German chamomile extract for improving appearance of blemish skin

Abstract

Disclosed herein is a method for improving the appearance of blemish skin comprising applying to the skin a composition comprising a German chamomile essence component as an active ingredient and a physiologically acceptable carrier. In one embodiment, the method involves reducing comedo, sebum secretion, pore oversize, post-acne scars and expression lines, improving skin barrier function, and improving complexion by reducing redness, hyperpigmentation and post-acne hyperpigmentation.

Description

Technical Field

[0001] This disclosure relates to the fields of cosmetics, personal care, beauty cosmetics, and skin cosmetics. More specifically, this disclosure relates to using ingredients derived from living plants to improve the appearance of blemished skin associated with oily and acne-prone skin. Background Technology

[0002] Imperfect skin is a common skin condition, usually associated with excessive sebum production, leading to oily skin, enlarged pores, blackheads, post-acne scars, post-acne hyperpigmentation, blemishes, and redness. While most skin imperfections are harmless to health, they can affect appearance and potentially have a psychological impact on daily life.

[0003] Seborrhea, or oily skin, primarily affects the facial and scalp and is caused by excessive sebum secretion from the sebaceous glands. Sebum is secreted by sebaceous cells through a process of cell differentiation and lipid synthesis, also known as lipogenesis.

[0004] Sebum secretion can be negatively influenced by personal, biological, and environmental factors, such as diet, age, sex, race, life stress, and hot and humid climates (Yang J, Yang H, Xu A, He L. A Review of Advancement on Influencing Factors of Acne: An Emphasis on Environment Characteristics. Front Public Health. 2020 Sep 17; 8:450).

[0005] Acne vulgaris is another common skin condition. It causes chronic inflammation of the sebaceous glands, leading to the formation of inflammatory bumps (papules, pustules, nodules, and cysts), comedones, and scars. The pathogenesis of acne involves several interrelated factors, including increased sebum production, hyperkeratosis, skin inflammation, and the proliferation of Propionibacterium acnes (C. acnes). Inflammation caused by C. acnes infection leads to hyperpigmentation and scarring (Dagnelie, M‐A., et al. "Cutibacterium acnes phylotypes diversity loss: a trigger for skininflammatory process." Journal of the European Academy of Dermatology and Venereology 33.12 (2019): 2340-2348.). Regardless of skin color, pigmentary complications and scarring are significant cosmetic concerns for all patients.

[0006] Gamma-aminobutyric acid (GABA) is an amino acid that acts as an inhibitory neurotransmitter in the brain but also functions in the skin. GABA(A) and GABA(B) receptors have been identified in human skin cells, and recent studies have shown that activation of the GABA(A) receptor improves skin barrier function and protects the skin from pathogens such as Propionibacterium acnes. Inhibition of melanin into human melanocytes via GABA(A) and GABA(B) receptors has also been described (Molagoda, Ilandarage Menu Neelaka, et al. "Gamma-Aminobutyric Acid (GABA) Inhibits α-melanocyte-stimulating hormone-induced melanogenesis through GABA(A) and GABA(B) receptors." International Journal of Molecular Sciences 22.15 (2021):8257). GABA has also shown immediate relief of expression lines and long-term improvement in skin barrier function (Nguyen, ThuQ., et al. "A randomized, double-blind, placebo-controlled clinical study investigating the efficacy and tolerability of a peptide serum targeting expression lines." The Journal of Clinical and Aesthetic Dermatology 14.5(2021): 14). Based on these results, GABA can be considered a promising compound for providing significant cosmetic effects.

[0007] Many active agents have been disclosed for improving imperfect skin, oily skin, and acne-prone skin. Among them, the following can be cited: polyphenols, retinoids, isotretinoin, salicylic acid, apigenin, niacinamide, and botulinum toxin (Endly DC, Miller RA. Oily Skin: A review of Treatment Options. J Clin AesthetDermatol. 2017 Aug;10(8):49-55).

[0008] However, these products have moderate or limited efficacy, and there is still a need to provide new, naturally derived active agents to offer comprehensive solutions for improving the appearance of blemished skin.

[0009] German chamomile extract has been described for the treatment of various inflammatory skin conditions and for its anti-aging, pigmentation-fading, soothing, antibacterial, antispasmodic, astringent, firming, revitalizing and deodorizing properties (Kotnala, Astha, et al. "Indian Medicinal Plants for skin care and cosmeceuticals: A review." Journal of Biomedical and Therapeutic Sciences 6.2 (2019): 24-60).

[0010] Patent application MX2009001657 discloses a chamomile extract obtained from dried chamomile flower heads through alcohol impregnation, and is rich in flavonoids and polysaccharides, α-bisabolol, bisabolol, chamomile azerosin, bicyclic ethers, and spirethers. This extract is used for its soothing and anti-inflammatory properties.

[0011] US Patent No. 11,154,493 discloses a German chamomile extract obtained from fresh German chamomile flowers for reducing the adverse effects of sun exposure on the skin and improving skin appearance associated with skin aging. When tested at 100% (confirmed by in vitro skin studies), this German chamomile extract is water-soluble / water-miscible, non-irritating, and very well-tolerated. It exhibits broad-spectrum UVA and UVB photostability, inhibition of sun-induced interleukin-8 (IL-8) and interleukin-6 (IL-6) involved in inflammation, antioxidant properties (DPPH quenching capacity and free radical scavenging properties), and inhibition of human elastase activity involved in skin aging.

[0012] Therefore, to date, no existing technology mentions that German chamomile extract obtained from fresh plants can provide benefits for blemished or acne-prone skin. Summary of the Invention

[0013] The inventors have now discovered that the German chamomile extract component rich in GABA ( Chamomilla recutita The field of beauty care (serum fraction) for improving blemish-looking skin has interesting biological characteristics.

[0014] In a first aspect, this disclosure relates to a cosmetic treatment method for improving the appearance of blemished skin, the method comprising applying a composition to the skin comprising a German chamomile flower extract component as an active ingredient and a physiologically acceptable carrier.

[0015] The German chamomile extract used in this disclosure is obtained by a fractional separation method comprising exposing plant cell sap (6) derived from fresh German chamomile plant biomass containing flowers and a small amount of leaves to an electromagnetic field with a frequency equal to or greater than 2.45 GHz for a period of time, said time effectively destabilizing the plant cell sap (14) to produce a coagulated cell sap mixture, removing the coagulated membrane component (20) from said coagulated cell sap mixture to obtain a cytoplasmic / cytosol component (30), followed by further processing (32 or 33) to separate the cytoplasmic component (36) from the cytosol component (38), said cytosol component being stabilized (37) to obtain the extract component (39).

[0016] In one aspect, the method includes reducing acne, sebum secretion, enlarged pores, and expression lines, smoothing the skin, improving skin barrier function, and improving skin tone by reducing redness and pigmentation spots.

[0017] This disclosure also relates to a cosmetic treatment method for improving the appearance of blemished skin associated with acne-prone skin and for protecting the skin from excessive lipid production by sebaceous cells caused by stress selected from air pollution, UVA radiation, and lifestyle stress.

