PL246900B1 - Cosmetic composition in the form of a nourishing emulsion for application to the skin - Google Patents

Abstract

The subject of the application is a cosmetic composition in the form of a W/O nourishing emulsion for application to the skin, which contains: as an active ingredient, dry extract of Ajuga Reptans root, in an amount of 0.1 - 1.0% by weight, and in addition: evening primrose oil, in an amount of 7 - 10% by weight; capuacu butter, in an amount of 2 - 3% by weight; isopropyl myristate, in an amount of 5 - 6% by weight; beeswax, in an amount of 3 - 5% by weight; triglycerides of palmitic and stearic acids, in an amount of 2.5 - 4.5% by weight; glycerin, in an amount of 3 - 5% by weight; a mixture consisting of esters of fatty acids derived from olive oil and polyglycerol-6 in an amount of 50 - 70% by weight. and polyglycerol-6 pentaoleate in an amount of 30-50% by weight, in an amount of 6-8% by weight; xanthan gum, in an amount of 0.5-1.5% by weight; sodium benzoate, in an amount of 0.2-0.5% by weight; magnesium sulfate, in an amount of 0.5-1.0% by weight; water in the balance to 100% by weight of the composition.

Description

Description of the invention

The subject of the solution is a cosmetic composition in the form of a nourishing emulsion for application to the skin, being a W/O emulsion containing Ajuga Reptans root extract as an active ingredient.

From the work of A. Toiu et al., "Comparative Phytochemical Profile, Antioxidant, Antimicrobial and In Vivo Anti-Inflammatory Activity of Different Extracts of Traditionally Used Romanian Ajuga genevensis L. and A. reptans L. (Lamiaceae)", published in Molecules, vol. 24, no. 1597, pp. 2-21,2019 it is known that the above-ground part (herb) of bugleweed is a rich source of active substances such as: chlorogenic acid, caffeic acid, quinic acid, numerous iridoids and phytosterols, diterpenes, catechins, flavonoids, anthocyanins, phytoecdysteroids, as well as vitamin C, thanks to which the bugleweed herb has antioxidant, antipyretic, antibacterial, allergic reaction-inhibiting effects, accelerating epithelium renewal and wound healing, and alleviating symptoms of inflammation.

S. Vertuani, P. Ziosi, R. D. Toso, C. B. Vicentini, and S. Manfredini, in the article “Dualistic Properties of Cosmetic Formulations Based on Phenylpropanoids from Ajuga Reptans”, J. Cosmet. Dermatological Sci. Appl., vol. 3, no. November, pp. 64-72, 2013, described the antioxidant activity of Ajuga Reptans herb extracts and their ability to protect the skin against UVA and UVB radiation.

The protective effect against UV radiation and antioxidant properties of Ajuga Reptans extracts result from the presence of phenylpropanoids, while thanks to the iridoids present in them, they also have anti-cancer effects (T. Esposito et al., "Study on Ajuga Reptans Extract: A Natural Antioxidant in Microencapsulated Powder Form as an Active Ingredient for Nutraceutical or Pharmaceutical Purposes", Pharmaceutics, vol. 12, no. 671, pp. 1-23, 2020).

Patent application CN106176542A presents a method of obtaining a moisturizing, anti-aging composition containing a number of plant extracts, including Ajuga Reptans extract.

Patent application WO2020008117A1 describes the use of HGRP (Histidine-rich glycoprotein) extracts obtained from Ajuga Reptans plant cells for use in cosmetic compositions with a rejuvenating effect.

Patent application KR20090104223A discloses an anti-aging cosmetic composition containing in its composition an extract of Ajuga Reptans obtained using a plant stem cell culturing technique.

Preparations based on Ajuga Reptans extracts for supporting hair growth and stimulating hair bulbs, particularly suitable for preventing and treating androgenic alopecia, in the form of preparations for oral administration or topical use are described in patent description EA013502B1.

