WO2017044050A1

WO2017044050A1 - Extract from wood trees of genus abies

Info

Publication number: WO2017044050A1
Application number: PCT/SI2016/000020
Authority: WO
Inventors: Katja ZMITEK; Nataša TAVCAR; Tina POGACNIK; Petra KERSMANC; Tadej REJC; Uroš PETRIC; Borut Strukelj; Samo Kreft; Janko Zmitek
Original assignee: Abies Labs D.O.O.
Priority date: 2015-09-10
Filing date: 2016-09-05
Publication date: 2017-03-16
Also published as: SI25053A

Abstract

The invention refers to the extract from the wood of the fir tree (Abies) genus, preferentially silver fir (Abies alba) wood, in particular from knots, which contain a compound of natural polyphenols with a reduced share of high molecular polyphenols, namely by 15-100% considering the total content of natural polyphenols, and with an analogously increased share of low molecular polyphenols with a molar mass below 1,000 Da, used to prevent, reduce and treat unwanted skin changes in cosmetic and dermatological use, namely the extract alone or in the form of formulated cosmetic and pharmaceutical preparations for cosmetic and dermatological use. Since the share of high molecular polyphenols in the extract is significantly lower and the share of low molecular polyphenols significantly higher than in the compounds or extracts of fir trees known until now, skin penetration is substantially improved. The extract is used either in the form of a concentrate or a solid, a dry compound for dermal use directly or in the form of liquid, semi-solid and solid preparations, such as dermal liquids, including sprays, drops, shampoos and compresses, dermal powders (dusting powders), hydrophilic gels, hydrophilic creams, hydrophobic creams, ointments, pastes, plasters and others.

Description

EXTRACT FROM WOOD TREES OF GENUS ABIES

The subject of the invention is the use of a fir tree wood extract and preparations containing such an extract, for preventing, reducing and treating undesired skin changes, enabling an effective use for cosmetic and dermatologic purposes.

The invention refers to the extract from the wood of the fir tree (Abies) genus, preferentially silver fir (Abies alba) wood, and in particular from knots, which contain a compound of natural polyphenols with a reduced share of high molecular polyphenolic ingredients, to be used in the prevention, reduction and treatment of undesired skin changes, and to formulated pharmaceutical and cosmetic preparations containing such an extract, as well as to the use of such preparations for the indicated purposes.

The following synonyms and abbreviations will be used in the continuation:

- Polyphenolic compounds: polyphenols or polyphenolic substances

- Transepidermal water loss: TEWL

- Minimal erythema dose: MED

The skin is the largest organ of the human body, covering almost its entire surface. It consists of the epidermis, dermis and subcutis. On the basis of various keratinocyte maturing phases, the epidermis is broken down into the stratum basale, stratum spinosum, stratum granulosum, stratum lucidum, and stratum corneum. In addition to basal cells - keratinocytes - the epidermis contains pigment cells (melanocytes), antigen-presenting cells (Langerhans cells), and mechanoreceptors (Merkel cells). The dermis is a firm connective layer providing the skin with mechanical strength (collagen) and elasticity (elastin, partly collagen) or both (hyaluronic acid). The upper layer of the dermis (the papillary layer) furthermore contains blood vessels, lymphatic vessels, nerves and soft connective tissue, excretory ducts of sweat glands, and the central parts of hair follicles, including the sebaceous glands. The dermis is made up of two important vascular plexuses: at the lower border, the reticular layer, is a deep plexus made up of larger vessels, and right below the epidermal-dermal border (papillary layer) is the upper plexus that consists of venules and capillaries. The skin also includes adnexa - appendages (sweat glands, scent glands, hair follicles, sebaceous glands, hairs and nails).

The skin is quite a complex organ; individual layers, cells, extracellular substances, individual components - from various molecules with different functions, many enzymes, to genetic materials - are interconnected through many co-dependent signal processes, biochemical and physiological processes, which regulate the skin functions and its responses to intrinsic and extrinsic influences.