[0018] According to another embodiment of this disclosure, a composition comprising a German chamomile extract component as an active ingredient and a physiologically acceptable carrier is provided for its use in treating acne-associated blemished skin.

[0019] These and other objects, features and advantages of the present invention will become apparent to those skilled in the art upon reading this disclosure. Attached Figure Description

[0020] To illustrate aspects of the invention, these aspects are depicted in the accompanying drawings of certain embodiments of the invention. However, the invention is not limited to the embodiments depicted in the drawings. Furthermore, if provided, the reference numerals included in the drawings are intended to identify similar or identical elements.

[0021] Figure 1 This is a schematic diagram illustrating one embodiment of a method for preparing the essence components of this disclosure.

[0022] Figure 2This is a schematic diagram showing the effect of German chamomile extract on lipid secretion by human primary sebaceous cells after cortisol stress.

[0023] Figure 3 This is a schematic diagram showing the effect of German chamomile extract components on lipid secretion by human primary sebaceous cells after glucose stress.

[0024] Figure 4 This is a schematic diagram showing the effect of German chamomile extract components on lipid secretion by human primary sebaceous cells after pollution (PM10) stress.

[0025] Figure 5 This is a schematic diagram showing the effect of German chamomile extract on lipid secretion by human primary sebaceous cells after UVA irradiation.

[0026] Figure 6 This is a schematic diagram showing the effect of German chamomile extract components on melanin expression after Propionibacterium acnes stress.

[0027] Figure 7 This is a schematic diagram showing the effect of German chamomile extract components on the expression of GABA(A) receptors in isolated skin.

[0028] Figure 8 This is a schematic diagram showing the effect of German chamomile extract on the lipase activity of Propionibacterium acnes at concentrations of 0.5%-5%.

[0029] Figure 9 This is a schematic diagram showing the effect of German chamomile extract components on lipid expression in isolated skin.

[0030] Figure 10 This is a schematic diagram showing the effect of German chamomile extract on the volume of pores in the body.

[0031] Figure 11 This is a schematic diagram showing the effect of German chamomile extract on the area of ​​pores in the body. Detailed Implementation

[0032] definition As used in this article, the terms “fresh,” “live,” or “fresh German chamomile” refer to the harvesting of fresh German chamomile flowers and a small amount of leaves and storage at 4°C until sufficient biomass is collected, storage for less than 8 hours, and then the preparation of the essential components begins.

[0033] The terms “plant biomass,” “German chamomile,” “German chamomile flower,” and “Chamomilla recutita (German Chamomile) flower” are used interchangeably in this text with the meaning of German chamomile flower, which includes a small number of leaves.

[0034] As used herein, the term "maceration" refers to the mechanical crushing of flowers to improve efficiency during pressing. It does not include the addition of any liquids or solvents.

[0035] The compositions described and used in this disclosure may comprise, consist substantially of, or consist of the following: essential active ingredients and physiologically acceptable carriers and optional components as described herein.

[0036] As used herein, “consistently of” means that a composition or component may include additional ingredients, but only if the additional ingredients do not substantially alter the essential and novel characteristics of the claimed composition or method.

[0037] As used herein, the term "physiologically acceptable" means that the composition or component is suitable for contact with human skin tissue without excessive toxicity, incompatibility, instability, allergic reactions, etc.

[0038] As used herein, the term “effective amount” means an amount of compound or composition sufficient to significantly induce appearance improvement and / or skin feel benefit, but low enough to avoid serious side effects (i.e., providing a reasonable benefit-risk ratio to the extent that a person skilled in the art can reasonably judge).

[0039] As used herein, the term "phenolic compound" means any molecule having one or more aromatic rings that carry one or more hydroxyl groups, such as phenolic acids, flavonoids, or any other polyphenols.

[0040] As used herein, the term “blemished skin” refers to skin exhibiting at least one of the following skin defects: acne, oily skin due to excessive sebum production, enlarged pores, pigmentation, post-acne scars, post-acne hyperpigmentation, redness, and expression lines.

[0041] As used in this article, the term "acne-prone skin" refers to skin that is prone to developing pimples due to excessive sebum production, and which may develop into acne if not properly treated. However, the term "acne-prone skin" also refers to healthy skin without any pathological conditions.

[0042] As used in this article, the term "postacne hyperpigmentation" refers to dark spots or patches that appear after acne lesions. This is a result of inflammation caused by Propionibacterium acnes infection, which leads to the release of factors that increase melanin production in the skin.

[0043] As used in this article, the term "smooth skin" refers to reducing at least one of the following signs: wrinkles, expression lines, acne scars, and pore size and volume.

[0044] As used in this article, the term "expression lines" refers to creases on the face caused by repetitive or excessive contraction of facial muscles. Expression lines are prone to becoming permanent as the skin ages.

[0045] All numerical ranges described herein encompass their subranges; explicitly listed upper and lower limits can be combined to form other ranges not explicitly listed.

[0046] The inventors have now discovered that the German chamomile extract component according to this disclosure has biological properties for improving blemished skin, more specifically for reducing acne, sebum secretion, enlarged pores, post-acne scars and expression lines caused by excessive facial muscle tension, and for improving skin barrier function and skin tone by reducing redness, pigmentation and post-acne hyperpigmentation.

[0047] This disclosure gains advantages from these new features.

[0048] In a first aspect, this disclosure relates to a cosmetic treatment method for improving the appearance of blemished skin, the method comprising applying a composition to the skin comprising a German chamomile extract component as an active ingredient and a physiologically acceptable carrier, wherein the German chamomile extract component is separated using a fractional separation method comprising exposing plant cell sap (6) derived from fresh German chamomile plant biomass containing flowers and a small amount of leaves to an electromagnetic field with a frequency equal to or greater than 2.45 GHz for a period of time, said time effectively destabilizing the plant cell sap (14) to produce a coagulated cell sap mixture, removing a coagulated membrane component (20) from said coagulated cell sap mixture to obtain a cytoplasmic / cytosol component (30), followed by further processing (32 or 33) to enable separation of the cytoplasmic component (36) from the cytosol component (38), said cytosol component being stabilized (37) to obtain an essence component (39).

[0049] The above-mentioned hierarchical separation method in Figure 1 As described in the text.

[0050] In one aspect, the method of this disclosure provides the step of applying the above composition to blemished skin that exhibits at least one of the following signs: acne, oily skin due to excessive sebum secretion, enlarged pores, expression lines, redness, and pigmentation.

[0051] According to this disclosure, the method is applicable to reducing acne, sebum secretion, enlarged pores, expression lines, smoothing the skin, improving skin barrier function, and improving skin tone by reducing redness and pigmentation spots.

[0052] In another embodiment, this disclosure relates to a method for smoothing the skin, more specifically for reducing at least one of the following signs: wrinkles, expression lines, and enlarged pores.

