RO138083A2 - Plant-extract-based topical product for skin conditions that cumulate inflammatory and infectious processes - Google Patents

Abstract

The invention relates to a process for preparing a topical product with effect on skin conditions that cumulate an inflammatory and infectious process, such as acne vulgaris and folliculitis. According to the invention, the process comprises the following stages: making the hydrogel base by combining 0.50% carbopol and 0.25...0.50% usual emulsifier with distilled water, at the ambient temperature, adding, as drops, a mixture of 1.5% vitamin E, 1.00...5.50% tamanu oil, 0.25...0.50% usual surface-active agent and 0.5% triethanol amine, adding the hydrophilic component consisting of 5% standard active extract of hayflower, possibly hayflower tea, in ethanol 40%, to result in a yellowish-white homogeneous product as light hydrogel or cream which provides a content of 100 mg GAE/100 g of hydrogel or cream, with a pH of 5.3...5.5 and specific plant fragrance.

Description

STATE OFFICE FOR INVENTIONS $1 TRADEMARKS Application for a patent for Invention

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Deposit date.........(1.7..7.10.-...¾¾

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DESCRIPTION OF THE INVENTION

Title "Topical product for skin conditions that combine an inflammatory process with an infectious process, based on plant extracts"

Authors: Pîrvu Lucia Camelia, Bâzdoacă Cristina, Rasit luksel

The invention refers to a topical product that is based on an active extract summing up natural compounds from 51 plant species, with action on skin conditions that combine an inflammatory process with an infectious process, such as acne vulgaris and folliculitis, and to the process of its preparation in two forms.

As is known, acne vulgaris and folliculitis are similar in that both conditions occur with the appearance of inflamed pustules, associated or not with an infectious process, located at the level of the hair follicle in the case of folliculitis, respectively at the level of the sebaceous gland in the case of acne, the main difference between these two dermatological conditions being the mechanism of occurrence. Thus, folliculitis is frequently caused by infection with Staphylococcus aureus, but it can also occur outside of a microbial infection; juvenile acne is caused by the clogging of pores with dead skin cells, the excessive formation of sebum by the sebaceous glands, followed or not by infection with Propionibacterium acnes. Both conditions develop with inflamed pustules that, if infected, can be complicated by the spread of inflammation and infection in the deep tissues of the epidermis. In the case of deepening of the inflammation and infection, boils result, which are much more difficult to heal, invariably leave permanent, visible and unsightly scars, it can also lead to the destruction of the hair follicle with hair loss, in the particular case of folliculitis. In this common context of location and unfolding mechanism, two major measures are required with the effect of prevention and treatment of inflamed pustules: increased hygiene from the moment of the appearance of the first signs of dermatological damage, and local intervention with a specialized product with an anti-inflammatory synergistic effect, soothing and antimicrobial, to prevent acne and folliculitis-type lesions from deepening and becoming chronic. As is known, the healing of an injury is a complex process that includes the re-formation of skin tissue layers and the control of infection, to minimize time, appearance and inconvenient scars, therefore a product with a synergistic effect is required.

In Romania, the products dedicated to dermatological conditions such as acne or folliculitis from the adjuvant category (dermatocosmetic products) are found in formulations such as cream, ointment, lotion, hydrogel, stick, patch, etc., and usually have plant extracts as active ingredients, frequently from calendula, plantain, chamomile, amica, or different opotherapy in combination with fatty and volatile oils from different plant sources. Specialized products, from the medicine category, predominate in ointment, cream or hydrogel formulations, and usually have synthetic substances as active ingredients, from the anti-inflammatory or antimicrobial category, rarely synergistic combinations of both classes.

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Apart from the actual active substance, the usual topical formulas contain a variable number of excipients intended to make the product base, to solubilize the active compounds in the proposed formulation, to provide emollient, to facilitate absorption, respectively to offer different advantages in transdermal penetration and target release of active compounds.

Here are two examples from the specialized literature, topical formulations based on combinations of natural compounds and active plant ingredients with anti-inflammatory, anti-algic and cicatrizing action. Thus, the patent RO 110906 Bl describes the composition of an ointment with anti-inflammatory and anti-algesic action: the product, according to the invention, consists of 20...72 parts Flos Amica sii Folium Betulae oil extract, 0.80...1.20 parts Oleum Pini , 26...50 parts ointment base, which is composed of 38.50 parts wax, 46 parts cetyl alcohol, 13 parts coconut oil and 2.50 parts emulsifying agent, to which 50 parts alcoholic extract of Flos Arnicae and Folium Betulae (weight ratios). Patent RO 131198B1 describes the composition for an anti-inflammatory gel based on chitosan and plant extracts: the product, according to the invention, consists of 15...20% chitosan, 30...40% carbopol, 0.8% triethanolamine, 2.. .3% glycerin and 8.25% combined extracts of sulfina, birch, three spotted brothers, rowan and cressora, having a total amino acid content, expressed in glutamic acid, of 0.5...0.07%, and distilled water up to 100%.

