Cosmetic use of hyacinth extract

Abstract

The present invention relates to the cosmetic use of an aqueous extract of Hyacinthus orientalis bulbs as a cutaneous neurosensory agent, for example for improving cutaneous perception, for rebalancing or improving the neurosensory responsiveness of the skin to stimuli. The invention also relates to a cosmetic composition comprising an aqueous extract from bulbs of the genus Hyacinthus, preferably from dormant Hyacinthus orientalis bulbs, as an active cosmetic ingredient, and at least one cosmetically acceptable excipient. The invention also relates to a cosmetic ingredient comprising an aqueous extract from bulbs of the genus Hyacinthus, said aqueous extract being in a suitable vehicle for incorporation into a cosmetic composition or product.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, and in particular to cosmetic agents that function as skin neurosensors, such as those that can improve skin perception, reduce harmful sensations and / or improve skin health, resulting in a more youthful, relaxed and radiant appearance. Background Technology

[0002] The skin is a complex organ with dense nerve innervation, playing a crucial role in sensory perception and communication with the nervous system. Sensory nerve endings in the skin respond to a variety of stimuli, such as heat, cold, and touch. Some fibers can even be activated by a variety of chemicals, including substances secreted by microorganisms, such as those present in the skin's microbiome. The connection between nerves and the skin occurs at the cellular level via neurotransmitters such as substance P or calcitonin gene-related polypeptides. The related polypeptide (CGRP) provides these signaling molecules. These molecules are released from nerve endings and also from the skin or immune cells within the skin. This communication between skin cells and sensory neurons influences many biological processes, including healing, inflammation, and immune responses, and is involved in the regulation of skin health and homeostasis.

[0003] However, sensory nerve endings in the skin are highly exposed to environmental challenges and stressors (such as weather conditions, chemicals, pollution, and certain lifestyle habits). Such overexposure can alter the skin's sensitivity and responsiveness to environmental aggressors and tactile stimuli. Similarly, certain physiological changes (such as aging and menopause) can also affect skin perception, for example, by impairing peripheral sensory function or reducing the number of stimuli receptors in the skin. Alterations in the skin's detection and perception of external stimuli can lead to skin... Impaired communication within the peripheral sensory system leads to inappropriate biological responses to stimuli (such as inappropriate immune responses in the skin). Skin homeostasis can be altered, resulting in skin sensitivity and discomfort, and ultimately, changes in the skin's visual appearance.

[0004] Therefore, there is a need for cosmetic neurosensory agents that can reduce or prevent non-pathological changes in skin perception. Summary of the Invention

[0005] This application relates to an aqueous extract of bulbs from the genus *Hyacinthus*, preferably for cosmetic use as a dermal neurosensor. Preferably, the extract is obtained from the dormant bulbs of *Hyacinthus*.

[0006] In some implementations, the extract can be obtained by a method including the following steps:

[0007] (a) Incubate the peeled and crushed bulbs in water to obtain the sap.

[0008] (b) Recover the aqueous phase from the slurry obtained in step (a).

[0009] (c) The aqueous phase obtained in step (b) is heated at a temperature above 95°C and then cooled to promote precipitation.

[0010] (d) Recover the aqueous phase from step (c), and

[0011] (e) Optionally filter and / or concentrate the aqueous phase obtained in step (d) to obtain the aqueous extract.

[0012] By acting as a neurosensory agent, aqueous extracts of hyacinth bulbs can be used to improve skin perception and / or to rebalance or improve the skin's neurosensory responsiveness to stimuli and / or to reduce or prevent skin noxious sensations.

[0013] In one specific implementation, hyacinth bulb extract is used to prevent or reduce age-related skin perception impairment, particularly skin sensitivity.

[0014] In another aspect, hyacinth bulb extract acts as an agonist of OXT receptors, particularly those present on sensory nerve endings in the epidermis.

[0015] The extracts of this invention can also be used to improve the visual appearance of the skin, particularly to alleviate, reduce, or treat one or more non-pathological changes in skin appearance, such as roughness, redness, loss of luster or radiance, dull or uneven skin tone, tired appearance, and combinations thereof. The extracts of this invention can also be used to relieve skin discomfort and / or skin sensitivity and / or to provide a comfortable effect on the skin.

[0016] The extracts of this invention can also be used to reduce the appearance of wrinkles on the skin and / or improve skin elasticity, particularly in subjects with mature skin.

[0017] When applied to the skin of an object, the extracts of the present invention can also be used to enhance the skin’s sensitivity to pleasant and / or mild tactile stimulation.

[0018] In one particular aspect, the present invention relates to the topical use of hyacinth bulb extract as a cosmetic agent for improving skin sensation and / or for reducing or preventing skin abrasive sensations and / or for alleviating skin discomfort.

[0019] In some implementations, the extract is applied topically to the skin of the intimate body area. Accordingly, the extract is applied topically to the skin of the intimate body area to reduce discomfort and / or increase sensation (e.g., sensitivity to emotional touch or pleasurable sensations) and / or enhance the sense of well-being of the skin in the intimate body area.

[0020] In some implementations, the extract is applied topically to the scalp. Therefore, the extract can be used to relieve scalp discomfort.

[0021] The extract is applied topically to the skin, preferably to sensitive and / or mature skin.

[0022] Extracts can be applied to the skin in the form of cosmetic compositions.

[0023] Cosmetic compositions can be of any type. They can be skin care products, skin hygiene products, hair care products, feminine care products, or cosmetic products, such as creams (e.g., night creams or day creams or massage creams), balms, ointments, serums, gels, masks, foundations, soaps, shower gels, shampoos, conditioners, or aftershaves.

[0024] In another aspect, this application relates to cosmetic compositions for improving skin sensation or providing a sense of well-being in a subject, the compositions comprising an aqueous extract from hyacinth bulbs, preferably from dormant hyacinth bulbs, as an active cosmetic ingredient, and at least one cosmetically acceptable excipient.

[0025] In one aspect, this application relates to a cosmetic method for improving skin perception in a subject, and / or for enhancing skin sensitivity (particularly sensitivity to pleasurable stimuli), and / or for reducing skin discomfort, and / or for improving the visual appearance of the skin, and / or for providing a sense of well-being, the method comprising topically applying an aqueous extract or cosmetic composition obtained from hyacinth bulbs to the skin of the subject, wherein the aqueous extract is preferably as defined herein, and the cosmetic composition is as defined herein.

[0026] In some implementations, the subject has mature skin and / or sensitive skin and / or the subject is a perimenopausal or postmenopausal woman.

[0027] In one specific aspect, this application relates to a cosmetic ingredient comprising an aqueous extract from hyacinth bulbs, preferably from dormant hyacinth bulbs, said aqueous extract being an active cosmetic agent in a cosmetically acceptable carrier, preferably in a glycerin / water mixture.

[0028] Cosmetic ingredients may also contain one or more cosmetically acceptable excipients selected from antioxidants, preservatives, and pH adjusters.

[0029] In another aspect, this application relates to the use of an aqueous extract of hyacinth bulb as a dermal neurosensor in the preparation of compositions for improving skin perception and / or for reducing or preventing skin nociceptive sensations and / or for alleviating skin discomfort.

[0030] The composition may be a skin care product, a feminine care product, or a hair care product. Attached Figure Description

[0031] Figure 1 The image shows that the extract of the present invention (right side) effectively activated the OXTR receptor in a chimeric CHO model, achieving over 70% receptor activation at 0.2% extract concentration. Left side: Activation response curve for the oxytocin positive control.

[0032] Figure 2 The effects of the extract of the present invention at 0.1% (Hyacinth. o at 0.1%) on nociceptive signal transduction upon capsaicin stimulation, in neurons, are demonstrated. The effects of CGRP release in co-cultured keratinocytes were determined by quantification. The antinociceptive effect of the extract at 0.1% completely offset the increase in signal transduction induced by capsaicin stimulation, reaching levels slightly lower than those observed in the two control pathways: oxytocin and capsaicin, 10 µM.

[0033] Figure 3A The extract of the present invention was shown to improve neurons at 0.1% (Hyacinth. o. at 0.1%). Neuronal survival in co-culture of keratinocytes was improved, while capsaicin reduced neuronal survival, both compared to the untreated control group.

[0034] Figure 3B The extract of the present invention was shown to significantly improve neuronal function at 0.1% (Hyacinth. at 0.1%) compared to the untreated control group. Dendriticity of neurons in co-culture of keratinocytes (shown here on a per-neuron basis).

[0035] Figure 4 This demonstrates the effects of hyacinth bulb extract treatment on neurons. The role of electrical signal transduction in organ-on-a-chip co-culture of keratinocytes in a multi-compartment chip. HO = Hyacinth bulb extract.

[0036] Figure 5AThe study demonstrated that 1% of the active ingredient of the invention in the formulation increased EEG signals measured in the left frontal region during mild feather touch stimulation, an effect associated with feelings of pleasure or enjoyment. This effect was evident when comparing the initial state (D0) to any of the subsequent time points shown (D0, D7, and D28 after product use). In contrast, the placebo control group showed no significant change. Figure 5B The mean alpha wave activation at electrode AF3 (left frontal lobe) is shown at day 0 (D0), (30 minutes after D0 + application), D7, and D28. Figure 5B It was determined that 1% of the component of the present invention (hyacinth bulb extract) in the formulation enhanced skin tactile sensitivity 30 minutes after application, and this effect was present on days 7 and 28 compared with the placebo cream.

[0037] Figure 6 It was demonstrated that after 28 days of use, in a clinical setting, 1% of the ingredients of the present invention in the formulation reduced nociceptive sensations in a significantly more effective manner than the effects recorded in the placebo control group, as evaluated by the capsaicin sting test.

[0038] Figure 7 The results show that after 28 days of product use, 1% of the ingredients of this invention in the formulation significantly enhanced the visible healthy radiance of the skin in a manner significantly stronger than that observed in the placebo control group, as evaluated by clinician grading (left) and panel member self-assessment (right). In each case, D28 results are shown immediately before product application ('D28', middle pair of columns) and immediately after product application ('D28Timm', right pair of columns).

[0039] Figure 8 It was shown that after 28 days of treatment, 1% of the component of the invention in the formulation significantly enhanced skin color in a manner superior to that observed in the placebo control group, such as brightness (L) as observed by chromaticity measurements. Significantly improved and reddened (a () significantly reduced.

[0040] Figure 9 It was shown that after 28 days of treatment, 1% of the ingredients of the present invention in the formulation enhanced skin elasticity in a manner significantly superior to that observed in the placebo control group.

[0041] Figure 10The results show that after 28 days of product use, 1% of the ingredients of this invention in the formulation reduced skin wrinkles in a manner significantly superior to that observed in the placebo control group, as evaluated by image analysis (left) and clinician grading (right). In the case of clinician grading, D28 results are shown immediately before product application ('D28', the middle pair of columns) and immediately after product application ('D28Timm', the right pair of columns).

[0042] Figure 11 Self-evaluation scores of skin comfort and pleasure are shown immediately after application on day 0 (D0) and day 28 (D28). Skin application of the cream (hyacinth bulb extract) of the present invention significantly improved skin comfort / pleasure compared to a placebo cream. Percentage changes are expressed as a comparison to the D0 score before cream application. Detailed Implementation

[0043] Oxytocin (OT) is a nonapeptide that is released into the central nervous system primarily from the paraventricular nucleus and supraoptic nucleus of the hypothalamus.