Various external and internal factors can contribute to the production of a large number of free radicals, which results in oxidative stress. A result of oxidative stress can be the destruction of complex actions in the form of cascades (through which the cell differentiation is carried out, for example), where many so-called signal molecules are involved. Damage to the cell structures (cell walls, organelle, nuclear DNA etc.) may also be caused, which appears as an insufficient ability of the cells to produce the key components necessary to ensure a suitable histological skin structure, and consequently poorer regeneration processes, followed by a deterioration in the properties of the skin. The structure and role of the skin is closely connected with skin conditions and changes to the skin. Various factors can lead to many unwanted skin changes that appear as, for example: a reduced activity of the fibroblasts leading to a decrease in the quantity of the basic structural components of the extracellular matrix of the dermis. Due to the weakened function of these components, especially in combination with direct injuries due to free radicals, for example, changes in the connecting tissue appear in the form of wrinkles, uneven skin texture, poorer elasticity, decreased skin firmness, a greater likelihood of developing scars, stretch marks, etc. Structural and physiological changes of the epidermis can also lead to the decreased protective property of the skin, increased permeability and consequently visible peeling, the feeling of tight skin, redness, itching and distinct dryness of the skin. The consequences of unwanted changes of the skin structure or physiological processes of the skin can also be seen as various forms of hyperpigmentation, telangiectasia etc.

The above-mentioned factors may contribute to the development of new or the worsening of existing skin diseases such as various types of dermatitis, psoriasis, rosacea, acne, scalp problems such as dandruff, hair loss, etc. Wound-healing, microcirculation, the inflammatory skin response and others are also weakened.

Various substances and preparations are used for prevention, soothing and treatment, which shows how widespread these problems are and a great need for such preparations.

The problem that has not been properly solved until now, and which is being solved with this invention is the elimination or reduction of unwanted skin changes caused by various intrinsic and extrinsic causes, such as biochemical, physiological and structural changes in the dermis and epidermis, which are reflected in reduced elasticity, smoothness, softness, moisture, skin tightness, changes in pigmentation, microrelief, the number and size of pores, glow, increased wrinkles, weakened skin barrier, changed pH of the skin surface, and other signs, increased sensitivity to UV radiation, changes in the structure and functions of the scalp, as well as the development or worsening of skin diseases such as various types of dermatitis, psoriasis, ulcers, acne, rosacea and other.

Although there is a range of ingredients and cosmetic and pharmaceutical preparations available to treat the mentioned problems, there is a constant need for new, more effective, and in particular natural substances and preparations based on them to prevent, reduce or eliminate the unwanted changes. Despite the trend of increasing needs for natural substance-based medicines for skin diseases, there are a relatively small number of such preparations available on the market.

A problem is also in the fact that individual known ingredients show effects on individual problems or a small number of problems; hence various ingredients are required for various purposes, whereas a complex treatment of the skin requires that the pharmaceutical and cosmetic preparations contain a combination of various ingredients for their topical use. There is therefore a constant need for new, particularly natural substances with a wide range of activities or effects. But here lies a constant problem that, in addition to the effectiveness of the ingredient as such, proved in vitro, the substance must prove the actual effectiveness in the skin - in vivo - and also exhibit the ability to penetrate into the skin to the target area.

The substances used in topical preparations for skin problems include antioxidants, and lately attention has been focused particularly on polyphenols.

Polyphenols are a large group of substances whose basic structural element is the aromatic ring with a minimum of one hydroxyl group. Hydroxyl groups attached to the basic aromatic ring can be free, esterified, etherified or linked to sugar. Structurally, these are highly diverse compounds, which include flavonoids, phenolic acids, lignans, stilbenes, pirones, xantones, quinones, furanocumarins and piranocumarins. They also include tannins which have an astringent effect.

