycin, at 37ºC and 5% O2, with the aim of promoting cell growth. After establishing the cellular medium, bacteria of the C. acnes ATCC 11827 strain were added to the medium with the aim of stimulating the release of specific interleukins present in acne condition.

[0072] At the end of the experiment, it was observed that the main interleukins produced in the inflammation caused by the C. acnes bacteria were IL-6 and IL-8.Cordiaverbenaceaoilwas tested in 3 concentrations (11 / 33 / 100 µg / mL) and the levels of interleukins released in the presence and absence of treatment with Cordia oil were quantified, as described in the following Table.

[0073] [Table 2] Inhibition of IL-6 and IL-8 release after treatment of fibroblasts withCordiaverbenaceaoilNegative ControlPositive ControlCordia verbenaceaoil 11 µg / mLCordia verbenaceaoil 33 µg / mLCordia verbenaceaoil 100 µg / mLIL-6(pg / mL ± SEM)152±1128±0*180±2158±1844±1*IL-8(pg / mL ± SEM)475±13156±0*432±4376±22*169±9**Statistically significant (p<0.05)

[0074] It was observed that the specific mechanism of action through TLR-II receptors, activation of MHC-II, generation of a cellular cascade and production of IL-6 and IL-8 through a specific intracellular pathway.Keratolytic effect of Cordia verbenacea oil

[0075] Another relevant pillar for acne treatment is the promotion of constant cell renewal, which is measured by the amount of keratin that is peeled from the skin (keratolytic effect).

[0076] In a study that sought to evaluate the potential ofCordiaverbenaceaessential oilon keratin, human keratinocyte cultures were maintained in an appropriate growth medium containing growth factor, gentamicin and pituitary extract and the cell media were treated with different concentrations ofCordiaverbenaceaoil. The measurement of the keratolytic effect is performed through the levels of inhibition of the incorporation of the BrdU substance in the keratinocytes, that is, the higher the levels of inhibition of the incorporation of the BrdU substance, the greater the keratolytic effect.

[0077] The table below shows thatCordiaverbenaceaessential oil was able to promote a keratolytic effect, with a statistically significant result, starting at a concentration of 30 μg / mL.

[0078] This is the first time that the keratolytic effect associated withCordiaverbenaceaoil has been reported.

[0079] [Table 3] Keratolytic effect ofCordiaverbenaceaoil evaluated by inhibition of BrdU incorporation in keratinocytesCompoundConcentrationResult (% inhibition ± SD)Control (no stimulus)-28% ± 6Negative control-0% ± 5Positive control (Cytosin β-D-arabinofuranoside3 μM70% ± 1*Cordia Verbenaceaoil3.3 μg / mL0% ± 710 μg / mL3% ± 330 μg / mL49% ±4**Statistically significant result (P<0.001)

[0080] Clinical efficacy data ofCordiaverbenaceaoil in acne condition

[0081] Volunteers with skin phototypes I-IV and different degrees of acne (IGA 0, I and II) were included and randomized into 3 different groups, which received the composition according to example 1 containing different concentrations ofCordiaverbenaceaoil(0.25% / 0.50% / 2.50%). Patients were instructed to wash their face before application and to use the products for 28 consecutive days, twice a day (morning and evening). To evaluate the efficacy of the products, equipment such as Visia, Sebumeter, clinical evaluation performed by a dermatologist, in addition to a sample collection of the skin surface, with a swab, to evaluate the lipid profile and levels of inflammatory cytokines before and after the study period were used.

[0082] After the end of the treatment, about 50% of the participants showed a reduction in the IGA level (IGA II > IGA I / IGA I > IGA 0), thus demonstrating that the composition and the product according to the present invention, as well asCordiaverbenaceaoil, are capable of acting to improve the general condition of acne.

[0083] From the point of view of inflammatory lesions (papules with reddish edges) and non-inflammatory lesions (closed and open comedones), the volunteers presented a significant reduction in both parameters after 28 days of using the products, with the reduction in non-inflammatory lesions being around 55%, while the reduction in inflammatory lesions reached a reduction of up to 75% in the concentrations tested (figures 1 and 2).

[0084] From the point of view of oiliness, after 28 days of using the products, the volunteers showed a reduction of around 25% in oiliness levels, and from the first day of use, the participants already showed a tendency for reduction, which means a contribution to the improvement of the acne condition ().

[0085] Additionally, the products according to the present invention, in all concentrations, promoted a reduction in the amount of porphyrins in the skin, a result that represents an action in the keratolytic and bactericidal pillars, since porphyrins are byproducts of the metabolism of keratin by the bacteria C. acnes, therefore, a reduction in porphyrin levels demonstrates a clinically evaluated bactericidal effect, in addition to a keratolytic effect ().Safety data of Cordia verbenacea oil

[0086] Toxicological warnings on products containing traditional active ingredients such as benzoyl peroxide and adapalene were considered.