[0044] This peptide is involved in various physiological functions, such as cardiovascular activity, respiration, eating, and social interaction. Recently, it has also been shown that oxytocin is involved in nociceptive signaling.

[0045] Oxytocin works through its receptor, a seven-transmembrane receptor that is coupled to different signaling pathways via various G protein subtypes.

[0046] Recent research has focused on the peripheral effects of oxytocin in the skin. It has been shown that the oxytocin receptor (OXTR) is expressed on skin fibroblasts, while oxytocin and its carrier protein, neurohypophyseal hormone carrier protein I, are synthesized by keratinocytes.

[0047] Cho et al. (British Journal of Dermatology, 2019, 181, p. 1216) Page 1225 shows that oxytocin inhibits SASP (senescence-associated secretory phenotype) in normal human dermal fibroblasts (NHDF) by binding to its receptors on fibroblasts. Associated secretory phenotype (OT)-induced aging, and this effect depends on the age of the donor NHDF. Cho et al. concluded that OT and OXTR agonists may be promising agents for improving age-related skin aging.

[0048] In addition, Hayre (Journal of drugs in Dermatology, 2020, 19, 1146 (1147) confirmed the correlation between OT levels and skin age score (SAS). High OT levels were associated with younger-looking skin and significantly lower-than-expected SAS scores. Hayre concluded that OT may provide a protective mechanism against photoaging, endogenous aging, or both.

[0049] Grinevitch et al. (2017, www.painjournalonline.com) and Gonzalez Hernandez et al. (2017, www.painjournalonline.com) demonstrated that oxytocin is secreted by keratinocytes, while OXTR is expressed on nociceptive fibers in the skin, and that peripheral oxytocin (e.g., released by keratinocytes) can inhibit the excitation of nociceptive fibers in the skin. Specifically, Gonzalez... Hernandez et al. demonstrated that subcutaneous injection of OXT induced nociceptive responses in animal models, suggesting a novel potential role for peripheral OT in pain modulation.

[0050] In addition, in the nervous system and nerves In the context of the skin interface, the literature demonstrates the pleiotropic effects of OT, including in neurogenesis and neurite growth.

[0051] Given the emerging evidence of OT involvement in skin homeostasis and perception, the inventors hypothesize that compounds capable of mimicking the effects of OT could act as cosmetic neurosensors.

[0052] Unexpectedly, the inventors identified an aqueous extract from hyacinth bulbs, specifically an aqueous extract obtained from dormant hyacinth bulbs, that can activate OXTR in vitro. Figure 1 Furthermore, the inventors have demonstrated that aqueous extracts from hyacinth bulbs can be used in neurons. Inhibition of nociceptive signaling during capsaicin stimulation in co-culture of keratinocytes (quantification via CGRP release) Figure 2 Notably, the extract of the present invention, at a concentration of 0.1%, is more effective than 10 µM oxytocin in reducing CGRP release induced by capsaicin attack. The inventors have also demonstrated that the extract of the present invention is at least as effective as 10 µM OT in improving neuronal survival and dendrite density during capsaicin attack. Furthermore, the inventors have shown that the extract of the present invention can induce NHDF proliferation in vitro in a dose-dependent manner. Figure 3A and Figure 3B ).

[0053] The inventors conducted a placebo-controlled clinical study to determine in vitro results. This study showed that a cream containing 1% of the component of this invention based on hyacinth bulb extract enhanced the skin's pleasant sensitivity to gentle skin touch, as evaluated by EEG readings obtained when the skin was stimulated with a feather. Figure 5A and Figure 5B The cream of the present invention also shows that it reduces skin irritation, as modeled by the capsaicin sting test. Self-evaluation using a rating scale clearly shows that, compared with a placebo cream, use of the cream containing 1% of the ingredients of the present invention significantly improves skin pleasure and comfort, indicating an improvement in skin sensation and well-being.

[0054] Furthermore, treatment with the cream of this invention for 28 days resulted in significant improvements in skin appearance, specifically in the significantly improved assessments of visible healthy radiance by both study participants and trained clinicians; a significant reduction in redness and skin lightening, both measured by Hunter L,a,b chromatic acuity; a reduction in wrinkles, assessed by both instrumental analysis (using PRIMOS image analysis) and grading by trained clinicians at the corners of the eyes (crow's feet area); and improved skin elasticity, as measured by a cutometer. Finally, self-assessments by panel members showed significant improvements in the levels of perceived parameters, including: visible skin health, overall skin aesthetics, healthy radiance, and rosy / pleasant radiance (Figures 5 to 10). In summary, this clinical trial demonstrates that the extract of this invention functions as an effective sensory agent. In particular, the extract of this invention exhibits a rebalancing of the skin's neurosensory responsiveness to external stimuli, reducing skin discomfort and promoting pleasurable tactile stimulation.

[0055] The extracts of this invention also show improved skin youthful and healthy appearance, providing skin rejuvenation.

[0056] The extracts of this invention also show enhanced positive skin-touch sensations, thereby generating a sense of skin health and overall well-being in the subject.

[0057] In summary, by activating the OXTR signaling cascade, the extracts of this invention act as a safety agent for improving skin well-being. In other applications, the extracts of this invention can be used as cosmetic sensory agents to improve skin perception (reducing noxious sensations and increasing sensitivity to pleasurable stimuli), alleviate skin discomfort, and promote a youthful appearance.

[0058] The extract and preparation method of the present invention

[0059] In a first aspect, the present invention relates to extracts from the genus Hyacinthus, and more particularly to aqueous extracts from the bulbs of the genus Hyacinthus.

[0060] As used in this article, the term "hyacinth genus" refers to any species belonging to the genus Hyacinth. Hyacinth plants are hardy, perennial bulbous herbaceous plants, also known as hyacinths, and are famous for their brightly colored and fragrant flowers. These plants typically bloom for two to three weeks in mid-spring.

[0061] The three main species in the genus *Hyacinthus* are *Hyacinthus litwinovii* and *Hyacinthus transcaspicus*. *Hyacinthus* (also known as common hyacinth, garden hyacinth, or Dutch hyacinth) is the most common species. Native to Southwest Asia, Turkey, Syria, Lebanon, and northern Israel, it was introduced to Europe in the 16th century. Hyacinths are widely cultivated in temperate regions. To date, there are thousands of hyacinth varieties.

[0062] The term "hyacinth" as used in this article includes any variety or variant belonging to the species "hyacinth". For example, extracts can be prepared from the hyacinth variety "White Pearl".

[0063] In a preferred embodiment, the extract of the present invention is obtained from hyacinth bulbs, preferably from dormant hyacinth bulbs, and more preferably from fresh dormant hyacinth (H. orientalis) bulbs.

[0064] For example, the extract of the present invention is obtained from the dormant bulbs of the hyacinth variety "White Pearl".

[0065] The term "dormancy" as used in this article refers to a physiological state in which the metabolic rate of cells or tissues is significantly reduced, and the growth of meristematic tissues is reversibly slowed or stopped. Dormancy can be induced by providing specific environmental and / or physical conditions to the bulb.

[0066] As used herein, the term "aqueous extract" refers to an extract obtained by water treatment of plant material (e.g., hyacinth bulbs and any fraction thereof), which enables the extraction of the target compound from the plant material. Typically, the plant material is incubated in water. Incubation can be carried out at room temperature or under heating. Ultrasonic treatment and / or stirring may also be applied. Incubation (also known as immersion) can last from several minutes to several hours. The plant material (here, hyacinth bulbs, preferably hyacinth bulbs) can be peeled and sliced, the size and shape of which may vary depending on the method used. For example, the plant material can be cut, chopped, crushed, ground, freeze-ground, etc., and then placed in water. Alternatively, peeled bulbs can be mixed or crushed in water to form a slurry.

[0067] The weight ratio of water to plant material is generally 0.5 to 10, for example, 1 to 5.

[0068] Plant materials can undergo several water extraction steps at the same or different temperatures.

[0069] After incubation, the plant material and the aqueous phase containing the target compound are separated. This separation can be carried out by any method known in the art (e.g., pressing, decanting, filtration, or centrifugation). Centrifugation is the preferred method.

[0070] The recovered aqueous phase may undergo one or more additional steps (e.g., filtration, ultrafiltration, or heat treatment followed by cooling) to remove proteins and other unwanted substances.

[0071] In some embodiments, the present invention relates to a method for obtaining an aqueous extract of Hyacinth bulbs, preferably an aqueous extract of Hyacinth bulbs, comprising the following steps:

[0072] (a) Incubate the peeled and crushed bulbs in water to obtain the sap.

[0073] (b) Recover the aqueous phase from the slurry obtained in step (a).

[0074] (c) The aqueous phase obtained in step (b) is heated at a temperature above 95°C and then cooled to promote precipitation.

[0075] (d) Recover the aqueous phase from step (c), and

[0076] (e) Optionally filter the aqueous phase obtained in step (d) to obtain the aqueous extract.

[0077] Prior to step (a), the bulb may undergo one or more treatments (e.g., washing, peeling, drying, grinding, freeze-grinding or freeze-pulverizing, freezing, freeze-drying, thawing, and combinations thereof).

[0078] In some embodiments, the aqueous extract of the present invention is prepared from fresh, peeled bulbs.

[0079] The bulbs can be crushed or ground in any suitable manner before or during step (a). For example, peeled bulbs can be crushed directly in water using a mixer to obtain a slurry. The weight ratio of water to plant material is typically 0.5 to 10. Once obtained, the slurry is allowed to stand, for example, 5 to 120 minutes, or for example, 10 to 60 minutes, preferably at room temperature, to enable the extraction of the target compound from the crushed plant material. Ultrasonic treatment and / or stirring can be applied to facilitate extraction.

[0080] In step (b), the slurry is separated into two phases: (i) an aqueous phase (containing the target compound) and (ii) solid particles initially present in the slurry. The separation can be performed by any method known to those skilled in the art, such as centrifugation.

[0081] Step (c) aims to remove at least a portion of the protein extracted from the plant material. The aqueous phase is heated to a temperature above 95°C, preferably 100°C to 130°C, for at least 10 minutes, preferably 15 minutes to 150 minutes, and then cooled, preferably to room temperature. As a result, the insoluble particles at room temperature are precipitated.

[0082] In step (d), the aqueous phase and precipitate are separated, for example by centrifugation and / or filtration.

[0083] If necessary, steps (c) and (d) can be repeated.

[0084] The aqueous phase is recovered, and it can generally be used as an aqueous extract according to the invention.

[0085] The aqueous extract of the present invention typically contains 20 mg / g to 160 mg / g of dry matter, preferably 40 mg / g to 85 mg / g, for example 45 mg / g to 70 mg / g.

[0086] If necessary, in step (e), the recovered aqueous phase may be concentrated, for example under reduced pressure, to obtain the aqueous extract of the invention in liquid form having the above-described dry matter, or in dried form (e.g., in powder form).

[0087] In some embodiments, the extract of the present invention is in liquid form and has a dry matter content of 20 mg / g to 160 mg / g, preferably 40 mg / g to 85 mg / g, for example 45 mg / g to 70 mg / g.