Polyphenols typically exhibit antioxidant, and also some antimicrobial activity, whereas tannins also have an astringent effect. Antioxidant activity is based on the phenolic group which can take an electron, producing a stable phenoxy radical to prevent the oxidation of cell components. Polyphenol compounds therefore act as free radical catchers, particularly those of the reactive oxygen species (ROS), thereby protecting cell lipids, proteins, and DNA against oxidative damage. They can also chelate metal ions present in the production of radicals, and moreover they can increase the activity of cell detoxification systems (such as catalase, superoxide dismutase) and block ROS-generating enzymes. Through the antioxidant activity or through the inhibition of specific proteins (NF-kappaB, AP-1 ) they can exhibit an anti-inflammatory effect, and through the inhibition of certain interleukins they can indirectly affect the normalization of the process of the proliferation and differentiation of the epidermal basal cells. Due to the chelating ability, they have a sequestrating effect and can therefore balance the activity of proteases in the dermis, especially matrix metalloproteinases, and balance the oxidation of tyrosine which is a part of the melanogenesis process, meaning they affect skin pigmentation. By affecting the activity of enzymes from the group of cytochromes P450, cyclooxygenases and lipoxygenases certain polyphenols can affect the inhibition of inflammatory processes or the regulation of proliferative processes and the differentiation of keratinocytes in the epidermis. Through various mechanisms they also help strengthen the blood vessel walls. Moreover, they have an antimicrobial effect.

Due to their antioxidant properties, polyphenols are lately becoming recognized as antioxidants in the form of food supplements, primarily the polyphenols in green tea, soft fruit (raspberries, blackberries, blueberries), grapes and red wine; polyphenol substances obtained from the bark of certain coniferous trees have proven as a good source of antioxidants. The best known among them is pycnogenol (Pycnogenol^®), which is an extract from maritime pine bark (Pinus pinaster). It contains polyphenolic components (catechin, taxifolin, procyanidins with chains of various lengths, of catechin and epicatechin units and phenolic acid), which in oral use show the effects in the fight against chronic and degenerative diseases, while there has been relatively little research - only on animals - done on the effects of the topical use on the skin, whereas in vivo clinical studies of the effects of the topical use of pycnogenol or other similar fir tree extracts are not known.

Typical of pycnogenol is that it contains a relatively high share of high molecular polyphenols substances which are poorly or not absorbed from the intestines at all, and similarly, they only poorly or do not penetrate the skin at all, which is obviously the reason why the use of fir tree wood or bark extracts for the cosmetic and dermatologic industry in the form of preparations for topical use is not known yet or why its effects on the skin have not been proven in vivo yet.

The antioxidant effects of the silver fir tree extract are known and described in the patent application SI P-201400392, and its use in decreasing the concentration of postprandial glucose is also known and described in the patent application SI P-201500116, but so far its effects on the skin are not yet known, since no research in vivo on the activity of silver fir tree extract on the skin of animals or people has been done or published yet.

The invention is explained below and shown in the figures, as follows:

Figure 1 shows the lightening of skin hyperpigmentation, namely hyperpigmentation before the start of use (upper left) and after 12 weeks of using the test cream (upper right), and before the start (bottom left) and after 12 weeks of using the placebo cream (bottom right); the ends of the lines show the compared areas.

Figure 2 shows the reduction in the area of skin hyperpigmentation, namely, the

hyperpigmentation before the start of use (upper left) and after 12 weeks of using the test cream (upper right), and before the start (bottom left) and after 12 weeks of using the placebo cream (bottom right).

The invention refers to the extract from the wood of the fir tree (Abies) genus, preferentially silver fir (Abies alba) wood, in particular from knots, which contain a compound of natural polyphenols with a reduced share of high molecular polyphenols, namely by 15-100% considering the total content of natural polyphenols, and with an analogously increased share of low molecular polyphenols with a molar mass below 1 ,000 Da, used to prevent, reduce and treat unwanted skin changes in cosmetic and dermatological use, namely the extract alone or in the form of formulated cosmetic and pharmaceutical preparations for cosmetic and dermatological use.