Ajuga Reptans occurs commonly throughout Poland and is a widely available raw material.

Due to its properties, Ajuga Reptans herb extracts have found application in cosmetic preparations, both for skin and hair care. Bugleweed herb extracts can be found in shampoos, rinses, and acne skin preparations.

However, both the literature and industrial practice do not reveal the possibility of using Ajuga Reptans root extracts in cosmetic or medical preparations.

The aim of the present invention is to develop a cosmetic composition in the form of a nourishing emulsion for application to the skin, being a W/O emulsion containing, as an active ingredient, a dry extract of the root of Ajuga Reptans.

According to the invention, a cosmetic composition in the form of a W/O type nourishing emulsion for application to the skin, consisting of a water phase containing a humectant, a stabilizer, a preservative, a rheology modifier and an active ingredient, an oil phase containing an emulsifier and an emollient and optionally additional ingredients, characterized in that:

- as an active ingredient it contains dry extract of the root of Ajuga Reptans, in the amount of 0.1-1.0% by weight, and also contains:

- evening primrose oil, 7-10% by weight, as an emollient,

- capuacu butter, 2-3% by weight, as an emollient,

- isopropyl myristate, 5-6% by weight, as an emollient and consistency modifier,

- beeswax, 3-5% by weight, as an emollient, consistency regulator and auxiliary emulsifier,

- triglycerides of palmitic and stearic acids, 2.5-4.5% by weight, as an emollient, - glycerin, 3-5% by weight, as a humectant,

- a mixture consisting of fatty acid esters derived from olive oil and polyglycerol-6 in the amount of 50-70% by mass and polyglycerol-6 pentaoleate in the amount of 30-50% by mass, in the amount of 6-8% by mass, as an emulsifier,

- xanthan gum, 0.5-1.5% by weight, as a theology modifier,

- sodium benzoate, 0.2-0.5% by weight, as a preservative,

- magnesium sulphate, 0.5-1.0% by weight, as an emulsion stabilizer,

- water to make up 100% by weight of the composition.

In the composition according to the invention, the active ingredient in the form of a dry extract of the root of the bugle Ajuga Reptans is a product obtained by extraction of freeze-dried, crushed root of Ajuga Reptans with an aqueous solution of ethyl alcohol with a concentration of 70 to 96% by volume, for 5 hours, at a mass ratio of plant material to solvent of 20-100:1, where the solvent is repeatedly recirculated through the plant material, preferably in an overflow extractor, and the extract separated from the solid parts is dried to a constant mass.

The active ingredient is a dry extract of the root of the bugle Ajuga Reptans, preferably obtained by extraction of freeze-dried Ajuga Reptans root with a 70% by volume aqueous solution of ethyl alcohol.

The qualitative composition of Ajuga Reptans root extracts was analyzed using thin layer chromatography (TLC), UV/VIS spectrophotometry, infrared spectroscopy (IR), nuclear magnetic resonance (HNMR and CNMR), and high-performance liquid chromatography-mass spectrometry (HPLC-MS) methods to compare the composition of the extracts obtained with the extracts from the state of the art. Spectroscopic methods confirmed the presence of compounds such as luteolin, rutin, gallic acid, aucubin, and chlorogenic acid in the extract, described by Toiu A, et al. "Comparative phytochemical profile, antioxidant, antimicrobial and in vivo anti-inflammatory activity of different Extracts of traditionally used Romanian Ajuga genevensis L. and A. reptans L. (Lamiaceae)", Molecules. 2019; 24: 2-6, 17, doi:10.3390/molecules24081597. The results of qualitative analysis using high-performance liquid chromatography coupled with mass detector confirmed the presence of steroids such as: Reptansterone, 28-epi-sengosterone, 5,29-Dihydroxycapitasterone, 2-Dehydroajugalactone, 3-Dehydroajugalactone, which are the subject of the works: Qing X, Yan HM, Ni ZY, et al. "Chemical and pharmacological research on the plants from genus Ajuga". Heterocycl Commun. 2017; 23(4): 2-3, 5-9, 15. doi:10.1515/hc-2017-0064 and Goger G, Kose YB, Demirci F, Goger F. "Phytochemical Characterization of Phenolic Compounds by LCMS/MS and Biological Activities of Ajuga reptans L., Ajuga salicifolia (L.) Schreber and Ajuga genevensis L from Turkey”, Turkish J Pharm Sci. 2021: 10, doi:10.4274/tjps.galenos.2021.33958, as well as compounds from the phytoecdysteroid group: 20-hydroxyecdysone, norcyasterone, cyasterone, isocysterone and ajugasterone, which is consistent with the results obtained by Matsumoto T, Tanaka N. "Production of Phytoecdysteroids by Hairy Root Cultures of Ajuga reptans var. Atropurpurea", Agric Biol Chem, 1991; 55(4): 1022-1024, doi:10.1271/bbb1961.55.1033.