[0088] In some specific embodiments, the extracts of the present invention contain marker molecules such as carbohydrates (especially sucrose and fructose), imino sugars (e.g., kanojirimycin, deoxynojirimycin, or deoxymannojirimycin) and amino acids (e.g., aspartic acid and glutamic acid).

[0089] In one specific embodiment, the extract of the present invention is in liquid form, having a dry matter content of 20 mg / g to 160 mg / g, preferably 40 mg / g to 85 mg / g, and comprising...

[0090] At least 0.5%, preferably 1% to 6% carbohydrates, as determined by GC. FID was evaluated after preliminary acid hydrolysis and derivatization.

[0091] At least 50 ppm, preferably 60 ppm to 1500 ppm, of iminosaccharides, such as by using 1 HPLC analysis of deoxynojirimycin as a standard UV quantification

[0092] and

[0093] At least 0.02%, preferably 0.04% to 2% amino acids, as quantified by ninhydrin spectrophotometry using aspartic acid as a standard.

[0094] In some embodiments, the extract of the present invention contains 3 ppm to 40 ppm of 1 Deoxynojirimycin (1) deoxynojirimycin (DNJ), for example 5 ppm to 30 ppm, for example 9 ppm to 25 ppm.

[0095] The aqueous extract of the present invention obtained in step (e) may undergo additional steps, such as filtration, ultrafiltration, sterilization (e.g. by nanofiltration) and / or formulation steps (e.g. by mixing the aqueous extract with a cosmetically acceptable carrier).

[0096] For example, the aqueous extract of the present invention can be diluted or dissolved in a liquid cosmetic-acceptable carrier, such as triethyl citrate, C2 C5 alkyl diols (e.g., propylene glycol, butanediol, pentanediol and their isomers), ketal esters (e.g., methyl levulinate or ethyl levulinate), glycerol, water, and mixtures thereof. Alternatively, the aqueous extract of the present invention can be adsorbed onto a solid carrier (e.g., maltodextrin).

[0097] In some preferred embodiments, the aqueous extract of the present invention is formulated with glycerol / water or C2. C5 alkyl diols in water, preferably prepared in water / glycerol.

[0098] One or more cosmetically acceptable excipients can be added, such as antioxidants, preservatives, or pH adjusters.

[0099] Some examples of antioxidants include tocopherol, ascorbic acid, its salts, and combinations thereof.

[0100] Some examples of preservatives include organic acids (such as sorbic acid, benzoic acid, dehydroacetic acid and their salts), benzyl alcohol, methylparaben, propylparaben, carvacrol, caprylyl glycol, caprylic acid glyceride and combinations thereof.

[0101] Some examples of pH adjusters include organic acids (such as citric acid or acetic acid), phosphate buffers, TRIS-based buffers (such as TRIS.HCl or TRISglycine), and combinations thereof.

[0102] For illustrative purposes only, the extract of the present invention can be formulated in a water / glycerin mixture to obtain a cosmetic ingredient that can be incorporated into a topical composition.

[0103] Cosmetic uses according to the present invention

[0104] The present invention also relates to the non-therapeutic cosmetic use of aqueous hyacinth extracts, preferably the aqueous hyacinth bulb extracts described herein, on the skin as a neurosensory agent.

[0105] As used herein, "skin-sensory agent" or "skin-neurosensory agent" refers to an active agent capable of rebalancing, normalizing, or improving the sensory perception of the skin (e.g., tactile perception) and / or the responsiveness of the skin's neurosensory system to stimuli. It also refers to an active agent capable of promoting feelings of well-being or pleasure in an object. Furthermore, it refers to a reagent capable of reducing the skin's sensitivity to nociceptive stimuli and / or increasing the skin's sensitivity to pleasurable stimuli. Preferably, such a reagent is applied topically to the skin.

[0106] For example, a "skin-sensing agent" can be a reagent that, when applied topically, can improve sensations arising from skin irritation and / or promote overall skin well-being.

[0107] As used in this article, the term "stimuli" refers to any stimulus that is directly detected on the skin. Stimuli can be of any type, such as thermal, mechanical, or chemical. This includes stimuli such as those caused by meteorological conditions (wind, dryness, humidity, sunlight, temperature (cold, hot)), stimuli induced by other external factors (electromagnetic radiation (e.g., blue light), pollution, chemicals, fumes, cosmetic treatments, etc.), vibration, hair movement, and skin contact (e.g., skin pressure or friction).

[0108] In some implementations, the irritant is skin contact.

[0109] In one specific implementation, "skin perception" refers to "tactile or tactile" skin perception.

[0110] The ability of a compound to act as a neurosensory agent can be demonstrated, and even objectively quantified, by any suitable in vitro or in vivo assay known to those skilled in the art. For example, the ability of a compound to reduce nociceptive sensation can be assessed by an in vitro capsaicin challenge as described in Example 3 or Example 4, or by an in vivo assessment as described in Example 5. The ability of a compound to enhance skin sensitivity to pleasurable stimuli and / or promote skin well-being or skin comfort can be assessed in vivo by EEG recordings during mild stimulation as described in Example 5, or by self-assessment of skin comfort using a visual rating scale before or after application of the agent to the skin as described in Example 5.

[0111] As used in this article, the term "skin" refers to any part of the human body, particularly the skin of the face, including the lips and eyelids, the skin of the neck, the skin of the hands, and the scalp.

[0112] The term “skin” as used in this article also includes the private external parts of the body, such as the external genitalia, buttocks, perineum, and breasts.

[0113] The extract of this invention is to be used topically, i.e., applied to the skin.

[0114] Preferably, the skin is healthy, meaning that the skin does not show any wounds or skin conditions (such as skin infections, eczema, or dermatitis).

[0115] In some implementations, the skin is mature. Therefore, the skin may show one or more signs of aging (such as wrinkles, fine lines, loss of elasticity and / or firmness, dull skin tone, dark circles and / or roughness).

[0116] In some alternative implementations, the skin is sensitive skin or atopic skin.

[0117] The term "sensitive skin" (also known as reactive skin) used in this article is characterized by a disproportionate responsiveness to external factors, leading to skin discomfort. This increased reactivity may result from non-pathological alterations in the skin barrier function or the cutaneous neurosensory system. Such alterations can lower the skin's tolerance threshold to stimuli.

[0118] This non-pathological change in the skin barrier or the skin's neurosensory system can be caused by physiological changes (e.g., due to aging and / or hormonal changes, such as during menopause or pregnancy), lifestyle habits (e.g., stress, lack of sleep, poor diet, alcohol, tobacco, jet lag, working at night, etc.) and / or external factors (e.g., skin exposure to climatic conditions (e.g., wind, dryness, humidity, cold, heat, sunlight), pollution (nanoparticles, ozone), and certain cosmetic treatments (e.g., peeling, shaving, or hair removal)).

[0119] As used in this article, “atopic-prone skin” refers to skin in subjects who have a history of atopic dermatitis (e.g., in childhood or infancy) and / or have an allergic background (i.e., a history of allergy and / or a family history of allergy) and who overreact to external stimuli.

[0120] The term "cosmetic active ingredient or reagent" as used in this article refers to a compound that exerts one or more cosmetic effects on the skin.

[0121] As used in this article, “cosmetic effect” refers to any non-therapeutic effect intended to improve and / or alter the appearance, mechanical properties, and / or response / perception of the skin to stimuli.

[0122] "Cosmetic effects" include the prevention, mitigation, delay of onset and / or reduction of any non-pathological changes in the skin caused by aging, lifestyle factors (such as jet lag, lack of sleep, working at night, poor diet (such as high-fat diet), alcohol consumption and / or tobacco) or exposure to external stressors (such as climatic conditions (wind, cold, heat, sun, dryness), pollution, smog, chemicals, etc.).

[0123] The extract of this invention can be used on anyone. The recipient can be of any gender or age.

[0124] In some implementations, the subjects are adults. In some implementations, the subjects are at least 25 years old, and more specifically at least 40 years old.

[0125] In some specific embodiments, the subjects of the present invention do not suffer from any neurological disorders or injuries, particularly those that cause hypersensitivity or hyposensitivity.

[0126] As mentioned above, the subject may have mature skin and / or sensitive skin.

[0127] In one specific implementation, the target group is women with mature and / or sensitive skin.

[0128] In some implementations, the target group is women, such as perimenopausal or postmenopausal women.

[0129] In one specific aspect, the extracts of the present invention are used as sensory agents capable of mimicking the peripheral effects of oxytocin in the skin. For example, the extracts of the present invention are used to activate OXT receptors expressed on sensory nerve endings in the epidermis. Without wishing to be bound by any theory, the inventors believe that such peripheral activation can rebalance the neurosensory response of the skin to stimuli.

[0130] Therefore, in one specific embodiment, the extract of the present invention is used to rebalance or normalize skin perception, particularly skin perception of tactile stimuli and / or stimuli that cause noxious sensations.

[0131] As used herein, "compounds capable of rebalancing or normalizing skin perception" refers to compounds that can improve skin's detection of stimuli and / or the skin's neurosensory response to said stimuli. Specifically, compounds can reduce and / or normalize the skin's neurosensory response to a standard threshold corresponding to the response of healthy, young, and non-sensitive skin to the same stimuli.

[0132] For example, skin sensitivity in a subject can increase due to external stress exposure or lifestyle habits. In such cases, the extracts of the present invention are designed to reduce skin sensitivity to external stimuli, particularly those that cause noxious sensations.

[0133] As shown in the examples, the extract of the present invention can counteract the effects of capsaicin, a known irritant and toxin that acts on unmyelinated sensory C fibers, particularly those that transmit nociceptive information. The inventors have also demonstrated that in neurons attacked by capsaicin... In a keratinocyte co-culture model, the extract of this invention improved neuronal survival and dendrite density to a degree similar to or even higher than that of 10 µM OT.

[0134] Therefore, in one specific aspect, the extracts of the present invention can be used to reduce and / or prevent nociceptive sensations in the skin. In other words, the present invention also relates to the use of the extracts described herein for reducing the sensitivity of nociceptive nerves.

[0135] In one specific embodiment, the present invention relates to the use of the extract of the invention for enhancing sensitivity to emotional touch and pleasurable sensations.

[0136] In some embodiments, the present invention relates to the use of the extract of the invention for increasing skin sensitivity to emotional touch and pleasurable sensations while reducing the sensitivity of nociceptive nerves.

[0137] Unbound by any theory, the applicant believes that this dual effect arises specifically from the extract’s ability to activate OXT receptors and thus activate OXT signaling pathways in sensory nerve endings in the epidermis.

[0138] In another specific aspect, the extracts of the present invention can be used to reduce, alleviate, delay the onset and / or prevent skin discomfort, particularly in subjects with sensitive and / or mature skin.

[0139] As used herein, “skin discomfort” or “skin discomfort sensation” encompasses a variety of unpleasant skin sensations (e.g., tingling, itching, numbness, burning or flushing, redness, tightness, or dryness). “Skin discomfort” can be observed in aging and / or sensitive skin. Results from the clinical trials described in Example 5 also showed that creams containing the extract of this invention significantly improved skin comfort and pleasure compared to placebo creams. Therefore, the extract of this invention can be used as a reagent to provide skin comfort, and more generally as a reagent to enhance skin comfort.