The extract is obtained with the extraction from ground or crushed wood of the fir tree (Abies) genus, preferentially silver fir tree wood (Abies alba), preferentially from the wood of the branches, and even more preferentially from knots, meaning from an easily accessible waste ingredient. The ground or crushed wood is heated whilst being stirred in water, preferentially at a temperature above 80°C, and even more preferentially at the temperature of boiling point at an external pressure or at an increased pressure at the temperature between the temperature at boiling point and 150°C. The water phase is separated using filtration; the water is partly evaporated under a reduced pressure to 1/10-1/3 of volume, to obtain a concentrate with an extract concentration of 2-10 wt. %. The concentrate can be added with a solid carrier, such as maltodextrin, cyclodextrin, cellulose, starch or their derivatives to decrease the agglutination and improve the consistency and stability, and the remaining water is entirely evaporated and dried to obtain a dry amorphous compound or a solid, dry compound. The solid carrier is added 0- 30 wt. % compared to the dry substance of the extract, preferentially 5-30 wt. %, and even more preferentially 10-20 wt. %. The extract obtained in such a way, which includes a compound of natural polyphenols and is either in the form of a concentrate or in the form of a solid substance, contains 10-80 wt. %, preferentially 50-80 wt. % of natural polyphenols compared to the dry substance of the extract. The share of the high molecular polyphenols in the extract is reduced or high molecular polyphenols are not present in the extract. The extract contains an increased share of low molecular polyphenols with a molar mass below 1000 Da, such as flavonoids, phenolic acids, lignans and others, and namely, between 50 and 100% of low molecular polyphenols compared to the total share of all natural polyphenols, preferentially 80-100%.

Since the share of high molecular polyphenols in the extract is significantly lower and the share of low molecular polyphenols significantly higher than in the compounds or extracts of fir trees known until now, skin penetration is substantially improved.

The extract is used either in the form of a concentrate or a solid, a dry compound for dermal use directly or in the form of liquid, semi-solid and solid preparations, such as dermal liquids, including sprays, drops, shampoos and compresses, dermal powders (dusting powders), hydrophilic gels, hydrophilic creams, hydrophobic creams, ointments, pastes, plasters and others.

If the extract is used in the form of liquid, semi-solid and solid preparations, the indicated preparations contain 0.5-30 wt. % of the extract compared to the total mass of the preparation, preferentially 1-10 wt. %, and even more preferentially 1-5 wt. %.

In addition to the extract, preparations or cosmetic and dermatologic formulations contain various auxiliary substances, which are common in cosmetic and dermatologic preparations, and other biologically important or active substances for the neutralization of free radicals, substances for the regeneration of components of the epidermis and dermis, substances for the prevention of the production of senescent cells, substances for the relaxation of the small muscles that cause skin wrinkles, and substances that prevent or balance the production of melatonin. These substances are, for example, vitamins, minerals, other antioxidants, amino acids and other acids, peptides, protein, saccharides, fats and extracts and secretions from plants, animals and their tissue cultures, which work synergistically with the extract, thus supplementing or reinforcing the effects of the relevant extract. We conducted in vivo studies on the effects of the extract which contains natural polyphenols; we found many surprisingly positive effects on the changes in the dermis, epidermis and skin surface, which are reflected primarily in the stimulation of the production and regeneration of structural proteins such as collagen and elastin, and consequently in the improvement of the structure of the dermis, and the stimulation of the regenerative processes that result in an improved condition of the epidermis. This is shown in:

- Improved microcirculation, elasticity, smoothness, softness, moisture, skin tightness, microrelief, glow, pH, protection barrier and skin function, skin resilience, e.g. reduced erythemic response to UV rays, i.e. increased resistance against the effects of UV radiation, wound healing and scarring, stretch marks, the condition of the scalp and hair, swellings and puffiness, etc.,

- Reduced acne, wrinkles, hyperpigmentation such as lentigo senilis, lentigo Solaris, melasma, post-inflammatory and post-traumatic hyperpigmentation, sun freckles, the expressiveness of telangiectasia, inflammatory changes, certain allergy and reactive skin diseases, etc.