The technical problem proposed to be solved by the present invention consists in obtaining a topical product in two formulations, cream and light hydrogel, accumulating natural active compounds from 51 plant species, therefore with multiple active components that converge to stop inflammation and infection in the skin areas and underlying tissues affected by acne and/or folliculitis, or for other skin conditions that combine inflammation with infection; the two formulas can be approved by the patent applicant, either as dermatocosmetic products or as medical devices.

The specific solution consists in combining the active components from a standardized extract obtained from *Ceai Flori de fenugreek with oil from tamanu fruits (Calophyllum inophyllum), in a cream-type formulation, respectively a light hydrogel, based on carbopol - pemulen.

*Hay Flower Tea is a complex vegetable raw material marketed by specialized units in Romania, which reconstitutes a product from folk medicine, namely the set of plant species found in meadows and pastures. Hay flowers are recommended and used in folk medicine for multiple conditions that have as a common denominator inflammation, infection, decreased resistance and immunity, as well as for the purification, rejuvenation and beauty of the skin.

The marketed product *Teai Flori de fan (from Hypericum Impex SRL) is presented as a standardized mixture of 51 plant species, including trees from the spontaneous flora of Romania, plus a series of cultivated plants with very high phytotherapeutic value.

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Scientific arguments regarding the convergence of the anti-inflammatory - antimicrobial calming effect of the set of plant species that make up the plant product *Fanflower tea:

- St. John's wort (Hypericum perforatuni) demonstrated an effect of reducing the level of the pro-inflammatory cytokine TNF-alpha (tumor necrosis factor), and an inhibitory effect on activated lymphocytes from the dermis (P Mansouri et al, 2017), at the same time proven with activity to accelerate wound healing (Ipek Peșin Siintar et al, 2009) and definite antimicrobial effect (Zeb Saddiqe et al, 2010), which supports the use of St. John's wort extracts in topical formulas in which the suppression of inflammation and infection represent the two poles necessary for the correct and rapid healing of the injured tissue;

- mint (Mentha piperita) increased the expression of the TGF-beta gene (a tissue growth and transformation factor that controls cell growth, proliferation, differentiation and apoptosis), which acts specifically in the wound healing process, at the same time it was shown to have an antimicrobial effect cert (Mohammad Ho sein Amirzade-Iranaq et al, 2022), so the extracts are beneficial in topical lesions;

- mouse tail (Achillea millefolium) significantly reduced the expression of inducible nitric oxide synthase (INOS), cyclooxygenase 2 (COX-2) and interleukin 6 (IL-), in studies on Raw264.7 cells stimulated with lipopolysaccharide (LPS), a fact that indicates a definite anti-inflammatory effect, but also the benefit in the treatment of atopic dermatitis and soothing properties on skin lesions (David R Burk et al, 2010);

- horsetail (Equisetum arvense) has a marked cicatrizing effect on the skin and mucous membranes due to its high content in minerals and organic silicon, at the same time it interferes with the function of immunocompetent cells, which gives its extracts a sustained anti-inflammatory effect (Carsten Grundemann et al, 2014);

- hawthorn {Crataegus monogynă) is proven with photo-protective and anti-inflammatory activity by opposing the damage of the extracellular matrix and the disruption of cell proliferation, at the same time by increasing cell survival, which brings benefits in wound healing (Federico Martinelli et al, 2021);

- aloe (Aloe arborescens) and derived products are indisputable in terms of the calming effect and speeding up the healing of skin and mucous membrane injuries, and clinical studies show effectiveness including in addressing chronic and/or (auto)immune skin injuries (Davood Hekmatpou et al., 2021);

- the hazel leaf (Corylus avellana) improves the bactericidal response of macrophages in humans, precisely by acting against Staphylococcus aureus, by increasing the expression of anti-inflammatory genes and iron metabolism (Giulia Cappelli et al, 2018);

- the bear's paw (Heracleum spondylium) has been proven to have specific antifungal and antimicrobial activity on Staphylococcus aureus strains (Daniela Benedec et al, 2017);

- field basil (Prunella vulgaris) has been shown to have an anti-inflammatory and calming effect, as well as an anti-complement effect, which indicates extensive benefits in atopic dermatitis and skin allergies,

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at the same time, it was active on strains involved in skin infections, such as Staphylococcus aureus, Staphylococcus epidermidis and Propionibacterium acnes (Junying Pan et al, 2022);

- chestnut leaf (Castanea sativa) blocked the pathogenicity and virulence of Staphylococcus aureus strains, without allowing the installation of microbial resistance (Cassandra L. Quave et al, 2015);

- chicory {Cichorium intybus) is known for its beneficial effects on the digestive system, but it also has a marked antioxidant, anti-inflammatory and antimicrobial effect (Katarzyna Janda et al, 2021);

- field thyme (Thymus serpyllum) is a reducible antimicrobial, active on strains of Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Yersinia enterocolitica, Listeria monocytogenes, Enterococcus faecalis, Salmonella typhiriwn, Salmonella enteriditis, Actinobacillu spleuropneumoniae, Streptococcus suis, Actinobacillus suis, Haemophilus parasuis, Pasteurella multocida and Bordetella bronchiseptica (Shashank M. Patil et al, 2021);