[0140] The results of the clinical trials described in Example 5 also showed that the cream containing the extract of the present invention was more effective than the placebo cream in improving skin appearance. Skin treated with the test cream of the present invention had an improved, younger, and healthier appearance compared to skin treated with the placebo.

[0141] Without wishing to be bound by any theory, the inventors believe that the improvement in skin perception, and particularly the reduction in nociceptive sensation and / or sensitivity, resulting from the peripheral effects of the extracts of this invention, has a particular impact on skin appearance: the skin looks more relaxed, radiant, healthy, and / or younger.

[0142] Therefore, in one specific embodiment, the present invention relates to the use of aqueous extracts of hyacinth bulbs, particularly aqueous extracts of hyacinth bulbs, for improving skin appearance, particularly for making skin look and / or feel healthier and / or younger.

[0143] In one specific embodiment, the present invention relates to the use of aqueous extracts of hyacinth bulbs, particularly aqueous extracts of hyacinth bulbs, as agents for providing skin rejuvenation.

[0144] In some specific embodiments, the extracts of the present invention are used as cosmetic surfactants to prevent (e.g., slow down or delay), alleviate, reduce, or treat one or more non-pathological changes in skin appearance, particularly in aging and / or sensitive skin. Non-pathological changes in skin appearance encompass, but are not limited to: roughness, redness, loss of radiance or shine, dull or uneven skin tone, tired appearance, and combinations thereof.

[0145] In another embodiment, the extract of the present invention can be used for at least one (e.g., 1, 2, 3 or more) of the following cosmetic purposes:

[0146] To prevent, treat, or reduce irregular skin pigmentation, including pigmentation spots caused by fatigue or stress.

[0147] Reduce signs of aging, including wrinkles and loss of tissue elasticity.

[0148] To make facial skin tone even or uniform.

[0149] To make facial skin brighter and / or more radiant.

[0150] To prevent or treat dull skin tone, uneven skin tone, and / or signs of a tired appearance.

[0151] It makes the skin, especially the face, fresher and brighter.

[0152] Soothe facial features,

[0153] Makes skin look less tired and more relaxed.

[0154] Revitalizes the skin, especially the face.

[0155] Improves the healthy glow of the skin.

[0156] Improve skin radiance

[0157] To prevent, treat, or reduce the appearance of redness.

[0158] To prevent, treat or relieve skin discomfort, and / or

[0159] Promotes a pleasant skin sensation.

[0160] As described above, the extract of the present invention is preferably used topically on subjects with sensitive skin and / or mature skin.

[0161] In another aspect, the present invention relates to the use of the extracts described herein, particularly aqueous extracts from hyacinth bulbs, as active cosmetic ingredients to provide cosmetic effects selected from relieving skin discomfort or hypersensitivity, improving tactile sensation of the skin, and / or improving the appearance of healthy and youthful skin (particularly radiance and luster).

[0162] As shown in Example 5, the extract of the present invention also promotes positive perception of the stimulation of feather touch, thereby generating a pleasant feeling and thus bringing a sense of well-being.

[0163] In another aspect, the present invention relates to the use of the extracts described herein, particularly aqueous hyacinth extract, for providing a feeling of well-being in a subject. Specifically, the extracts of the present invention can be used to promote or enhance a pleasurable skin sensation in a subject.

[0164] In the uses and methods described herein, the extract of the present invention is applied topically, i.e., applied to the skin. Generally, the extract is applied via a cosmetic composition. In other words, the extract is present in a cosmetic composition applied to the target skin. The intended cosmetic composition is further described below.

[0165] In some embodiments, the extracts of the present invention can be directly incorporated into cosmetic compositions.

[0166] In other embodiments, the extract of the present invention is preformed as a cosmetic active ingredient before being incorporated into a cosmetic composition.

[0167] The active ingredients of the target cosmetic are further described below.

[0168] The aqueous extract of the present invention, preferably an aqueous extract of hyacinth bulb, is typically present in cosmetic compositions in an amount of 0.005% to 10.0% by weight, preferably 0.01% to 5.0% by weight, for example, 0.5% to 2.0% by weight.

[0169] Another object of the present invention is a cosmetic method for providing one or more cosmetic effects as described herein, the method comprising applying an extract according to the invention, particularly an aqueous extract of hyacinth bulb, to the skin of a subject.

[0170] The extract is preferably applied in the form of the cosmetic composition described herein.

[0171] The cosmetic method of the present invention is preferably used to improve skin tactile perception and / or reduce skin discomfort, and / or improve the appearance of healthy and youthful skin.

[0172] In one specific aspect, the makeup method of the present invention can achieve one or more of the following effects on the object:

[0173] To prevent, treat, or reduce irregular skin pigmentation, including pigmentation spots caused by fatigue or stress.

[0174] Reduce signs of aging, including wrinkles and loss of tissue elasticity.

[0175] To make facial skin tone even or uniform.

[0176] To make facial skin brighter and / or more radiant and / or clearer.

[0177] To prevent or treat dull skin tone, uneven skin tone, and / or signs of a tired appearance.

[0178] It makes the skin, especially the face, feel fresher, more relaxed, and brighter.

[0179] Soothe facial features,

[0180] Makes skin look less tired and more relaxed.

[0181] It revitalizes the skin, especially the face.

[0182] Promotes healthy, radiant skin.

[0183] To prevent, treat, or reduce the appearance of redness.

[0184] To prevent, treat, or alleviate skin discomfort.

[0185] Promotes pleasurable skin sensations in the subject, and / or

[0186] Promote a sense of health among the individuals.

[0187] As mentioned above, the preferred subjects are adults with sensitive and / or mature skin.

[0188] In the cosmetic methods and uses described herein, the dosage to be applied and the frequency of application according to the invention vary depending on the desired cosmetic effect and the individual's characteristics (particularly their sex, age, and skin type). Generally, the extract of the invention or the cosmetic composition containing it can be applied once or twice daily to the skin area to be treated, for example, after skin cleansing, for several weeks or even months, for example, for at least 3 months. For example, an object can apply a dose of 1 g to 2 g of the cosmetic composition to its face in the morning and / or evening. Preferably, the extract or cosmetic composition of the invention is applied to the skin by massage. Application by massage can last from a few seconds to several minutes, for example, from 1 minute to 30 minutes.

[0189] Without being bound by any theory, the applicant believes that the results shown in Example 5 for skin areas (e.g., face and arms) can be extrapolated to any skin area, including the scalp and intimate skin areas.

[0190] Therefore, the present invention also relates to the use of aqueous extracts obtained from hyacinth bulbs (such as those described herein) as active cosmetic ingredients in hair care and feminine care products.

[0191] In one specific aspect, the present invention relates to the use of an aqueous extract from hyacinth bulbs (such as that described herein) as a neurosensory agent for the scalp. In one specific embodiment, the aqueous extract from hyacinth bulbs (such as that described herein) can be used to rebalance or normalize the scalp's neurosensory perception of external stimuli, particularly those causing nociceptive sensations. In some embodiments, the present invention relates to the use of an aqueous extract from hyacinth bulbs for relieving scalp discomfort and / or improving scalp comfort.

[0192] Unbound by any theoretical framework, the applicant believes that rebalancing scalp neurosensory perception (particularly by reducing nociceptive sensitivity) can improve scalp comfort. Furthermore, activation of the OXT signaling pathway may lead to cell proliferation in the scalp, and thereby potentially enhance hair renewal and hair growth.

[0193] Accordingly, the present invention also relates to the use of aqueous extracts obtained from hyacinth bulbs (such as those described herein) as agents to promote hair growth and / or renewal. The extracts of the present invention can be applied directly to the scalp or incorporated into any hair care product (e.g., shampoo, mask, conditioner, spray, serum, foam, etc.).

[0194] In one particular aspect, the present invention relates to methods for rebalancing or normalizing scalp neurosensory perception and / or for alleviating scalp discomfort and / or for promoting hair growth and / or hair renewal, wherein an aqueous extract obtained from hyacinth bulbs (e.g., as described herein) is applied topically to the scalp.

[0195] Generally, the extracts of the present invention or cosmetic compositions containing them (e.g., shampoos, films, or conditioners) can be applied to the scalp at least once a week for several weeks or even months, for example, for at least 3 months. The extracts or cosmetic compositions of the present invention can also be applied to the scalp by massage. Massage application can last from a few seconds to several minutes, for example, 1 to 5 minutes.

[0196] Unbound by any theoretical constraints, the applicant believes that the extract of the present invention could be beneficial as a neurosensory agent for intimate skin areas, for example, in women experiencing hormonal changes, such as perimenopausal and postmenopausal women. For instance, sensory skin perception in intimate areas may change with age and / or hormonal changes. Increased discomfort, nociceptive sensations, and / or decreased sensitivity to emotional touch can be observed in intimate skin areas. Furthermore, hair removal in the intimate area may also alter skin perception.

[0197] Therefore, in one specific aspect, the present invention relates to the use of an aqueous extract obtained from hyacinth bulbs (such as those described herein) as a neurosensory agent for use in intimate skin areas. The aqueous extract of the present invention can be incorporated into any type of intimate care product (e.g., intimate hygiene products, intimate creams, intimate gels, etc.).

[0198] In one specific aspect, the extracts of the present invention are used to reduce intimate discomfort and / or increase skin responsiveness (e.g., to pleasurable stimulation) and / or reduce sensitivity to noxious sensations in the intimate area. In some embodiments, the extracts of the present invention are used to reduce uncomfortable skin sensations (e.g., tingling, itching, numbness, tightness, or dryness) in the intimate area, particularly in the external genitalia, buttocks, and / or perineum.

[0199] In one specific aspect, the present invention relates to the use of an aqueous extract from hyacinth bulbs (such as that described herein) as an agent for enhancing sensitivity to emotional touch and pleasurable sensations in intimate skin areas. In another specific aspect, the aqueous extract from hyacinth bulbs (such as that described herein) can be used as a topical agent to enhance intimate well-being and / or enhance sexual sensitivity.

[0200] In one particular aspect, the present invention relates to a method for rebalancing or normalizing neurosensory perception of the skin in a genital area, for example for reducing genital discomfort and / or increasing skin comfort in the genital area and / or increasing sensitivity to emotional touch and pleasurable sensations in the genital skin area, wherein an aqueous extract obtained from hyacinth bulbs (e.g., as described herein) is applied topically to the genital area, such as the external genitalia, buttocks, and / or perineum.

[0201] Generally speaking, the extracts of the present invention can be used routinely, such as daily use, as a feminine hygiene product or daily feminine care product, or before or during sexual intercourse, for example by application in the form of a cream or gel through massage.

[0202] Cosmetic active ingredients and cosmetic compositions according to the present invention

[0203] In one specific aspect, the present invention relates to a cosmetic ingredient comprising (i) an extract of the present invention, preferably an aqueous extract of hyacinth bulb, and (ii) a cosmetically acceptable carrier as described above. For example, the carrier may be selected from triethyl citrate, C2... C5 alkyl diols (e.g., propylene glycol, butanediol, pentanediol and their isomers), ketal esters (e.g., methyl levulinate or ethyl levulinate), glycerol, water, and mixtures thereof). A preferred carrier is glycerol or a glycerol / water mixture. The glycerol / water weight ratio is generally from 0.25 to 4.0, for example from 0.5 to 2.0 or from 0.75 to 1.25.