Total antioxidant activity was determined for the alcohol extracts used in the invention using a modified Brand-Williams method (using a probe molecule - the stable 2,2-diphenyl-1-picrylhydrazyl - DPPH radical) described in "Use of a Free Radical Method to Evaluate Antioxidant Activity", ME Cuvelier and C. Berset, Leb. Technol., vol. 28, pp. 25-30, 1995. The method was modified in such a way that 3.0 cm ^{3 of} 0.1 mM DPPH solution was added to 0.4 cm ³ of the tested extract with a concentration of 0.5 wt.% (obtained by dissolving 0.5 g of dry Ajuga Reptans root extract in 99.5 g of 70% by volume ethyl alcohol). Absorbance was measured 30 minutes after sample preparation at a wavelength of 517 nm. Total antioxidant activity was presented as a percentage of inhibition of oxidation processes initiated by the DPPH radical with respect to the blank. It is 97.52% for extraction with 96% vol. ethyl alcohol and 90.48% for extraction with 70% vol. ethyl alcohol, respectively.

Dry root extracts were evaluated for their safety for application to the skin by testing for irritation potential using the Epiderm® reconstructed epidermal model, as described in H. Kandarova et al., “Optimisation of the EpiDerm Test Protocol for the Upcoming ECVAM Validation Study on In Vitro Skin Irritation Tests”, 2001. The results of the optical density measurement of the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) solution used to determine irritation potential, expressed as cell viability in the reconstructed epidermal model, are summarized in Table 1.

PL 246900 Β1

Table 1

Irritating properties of dry extracts of Ajuga Reptans root

Solvent Relative cell viability [%] 96% vol. ethyl alcohol 100.79 70% vol. ethyl alcohol 120.00 Positive control (5% SDS*) 6.03 Negative control (phosphate buffer pH=7.4) 100.00

*SDS - sodium dodecylsulfonate (sodium lauryl alcohol sulfate)

Ajuga Reptans root extracts obtained by the extraction method presented in the invention do not exhibit irritating potential for epidermal cells and can be safely used in formulations applied to the skin. In addition, in the case of extracts obtained using 70% vol. ethyl alcohol, an increase in the degree of skin cell proliferation by 20% was observed. High metabolic activity of cells in the test on the reconstructed epidermis model Epiderm® indicates an increase in the number of keratinocytes and fibroblasts, which indicates their increased proliferation.

UV/VIS spectra (fig. 1) of extracts from the root of bugleweed, where the eluent is 96% alcohol by volume and 70% alcohol by volume, clearly indicate that these extracts have the ability to absorb radiation in the entire UV range, with the highest absorbance in the UVC range (200-280 nm) and 280 to 350 (UVB). This indicates the protective properties of the extracts obtained against the action of ultraviolet radiation in the entire range (200-400 nm), as well as against the action of blue radiation, from computer monitors and mobile phones, in the wavelength range of 380-500 nm.