In vivo studies were carried out with the cream after the invention and with a placebo cream. The relative average results with the cream after the invention compared to the placebo cream show:

- Reduced wrinkles, namely the volume by 33% and the area of wrinkles by 29%;

- 17% improvement in moisture;

- 19% decrease in TEWL, pointing to a significant improvement of the skin barrier;

- 13% decrease in retraction time and 16% increase in viscoelasticity, pointing to a significant increase in skin elasticity;

- 31 % increase in the intensity of the ultrasonic signal rebound of the dermis, pointing to an improvement in the structure of the dermis or the condition of the structural proteins - collagen and elastin;

- 2% increase in dermis thickness; - 8% increase in the minimal erythema dose (MED), pointing to an improvement in the photoprotective function of the skin and its response to UV radiation and a decrease in the possibility of inflammatory reactions;

- Female volunteers positively assessed the effects of the test cream in comparison to the placebo for virtually all parameters that were not within the limits of the normal condition;

- Expert evaluations of the test cream effects were positive for virtually all parameters that were not within the limits of the normal condition.

Individual responses were significantly higher compared to the initial state.

The obtained results therefore prove that the extract and preparations containing the extract surprisingly affect many properties and functions of the skin by significantly improving the negative skin changes, whilst not affecting the normal skin condition. At the same time, it is important that the extract is natural, which makes the invention follow the trend of using natural ingredients, and moreover, the ingredient is actually a wood processing waste material and as such easily accessible and inexpensive.

The invention solves the problem of the effective prevention, soothing and treatment of skin changes due to various intrinsic and extrinsic causes, such as biochemical, physiological and structural changes in the dermis and epidermis, which are reflected in reduced elasticity, smoothness, softness, moisture, skin tightness, pigmentation changes, microrelief, the number and size of the pores, glow, increased wrinkles, weaker protective barrier, change of the skin surface pH and other signs, increased sensitivity to UV radiation, changed structure and functions of the scalp, as well as in the development or worsening of skin diseases, such as various types of dermatitis, psoriasis, ulcers, acne, rosacea and others.

The invention is explained, yet not limited by the implementing cases. The following apparatuses were used to analyse the effects of the compound and the preparations on the skin:

- Cortex DermaLab Combo SkinLab - Cortex Technology ApS, Smedevaenget 10, 9560 Hadsund, Denmark; probes for the analysis of the dermis (structure/intensity and thickness of the dermis), elasticity, transepidermal water loss, moisture, colour/redness/pigmentation (CIE LAB)

- VisioFace Quick (VFQ), round shaped model - Courage + Khazaka electronic GmbH, Mathias Bruggen-Str. 91 , 50829 Cologne, Germany (for photoanalyses)

- Dr. Hoenle Dermalight 80 MED-Tester (for the determination of the minimal erythema dose - response to UV rays)

- DermaScope MEDL4D, Dino-Lite, Netherlands (visual analyses)

- Multi Skin Test centre - Courage + Khazaka electronic GmbH, Mathias Bruggen- Str. 91 , 50829 Cologne (probes for moisture, elasticity, pH, TEWL, redness, pigmentation/melanin)

Case 1 - preparation of the extract concentrate and the solid, dry compound of the extract from silver fir wood

5 I of boiling deionized water were poured over 1 kg of crushed branches of the silver fir (Abies alba), which is one of the species of the fir genus (Abies sp.); while stirring, the mixture was heated to boiling point for 20 min. The samples were then filtered under a lowered pressure and the water extract was dried with a combination of drying with a rotary evaporator and lyophiliser. Once the volume was reduced to approx. 1/3, 0.5 I of the solution was removed to obtain a concentrated solution of the extract with a concentration of approx. 3%, and approx. 10% of maltodextrin was added to the rest (expressed as a share of maltodextrin to the dry mass of the extract) and the compound was dried. The prepared concentrated solution of the extract and the solid, dry compound of the extract were used for analyses, the preparation of the formulated preparations and the implementation of in vivo studies. We carried out analyses with colour reactions to determine polyphenols, the method of skin powders (the determination of the share of high molecular polyphenols), and with HPLC (mobile phase A: water, 0.1 % TFA, mobile phase B: acetonitrile; gradient: B (0-1 min 5% B, 1-10 min 5-30% B, 10-15 min 100% B, mobile phase flow 2 ml/min; 20 μΙ of the sample). The analyses showed approx. 60% of the polyphenols in the dry substance, of which approx. 90% were low molecular polyphenols with the molar mass below 1 ,000 Da, predominantly phenolic acids, flavonoids and lignans.