[0204] In some embodiments, the cosmetic ingredients of the present invention further comprise one or more cosmetically acceptable excipients, preferably selected from antioxidants, preservatives, pH adjusters, and combinations thereof. The antioxidants, preservatives, and pH adjusters may be as described above. The one or more cosmetically acceptable excipients comprise from 0.01% to 10.00% by weight of the total weight of the cosmetic ingredients of the present invention, preferably from 0.05% to 1.00% by weight.

[0205] In one specific embodiment, the cosmetic ingredient comprises (i) an aqueous extract of the present invention as a cosmetic active ingredient, (ii) glycerol / water as a cosmetically acceptable support, (iii) citric acid as a pH adjuster, and (iv) benzoate and / or sorbate as a preservative.

[0206] In another aspect, the cosmetic ingredients of the present invention consist of or are substantially composed of: (i) an aqueous extract of the present invention as a cosmetic active ingredient, (ii) citric acid as a pH adjuster, and (iii) benzoate and / or sorbate as preservatives, in a glycerol / water mixture as a cosmetically acceptable carrier.

[0207] In some embodiments, the active ingredient of the present invention may comprise one or more additional cosmetic surfactants in addition to the extract of the present invention. Preferred additional cosmetic surfactants are further described below. For example, preferred additional cosmetic surfactants include: Rosa rugosa extract, for example under the brand name IBR. Chill™ is sold; fat-soluble extracts of lavender hybrids, such as those sold under the trade name Immunight™; fat-soluble extracts of Melaleuca alternifolia, such as those sold under the trade name Regenight™; quinoa starch and its derivatives, such as those sold under the trade name Pickmulse™; and combinations thereof.

[0208] In some embodiments, the extract of the present invention, preferably an aqueous extract of hyacinth bulbs, is the only cosmetic active agent present in cosmetic ingredients.

[0209] The cosmetic ingredients of the present invention are typically incorporated into the cosmetic composition in an amount of 0.1% to 10% by weight, for example, 0.5% to 5% by weight, of the total weight of the cosmetic composition.

[0210] In another aspect, the invention also relates to cosmetic compositions comprising extracts of the invention or cosmetic ingredients of the invention, as described herein.

[0211] Generally speaking, the cosmetic compositions of the present invention comprise (i) the extracts of the present invention, particularly an aqueous extract of hyacinth bulbs, and (ii) one or more cosmetically acceptable excipients.

[0212] Cosmetic-acceptable excipients can be selected from standard excipients based on the cosmetic composition to be formulated.

[0213] Cosmetic excipients acceptable include, but are not limited to: diluents, dispersants, gelling agents, emollients, carriers (e.g., polycationic polymers, phospholipids, liposomes, layered systems, lipid or polymer vesicles, nanospheres, micron or nanoparticles of natural or non-natural polymers, hydrogels), gums, resins, solvents, especially water and C2... C6 alcohols or polyols, including ethanol, isopropanol, dipropylene glycol, butylene glycol, propylene glycol, glycerin, sorbitol, propylene glycol and combinations thereof, fillers (e.g., modified and polymerized starch), preservatives, pearlescent agents, odor absorbers, pH adjusters, lubricants, thickeners, surfactants, including anionic surfactants, cationic surfactants, amphoteric surfactants or nonionic surfactants, humectants, wetting agents, stabilizers, bulking agents, dispersants, fragrances, dyes, organic or mineral pigments (e.g., iron oxide), oiling agents (e.g., vegetable or animal oils or fats, such as triglycerides, fatty alcohols, fatty acids or their salts), synthetic oils (e.g., paraffin or isoparaffin), silicone oils (cyclomethyl silicone oils), fatty alcohol esters, fluorinated oils, waxes and / or other substances commonly used in cosmetic formulations.

[0214] In one specific embodiment, the cosmetic composition comprises 0.005% to 10.0% by weight, preferably 0.01% to 0.5% by weight, such as 0.05% to 2.0% by weight of the aqueous hyacinth bulb extract described herein, where percentages refer to the total weight of the cosmetic composition.

[0215] More specifically, the cosmetic composition may comprise:

[0216] From 0.005% by weight to 10.0% by weight of the aqueous hyacinth bulb extract described herein, particularly the aqueous hyacinth bulb extract, and

[0217] 70% to 99.995% by weight of one or more cosmetically acceptable excipients.

[0218] In some embodiments, the cosmetic composition further comprises one or more active agents having additional cosmetic effects. The one or more active agents may comprise up to 29.995% by weight of the total composition. When present, one or more additional cosmetic active ingredients comprise from 0.0001% to 29.995% by weight of the total cosmetic composition.

[0219] The term "active agent or cosmetic active ingredient with cosmetic effect" refers to an agent capable of exerting at least one cosmetic effect on the skin or its appendages. "Cosmetic effect" means any non-therapeutic effect intended to alter and / or improve the visual appearance and mechanical properties of skin or mucous membranes (e.g., lips), protect them from external aggressors (sunlight, wind, moisture, dryness, chemicals), prevent and / or correct aging-related phenomena, or prevent or treat the effects of stress or fatigue on the skin.

[0220] Examples of such agents include, in particular, anti-wrinkle agents, anti-aging agents, antioxidants, moisturizers, soothing agents, anti-redness agents, exfoliants, sebum regulators, brightening agents, concealers, anti-dark spot agents, anti-pollution agents, anti-stress agents, sunscreens, and combinations thereof.

[0221] In one specific embodiment, the cosmetic composition of the present invention may contain one or more additional cosmetic active agents selected from anti-wrinkle agents, anti-redness agents, antioxidants, moisturizing active agents, soothing agents, sebum regulators, brightening agents, anti-stress agents, anti-fatigue agents, anti-pollution active agents, concealing agents, sunscreens, and combinations thereof.

[0222] Some examples of antifouling agents include extracts of Chrysanthellum Indicum polysaccharides, particularly marine extracts from Alteromonas fermentation media, and Nigari salts.

[0223] Some examples of concealers include extracts of golden chamomile or algae (including Ascophyllum nodosum or Asparagopsis Armata) and sulfated polysaccharides.

[0224] Some examples of humectants include urea, pidolic acid (PCA) and its derivatives (especially its salts, such as arginine PCA, chitosan PCA, its copper salt (copper PCA), magnesium (magnesium PCA), sodium (sodium PCA) or zinc, ethylhexyl PCA), calcium gluconate, hyaluronic acid and its salts and other glycosaminoglycans, trehalose, polydextrose, sucrose, maltitol, mannitol, sorbitol, xylitol and other sugars and their derivatives, and polyethylene glycol (e.g., PEG). 7. PEG 8. PEG 10. PEG 12 or PEG 14) Glycerin, propylene glycol, butylene glycol, betaine, citrulline, collagen and its derivatives, histidine, serine, keratin or soybean hydrolysate, plant extracts rich in polysaccharides and / or polyphenols (e.g., aloe vera, blueberry (Centaurea cyanus), burdock extract), allantoin, extracts rich in polysaccharides, especially extracts from fermentation media of marine microorganisms (e.g., Porphyridium or Porphyridium), and combinations thereof.

[0225] Some examples of anti-aging agents include ascorbic acid and its derivatives (e.g., magnesium ascorbate phosphate), glycosaminoglycans and their derivatives (e.g., hyaluronic acid), retinoids (e.g., retinol), Cyathea polysaccharides, collagen, flaxseed extract (Linum usitatissimum), and peptides (e.g., hexanoyl tetrapeptide). 3 and trifluoroacetyl tripeptide 2) Extracts of Polygonum aviculare, extracts of brown algae, especially Ascophyllum nodosum and Cyathea cumingii, and extracts under the trade name Skinectura. ™ Extracts from the paws of the tall kangaroo (Anigozanthos flavidus) are sold.

[0226] Some examples of anti-stress agents include rose extract, such as Rosality. ™ It is a combination of rose water and rose essential oil, as well as pink rock rose extract, for example, under the brand name IBR. Chill ™ Sale.

[0227] Some examples of anti-fatigue agents include fat-soluble extracts of lavender hybrids and Melaleuca alternifolia, marketed under the trade names Immunight™ and Regenight™.

[0228] Some examples of soothing agents include allantoin, aloe vera, birch (e.g., white birch (Betula alba)), willowherb (e.g., willowherb (Epilobium angustifolium)), chestnut (e.g., European chestnut (Castenea sativa)), blueberry (e.g., cornflower (Centaurea cyanus)), centella (e.g., centella asiatica)), horsetail (e.g., horsetail (Equisetum arvense)), fennel (e.g., fennel (Foeniculum vulgare)), witch hazel (e.g., North American witch hazel (Hamamelis virginiana)), ivy (e.g., American ivy (Hedera helix)), hibiscus (Hibiscus sabdariffa), lily (e.g., Madonna lily (Lilium candidum)), mallow (e.g., European mallow (Malvasylvestris)), and lemon balm (e.g., lemon balm (Melissa)). Extracts of *Scutellaria baicalensis*, *Mimosa tenuiflora*, and *Potentilla erecta*, oligosaccharide extracts or oligosaccharides (e.g., flaxseed), peptides (e.g., palmitoyl tripeptide) from *Scutellaria baicalensis*, *Mimosa tenuiflora*, and *Potentilla erecta*. 8) Extracts of polysaccharides, especially extracts of extracellular polysaccharides from fermentation media of Alternaria alternifolia or Porphyromonas spp.

[0229] Some examples of antioxidants include HMR (hydroxymethylresorcinol), ascorbic acid and its derivatives, vitamin B9, histidine hydrochloride or Epilobium augustifolium extract, tocopherol, carotenoids and carotenoid-containing extracts (such as Dunaliella salina extract).

[0230] Some examples of sebum regulators include linoleans, rice flour, zinc gluconate, sarcosine, extracts of Cinnamomum zeylanicum bark, avocado extracts, extracts of Backhousia citriodora leaves, and combinations thereof.

[0231] Some examples of anti-redness agents include saponins, flavonoids, ruscogenins, esculoside and extracts containing them, such as extracts of Ruscus, as well as certain essential oils, such as lavender or rosemary.

[0232] Some examples of anti-spot agents include extracts such as licorice (Glycyrrhyza glabra), jackfruit extract (Artocarpus heterophyllus), sorrel extract (R. occidentalis), extracts from plants belonging to the Citrus genus, resveratrol, and peptides (such as oligopeptides). 68. Nonapeptide 1) Kojic acid, magnesium ascorbate phosphate and their combinations.

[0233] Other preferred cosmetic agents include: rosehip extract, for example, under the brand name IBR Chill™ is sold; fat-soluble extracts of hybrid lavender species, such as those sold under the trade name Immunight™; fat-soluble extracts of Melaleuca alternifolia, such as those sold under the trade name Regenight™; quinoa starch and its derivatives, such as those sold under the trade name Pickmulse™; and combinations thereof.

[0234] Needless to say, the additional active ingredients and excipients used depend on the type of composition to be formulated and the skin area to which the composition will be applied.