[0087] Acute exposure to benzoyl peroxide can cause moderate to severe irritation to the eyes, nose, and throat, with redness of the eyes, runny nose, coughing, and difficulty breathing. On contact with the skin, benzoyl peroxide causes redness and swelling.

[0088] Adapalene is known to cause mild side effects, such as photosensitivity, irritation, redness, dryness, itching, and burning. It is common (between 1% and 10% of users) to experience a brief sensation of heat or stinging, as well as dry skin, peeling, and redness during the first 2 to 4 weeks of medication use.

[0089] In view of these data of conventional products, the safety ofCordiaverbenaceaoil regardinggenotoxic potential was evaluated.

[0090] One of the most widely used studies by the scientific community to assess the mutagenic potential of substances is the Ames test, described in the OECD 471 protocol. In this study, different bacterial strains are incubated with the chemical substance to be tested under different experimental conditions and the number of changes in the DNA of these bacteria is assessed. High mutation rates indicate that the substance has the potential to cause damage to genetic material.

[0091] In a study that aimed to evaluate the mutagenic potential ofCordiaverbenaceaoil, according to OECD protocol 471, bacteria from theSalmonellathyphymuriumandE. colistrainswere incubated with different concentrations ofCordiaverbenaceaoil,in the presence and absence of metabolic activation (mouse S9 enzyme system).At the end of the study, it was possible to demonstrate that the different concentrations ofCordiaverbenaceaoil were not capable of altering the bacterial replication profile, thus demonstrating an absence of mutagenic effect.

[0092] Another study conducted with the aim of evaluating the safety profile of substances that will be applied topically is the ocular irritation test according to the OECD 492 protocol. In this protocol, reconstructed human corneal epithelia are incubated with different concentrations of the test substance and an assessment of the cytotoxicity of the reconstructed epithelium is performed. Substances that present low levels of cytotoxicity present a satisfactory safety profile.

[0093] In a study in whichCordiaverbenaceaoil was applied to reconstructed human corneal epithelium, it was observed that this oil presents low levels of cytotoxicity, thus demonstrating a low potential to cause toxicological effects in the eye region.

[0094] Considering that the use of products for the treatment and prevention of acne have safety warnings regarding phototoxicity, a non-clinical study was carried out with the aim of evaluating the potential ofCordiaverbenaceaoilto cause phototoxicity reactions.

[0095] Phototoxicity occurs when a certain substance, in the presence of UV radiation, has its chemical structure altered and the newly formed substance interacts with skin proteins, resulting in an allergic reaction. These phototoxic substances, such as adapalene and benzoyl peroxide, for example, cannot be used during the day, limiting their treatment to nighttime periods.

[0096] In view of this need, a study was designed in accordance with OECD guide 432, in which 3T3-NRU cell line taken from mice were incubated in the presence and absence ofCordiaverbenaceaoil(1,000 µg / mL) and subjected to a standardized amount of UV radiation (5 J / cm2) in the light range of 320 to 400 nm. At the end of the study, the level of cytotoxicity caused by the test substance in the presence and absence of UV radiation was evaluated by calculating the PIF (Photo Irradiation Factor), which is calculated using the following formula:

[0097]

[0098] According to OECD guide 432, the reference values for assessing phototoxicity are demonstrated in the following table.

[0099] [Table 4] Phototoxicity dataPIF < 2Non-phototoxic2 ≤ PIF ≤ 5Potentially phototoxicPIF > 5Phototoxic

[0100] The results are shown below in the following table.

[0101] [Table 5] Phototoxic effect ofCordiaverbenaceaoil on 3T3-NRU cellsSubstanceIC50 (-UVA) μg / mLIC50 (+UVA) µg / mLPIFPositive control7 – 900.1 – 2>6Cordia verbenaceaoil 1,000 µg / mL90.9882.031.11

[0102] From the results obtained in this study, it is possible to conclude thatCordiaverbenaceaoildoes not have a phototoxic profile and can be used to treat acne at any time of the day.

[0103] Clinically,Cordiaverbenaceaoil was evaluated through the HRIPT (Human Repeated Insult Patch Test), a protocol recognized and required by different regulatory bodies, which aims to evaluate the safety profile of substances that will be used topically before long-term exposure. This study consists of applying a determined amount of the substance to the volunteer’s skin and occluding it with a topical patch. The volunteers remain with the occlusive patchfor 48 hours straight, followed by a 10-day interval between applications. Three application cycles are performed on each volunteer. At the end of the exposure time, an assessment is made of the amount of erythema formed on the volunteers’ skin. The erythema scale ranges from 0 (no erythema) to 5 (severe erythema). When evaluated through the HRIPT study,Cordiaverbenaceaoilpresented erythema on a scale of 0, thus demonstrating the absence of irritation potential of this substance.