[0053] According to this disclosure, the method also involves protecting the skin from excessive lipid production by sebaceous gland cells caused by various stresses such as air pollution, UVA radiation, and lifestyle stresses such as a high glycemic load diet or mental stress.

[0054] According to this disclosure, the method also involves reducing the amount of sebum on the skin surface, the number of enlarged pores, and the volume and area of ​​the pores.

[0055] According to this disclosure, the method also relates to improving the appearance of blemished skin associated with acne-prone skin.

[0056] According to this disclosure, the method also involves increasing the expression of GABA(A) receptors in skin cells.

[0057] It has been shown that, when applied for three to four weeks, compositions containing German chamomile extract as an active ingredient and a physiologically acceptable carrier significantly reduce the amount of sebum on the skin surface.

[0058] For example, clinical studies showed that volunteers had a 178.5% reduction in sebum on their skin surface.

[0059] When applied for three to four weeks, the composition containing German chamomile extract as an active ingredient and a physiologically acceptable carrier significantly reduced the number, volume, and area of ​​enlarged pores.

[0060] For example, clinical studies have shown a 113% reduction in the number of enlarged pores, a 90% reduction in average pore volume, and a 101.5% reduction in pore area.

[0061] In another embodiment, when applied for three to four weeks, the composition containing German chamomile extract as an active ingredient and a physiologically acceptable carrier significantly reduces the number of fine lines.

[0062] For example, clinical studies have shown a 140% reduction in the number of fine lines.

[0063] According to this disclosure, the above-described composition shows and improves skin imperfections in all racial skin types (Caucasian, Asian, Latino, and dark-skinned).

[0064] In another embodiment, this disclosure relates to a method for improving the appearance of acne-related blemished skin, wherein the skin of the subject in need exhibits at least one indication selected from: post-acne scarring or post-acne hyperpigmentation.

[0065] In another embodiment, this disclosure relates to a composition comprising a German chamomile extract component as an active ingredient and a physiologically acceptable carrier for its use in treating acne-associated blemished skin, wherein the German chamomile extract component is separated using a fractional separation method comprising exposing plant cell sap (6) derived from fresh German chamomile plant biomass containing flowers and a small amount of leaves to an electromagnetic field with a frequency equal to or greater than 2.45 GHz for a period of time, said time effectively destabilizing the plant cell sap (14) to produce a coagulated cell sap mixture, removing the coagulated membrane component (20) from said coagulated cell sap mixture to obtain a cytoplasmic / cytosol component (30), followed by further processing (32 or 33) to enable separation of the cytoplasmic component (36) from the cytosol component (38), said cytosol component being stabilized (37) to obtain an essence component (39).

[0066] Advantageously, in this particular embodiment, the blemished skin exhibits at least one of the following signs: post-acne scars or post-acne hyperpigmentation.

[0067] Advantageously, in this particular embodiment, the composition can be used for treatment to prevent melanin overexpression induced by Propionibacterium acnes and to reduce the virulence of Propionibacterium acnes by reducing its lipase activity.

[0068] For example, it has been demonstrated that compositions containing German chamomile extract components and physiologically acceptable carriers can reduce the virulence of Propionibacterium acnes by decreasing its lipase activity and prevent melanin overexpression induced by Propionibacterium acnes, which causes post-acne hyperpigmentation.

[0069] According to this particular embodiment, the composition also relates to increasing the expression of GABA(A) receptors in skin cells.

[0070] According to the above method, using the fractionation separation method described in U.S. Patent Nos. 7,442,391, 11,154,493 and European Patent No. 2919757, the German chamomile extract component was separated from fresh German chamomile flowers and a small amount of leaves, all of which are incorporated herein by reference.

[0071] The active ingredients used in the methods disclosed herein are not naturally occurring, but are produced using artificial manufacturing processes.

[0072] The entire method for preparing the bioactive essence component of the present invention is as follows: Figure 1 As shown.

[0073] Fresh German chamomile plants containing flowers and a small number of leaves were harvested, collected, and washed to obtain fresh plant biomass (2). The fresh plant biomass was ground, crushed, and pressed (4) to obtain intracellular substances (cell juice) (6) and fiber-rich substances (press cake) (8). The cell juice (6) was then filtered through a nylon filter (10) to remove solid particles, resulting in filtered plant cell juice (12). The filtered cell juice was exposed to electromagnetic waves at a frequency equal to or higher than 2.45 GHz (14) for a period of time, which effectively triggered its destabilization. The destabilized cell juice was then centrifuged (18) to remove precipitates (20) containing membrane components, and the supernatant, i.e., the cytoplasm / cytosol component (30), was collected. The cytoplasm / cytosol component was further processed to separate the cytoplasm component (36) from the cytosol component (38).

[0074] As a non-limiting example, the treatment may include (i) isoelectric precipitation (32) or alternatively (ii) additional electromagnetic treatment (33) at a frequency >2.45 GHz, followed by centrifugation (34) to separate the precipitated cytoplasmic components from the supernatant containing cytosolic components.

[0075] The stabilization step (37) may include incubating the cytosol component (38) in a mixture of at least one preservative, at least one chelating agent and at least one antioxidant to produce a stable cytosol component (39).

[0076] Advantageously, after the washing step (2), excess water is removed from the plant biomass to keep the dry matter content close to the natural level.

[0077] Advantageously, in step (4), the fresh plant biomass is not ground, but only crushed for 1 to 5 minutes, and then pressed.

[0078] Advantageously, a destabilization treatment (14) is carried out such that the temperature of the cell juice does not exceed 30°C during the treatment.

[0079] After centrifugation (16), the precipitate containing membrane components is removed, and the supernatant cytoplasm / cytosol component (30) is substantially free of membrane components.

[0080] The isolated cellular solute components contain low molecular weight water-soluble components dissolved in intracellular water.

[0081] The cytosol component is a transparent liquid with a yellow color and a slightly characteristic odor.

[0082] Reagents suitable for the stabilization steps of this disclosure include, but are not limited to, potassium sorbate, sodium benzoate, ascorbate glucoside, and sodium phytate.

[0083] An example of an antioxidant suitable for use in this invention is ascorbate glucoside.

[0084] A suitable chelating agent is sodium phytate.

[0085] The obtained cell extract components can be frozen at -18°C for storage.

[0086] According to the method disclosed herein, the obtained German chamomile extract component is rich in protein, amino acids (including asparagine, GABA and proline), vitamin B5 and organic acids (including malic acid and quinic acid).

[0087] The obtained German chamomile extract component has 30 to 80 g / kg (3 to 8%) of dry matter and contains at least 0.5 g / kg of amino acids, including at least 50 mg / kg of GABA, at least 0.5 g / kg of phenolic compounds and at least 1 g / kg of organic acids.

[0088] The content of German chamomile extract components is shown in Table 1.