[0235] The cosmetic compositions of the present invention are generally applied to the skin. The cosmetic compositions can be any form suitable for such topical application. In one specific embodiment, the cosmetic compositions of the present invention are selected from aqueous solutions, hydroalcoholic solutions, oil-in-water emulsions (O / W) or water-in-oil emulsions (W / O) or multiple emulsions, including nanoemulsions, aqueous gels (also called hydrogels), latexes, suspensions, preferably in aqueous or hydroalcoholic media, liposome suspensions, oily compositions, serums, powders, lotions, emulsions, creams, ointments, gels, foams, balms, and aerosols.

[0236] In some embodiments, the cosmetic composition according to the invention is selected from creams, balms, serums, or lotions.

[0237] More generally, the compositions according to the invention can also be in the form of any cosmetic or skin care product. It can be a skin care product, makeup product, or body hygiene product, such as a lotion, emulsion, serum, aqueous or oily gel, emulsion, cream, or gel. Creams, ointments, balms, foundations, sprays, mascaras, stick formulations, lipsticks, lip glosses, deodorants, moisturizing masks, shower gels, aftershaves, cleansing oils, soaps, shampoos or conditioners, hair masks or hair oils (including massage oils). In some preferred embodiments, the compositions of the present invention are skin care products, such as creams, balms, serums, gel creams, or ointments.

[0238] In other embodiments, the compositions of the present invention are hair care products, such as shampoos, conditioners, hair masks or balms, serums, hair oils or hair forms, and this list is not exhaustive.

[0239] In other embodiments, the compositions of the present invention are intimate care products, such as intimate hygiene (cleaning) care products (gels, soaps), intimate wipes, or intimate creams or gels.

[0240] Other aspects and advantages of the invention will become apparent from the following embodiments, which should be considered illustrative rather than limiting.

[0241] Example

[0242] Example 1: Preparation of extracts and cosmetic ingredients according to the present invention

[0243] Preparation and characterization of extracts

[0244] The aqueous extract of hyacinth bulbs was prepared as follows:

[0245] Fresh, dormant hyacinth bulbs are peeled. Water is added to the peeled bulbs, and the mixture is processed into a slurry (water / bulb weight ratio: about 0.5 to 1.0). The resulting slurry is allowed to stand for at least 20 minutes with intermittent stirring. The slurry is then centrifuged. The supernatant is recovered and heated at a temperature of about 100°C to 125°C for at least 30 minutes. The supernatant is cooled at room temperature to form a precipitate, which is then centrifuged. The supernatant is heated at a temperature of about 100°C to 125°C for at least 10 minutes and then cooled at room temperature. After cooling, the mixture is centrifuged and filtered down to 0.45 μm, thereby obtaining the aqueous extract of the present invention.

[0246] The dry matter content of the obtained extract is approximately 45 mg / g to 75 mg / g.

[0247] The resulting extract is directly formulated to obtain the cosmetic active ingredient according to the present invention.

[0248] Chemical characterization of the extract of the present invention

[0249] The compositional characterization of the obtained extract is as follows:

[0250] The total carbohydrate content (including sucrose and fructose) is approximately 1% to 2%, as measured by GC. FID was evaluated by preliminary acid hydrolysis and derivatization.

[0251] Iminosaccharides (such as takanojirimycin, deoxynojirimycin, deoxymannojirimycin, etc.) are present at concentrations ranging from approximately 60 ppm to 1200 ppm. For example, by using 1... HPLC analysis of deoxynojirimycin as a standard UV quantification (according to Rodríguez) Sánchez, S.; Quintanilla López, JE; Soria, AC; Sanz, MLEvaluation of different hydrophilic stationary phases for the simultaneous determination of iminosugars and other low molecular weight carbohydrates invegetable extracts by liquid chromatography tandem mass spectrometry, J.Chromatography A, 2014, 1372, 81 90.

[0252] The amino acid content (including aspartic acid and glutamic acid) is 0.05% to 1%, as quantified by ninhydrin spectrophotometry using aspartic acid as a standard (according to Michel, MC Dosage des acides amines etamines par la ninhydrin. Amelioration pratique. Ann. Biol. Anim. Bioch. Biophys. 1968, 8(4), 557). 563).

[0253] Color: Transparent to translucent, colorless to yellow.

[0254] pH: 5.0 to 6.5.

[0255] Preparation of cosmetic active ingredients of the present invention

[0256] The aqueous extract obtained as described above was formulated with glycerin and preservatives (potassium sorbate, sodium benzoate). The pH was adjusted to approximately pH 5.5 (using citric acid).

[0257] The composition (by weight %) of the active ingredients in the cosmetic of this invention:

[0258] The aqueous extract of this invention: 49%

[0259] Glycerin: 50%

[0260] Potassium sorbate, sodium benzoate, and citric acid: 1.0%

[0261] pH: 5.8.

[0262] Example 2: The extract of the present invention activates oxytocin receptors

[0263] Protocol: This study used CHO inhibitors expressing the human oxytocin receptor OXTR (accession number CAA46097.1). K1 (hamster ovary) cells. IP was used at 1 hour, i.e., at equilibrium. One HTRF assay evaluates activity (monitoring downstream products of target receptor activation; in this case, it detects the accumulation of inositol monophosphate, which is a stable downstream metabolite of IP3 induced by activation of the phospholipase C (PLC) cascade).

[0264] The extract of the present invention (prepared as in Example 1) was tested at four concentrations (0.025%, 0.05%, 0.1%, and 0.2%), where n=2. Oxytocin (Bachem H) 2510, Bubendorf, Switzerland) was used as a positive control (10 concentrations, from 0.01 nM to 1000 nM).

[0265] Results: The extract of the present invention showed activation of OXTR at concentrations of 0.1% or 0.2% by weight. At a concentration of 0.1% extract, receptor activation was 53.14%; at 0.2%, activation was 71.49%. Figure 1 ).

[0266] Example 3: Effects of the extract of the present invention on co-culture of human sensory neurons and human skin keratinocytes under capsaicin attack.

[0267] Materials and Methods:

[0268] a) Co-culture of human sensory neurons and keratinocytes

[0269] Human sensory neurons originate from hiPS cells (human induced pluripotent stem cells), which are generated by the use of cells encoding KLF4, OCT4, SOX2, and c. Human fibroblasts (foreskin fibroblasts, Coriell Institute AG08498, Camden, NJ, USA; cells obtained from neonatal circumcision remnants at the Coriell Institute, fully compliant with local (NJ, USA) guidelines and ethical standards, which do not require specific ethical review or approval for such cases) were obtained by transfecting the MYC gene with a lentiviral agent. Cell plates were seeded in 96-well plates coated with a thin layer of Matrigel (Corning 354277, New York, NY, USA) and placed in differentiation medium (DMEM / F12, Panbiotech P04). 41450, Aidenbach, Germany + Knockout™ serum substitute synthetic serum, Fisher Scientific 10828010, Waltham, MA, USA + SU 5402 Fibroblast growth factor receptor inhibitor, Sigma Aldrich SML0443, St Louis, MO, USA + LDN193189 ALK2 / 3 hydrochloride (type I receptor serine) The medium was incubated with a threonine kinase inhibitor (Sigma Aldrich SML0559 + retinoic acid, Sigma Aldrich R4643) at 37°C and 5% CO2 for 6 days (with the medium changed every 2 days). After 9 days of differentiation, the medium was changed to human sensory neuron maturation medium (DMEM / F12 + N). 2 supplements, Fisher Scientific 17502048 + PS (penicillin / streptomycin), Panbiotech P06 07100 + NGF (nerve growth factor), Sigma Aldrich N1408). Incubate cells at 37°C and 5% CO2. Change the medium every 2 to 3 days.

[0270] Adult keratinocytes (PromoCell C12013, Heidelberg, Germany) (from a donor approximately 30 years old) were cultured in a keratinocyte growth medium (PromoCell C12013). After one cycle of amplification in 20111), the keratinocytes were then dissociated using trypsin and frozen. These keratinocytes were then thawed and amplified again in a keratinocyte growth medium, followed by trypsin dissociation. Cell viability was determined by cell counting.

[0271] After 14 days of culture, keratinocytes were combined with the aforementioned sensory neurons at an initial ratio of 5:1 (keratinocytes / neurons) by first seeding hiPS differentiated cells, followed by seeding keratinocytes on top of the hiPS layer in the same culture well. The co-culture was maintained at 37°C and 5% CO2 in a medium consisting of a mixture of neuron maturation medium and keratinocyte growth medium. The medium was changed every 2 to 3 days.

[0272] b) Co-culture treatment for evaluating the activity of the extract against nociceptive responses

[0273] Sensory neurons and keratinocytes were co-cultured as described above. On day 17 of culture, the culture medium was removed and replaced with fresh medium. After 2 hours of incubation, the culture medium was removed and replaced with fresh medium containing 0.1% of the extract of this invention, in addition to: an untreated control (culture medium only); a positive control (10 μM oxytocin); and a secondary control (10 μM capsaicin, Sigma C191). 5 mg – capsaicin antagonist. After 24 hours of incubation, cells were activated for 30 minutes with 10 μM capsaicin (Sigma M2028) in parallel with the unstimulated control. The culture medium was then recovered for ELISA quantification of CGRP and replaced with capsaicin-free medium.

[0274] After an additional 24 hours of incubation (in the absence of capsaicin), the supernatant was collected and stored. At 80°C, cells were fixed in 2% paraformaldehyde solution (Fisher Scientific J19943, Waltham, MA, USA) and stored at +4°C. One culture was performed with 6 wells for each condition.

[0275] c) CGRP ELISA quantification

[0276] The culture supernatant was recovered 30 minutes after cell activation and thawed. The amount of CGRP released was quantified by ELISA (Antibodies online, Aachen, Germany, ABIN2542823; Multiskan FC plate reader, Thermofisher Scientific, Waltham, MA, USA).

[0277] Statistical analysis was performed using one-way ANOVA tests, including Brownian analysis. The Forsythe test and Bartlett test were used, with the multiple comparison test option to compare all means, Dunnett correction was applied, and the significance threshold for all tests was p < 0.05.

[0278] d) Immunofluorescence staining (assessment of neuronal plasticity):

[0279] Cells were combined with an anti-β antibody Tubulin (Abcam ab41489, Cambridge, UK), anti-OXTR (Abcamab300443) 1004) and antisynaptoxin (Abcam AB8049) The antibodies were incubated together with 1001. These antibodies were visualized via secondary antibodies conjugated with fluorescent dyes (Fisher Scientific A21449, A11011, and A11001). Twenty images per well were captured using an InCell 2200 automated microscope (GE Healthcare, Piscataway, NJ, USA). Images were analyzed using the INCell Developer toolbox software for automated detection of cell bodies, including neurons, neurites, and receptor expression. The number of sensory neurons, their neurite length, and their dendritic density (via β-ray diffraction) were measured. (Microtubule staining). The expression rates of OXTR receptor and synaptophysin were also determined.

[0280] Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparison test (using Prism 8.4.3, GraphPad, Boston, MA, USA), with p < 0.05 as the significance threshold.

[0281] result:

[0282] The effect of the extract of this invention on in vitro nociceptive signal transduction

[0283] The result is Figure 2 As shown in the figure. The extract of the present invention shows reduced nociceptive signaling in a co-culture of neurons derived from human keratinocytes and iPSCs, said co-cultures attacked with capsaicin (a widely accepted model of nociceptive stimulation that produces discomfort such as stinging, burning or even pain).