In addition, the root extract of Ajuga Reptans in 70% vol. ethyl alcohol was subjected to tests to assess the chelating capacity. The chelating activity was presented as the percentage capacity to chelate heavy metal ions and pollutants present in the air. The results obtained for the Ajuga Reptans root extract were compared to the chelating capacity of 0.1 mM EDTA solution (edetic acid), which is a commonly known chelating agent. The obtained measurement results clearly indicate the high capacity of the tested extract to chelate heavy metal ions. The root extract of Ajuga Reptans has a chelating capacity of 84.7%, which is slightly different from the chelating capacity of the standard solution (0.1 mM EDTA), which is 90.8%. Ajuga Reptans root extract obtained according to the procedure covered by the invention is an active ingredient that protects the skin against the effects of toxins present in the air by effectively binding them.

Example I

Obtaining dry extract of Ajuga Reptans root.

The dry extract of Ajuga Reptans root used in the preparation of the W/O type cosmetic composition was obtained according to the following procedure:

Fresh young creeping bugle seedlings 10-15 cm in size, before flowering, were cleaned under running water to remove any soil and impurities. Then, each plant was rinsed three times with 200 cm ^{3 of} deionized water and the root was separated from the herb. The root was then frozen, freeze-dried and mechanically crushed to obtain particles with a diameter of less than 2 mm. The dry, crushed material was subjected to extraction in a flow-through extractor, Soxhlet apparatus. A 70% vol. aqueous solution of ethyl alcohol was used as the solvent. The extraction was carried out for 5 hours. The ratio of plant material to solvent was 100 : 1 (m/m). The obtained extract was subjected to a filtration process to separate solid residues, then the solvent was pre-evaporated in a vacuum evaporator and dried using a freeze dryer to obtain a dry extract.

PL 246900 Β1

Example II

An example recipe for a nourishing emulsion for use on W/O skin type is presented in Table 2.

Table 2

Recipe for W/O nutrient emulsion

Lp- Phase Name of raw material Function [% wt.] 1 And Glycerin humectant 3.0 2 Magnesium sulfate stabiliser 0.5 3 Sodium benzoate preservative 0.2 4 Xanthan gum rheology modifier 0-5 5 Ajuga Reptans root dry extract obtained according to the procedure of example 1 active substance 0.2 6 Deionized water solvent up to 100.0 7 B Mixture of the following composition: Fatty acid esters derived from olive oil and polyglycerol-6 50-70¾ wt., polyglycerol-6 pentaoleate 30-50 wt.%. emulsifier 6.0 8 Palmitic/stearic triglyceride emollient 2.5 9 Beeswax emollient, consistency regulator, auxiliary emulsifier 3.0 10 Isopropyl myristate emollient, consistency modifier 5.0 11 Cupuacu Butter emollient 2.0 12 Evening Primrose Oil emollient 7.0 13 C Fragrance composition q.s.

The emulsion was prepared according to the following procedure:

The ingredients of the aqueous phase (A) were weighed into a beaker in the following order: glycerin and xanthan gum, then after mixing both ingredients, water, magnesium sulfate, sodium benzoate and dry bugleweed root extract obtained according to the procedure of Example I were stirred until dissolved.

In the second beaker, the components of Phase B were combined: olive oil polyglycerol-6 fatty acid esters, polyglycerol-6 pentaoleate, palmitic/stearic triglyceride, beeswax, isopropyl myristate, cupuacu butter, and evening primrose oil.

Phases A and B were heated to 60°C with continuous stirring to homogenize. Then, maintaining the temperature at 60°C, both phases were combined, introducing the water phase A into the oil phase B and dispersed using a mechanical stirrer at a speed of vi = 700 rpm, then, without stopping stirring, the temperature of the emulsion was lowered. At 40°C, the fragrance composition (C) was added and the temperature was lowered to room temperature, continuing stirring. Finally, the obtained emulsion was homogenized using a high-speed homogenizer, the rotor speed was V2 = 13,000 rpm.