Case 2 - cream

Ingredient Quantity of the ingredient in

100 g of the preparation

Phase a Lanette N 5.00

Stearic acid 3.50

Cetanol 0.50

Tegosoft CT 3.00

Apricot oil 8.00

Cetiol SN 2.00

Ti0₂ 0.20

Paraffin oil 1.00

Phase b Water, demineralized 68.10

Glycerol 3.00

Urea 1.00

Phenonip 0.50

Extract from Case 1 2.00

Phase c Water, demineralized 2.00

TEA 0.20

Phase d Fragrance 0.20 The ingredients of phase a) were weighted in a 400 ml cup, and the ingredients of phase b) in a 200 ml cup. Both phases were heated to 75°C. The heated phases were then combined by adding the water phase into the oil phase while mixing with an electrical mechanic blender. Demineralized water and TEA - phase c) were weighted in a 20 ml cup and stirred well to obtain a clear solution which was added to the phase a) and b) compound. After a few minutes of mixing, phase d) was added. The obtained emulsion was cooled to room temperature while constantly mixing it, and the obtained cream with a 2% extract content was put into a container. The cream was used for the in vivo study of effects.

Case 3 - in vivo study of skin effects

10 female volunteers, aged 45-65 years, phototypes II and III, with various changes or skin conditions, applied the cream from Case 2 to one half of the face and a half of the gluteal part twice daily, and to the other half of the face the placebo cream, i.e. the preparations that were not changed from the start of the study and regime, over a period of 12 weeks. The volunteers signed informed consents, and the study was approved by the competent Ethics Commission. We used measurements, photoanalyses, expert evaluations and self-evaluations to analyse the skin condition at the start (one day before starting with the cream application - Term 1 ), after 6 weeks (Term 2) and after 12 weeks (Term 3). The results are presented in Tables 1 , 2 and 3.

Table 1 : Average absolute and relative change of the volume and area of wrinkles in the lateral periorbital part of the eye (Crow's feet) between Terms 1 (before the use of the cream) and 3 (after 2 weeks of the use of creams) of the measurements on the right (test cream, 2% of the compound) and the left cheek (placebo cream)

Table 2: Average absolute and relative change of moisture, elastic module (E),

viscoelasticity (VE) and retraction (skin elasticity parameter), transepidermal water loss (TEWL), thickness and intensity of the dermis between Terms 1 (before the use of creams) and 3 (after 12 weeks of the use of creams) of measurements on the right (test cream, 2% of the compound) and left cheek (placebo cream).

RIGHT CHEE K - cream LEFT CHEE - placebo

Change Relative change Change between Relative change between between Terms 1 and Terms 1 and 3 between Terms 1 Terms 1 3 and 3 and 3

Moisture 42.9 μ$ 27.2% 15.6 μ≤ 9.6%

VE 0.3 MPa 38.2% 0.1 MPa 22.0%

Retraction -866.5 ms -23.2% -522.4 ms -9.8%

-2.6

TEWL g/cm²/h -19.4% -0.1 g/cm²/n -0.7%

Thickness of

dermis 38.8 μιη 3.6% 15.1 μηι 1.6%

Intensity of

dermis (index

0-100) 3.5 18.4% -3.2 -12.6% Table 3. Improvement in the photoprotective function of the skin - changes in the minimal erythema dose (MED)

Case 4 - reduced wrinkles

A female volunteer, aged 45 years, used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. The analysis of the selected wrinkles in the lateral periorbital part of the eyes (Crow's feet) showed a reduced depth of wrinkles by 50% and the surface area by 48% compared to the placebo.

Case 5 - improvement of elasticity and moisture and less expressive telangiectasia and couperose

A female volunteer, aged 51 years, with expressively dry skin in the area of the nose, the wider area of the cheeks and chin, with an expressive telangiectasia in the central area and an expressive couperose used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. The analysis of the skin elasticity measurement results showed an improvement in viscoelasticity (VE) by 58%, an improvement in the elastic module E by 5%, and a 30% improvement in the moisture compared to the placebo, and a visible reduction in the telangiectasia and couperose expressiveness.