[0284] Neuronal output (representing the signal generated by a stimulus) is evaluated by measuring CGRP release.

[0285] In the presence of 0.1% extract, nociceptive signals were significantly reduced (compared to the control group stimulated by capsaicin). (22%, p = 0.0005), restored signal transduction to slightly lower values ​​than the untreated, unattacked control, and showed a slightly stronger effect than the 10 μM OXT or 10 μM capsaicin (capsaicin antagonist) control group.

[0286] Effects of the extract on neuronal survival and dendrite density of neurons co-cultured with keratinocytes after capsaicin attack.

[0287] The result is Figure 3A and Figure 3B As shown in the image.

[0288] In the same co-culture model described above, the extract of the present invention also provides the following benefits:

[0289] Under capsaicin challenge, treatment with 0.1% extract significantly improved neuronal survival: +44% compared to untreated and unattacked controls (p<0.01); +89% compared to capsaicin challenge and untreated controls (p<0.01); and slightly better than the reference group treated with capsaicin and 10 μM oxytocin (+20%, p = 0.2).

[0290] The extract of this invention improves the dendritic density of neurons under capsaicin challenge: treatment with 0.1% extract resulted in a +25% improvement compared to untreated or unchallenged controls (p<0.05). A positive trend (+12%, p<0.20) was observed in cultures when treated with 0.1% extract under capsaicin challenge compared to untreated or unchallenged controls.

[0291] Therefore, the extract of the present invention provides protection against capsaicin attack: in co-culture with keratinocytes under capsaicin, it increases neuronal survival and dendrite density to a level similar to or even higher than that of 10 µM oxytocin.

[0292] Example 4: Organs and Nerves on a Chip Skin interface model

[0293] This study was conducted with two consecutive identical runs.

[0294] Cultivation Chip: This study utilized the Netri (Lyon, France) Neurofluidics™ MEA organ culture chip, which consists of two parallel channels. Neurons were seeded in the left channel, while keratinocytes were seeded in the right channel. These two channels were connected by two hundred 125×6×3 μm microchannels (length×width×height) spaced 20 μm apart (large enough to allow axons to grow from one compartment to the next, but not to allow cell migration). This layout was covered with 42 electrodes (15 in each cell compartment, 10 in the microchannel region, and 2 central reference electrodes) to measure electroactivity.

[0295] Culture chip using 0.1 mg / mL poly D The sample was coated with lysine (Thermofisher A38904, Waltham, MA, USA) and incubated overnight at 37°C, then washed three times with PBS. Finally, it was coated with SureBond. The chip was coated with XF solution (Axol Bioscience ax0053, Easter Bush, UK), diluted 200-fold, and incubated at 37°C for 4 hours. SureBond was then removed from the chip. XF solution was used, and then cells were inoculated.

[0296] Thawed AxoCells sensory neurons (Axol Bioscience ax0055) were seeded in neuronal compartments at a density of 2000 cells / mm². On days 1 and 3, the culture medium was completely replaced with 0.25% 1:200 Surebond supplementation. XF enhanced sensory neuron maintenance medium; on day 3, 75% of the medium was replaced with enhanced sensory neuron maintenance medium supplemented with 2.5 μg / mL mitomycin C (Sigma M4287, St Louis, MO, USA); on day 6, 75% of the medium was replaced with 0.25% 1:200 Surebond supplemented. XF enhanced sensory neuron maintenance medium; on days 9, 13, 16, 20, and 23, 50% of the medium was replaced with enhanced sensory neuron maintenance medium. The enhanced sensory neuron maintenance medium consisted of: sensory neuron maintenance medium (Axol Bioscience ax0060) + 1% sensory maturation maximizer (Axol Bioscience ax0058) + 25 ng / mL recombinant human glial cell-derived neurotrophic factor (Axol Bioscience ax139855) + 25 ng / mL recombinant human nerve growth factor (Axol Bioscience ax139789) + 10 ng / mL recombinant human brain-derived neurotrophic factor (Axol Bioscience ax139800) + 10 ng / mL recombinant human neurotrophic protein. 3 (Axol Bioscience ax139811).

[0297] On day 20, keratinocytes (PromoCell C12005, Heidelberg, Germany) were seeded at 25,000 cells / channel. The culture medium was completely replaced at 24 hours, and subsequently three times a week. The culture medium consisted of a keratinocyte supplement mixture supplemented with 2.5% keratinocytes (Promocell C12005). 39016) and 0.012% of 0.5 M calcium chloride (Promocell C) 34005) keratinocyte growth medium (Promocell C) Composed of 20011B).

[0298] Incubate at 37°C and 5% CO2.

[0299] Processing and measurement of electrical signal transmission: Treatments were applied on day 21, in four groups of four chips each, with each group treated with one of the following: 10 μM capsaicin; 10 μM oxytocin; or 0.1% or 0.2% HO extract. This was performed simultaneously with recording the immediate effects of the treatments on electrical signal transduction using a Maestro Pro (Axion Biosystems, Atlanta, GA, USA). After a 3-minute baseline recording, 20 μL of the appropriate sample vehicle was added through the keratinocyte channel inlet. Electrical activity was recorded for another 3 minutes, followed by removal of the culture medium from the keratinocyte channel inlet and the addition of the treatment (simulating real-world exposure through the stratum corneum and viable epidermis) in 20 μL of the keratinocyte channel inlet. After a further 55 minutes of incubation, electrical signal transduction was recorded over 10 minutes.

[0300] Recordings were extracted from AxIS Navigator 3.7.4 software (AxionBiosystems) using spike count output, in 3-second bins. For each chip, all data points were normalized using the mean firing rate (MFR) over the first three minutes of the fast-action recording. The activity fold variation was obtained by normalizing the MFR over the entire long-term recording length.

[0301] After checking for normality and homogeneity of variance, one-way ANOVA with Sidak post-hoc test was used to assess the effect; for non-normal data, Kurskal-Wallis analysis with Dun post-hoc test was used instead. Outliers were excluded.

[0302] Immunofluorescence staining of DAPI (nuclear marker) and peripheral proteins (axonal marker): Once electroactivity recording is complete, remove the cell culture medium and use D... Wash cell culture channels with PBS. Add D. A 4% formaldehyde solution (diluted from Thermofisher 28908) in PBS was used, followed by incubation at room temperature for 30 minutes. Then, D... The channel was washed three times with PBS. 0.1% Triton X100 (Sigma Aldrich X100) permeabilization solution was added, followed by incubation at room temperature for 10 minutes and then... Wash three times with PBS. Add 1% BSA blocking solution (Thermofisher 14190250), then incubate at room temperature for 30 minutes and remove the blocking solution. Add primary antibody solution (anti-PRPH (rabbit), diluted 1 / 50, Abcam ab4666, Cambridge, UK) to the neuronal compartment, then incubate overnight at 4°C. Use D... After washing three times with PBS, add secondary antibody solution (anti-rabbit AF647, diluted 1 / 1000, Abcam ab150075) in the dark, then incubate at room temperature in the dark for 2 hours, and use D... Wash three times with PBS. Finally, add 3 μM DAPI solution (Sigma D8417), then incubate at room temperature for 10 minutes and wash with D... Wash three times with PBS.

[0303] Images were acquired using an Axio observer 7 microscope (Zeiss, Oberkochen, Germany). Images were reconstructed using instrument software (Zeiss Zen 3.3.89).

[0304] Image analysis:

[0305] Image analysis is performed using Fiji and Cellpose, as implemented in the BIOP Fiji plugin.

[0306] Neuron nucleus segmentation: Noise correction was also performed on the original DAPI images of the neuronal compartments using a 2D Gaussian blur filter (1×1 kernel value). This was done to reduce noise in the DAPI images. To mitigate the blurring effect observed in neuron staining and improve kernel segmentation, a Fast Fourier Transform (FFT) containing only high-frequency information (corresponding to the kernel staining) was applied. Unsharpening masking (radius=15, mask=0.9) was performed, a blurred copy of the image was subtracted, and DAPI was sharpened. Nuclear staining. Automatic nuclear segmentation was performed using the stardist model (as implemented in Fiji), and the corresponding output segmentation map was used to calculate the total number of neurons in the neuronal compartment.

[0307] Quantitative analysis of peripheral proteins Peripheral protein staining was measured within neuronal compartments. Gaussian noise correction was applied to the original image using a 2D Gaussian blur filter (1×1 kernel value). Background correction was then performed using a rolling ball algorithm, which determines local background values ​​by averaging all values ​​within a large region of interest (ROI) around each pixel and subtracting this average from the original image. A score for the "tubularity" of each point in the image was then generated using the Tubeness Fiji plugin, allowing for specific focus on peripheral protein staining corresponding to neuronal axons. Finally, a k-means clustering algorithm (k=4) was applied to delimit these neuronal axons within the compartments. The peripheral protein density was quantified using this neuronal axon ROI, defined as the ratio of the area of ​​the neuronal axons within the compartment to the image area.

[0308] Statistical analysis was performed using Prism 10.2.3 (GraphPad, Boston, MA, USA). For parametric tests, one-way ANOVA with post-hoc Dunnett's or Sidak's tests was used; for nonparametric tests, Kruskal's test with post-hoc Dunn's was used. Wallis test. All analyses used a significance threshold of p < 0.05.

[0309] result:

[0310] In on-chip organ models equipped with separate compartments for neurons and keratinocytes, electrical signal transduction data ( Figure 4The results showed that capsaicin slightly increased signal transduction (+19%) compared to baseline, while oxytocin treatment decreased signal transduction (compared to capsaicin). 34%, p = 0.0147). Treatment with 0.1% and 0.2% hyacinth extract reduced the signal transduction activity to a similar degree, with a 41% reduction at 0.1% (p = 0.0036) and a 43% reduction at 0.2% (p = 0.0043) compared to capsaicin treatment.

[0311] Example 5: Clinical Trial

[0312] plan:

[0313] This study employed a double-blind, randomized, placebo-controlled design, with participants divided into two groups (active product and placebo). The product was applied to the face and forearms twice daily for 28 days. To capture immediate, short-term, and longer-term effects, measurements were taken immediately before and after product application on days 0 and 28 (approximately 30 minutes or 1 hour after application), and on day 7.

[0314] Sixty participants were recruited (divided into two groups of 30 each), aged 18 to 65, with both sexes represented (50% in each group); all skin types were included; Fitzpatrick patterns from I to IV were selected. Menopausal status was recorded.

[0315] All evaluations were conducted at one of two time periods (9:00 to 13:00 and 13:00 to 19:00) to minimize diurnal variation, in a climate-controlled room, and after at least 15 minutes of acclimatization (T = 21°C ± 2°C; RH = 55% ± 10%).

[0316] The following assessments were conducted:

[0317] The sensory response of the subjects to mild tactile stimulation of the skin was assessed by electroencephalography (EEG) (using a 32-channel Neuroelectrics Enobio). 32 apparatus (iMotions, Copenhagen, Denmark). Gentle touches using feathers on the forearm were used as model stimuli. Measurements were taken before and after product application at D0, at D7 (without product application), and before and after product application at D28. Power spectral decomposition of the EEG signals was performed using the instrument software as follows: signal preprocessing: bandpass filtering and notch filtering (using a zero-phase hysteresis bandpass Butterworth filter); rereference; artifact removal; fast Fourier transform (FFT).