[0089] Digital applications using various specialized databases can predict the potential biological effects of phytochemicals of interest contained in German chamomile extract components, such as ferulic acid, chlorogenic acid, malic acid, proline, GABA, and vitamin B5. Potential biological effects highlighted by Gene Ontology (GO) related terms include responses to oxidative stress, responses to UVA irradiation, regulation of lipid metabolism, regulation of porphyrin activity associated with potential antimicrobial effects, and the GABA signaling pathway. More specifically, digital analysis predicts the potential impact of GABA and proline contained in German chamomile extract components on the GABA receptor signaling pathway.

[0090] In a more specific embodiment, the composition contains 0.001 to 10%, preferably 0.005 to 5%, more preferably 0.01 to 2% of German chamomile extract as an effective amount of active ingredient, based on the total weight of the composition.

[0091] According to the methods of this disclosure, the compositions may be in the form of aqueous, hydroalcoholic, or oily solutions; and in the form of oil-in-water emulsions, water-in-oil emulsions, or a variety of emulsions; they may also be in the form of suspensions or powders suitable for application to skin, mucous membranes, lips, and / or hair.

[0092] These compositions can be more or less fluid and have the appearance of a cream, lotion, emulsion, serum, ointment, gel, paste, or foam. They can also be in solid form, such as sticks, or applied to the skin as an aerosol.

[0093] The composition may be administered by any suitable route, particularly by external topical route, and the formulation of the composition will be modified by those skilled in the art.

[0094] In a preferred embodiment, the composition used according to this disclosure is in a form suitable for topical application.

[0095] Obviously, this invention generally relates to mammals, and more specifically to humans.

[0096] These compositions may also include any additives commonly used in the envisioned application areas, as well as the auxiliaries required for their formulation, such as solvents, thickeners, diluents, antioxidants, colorants, sunscreens, self-tanning agents, pigments, fillers, preservatives, fragrances, odor absorbers, cosmetic or pharmaceutical actives, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0097] In each case, those skilled in the art will ensure that the adjuvants (excipients) and their proportions are selected so as not to interfere with the desired advantageous properties of the composition of the invention. These adjuvants may, for example, correspond to 0.01 to 20% of the total weight of the composition. When the composition of the invention is an emulsion, the fatty phase may comprise 5 to 80% by weight, preferably 5 to 50% by weight, relative to the total weight of the composition. The emulsifiers and co-emulsifiers used in the composition will be selected from those conventionally used in the field under consideration. For example, they may be used in proportions of 0.3 to 30% by weight, relative to the total weight of the composition.

[0098] Advantageously, the compositions applicable to this disclosure may contain, in addition to the active ingredient of German chamomile extract, at least one other active agent having effects similar to and / or complementary to those of the present invention. According to the present invention, this active agent will be defined as an "additional active agent".

[0099] For example, additional active agents may be selected from: sunscreen active ingredients, anti-UV agents, anti-VIS agents, anti-IR agents, light stabilizers, anti-aging agents, color toners, brightening agents, moisturizers, drainage agents and microcirculation promoters, drugs, exfoliants, scrubs, extracellular matrix stimulants, energy metabolism activators, antibacterial agents, antifungal agents, soothing agents, free radical scavengers and anti-acne agents, anti-inflammatory agents, anesthetics, warming agents, cooling agents and weight loss agents.

[0100] Such supplementary reagents may be selected from the following group: Vitamin A (especially retinoic acid), retinol, retinol propionate, retinol palmitate, Vitamin B3 (especially nicotinamide), tocopheryl nicotinate, Vitamin B5, Vitamin B6, Vitamin B12, Vitamin C (especially ascorbic acid), ascorbate glucoside, ascorbate tetrapalmitate, magnesium ascorbate phosphate and sodium ascorbate phosphate, Vitamins E, F, H, K, PP, Coenzyme Q10, metalloproteinase inhibitors, TIMP activators. DHEA, its precursors and derivatives; amino acids such as arginine, ornithine, hydroxyproline, dipalmitoyl hydroxyproline, palmitoyl glycine, hydroxylysine, methionine and their derivatives; N-acyl amino acid compounds; natural or synthetic peptides (including dipeptides, tripeptides, tetrapeptides, pentapeptides, hexapeptides and their lipophilic derivatives, isomers, and complexes with other substances such as metal ions (e.g., copper, zinc, manganese, magnesium); plant peptide extracts such as soybean, spelt wheat, grapevine, rapeseed, flaxseed, rice, corn. Rice and pea extracts, yeast extracts, Artemia extracts, dehydroacetic acid (DHA), synthetic or naturally derived sterols, salicylic acid and its derivatives, α- and β-hydroxy acids, amino sugars, glucosamine, D-glucosamine, N-acetylglucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetylmannosamine, galactosamine, N-acetylgalactosamine, polyphenol extracts, isoflavones, flavonoids such as grape extracts, pine extracts and olive extracts, lipids such as ceramides Amines or phospholipids, animal-derived oils such as squalene or squalane, vegetable oils such as sweet almond oil, coconut oil, castor oil, jojoba oil, olive oil, rapeseed oil, peanut oil, sunflower seed oil, wheat germ oil, corn germ oil, soybean oil, cottonseed oil, alfalfa oil, poppy seed oil, winter melon seed oil, evening primrose oil, millet oil, barley oil, rye oil, safflower oil, passion fruit oil, hazelnut oil, palm oil, almond kernel oil, avocado oil, and calendula oil; ethoxylated vegetable oils, and shea butter, as well as various UV shielding agents and broad-spectrum sunscreens.

[0101] The specific implementation of this cosmetic treatment method also arises from the above description. Further advantages and features of the invention will become more apparent after reading the embodiments provided for illustrative and non-limiting purposes.

[0102] Example The following embodiments are intended to illustrate specific implementations of the present invention, but are in no way intended to limit the scope of the invention.

[0103] Example 1. A hierarchical separation method as described in U.S. Patent No. 11,154,493 The entire method for preparing the bioactive essence component of the present invention is as follows: Figure 1 As shown.

[0104] Fresh German chamomile plants, including flowers and a few leaves, are harvested, collected, and washed to obtain fresh plant biomass (2). Excess water is carefully removed from the flowers to keep the dry matter content close to natural levels.

[0105] The fresh plant biomass was crushed and pressed using a screw press (4). The plant biomass (4) was rotated through the machine before pressing, crushing it for 1 to 5 minutes. This step produced intracellular plant material (cell juice) (6) and fiber-rich material (cake) (8). The cell juice (6) was then filtered through a nylon mesh (10) to remove solid particles and obtain filtered plant cell juice (12). The filtered cell juice was exposed to electromagnetic waves at a frequency equal to or greater than 2.45 GHz (14) for a period of time to effectively trigger its destabilization. The destabilized cell juice had the appearance of a coagulated cell juice mixture. The coagulated cell juice mixture was then subjected to centrifugation (16). The precipitate containing membrane components (20) was removed, and the supernatant, i.e., the cytoplasm / cytosol component (30), was collected.

[0106] Advantageously, a destabilization treatment (14) is carried out such that the temperature of the cell juice does not exceed 30°C during the treatment.