Case 6 - improved skin elasticity

A female volunteer, aged 54 years, with an expressively poor skin smoothness, expressive microrelief and poor skin elasticity used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. The analysis of the skin elasticity measurement results showed an improvement in the time needed for skin retraction by 13% compared to the placebo; the microrelief and skin smoothness improved considerably.

Case 7 - improved dermis structure and skin barrier

A female volunteer, aged 50 years, showing skin ageing signs and mimic wrinkles and a weakened skin barrier (increased TEWL), used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. An analysis of the dermis structure and the TEWL measurement results showed an increased intensity in the ultrasonic signal response (improved condition of structural proteins - collagen and keratin) by 84% and a reduced transepidermal water loss (improved protection function of the skin) by 26% compared to the placebo.

Case 8 - increased dermis thickness

A female volunteer, aged 64 years, with photodamaged skin, expressive sun freckles (ephelides) and baked apple morphotype wrinkles, smoker, with an expressively poor condition of the dermis used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. The analysis of the dermis structure measurement results showed an improved thickness of the dermis by 16% compared to the placebo.

Case 9 - increased minimal erythema dose

A female volunteer, aged 54 years, phototype III, used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. The analysis of the minimal erythema dose measurements showed that a 12% higher UV radiation dose was needed for the development of redness than before the use, while there was no difference in the case of the placebo cream.

Case 10 - lightened hyperpiqmentation

A female volunteer, aged 55 years, phototype II, with expressive hyperpigmentation - solar lentigo in the lateral periorbital part of the face used the cream from Case 2 and the placebo cream such as stated in Case 3, for 2 weeks. We measured the colour (E) and brightness (L) of the hyperpigmented area - solar lentigo, and the area next to it; the results are presented in Figure 1 and Table 4.

Table 4. The pigmentation measurement results: AL - difference in brightness L between the hyperpigmentation and the area next to it; relative change AL - relative change AL between terms 1 and 3 (after 12 weeks of using the cream); ΔΕ - difference in colour E between the hyperpigmentation and the area next to it; relative change ΔΕ - relative change ΔΕ between terms 1 and 3 (after 12 weeks of using the cream).

The analysis of the pigmentation measurement results showed significant lightening and change of colours, namely, the hyperpigmentation became lighter by approx. 63% compared to the area next to it, and compared to the placebo results by 100%; the difference in colour between the hyperpigmentation and the area next to it dropped by 66% (and in the case of the placebo the difference was only around 12%).

Case 1 1 - reduced area of hyperpigmentation

A female volunteer, aged 45 years, phototype III, with expressive age-related hyperpigmentation in the lateral periorbital part of the face used the cream from Case 2 and the placebo cream such as stated in Case 3, for 12 weeks. During the study, the volunteer was occasionally exposed to the sun, which resulted in darkened skin. The surface area of the selected hyperpigmented parts was measured. The results are presented in Figure 2 and Table 5.

Table 5. Surface areas of the selected hyperpigmented areas before and after the use of the cream and placebo.

Figure 2 and Table 5 show a considerable reduction in the surface area of the hyperpigmentation after the use of the cream (by 68%), while the use of a placebo even resulted in an increase by 38% as a result of exposure to the sun or UV rays. The differences in the results for the cream and placebo show, in addition to the effectiveness of the compound, an increase in the protective function of the skin and a reduced proneness to developing hyperpigmentation.