For the nourishing emulsion for application to the skin of the W/O type, obtained according to Example II and for the reference emulsion with the composition according to Table 2 but without the active ingredient, Ajuga Reptans root extract, measurements of total antioxidant activity and chelating capacity were performed.

PL 246900 Β1

For this purpose, a solution was prepared according to the following procedure. 1 g of the ready emulsion was mixed with 5 cm ³ of ethyl alcohol, then the whole was shaken on a Vortex shaker and placed in an ultrasonic bath for 10 minutes to thoroughly crush the emulsion particles in ethyl alcohol. The whole mixture was centrifuged at 3500 rpm for 20 minutes.

The solution above the centrifuged sediment was tested for antioxidant activity according to the modified Brand-Williams method described in "Use of a Free Radical Method to Evaluate Antioxidant Activity", Μ. E. Cuvelier and C. Berset, Leb. Technol., vol. 28, pp. 25-30, 1995. The antioxidant activity of the finished product was 58.73%, and in the case of the reference emulsion without Ajuga Reptans root extract it was 30.54%, which indicates the high ability of the preparation to inhibit oxidation processes by reducing reactive oxygen species.

The chelating capacity was measured using the procedure described by Kubica et al., in the publication “Production of Verbascoside, lsoverbascoside and Phenolic Acids in Callus, Suspension, and Bioreactor Cultures of Verbena officinalis and Biological Properties of Biomass Extracts.” Molecules 2020, 25, 5609. https://doi.org/10.3390/molecules25235609. The capacity to chelate metal ions for the product obtained according to Example II is 22.52%, and in the case of the reference emulsion without Ajuga Reptans root extract it is 0.25%. This proves the high capacity of the preparation containing Ajuga Reptans root extract to chelate heavy metal ions and pollutants present in the air.

Example III

An example recipe for another nourishing emulsion for use on W/O skin type is presented in Table 3.

Table 3

Recipe for W/O nutrient emulsion

No. - Phase Name of raw material Function [% wt.) 1 And Glycerin humectant 4.0 2 Magnesium sulfate stabiliser 0.7 3 Sodium benzoate preservative 0.4 4 Xanthan gum rheology modifier 1.0 5 Ajuga root dry extract Reptans obtained according to the procedure of example 1 active substance 0.5 6 Deionized water solvent up to 100.0 7 B Mixture of the following composition: Esters of fatty acids derived from olive oil and polyglycerol-6 50-70% wt., polyglycerol-6 pentaoleate 30-50% wt. emulsifier 7.0 8 Palmitic/stearic triglyceride emoBent 3.5 9 Beeswax emollient, consistency regulator, auxiliary emulsifier 4.0 10 Isopropyl myristate emollient, consistency modifier 5.5 11 12 13 Cupuacu Butter emollient 2.5 Evening Primrose Oil emollient 8.0 0 Fragrance composition q,s.

PL 246900 Β1

The emulsion according to the recipe in Table 3 was prepared according to the procedure presented in Example II.

For the nourishing emulsion for application to the skin of the W/O type, obtained according to Example III, and for the reference emulsion with the composition according to Table 3 but without the active ingredient, Ajuga Reptans root extract, measurements of total antioxidant activity and chelating capacity were carried out according to the procedure described in Example II.

The antioxidant activity of the product was 59.02%, and in the case of the reference emulsion without Ajuga Reptans root extract it was 30.54%, which proves the high ability of the preparation to inhibit oxidation processes.

The ability of the composition to chelate metal ions was 41.25%, and in the case of the reference emulsion without Ajuga Reptans root extract it was 0.25%. This proves the high ability of the preparation containing Ajuga Reptans root extract to chelate heavy metal ions and pollutants present in the air.