[0107] The cytoplasmic / cytosolic component (30) is further treated to enable separation of the cytoplasmic component from the cytosolic component. As a non-limiting example, the treatment may include (i) isoelectric precipitation (32) or alternatively (ii) additional electromagnetic treatment (33) at a frequency >2.45 GHz, followed by centrifugation (34) to enable separation of the precipitated cytoplasmic component (36) from the supernatant containing the cytosolic component (38).

[0108] The isolated cellular solute components (38) contain low molecular weight water-soluble components dissolved in intracellular water.

[0109] The cytosolic component is a clear liquid with a yellow color and a slightly characteristic odor. Within hours, the unstable cytosolic component irreversibly transforms into a dark brown suspension containing heavy precipitates and a strong, non-characteristic odor.

[0110] The stabilization step includes incubating the cytosol (38) component in a mixture of at least one preservative, at least one chelating agent and at least one antioxidant (37) to obtain a stable cytosol component (39).

[0111] Reagents applicable to the stabilization step (37) of this disclosure include, but are not limited to, potassium sorbate, sodium benzoate, ascorbate glucoside and sodium phytate.

[0112] An example of an antioxidant suitable for use in this invention is ascorbate glucoside.

[0113] A suitable chelating agent is sodium phytate.

[0114] The obtained cell extract components can be frozen at -18°C for storage.

[0115] Example 2: Analysis of the components of German chamomile extract relative to conventional flower extracts Extracts: Three batches of German chamomile extract components, prepared without preservatives and stored at -18°C as described in Example 1, were analyzed.

[0116] For comparative purposes, a conventional extract was prepared. A mixture of 1% dried German chamomile flowers (including a small amount of leaves) in distilled water was heated at 25°C for 1 hour, and then filtered through a filter with a large pore size of 30 μm to remove solid residual plant matter from the liquid fraction. The extract was then sequentially filtered through filters with gradually decreasing porosity to clarify it, until final sterile filtration was performed at a pore size of 0.2 μm to obtain the liquid fraction as the conventional extract.

[0117] Analysis plan: Total soluble sugar analysis was performed using a colorimetric method. 1 mL of each sample or standard was reacted with 5 mL of ice-cold 2000 ppm anthrone in 72% sulfuric acid. The reagent was heated to 100°C and held for 10 minutes. A four-point calibration curve was generated from the absorbance values ​​of the solution at 680 nm, and this curve was compared with the calibration curves for fructose and glucose to determine their total sugar concentration.

[0118] After reducing the Folin-Ciocalteu reagent with phenol, the total phenol content was measured spectrophotometrically at 760 nm. Quantification was performed using a standard gallic acid curve, and the results are expressed as gallic acid equivalents.

[0119] Following the colorimetric reaction of biuret with Folin-Ciocalteu reagent, the total protein content was measured spectrophotometrically at 550 nm. Quantification was performed using a standard BSA (bovine serum albumin) curve.

[0120] The total amino acid content was quantified using colorimetric determination at 570 nm. Free amino acids react with ninhydrin reagent to form a colored complex. The total amino acid content was determined using a standard curve of the amino acid cell.

[0121] The content of phenolic compounds was analyzed separately by liquid chromatography combined with a UV detector fixed at 254 nm. A solution of 0.1% formic acid in water and methanol was used as the mobile phase, and samples were separated on an UPTISPHERE CS EVOLUTION C18-AQ column using an Agilent 1200 HPLC system (Agilent Technologies).

[0122] Individual organic acid contents were determined by high-performance liquid chromatography combined with a mass spectrometer equipped with a negative ion mode electrospray ionization source (Acquity Qda, Waters). Samples were separated using an Agilent 1260 HPLC system (Agilent Technologies) on an EC 150 / 4.6 Nucleoshell RP 18plus-5μm column (Macherey Nagel: 763236.46). The flow rate was 0.3 mL / min. The mobile phase consisted of an aqueous solution of 0.01% formic acid and acetonitrile.

[0123] Individual amino acid content analysis was performed by liquid chromatography combined with a UV detector immobilized at 254 nm. After derivatization with phenyl isothiocyanate, the sample was separated using an Agilent 1200 HPLC system (Agilent Technologies) on a UPTIPHEREstrategy C18-2 column. The mobile phase consisted of an aqueous solution of 0.1% phosphoric acid and acetonitrile.

[0124] B vitamins were characterized and quantified by high-performance liquid chromatography combined with a mass spectrometer equipped with a negative ion mode electrospray ionization source (Acquity Qda, Waters). Samples were separated using an Agilent 1260 HPLC system (Agilent Technologies) on an EC 150 / 4.6 Nucleoshell RP 18plus-5μm column (Macherey Nagel: 763236.46). The flow rate was 0.5 mL / min. The mobile phase consisted of 20 mmol ammonium acetate and 0.1% formic acid in water and methanol / acetonitrile (65 / 35).

[0125] result Table 2: Quantitative results of German chamomile extract components relative to conventional extracts.

[0126] LQ: Limit of quantitation, which can vary depending on the method.

[0127] German chamomile extract contains 33 times more protein, 7 times more phenolic compounds (including 2 times more ferulic acid and its derivatives and at least 17 times more apigenin and its derivatives), 30 times more vitamin B5, and 80 times more organic acids (including 74 times more malic acid and at least 100 times more quinic acid) than conventional extracts. Amino acids (especially asparagine, proline, and GABA) are highly detected in German chamomile extract, but are absent in conventional extracts.

[0128] Example 3: Predicting the bioactivity of German chamomile extract components using digital applications Based on the content of the German chamomile extract components in Example 1 in biologically significant phytochemicals—ferulic acid, chlorogenic acid, malic acid, proline, GABA, and vitamin B5—various numerical tools were used to predict their bioactivity.

[0129] Target prediction was performed using SMILES files and the publicly available database SwissTargetPrediction (Daina A., Michielin O. and Zoete V, 2019. SwissTargetPrediction: updated data and new features for efficient prediction of protein targets of small molecules. Nucleic Acids Res, 47: W357–W364) based on the principle of target / ligand similarity. Subsequently, the predicted targets were aggregated and target enrichment analysis was performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database (Sherman BT, Hao M., Qiu J., Jiao X., Baseler MW, Lane HC, Imamichi T. and Chang W, 2022. DAVID: a web server for functional enrichment analysis and functional annotation of gene lists (2021 update). Nucleic Acids Research, 50(W1): W216-W221; Huang DW, ShermanBT, Lempicki RA, 2009. Systematic and integrative analysis of large genelists using DAVID Bioinformatics Resources. Nature Protoc., 4(1): 44-57). This process enabled the predicted target proteins to be classified according to the different biological functions they participate in. In fact, the DAVID database provides rich biological subject information for the overall predicted targets, especially gene ontology (GO) terminology.

[0130] Table 3

[0131] Digital applications using various specific databases allow for the prediction of seemingly plausible biological effects by identifying Gene Ontology (GO) related terms: responses to oxidative stress, responses to UVA irradiation, regulation of lipid metabolism, regulation of porphyrin activity associated with potential antimicrobial effects, and the GABA signaling pathway.