Case 12 - complex cream with added ingredients

Structure:

Ingredient Quantity of the ingredient in 100 g

of the preparation

Phase Myritol 318 7.00

a

Lanette O 2.50 Almond oil 5.00

DC 245 1.5

Microcapsules with squalene 0.015

Phase Demi water 60.10

b

EDTA 0.8

Emulgade F special 1.5

Glycerine 5.00

Phenonip 0.50

Phase Xanthan gum 0.20

c

Water 10.00

Phase Hydroviton 1.00

d

Aloe barbadensis extract 3.00

Extract from Case 1 1.50

Vitamin E acetate 0.10

Milk protein 1.00

Phase NaOH When necessary

e

Preparation of phase c): 10 g of distilled water is weighted in a 40 ml cup. Following this, 0.20 g of Xanthan gum is separately weighted, added into the cup, and stirred with a glass stick for approx. 10 minutes for the Xanthan gum to dissolve. Preparation of phase b): 60.10 g of distilled water is weighted in a 400 g cup. 0.8 g of EDTA is added, and stirred with a glass stick for approx. 10 minutes was allowed for the EDTA to dissolve. Then 1.5 g of emulgade F special is added. 5.00 g of glycerol and 0.5 g of phenonip is added to phase b). This is followed by stirring everything and starting to heat the ingredients in a water bath. Preparation of phase a): In a 250 ml cup, 7 g of myritol 318 (Tegosoft CT), 2.5 g of lanette O, 5 g of almond oil, 1.5 g of DC 245 and 0.015 g of microcapsules with squalene are weighted one after another. The cup with the mixture of all the added ingredients is heated in a water bath. While heating the water and oil phases in water bath, their temperature is constantly monitored. When both phases reach the temperature of 78-80°C, the cups are removed from the water bath and the content of the water phase in the cup is mixed with a mechanical blender. Afterwards, the oil phase is slowly added to the water phase at a constant mixing speed. While mixing the emulsion, phase d) is prepared. Preparation of phase d): When the temperature of the emulsion drops to 38°C, the above-mentioned ingredients of phase d) are added one after another. The emulsion is mixed at a constant speed for some time.

We now have the cream which contains the extract and other ingredients that work synergistically with the extract to increase its effects.

Claims

1 . The extract from the wood of the fir tree genus, preferably silver fir, containing natural polyphenols for use in the prevention, soothing and treatment of the changes on the skin of the face, body and scalp resulting from the biochemical, physiological and structural changes in the dermis and epidermis.

2. The extract according to claim 1 for use in the stimulation of the production and regeneration of structural proteins such as collagen and elastin, in the improvement of the dermis structure, in the stimulation of the regeneration processes resulting in the improved condition of the epidermis.

3. The extract according to claims 1 and 2 for use in the improvement of microcirculation, elasticity, smoothness, softness, moisture, skin tightness, microrelief, glow, pH, skin barrier and resilience, in the reduction of acne, wrinkles and hyperpigrnentation, and in the treatment of dermatitis, psoriasis, ulcers and rosacea.

4. The extract according to claims 1 to 3, characterized in that a content of natural polyphenols is between 0 and 80 wt. %, preferentially between 50-80 wt. % compared to the dry substance of the extract, and the content of low molecular polyphenols with a molar mass below 1 ,000 Da is between 50% and 100% compared to the total share of all the natural polyphenols, preferentially between 80-100%.

5. The extract according to claims 1 to 4, characterized in that low molecular polyphenols with a molar mass below 1 ,000 Da are preferentially flavonoids, phenolic acids and lignans.

6. The extract according to claims 1 to 5, characterized in that it is used in the form of a concentrated water solution with a concentration of the extract between 2% and 10% or in the form of a solid, dry compound.

7. Cosmetic and dermatological preparations characterized in that they contain the extract according to claims 1 to 6.

8. The preparations according to claim 7, characterized in that they contain from 0.5-30 wt. % of the extract compared to the total mass of the preparation, preferentially from 1-10 wt. %, and more preferentially from 1-5 wt. %.

9. The preparations according to claims 7 and 8, characterized in that they are liquid, semi-solid or solid.

10. The preparations according to claims 7 to 9, characterized in that they contain other biologically active and auxiliary ingredients that work synergistically.

11. The preparations according to claim 10, characterized in that biologically active substances are selected from vitamins, minerals, other antioxidants, amino acids and other acids, peptides, proteins, saccharides and fats, and extracts and excretions from plants, animals and their tissue cultures.

12. The use of the extract according to claims 1 to 6 and the preparations according to claims 7 to 1 for the topical application to the skin of the face, body and scalp.