[0318] The capsaicin sting test was used to assess nociceptive sensation and skin sensitivity, in which the skin was soaked in 3.16 × 10⁻⁶ capsaicin solution. 5% capsaicin solution (Sigma) Cotton pads from Aldrich (1003366230, St Louis, MO, USA) were applied to the nasolabial folds. Stinging sensation was assessed at 2, 5, and 10 minutes using a 7-point scale ranging from 'absent' to 'significant'. The final stinging score was the average of the three assessments. This assessment was conducted on days 0 and 28, each before and after product application.

[0319] The appearance of a visibly healthy, radiant skin was assessed by study participants and trained clinicians using a scale ranging from a score of 0 (no radiance) to a score of 4 (very healthy radiance). Grading was performed on the entire face at D0 and D28 (each time, both before and after product application). Half-scores were allowed to improve resolution.

[0320] A tristimulus colorimeter (CR) is used to measure reflected light. The Minolta, Japan (400) evaluated skin color and represented light as brightness (L) in the three-parameter Hunter color space. ), a (red (green) and b (blue (Yellow). Measurements were taken at D0 and D28 (without product application) in the periorbital area.

[0321] Use band 2 in D0 and D28 (no product applied). The Cutometer® dual MPA 580 (Courage & Khazaka, Germany) with a mm probe evaluates skin elasticity. This method uses negative pressure (suction) to measure the elasticity of the upper skin layer, which mechanically deforms the skin. The depth of tissue penetration into the probe (indicating the degree of skin deformation) is determined by an optical measurement system. Elasticity is evaluated as the skin's ability to return to its original position after deformation.

[0322] Skin wrinkles were assessed using image analysis, employing Visia with PRIMOS. CR (VISIA) (CR, Canfield, USA) Standardized photographic images of participants' faces were taken at D0, D7, and D28 (without product application) using normal, cross-polarized, and parallel-polarized light. The instrument assessed wrinkle counts through bias analysis of the fringe projection images. Additionally, skin wrinkles were graded by trained clinicians at D0 and D28 (before and after each product application) based on the Bazin scale (Bazin, R. and Doublet, E., 2007. Skin aging atlas. Med'com.).

[0323] Self-assessment of skin comfort / pleasure (skin sensation): Skin comfort / pleasure was quantified before and after application of the placebo cream or the cream of the present invention to the skin by self-rating using a visual five-point scale (from very unpleasant (0) to very pleasant (5)). The percentage change is expressed as a comparison with the D0 score before cream application.

[0324] For clinical trial purposes, the ingredients of this invention were formulated at 1% in a cream. In the gel matrix, the corresponding placebo products are listed side by side as detailed in the table below.

[0325]

[0326] result:

[0327] The main results of the clinical trial are shown in Figure 5 to 10. Figure 10 As shown in the figure, it can be summarized as follows:

[0328] EEG during feather touch stimulation: Selective left frontal alpha wave activation was observed in the formulation containing 1% hyacinth bulb extract; the same was not observed with the placebo product. This effect was evident when comparing EEG signal visualizations prior to D0 product application with visualizations recorded on D7 and D28 after D0 product application. This suggests that the extract exerts both short-term and long-term effects and promotes positive mood and well-being in users by enhancing the perception of pleasant skin touch. Figure 5A These results were determined by recording alpha wave activation at electrode AF3 (left frontal lobe) on day 0 (D0), (D0 + 30 minutes after administration), D7, and D28. In the literature, this alpha wave activation is associated with positive mood and well-being and indicates a dominance of positive emotional states. Figure 5BThe invention's 1% component (hyacinth bulb extract) in the formulation enhanced brain activity at AF3, and thus enhanced skin tactile sensitivity 30 minutes after application. This effect was also present on days 7 and 28 compared to placebo. These results can be interpreted as indicating a pleasant or pleasurable sensation during feather touch, which was more pronounced in the cream of the invention than in the placebo cream. It can be concluded that, as evaluated by EEG, the extract of the invention confers both short-term and long-term enhanced sensitivity to mild, pleasurable tactile stimuli (represented by feather touch). Therefore, the extract of the invention can promote positive emotions and well-being in subjects by enhancing pleasurable skin tactile perception. Capsaicin sting test: After 28 days, 1% hyacinth bulb extract in the formulation showed a significant reduction in capsaicin-induced stinging sensation compared to placebo. 24%, p < 0.01). This indicates that the extract reduces skin irritation and confirms the in vitro indications detailed in Example 3 above ( Figure 6 It is worth noting that similar results were obtained for heat-triggered nociceptive sensations (data not shown).

[0329] Self-evaluation of skin comfort / pleasure (skin sensation): Self-evaluation determined that the cream of the present invention significantly improved skin comfort in the short and long term compared with the placebo cream.

[0330] Grading of Healthy Skin Gloss Appearance: Both clinician grading and self-assessment showed a strong and statistically significant advantage over placebo in terms of healthy luster when the formulation contained 1% hyacinth bulb extract. Compared to D0, before product application, D28 showed a +19% increase in clinician grading (p < 0.05) and a +32% increase in self-assessment (p < 0.05); after product application, the increase was 30% (p < 0.01) and 36% (p < 0.05). Figure 7 ).

[0331] Skin color: After 28 days of treatment, compared with placebo, colorimetric data showed skin redness (α) when the formulation contained 1% hyacinth bulb extract. Significantly reduced ( 9%, p < 0.01), which supports the reduction in skin sensitivity. It also showed a small but significant brightening effect (L). (+2%, p < 0.01) Figure 8 ).

[0332] Skin elasticity: Cutometer data at D28 showed that skin elasticity was improved (+13%, p < 0.05) when the formulation contained 1% hyacinth bulb extract compared to placebo. Figure 9 ).

[0333] Skin wrinkles: Image analysis showed that the formulation containing 1% hyacinth bulb extract had an advantage in wrinkle counts compared to placebo at D28 vs. D0. 18% (p < 0.05). This was supported by a clinician grading system that showed that the formulation containing 1% hyacinth bulb extract, compared to placebo, resulted in a lower wrinkle grade before product D28 administration compared to D0. 5% (p < 0.05), and wrinkle grades decreased after application of product D28 compared to D0. 7% (p< 0.05).

Claims

1. The use of an aqueous extract of hyacinth (Hyacinthus orientalis) bulb in cosmetics as a skin neurosensor.

2. The cosmetic use according to claim 1, wherein the extract is obtained from the dormant bulbs of hyacinth.

3. The cosmetic use according to claim 1 or 2, wherein the extract is obtained by a method comprising the following steps: (a) Incubate the peeled and crushed bulbs in water to obtain the sap. (b) Recover the aqueous phase from the slurry obtained in step (a). (c) The aqueous phase obtained in step (b) is heated at a temperature above 95°C and then cooled to promote precipitation. (d) Recover the aqueous phase from step (c), and (e) Optionally filter and / or concentrate the aqueous phase obtained in step (d) to obtain the aqueous extract.

4. The cosmetic use according to any one of claims 1 to 3, wherein the extract is used to improve skin sensation.

5. The cosmetic use according to any one of claims 1 to 4, wherein the extract is used to rebalance or improve the neurosensory response of the skin to stimulation.

6. The cosmetic use according to any one of claims 1 to 5, wherein the extract is used to reduce or prevent skin irritation.

7. The cosmetic use according to any one of claims 1 to 6, wherein the extract is used to enhance the skin’s sensitivity to pleasant and / or mild tactile stimulation.

8. The cosmetic use according to any one of claims 1 to 7, wherein the extract is used to relieve skin discomfort and / or to provide a comfortable effect to the skin.

9. The cosmetic use according to any one of claims 1 to 8, wherein the hyacinth bulb extract is used to prevent or alleviate age-related skin-sensory damage, particularly skin sensitivity.

10. The cosmetic use according to any one of claims 1 to 9, wherein the extract is also used to improve the visual appearance of the skin, particularly to alleviate, reduce or treat one or more non-pathological changes in the appearance of the skin, such as roughness, redness, loss of luster or radiance, dull or uneven skin tone, tired appearance and combinations thereof.

11. The cosmetic use according to any one of claims 1 to 10, wherein the extract is applied topically to the skin, for example, to sensitive skin and / or mature skin.

12. The cosmetic use according to any one of claims 1 to 9, wherein the extract is applied topically to the skin of an intimate body area.

13. The cosmetic use of claim 12, wherein the extract is applied topically to the skin of the intimate body area to relieve discomfort and / or increase sensitivity and / or enhance well-being in the intimate skin area.

14. The cosmetic use according to any one of claims 1 to 9, wherein the extract is applied topically to the scalp.

15. The cosmetic use according to claim 14, wherein the extract is used to relieve scalp discomfort.

16. The cosmetic use according to any one of claims 1 to 15, wherein the extract is present as an active cosmetic ingredient in a cosmetic composition applied to the skin.

17. The cosmetic use according to claim 16, wherein the cosmetic composition is selected from skin care products, skin hygiene products, cosmetic products, hair care products or intimate care products, such as creams, such as night creams or day creams or massage creams, balms, ointments, serums, gels, films, foundations, soaps, shower gels, shampoos, conditioners or aftershaves.

18. A cosmetic composition for improving skin perception in a subject, for improving the visual appearance of the skin, and / or for enhancing skin sensitivity, and / or for reducing skin discomfort and / or for providing a sense of well-being, said cosmetic composition comprising an aqueous extract obtained from hyacinth bulbs and at least one cosmetically acceptable excipient.

19. The cosmetic composition of claim 18, wherein the aqueous extract is defined according to any one of claims 2 to 3.

20. The cosmetic composition of claim 18 or 19, wherein the aqueous extract is formulated into glycerin prior to incorporation into the cosmetic composition.

21. A cosmetic method for improving skin perception in a subject, and / or for enhancing skin sensitivity, and / or for reducing skin discomfort, and / or for improving the visual appearance of the skin, and / or for providing a sense of health, said method comprising topically applying an aqueous extract or cosmetic composition obtained from hyacinth bulbs to the skin of said subject, said aqueous extract preferably as defined in claim 2 or 3, said cosmetic composition as defined in claim 18.

22. The method of claim 21, wherein the subject has mature and / or sensitive skin and / or the subject is a perimenopausal or postmenopausal woman.

23. A cosmetic ingredient comprising an aqueous extract from hyacinth bulbs, preferably an aqueous extract from dormant hyacinth bulbs, said aqueous extract being a cosmetic active agent in a cosmetically acceptable carrier, preferably in a glycerol / water mixture.

24. The cosmetic ingredient of claim 23, further comprising one or more cosmetically acceptable excipients selected from antioxidants, preservatives and pH adjusters.

25. Use of an aqueous extract of hyacinth bulb as a dermal neurosensor in the preparation of compositions for improving skin perception and / or for reducing or preventing skin nociceptive sensations and / or for alleviating skin discomfort.

26. The use as claimed in claim 25, wherein the composition is a skin care product, a feminine care product, or a hair care product.

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