[0104] Therefore, the tests presented herein show that, in experiments performed in cell culture:1) TheCordiaverbenaceaoil according to the present invention acts directly on the microbial pillar of the pathology, since it has a bactericidal, bacteriostatic and biofilm formation inhibitory effect against the bacteria C. acnes.2) TheCordiaverbenaceaoil according to the present invention acts directly on the follicular hyperkeratosis pillar of the pathology, reducing proliferation of keratinocytes in a cell culture.3) TheCordiaverbenaceaoil according to the present invention acts directly on the inflammatory pillar of the pathology, as verified by the reduction of IL-6 and IL-8 in a keratinocyte culture stimulated by C. acnes.4) As microbial colonization by C. acnes and follicular hyperkeratosis contribute to the inflammatory condition of the pathology, theCordiaverbenaceaoil according to the present invention also acts indirectly on the inflammatory pillar of the pathology.

[0105] Furthermore, the effects observed in cell culture translated into clinical efficacy in a study conducted with 90 participants who applied a formulation containingCordiaverbenaceaoilaccording to the present invention to their faces for 28 days, where:1) There was an average reduction of 25% in the amount of IL-1 (inflammatory marker) on the volunteer’ faces.2) There was a reduction of up to 55% in the number of non-inflammatory lesions and up to 75% in the number of inflammatory lesions on the volunteers’ faces.3) There was a reduction of up to 25% in the amount of oil on the volunteers’ faces, effective from the first day of application.4) There was a reduction of up to 15% in the colonization of C. acnes and hyperkeratosis on the faces of volunteers.

[0106] As a consequence of effects 1-4, 50% of volunteers showed a clinical reduction in the overall intensity of acne, measured by the reduction in the IGA grade.

[0107] Those skilled in the art will appreciate the knowledge presented herein, which allows the invention to be reproduced in the embodiments presented and in other variants, covered by the scope of the attached claims.

Claims

Use ofCordiaverbenaceaessential oilcharacterizedby the fact that it is in the preparation of a stable dermocosmetic composition for the treatment of acne vulgaris.Use, according to claim 1,characterizedby the fact that it presents simultaneous anti-bacterial, anti-inflammatory, anti-oiliness and keratolytic effects.Use, according to claim 1,characterizedby the fact that it presents an anti-inflammatory effect by acting through the TLR-2 and MCH-II pathways.Use, according to any one of claims 1 to 3,characterizedby the fact that theCordiaverbenaceaessential oilcomprises alpha-pinene, beta-pinene, 1,8-cineole, trans-caryophyllene, alpha-humulene, beta-farnesene, allo-aromadendrene, germancrene d, bicyclogermancrene, beta-bisabolol, or mixtures thereof.Stable dermocosmetic compositioncharacterizedby the fact that it comprisesCordiaverbenaceaessential oilin a dermocosmetically acceptable silicone base.Composition, according to claim 5,characterizedby the fact that theCordiaverbenaceaessential oilcomprises alpha-pinene, beta-pinene, 1,8-cineole, trans-caryophyllene, alpha-humulene, beta-farnesene, allo-aromadendrene, germancrene d, bicyclogermancrene, beta-bisabolol, or mixtures thereof.Composition, according to any one of claims 5 or 6,characterizedby the fact that it comprises about 0.05 to about 0.5% w / w ofCordiaverbenaceaessential oil.Composition, according to any one of claims 5 to 7,characterizedby the fact that it comprises about 0.5% to about 3% ofCordiaverbenaceaessential oil with fragrance.Composition, according to claim 8,characterizedby the fact that the fragrance is present in an amount of about 0.05% to 0.2% w / w.Composition, according to claim 5,characterizedby the fact that the silicone base comprises cyclopentasiloxane, dimethicone, dimethiconol, dimethicone crosspolymer, peg / ppg-18 / 18-dimethicone, or mixtures thereof.Composition, according to claim 10,characterizedby the fact that the silicone base comprises about 2 to about 8% silicone.Composition, according to claim 11,characterizedby the fact that the silicone base comprises approximately 5.5% silicone.Composition, according to any one of claims 5 to 12,characterizedby the fact that it comprises excipients selected from the classes of solvents, anionic polyelectrolytes, glycol ethers, fat-soluble vitamins, high-density glycols, surface-active polymers, salts, non-ionic surfactants, or mixtures thereof.Product for treating acne vulgarischaracterizedby the fact that it comprises a cosmetic composition as defined in any one of claims 5 to 13.Method for treating acne vulgarischaracterizedby the fact that it comprises a step of applying, to the skin,Cordiaverbenaceaessential oil or a layer of a cosmetic composition as defined in any one of claims 5 to 13 or a product as defined in claim 13.Method according to claim 15,characterizedby the fact that it comprises application twice a day.

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