[0132] Predictions obtained using the Swiss target prediction tool demonstrate the potential impact of GABA and proline contained in the extract on the GABA receptor signaling pathway.

[0133] Example 4: German chamomile extract protects sebaceous gland cells from excessive lipid production induced by several stresses applied in vitro to sebaceous gland cells. Objective: To evaluate the benefits of German chamomile extract components on lipid overproduction in sebaceous gland cells induced by several stresses. Evidence suggests that various environmental stresses, such as exposure to air pollutants or UV exposure, and lifestyle stresses (including high glycemic load diets), induce excessive lipid production in sebaceous gland cells. Furthermore, cortisol is the main steroid hormone secreted by the adrenal cortex (HPA pathway) and is a potential indicator of mental stress. Lipid production was monitored using a standard procedure of Nile Red staining (a selective fluorescent dye for intracellular lipid droplets).

[0134] plan: Sebaceous gland cells: Primary sebaceous gland cells from normal individuals were isolated during plastic surgery. The epidermis and dermis were separated using enzymatic digestion with a dispersing enzyme. Sebaceous glands were isolated from the skin (dermis and epidermis) and their ducts were removed by microscopic dissection under a microscope. The isolated sebaceous gland cells were cultured for several days after secondary enzymatic digestion with trypsin.

[0135] After several days of culture, sebaceous gland cells were treated for 48 hours with the preservative-free German chamomile extract fraction (0.1% (vol. / vol.) diluted in PBS) from Example 1. Simultaneously, they were subjected to cortisol solution (10 μM), glucose solution (3 g / L), pollutants (PM10; 50 μg / mL), or UVA irradiation (2 J / cm²). 2 Stress was applied to sebaceous gland cells. An untreated control and a control without stress were established. After stress, sebaceous gland cells were stained with Nile Red solution, and lipid droplets were observed under a fluorescence microscope to quantitatively analyze lipid production.

[0136] Image Quantification: Nile red is a phenoxazinone dye that stains hydrophobic lipids with a strong red color. Staining intracellular lipid droplets with a Nile red fluorescent probe using a red filter (excitation 515-560 nm; emission >590 nm) confirmed the presence of all lipids. Images were acquired using a Nikon Eclipse Ni-E microscope equipped with a DS-F i3 Nikon camera, using NiS-AR (Nikon) acquisition software. Six images under each condition were analyzed using Volocity image analysis software (Improvision). Fluorescence intensity was used to select representative regions. The software quantified the level of fluorescence intensity, which was adjusted by taking into account the number of cells.

[0137] Statistical analysis: Statistical tests were used to assess the significance between the treatment group and the untreated group (100% expression reference). Student's t-test was used for independent samples comparisons, and a one-sided rejection region was defined. p ≤ 0.05 = * was considered significant, p ≤ 0.01 = ** was highly significant, and p ≤ 0.005 = *** was highly significant. Six images were analyzed for each condition.

[0138] Results: Compared with the negative control (no stress), as Figure 2 Cortisol (mental stress) Figure 3 Glucose (high glycemic load diet) Figure 4 PM10 (air pollution) or Figure 5 UVA irradiation showed that lipid production by sebaceous gland cells increased by +93%***, +41%***, +24%***, and +32%**, respectively. Treatment of sebaceous gland cells with German chamomile protected the cells from stress and reduced lipid production (compared to the stress condition, -35%*, -19%*, -17%***, and -38%***, respectively).

[0139] Conclusion: Treatment of sebaceous gland cells with 0.1% German chamomile extract protects cells from stress induced by cortisol (mental stress), glucose (high glycemic load diet), PM10 (air pollution), or UVA radiation, and reduces lipid production in human sebaceous gland cells.

[0140] Example 5: German chamomile extract prevents hyperpigmentation induced by Propionibacterium acnes. Objective: The primary bacteria involved in acne is *Propionibacterium acnes*, which also exists on the surface of normal skin. *Propionibacterium acnes* can induce inflammation, leading to hyperpigmentation due to the release of melanogen cytokines that stimulate melanin production. To evaluate the protective effect of German chamomile extract, an in vitro experiment was designed to simulate *Propionibacterium acnes*-induced hyperpigmentation.

[0141] plan: Ex vivo skin: Normal human skin obtained from plastic surgery (abdomen, female donor, 30 years old).

[0142] Skin biopsy tissue was prepared using a 6 mm diameter perforator (PFM medical) and held in organ culture medium (DMEM / Ham's-F12 50 / 50) supplemented with 10% FBS, 2 ml M-glutamine, and 100 μg / ml Primocin. The skin biopsy tissue was maintained at 37°C under a humidified atmosphere of 5% CO2. Propionibacterium acnes was inactivated by heating at 80°C for 30 minutes, and protein content was determined by the BCA method. Skin biopsy tissue was treated with 1% (v / v) of the preservative-free German chamomile extract component from Example 1 for 48 h, followed by stress with 100 μg / ml inactivated bacteria for 24 h. After stress, the skin biopsy tissue was stained with Fontana-Masson solution, and melanin was determined by reducing the ammonia-containing silver nitrate solution to metallic silver (brown) without the use of an external reducing agent.

[0143] Image quantification: Images were acquired using NiS-AR (Nikon) acquisition software with a Nikon Eclipse Ni-E microscope equipped with a DS-Fi3 Nikon camera. Six images were analyzed under each condition. ImageJ software allowed for counting the number of brown / dark pixels in manually selected epidermal regions for each image. The obtained pixel count was adjusted to the length of the epidermal region examined.

[0144] Statistical analysis: Statistical tests were used to assess the significance between the treated and untreated groups (100% expression reference). Student's t-test was used for independent samples comparisons, and a one-sided rejection region was defined. p ≤ 0.05 = * was considered significant, p ≤ 0.01 = ** was highly significant, and p ≤ 0.005 = *** was highly significant. Six images were analyzed for each condition.

[0145] Results: Stress-induced overexpression of melanin by Propionibacterium acnes was quantified by Fontana-Masson staining (+59%** compared to untreated conditions). Figure 6 Pretreatment of skin biopsy tissue with kojic acid (positive control) or 1% German chamomile extract prevented the overexpression of melanin induced by Propionibacterium acnes (-49%*** and -36%***, respectively, compared to the stress condition).

[0146] Conclusion: Treatment of skin biopsy tissue with 1% German chamomile extract can prevent the consequences of Propionibacterium acnes infection on melanocyte overexpression.

[0147] Example 6: German chamomile extract increases GABA(A) receptor expression. Objective: Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in the brain, but it also plays a role in the skin. In fact, GABA receptors are present in skin cells such as keratinocytes. In this experiment, the regulation of GABA(A) receptor expression in isolated skin was observed using immunoassay.