Example IV

An example recipe for another nourishing emulsion for use on W/O skin type is presented in Table 4.

Table 4

Recipe for W/O nutrient emulsion

No. Phase Name of raw material Function [% wt.] 1 And Glycerin humectant 5.0 2 Magnesium sulfate stabiliser 1.0 3 Sodium benzoate preservative 0.5 4 Xanthan gum rheology modifier 1.5 5 Ajuga Reptans root dry extract obtained according to the procedure of example 1 active substance 1.0 Deionized water solvent up to 100.0 7 B Mixture of the following composition: Esters of fatty acids derived from olive oil and polyglycerol-6 50-70% wt, polyglycerol-6 pentacleate 30-50% wt. emulsifier 8.0 8 9 Palmitic/stearic triglyceride emollient 4.5 Beeswax emollient, consistency regulator, auxiliary emulsifier 5.0 10 Isopropyl myristate emollient, consistency modifier 6.0 11 Cupuacu Butter emollient 3.0 12 Evening Primrose Oil emollient 10.0 13 C Fragrance composition

The emulsion according to the recipe in Table 4 was prepared according to the procedure presented in Example II.

For the nourishing emulsion for application to the skin of the W/O type, obtained according to Example IV, and for the reference emulsion with the composition according to Table 4 but without the active ingredient, Ajuga Reptans root extract, measurements of total antioxidant activity and chelating capacity were carried out according to the procedure described in Example II.

The antioxidant activity of the product was 61.56%, and in the case of the reference emulsion without Ajuga Reptans root extract it was 30.54%, which proves the high ability of the preparation to inhibit oxidation processes.

The ability of the composition to chelate metal ions is 57.76%, and in the case of the reference emulsion without Ajuga Reptans root extract it is 0.25%. This proves the high ability of the preparation, containing Ajuga Reptans root extract, to chelate heavy metal ions and pollutants present in the air.

Claims (4)

1. Cosmetic composition in the form of a W/O type nourishing emulsion for application to the skin, consisting of a water phase containing a humectant, a stabilizer, a preservative, a rheology modifier and an active ingredient, an oil phase containing an emulsifier and an emollient and optionally additional ingredients, characterized in that: - as an active ingredient it contains dry extract of the root of Ajuga Reptans, in the amount of 0.1-1.0% by weight, and also contains: - evening primrose oil, 7-10% by weight, - capuacu butter, 2-3% by weight, - isopropyl myristate, 5-6% by weight, - beeswax, 3-5% by weight, - triglycerides of palmitic and stearic acids, 2.5-4.5% by weight, - glycerin, 3-5% by weight, - a mixture consisting of fatty acid esters derived from olive oil and polyglycerol-6 in the amount of 50-70% by mass and polyglycerol-6 pentaoleate in the amount of 30-50% by mass and in the amount of 6-8% by mass, - xanthan gum, 0.5-1.5% by weight, - sodium benzoate, 0.2-0.5% by weight, - magnesium sulfate, 0.5-1.0% by weight, - water to make up 100% by weight of the composition. 2. Cosmetic composition according to claim 1, characterized in that the active ingredient in the form of a dry extract of the root of Ajuga Reptans, is a product obtained by extraction of freeze-dried, crushed Ajuga Reptans root with an aqueous solution of ethyl alcohol with a concentration of 70 to 96% by volume, for 5 hours, at a mass ratio of plant material to solvent of 20-100:1, wherein the solvent is repeatedly recirculated through the plant material, and the extract separated from the solid parts is dried to a constant mass. 3. Cosmetic composition according to claim 2, characterized in that the extraction of the active ingredient from the root of Ajuga Reptans is carried out in a flow-through extractor. 4. Cosmetic composition according to claim 2, characterized in that the dry extract of Ajuga Reptans root is obtained by extraction of freeze-dried Ajuga Reptans root with a 70% by volume aqueous solution of ethyl alcohol.