[0148] plan: Normal human skin was obtained from the same plastic surgery procedure as in Example 5. Skin biopsy tissue was obtained using a 6 mm diameter trephine (PFM medical) and maintained in organ culture medium (DMEM / Ham's-F12 50 / 50) supplemented with 10% FBS, 2 mM L-glutamine, and 100 μg / ml Primocin. The skin biopsy tissue was maintained in a humidified atmosphere at 37°C and 5% CO2. The skin biopsy tissue was treated for 48 h with 1% (vol. / vol.) of the preservative-free German chamomile extract component of Example 1 or with 1 mM, 10 mM, or 20 mM GABA. After treatment, GABA(A) receptors were detected using specific antibodies and quantitatively analyzed using fluorescence microscopy.

[0149] Image quantification: Images were acquired using a Nikon Eclipse Ni-E microscope equipped with a DS-Fi3 Nikon camera and NiS-AR (Nikon) acquisition software. Six images under each condition were analyzed using Volocity image analysis software (Improvision). This software quantifies the level of fluorescence intensity, adjusting the level of fluorescence intensity by taking into account the area of ​​the epidermal region examined.

[0150] Statistical analysis: Statistical tests were used to assess the significance between the treated and untreated groups (100% expression reference). Student's t-test was used for independent samples comparisons, and a one-sided rejection region was defined. p ≤ 0.05 = * was considered significant, p ≤ 0.01 = ** was highly significant, and p ≤ 0.005 = *** was highly significant. Six images were analyzed for each condition.

[0151] Results: Skin biopsy tissue was treated with 1% German chamomile extract or 1 mM, 10 mM or 20 mM GABA solution. Figure 7 ) induces an increase in GABA(A) receptor expression (+14%***, +8%***, +9%*** and +11%***, respectively).

[0152] Conclusion: Treatment of skin biopsy tissue with German chamomile extract can increase the expression of GABA(A) receptors in skin biopsy tissue.

[0153] Example 7: German chamomile extract reduces the lipase activity of Propionibacterium acnes. Objective: Propionibacterium acnes secretes enzymes such as lipases, which are involved in the pathogenesis of acne. Lipases from Propionibacterium acnes metabolize sebum and release inflammatory metabolites in the skin (Platsidaki, Eftychia, and ClioDessinioti. "Recent advances in understanding Propionibacterium acnes (Cutibacterium acnes) in acne." F1000Research 7 (2018). Ketoconazole, a well-known antifungal agent, induces a decrease in the activity of Propionibacterium acnes, thereby reducing lipase activity.

[0154] The protective effect of German chamomile extract was verified by measuring the lipase activity of Propionibacterium acnes.

[0155] plan: Propionibacterium acnes: Propionibacterium acnes strain NCTC 737 [VPI 0389] is a whole-genome sequenced bacterial strain (ATCC-6919) isolated from facial acne.

[0156] In 96-well plates, 100 µL of *Propionibacterium acnes* with a dosing of 0.5 DO was inoculated into enhanced Clostridium tumefaciens medium, with the medium supplemented with either ketoconazole (as a positive control antifungal drug) at a final concentration of 16 µg / mL, or with unpreserved German chamomile extract at final concentrations of 0.5%, 1%, 2%, and 5%. The plates were anaerobically incubated at 37°C for 24 hours. The plates were then centrifuged at 4700 rpm for 10 minutes. 50 µL of the supernatant (diluted 1:1) was mixed with 50 µL of 0.4 mM 4-methylumbelliferyl oleate (MUO) dissolved in 13 mM Tris-HCl, 0.15 M NaCl, and 1.3 mM CaCl2 (pH 8.0), and dropped into black 96-well plates. The mixture was incubated at 25°C for 30 minutes. The enzyme reaction was terminated by adding 100 µL of 0.1 M sodium citrate (pH 4.2). The content of 4-methylumbelliferone released by lipase was detected using a fluorescent microplate reader (Synergy 2, Biotek) at an excitation wavelength of 355 nm and an emission wavelength of 460 nm.

[0157] Lipase activity quantification: The fluorescence value of the blank was subtracted from the fluorescence value of the German chamomile extract or ketoconazole (containing Propionibacterium acnes). The blank corresponded to wells without Propionibacterium acnes but treated with German chamomile extract or ketoconazole (16 μg / mL). The relative percentage of fluorescence for each sample was calculated relative to the average untreated control wells without ketoconazole.

[0158] Statistical analysis: Statistical analysis was performed using Student's t-test for independent samples comparison, with a one-sided rejection region defined. p ≤ 0.05 = * was considered significant, p ≤ 0.01 = ** was highly significant, and p ≤ 0.005 = *** was highly significant. Three replicate experiments were conducted.

[0159] Results: Ketoconazole treatment induced a decrease in lipase activity (-64%*** compared to untreated conditions). Figure 8 Treatment with German chamomile extract at final concentrations of 0.5%, 1%, 2%, and 5% induced a decrease in Propionibacterium acnes lipase activity (compared to stress conditions: -5%, -18%***, -25%***, and -54%***, respectively).

[0160] Conclusion: Treatment of Propionibacterium acnes with different concentrations of German chamomile extract inhibited lipase activity. The inhibitory effect of 5% German chamomile extract on Propionibacterium acnes lipase activity was similar to that of ketoconazole.

[0161] Example 8: German chamomile extract improves barrier function Objective: The skin barrier function is crucial for protecting the skin from external aggressors and pathogens. In this study, lipid expression in the epidermis was measured to assess the effect of German chamomile extract components on barrier function.

[0162] Protocol: Normal human skin was obtained from the same plastic surgery procedure as in Example 5. Skin biopsies were obtained using a 6 mm diameter trephine (PFMmedical) and maintained in organ culture medium (DMEM / Ham's-F12 50 / 50) supplemented with 10% FBS, 2 mM L-glutamine, and 100 μg / ml Primocin. The skin biopsies were maintained at 37°C under a humidified atmosphere of 5% CO2. The skin biopsies were treated with 1% and 2% of the preservative-free German chamomile extract from Example 1 for 48 h. Following treatment, the effect on barrier function was assessed by Nile Red staining, and total lipid expression was quantitatively analyzed by observation under a fluorescence microscope.

[0163] Nile red is a phenoxazinone dye that stains hydrophobic lipids with a strong red color. Staining intracellular lipid droplets with a Nile red fluorescent probe using a red filter (excitation 515-560 nm; emission >590 nm) can confirm the presence of all lipids. Images were acquired using a Nikon Eclipse Ni-E microscope equipped with a DS-Fi3 Nikon camera and NiS-AR (Nikon) acquisition software. Six photographs under each condition were analyzed using Volocity image analysis software (Improvision). Fluorescence intensity was used to select representative areas. The software quantifies the level of fluorescence intensity, which is adjusted by taking into account the area of ​​the epidermal region examined.

[0164] Statistical Analysis: Statistical analysis was performed using Student's t-test for independent samples comparison, with a one-sided rejection region defined. p ≤ 0.05 = * was considered significant, p ≤ 0.01 = ** was highly significant, and p ≤ 0.005 = *** was highly significant. Six photographs were analyzed under each condition.

[0165] Results: Treatment with 1% or 2% German chamomile extract induced an increase in epidermal lipid expression (+40%*** and +33%***, respectively). Figure 9 ).

[0166] Conclusion: Treatment with different concentrations of German chamomile extract increased lipid expression in the epidermis, leading to improved skin barrier function.

[0167] Example 9: German chamomile extract improved blemished skin in clinical studies. Objective: To conduct a clinical study on Caucasian skin to evaluate the effects of a 1% German chamomile extract formulation on skin with blemishes.

[0168] Volunteers: 18 volunteers aged 21 to 67 with oily skin and enlarged pores; Toppan photography II to IV.

[0169] Protocol: Facial skin imperfections were assessed at different time points (D0, D7, D21, and D28) one month before and after using a cream containing 1% German chamomile extract from Example 1. This study was a randomized, double-blind, half-face controlled trial with a placebo as a control.

[0170] [Table 4]: Care formulas containing 1% German chamomile extract used in clinical trials

[0171] The placebo was formulated identically to the German chamomile extract component without Example 1.

[0172] The following parameters were evaluated during the study: - Use Sebumeter® SM 815 (Courage & Khazaka) to determine the amount of sebum on the skin surface. -Analyze pore size (0.35 to 2.5 mm) and the number of enlarged pores through image analysis. -Colored images of faces Statistical analysis: Depending on whether the data follows a normal distribution, Student's t-test or Wilcoxon test is used for analysis. A result is considered significant when p ≤ 0.05, highly significant when p ≤ 0.01, and highly significant when p ≤ 0.005.

[0173] result: After 3 and 4 weeks of application of a cream containing 1% German chamomile extract, a significant reduction in sebum production on the skin surface was observed compared to a placebo cream. These preliminary results suggest that the German chamomile extract reduces sebum secretion on the skin surface, resulting in a less oily appearance.

[0174] [Table 5]: Statistical results of skin surface sebum levels on the side treated with 1% German chamomile extract and the side treated with placebo.

[0175] Furthermore, after 3 and 4 weeks of using a cream containing 1% German chamomile extract, the number of enlarged pores, as well as the volume and area of ​​pores, were significantly reduced compared to the placebo side. Figure 10 and 11 The results indicate less pore enlargement, demonstrating the astringent (or pore-tightening) effect of the German chamomile extract.

[0176] Fine lines on the cheeks were also evaluated to assess the effect of the German chamomile extract on expression lines that may be associated with skin laxity. Lax skin is defined as skin with fewer expression lines and fine lines. After 4 weeks of using a cream containing 1% German chamomile extract, fine lines were significantly reduced compared to the placebo side. Skin was smoother, less lax, and fine lines were visibly reduced.

[0177] Conclusion: The cream containing 1% German chamomile extract significantly reduced the amount of sebum on the skin surface and decreased the number and size of pores, demonstrating the astringent (or pore-tightening) effect of the German chamomile extract. Furthermore, after using the cream containing 1% German chamomile extract, the number and size of fine lines were reduced. Under the conditions of this invention, the 1% German chamomile extract formulated in the cream can allow for the counteracting of skin imperfections and the improvement of the appearance and pore size of oily skin.

Claims

1. A cosmetic treatment method for improving the appearance of blemished skin, comprising applying a composition to the skin, the composition comprising a German chamomile flower extract component as an active ingredient and a physiologically acceptable carrier, wherein the German chamomile flower extract component is obtained by a fractional separation method comprising exposing plant cell juice (6) derived from fresh German chamomile plant biomass containing flowers and a small amount of leaves to an electromagnetic field with a frequency equal to or greater than 2.45 GHz for a period of time, said time effectively destabilizing the plant cell juice (14) to produce a coagulated cell juice mixture, removing a coagulated membrane component (20) from the coagulated cell juice mixture to obtain a cytoplasmic / cytosol component (30), followed by further processing (32 or 33) to enable separation of the cytoplasmic component (36) from the cytosol component (38), said cytosol component being stabilized (37) to obtain the essence component (39).

2. The method of claim 1, wherein improving blemished skin includes reducing acne, sebum secretion, enlarged pores and expression lines, smoothing the skin, improving skin barrier function, and improving skin tone by reducing redness and pigmentation.

3. The method of claim 1, wherein the skin is protected from excessive lipid production by sebaceous gland cells caused by stress selected from air pollution, UVA radiation and lifestyle stress.

4. The method of claim 1, wherein the expression of GABA(A) receptors is increased in skin cells.

5. The method according to claim 1, which is used to reduce the amount of sebum on the skin surface, the number of enlarged pores, and the volume and area of ​​pores.

6. The method of claim 1, which is used to improve the appearance of blemished skin associated with acne-prone skin.

7. The method according to claim 1, wherein the German chamomile extract component has 30 to 80 g / kg (3 to 8%) of dry matter and contains at least 0.5 g / kg of amino acids, including at least 50 mg / kg of GABA, at least 0.5 g / kg of phenolic compounds and at least 1 g / kg of organic acids.

8. The method according to claim 1, wherein the composition comprises 0.001% to 10%, preferably 0.005% to 5%, and more preferably 0.01% to 2% of German chamomile extract based on the weight of the total composition.

9. The method of claim 1, wherein the composition further comprises an additional ingredient selected from sunscreen active substances, anti-inflammatory agents, and skin-tone agents.

10. A composition comprising a German chamomile extract component as an active ingredient and a physiologically acceptable carrier for use in treating acne-associated blemished skin, wherein the German chamomile extract component is obtained using a fractional separation method comprising exposing plant cell sap (6) derived from fresh plant biomass containing flowers and a small number of leaves of German chamomile flower to an electromagnetic field with a frequency equal to or greater than 2.45 GHz for a period of time, said time effectively destabilizing said plant cell sap (14) to produce a coagulated cell sap mixture, removing a coagulated membrane component (20) from said coagulated cell sap mixture to obtain a cytoplasmic / cytosol component (30), followed by further processing (32 or 33) to enable separation of the cytoplasmic component (36) from the cytosol component (38), said cytosol component being stabilized (37) to obtain said extract component (39).

11. The composition of claim 10, wherein the blemished skin exhibits at least one of the following signs: post-acne scarring or post-acne hyperpigmentation.

12. The composition of claim 10, which is used to prevent melanin overexpression induced by Propionibacterium acnes and to reduce the virulence of Propionibacterium acnes by reducing its lipase activity.

13. The composition according to claim 10, wherein the expression of GABA(A) receptors is increased in skin cells.

14. The composition according to claim 10, wherein the German chamomile extract component has 30 to 80 g / kg (3 to 8%) of dry matter and contains at least 0.5 g / kg of amino acids, including at least 50 mg / kg of GABA, at least 0.5 g / kg of phenolic compounds and at least 1 g / kg of organic acids.

15. The composition according to claim 10, wherein the composition comprises, by weight of the total composition, 0.001% to 10%, preferably 0.005% to 5%, and more preferably 0.01% to 2% of German chamomile extract.

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