- maggots (Tagetes patula) have shown marked activity on multiple strains of bacteria, fungi, yeasts, viruses, and even parasites, being frequently used in specialized products (Muhammad Riaz et al, 2021);

- watercress (Alchemilla vulgaris) is very active in the treatment of wounds through its marked astringent properties, proven especially in hydrogel formulations, which underlines the need for a suitable vehicle for the incorporation of plant extracts in topical use (Marija Tasic-Kostov et al, 2019 );

- cretusca (Filipendula ulmaria) by extraction in concentrated ethanol indicated a marked antioxidant activity by suppressing reactive species of intracellular oxygen and mitochondrial superoxide, generated in HaCaT cell cultures exposed to proinflammatory cytokines (TNF-α and interferon-yamma/lY), it also proved active in reducing the synthesis of very strong pro-inflammatory cytokines, such as mitogen-activated protein kinase (MAPK) and nuclear factor kappa-amplifier (NF-κΒ), through the active flavonoids (+)-catechin, michelianin, scutelarin and quercitrin ( Xiao-JieMi et al, 2022);

- black beans and bean pods (Phaseolus vulgaris) have shown a marked antioxidant effect that recommends them punctually in cosmetic applications (David Fonseca-Hemăndez et al, 2021);

- the ash leaf (Fraxinus excelsior) showed synergistic anti-inflammatory, antioxidant, calming and antimicrobial effects, therefore it is suitable in formulas addressed to inflammatory conditions of the dermis, through the specific active compounds esculetin, esculin, fraxin and fraxetin (Iqra Sarfraz et al, 2017);

- marigolds (Calendula officinalis), in polar and non-polar extracts/formulas, are, most likely, with the greatest use in the treatment of skin conditions, the marked cicatrizing effect being the most important attribute of the resulting derived products; the effect is achieved through the accumulation of anti-inflammatory and antibacterial effects, to which are added angiogenic and fibroplastic effects, which act synergistically in the healing phases of wounds of different etiologies (Leila Maria Leal Parente et al, 2012);

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- thistle (Xanthium spinosum), in polar and non-polar extracts, showed a definite inhibitory effect on methicillin-sensitive and resistant strains of Staphylococcus aureus (Javad Sharifi Rad. et al, 2013);

- hops (Humulus lupulus) in the form of a 1% extract demonstrated an anti-inflammatory effect similar to that of hydrocortisone 1%, punctually in oil-in-water, O/W formulations (Zita Hurth et al., 2022), which argues the need for a topical product in 2 formulations, one hydrophilic and the other lipophilic, and their alternative application, to obtain the maximum efficiency of the active components;

- ivy (Hedera helix) is a remarkable anti-inflammatory, comparable to diclofenac; data show that intraperitoneally administered alcoholic extracts inhibit 88.89% of ear inflammation, in a model of mouse edema induced with formalin in vivo, compared to the reference drug, diclofenac, which inhibits the equivalent of 94.44% of inflammation (Anuradha Rai , 2013);

- juniper (Juniperus comunis) in hydro-alcoholic extraction is proven to have genoprotective, antioxidant, antifungal and anti-inflammatory effects (Irina Fierasco et al, 2018);

- deer basil (Mentha pulegium), through luteolin and apigenin, has shown strong antiradical and antibacterial activity, punctually on Staphylococcus aureus sensitive and resistant to methicillin, being thus recommended in combined therapy against pathogenic bacteria (Aires Alfredo et al, 2016);

- stone lichen (Cetraria islandica) in alcoholic extraction showed remarkable antioxidant, antimicrobial activity, especially on Bacillus cereus (Darko Grujicic at al, 2014);

- the poppies fruit (Rosa canina) and derived products are very rich in vitamins (especially Vitamin C) and polyphenols, which ensures an extensive antioxidant effect, both in polar and non-polar environments, being of great interest primarily for reducing oxidative stress (Ines Mărmol et al, 2017);

- birch leaf (Betula verrucosa), in crude extracts, selective fractions, or isolated active compounds have proven to have a very wide spectrum of pharmacological activities, including immunomodulatory, anti-inflammatory, antimicrobial, antiviral and antioxidant effects, which recommends them in products that address skin conditions and the field of dermatology (Subha Rastogi et al, 2014);

- chamomile (Matricaria chamomilla) is a species with very intense use in the dermatocosmetics industry; the derived extracts are preferably included in products aimed at babies and small children, the soothing effect of chamomile being the result of the synergy of antioxidant, anti-allergic, anti-inflammatory and antimicrobial effects (Janmejai K Sri vasta va et al, 2010);

- napraznicul (Geranium robertianum) indicated a strong anti-inflammatory effect and a mechanism of action that suggests an anti-inflammatory activity through the scavenger effect on the NO'f radical Marcelo D Catarino et al, 2017);

- barley (Hordeum vulgare) showed activity against all platelet agonists in humans, except adenine diphosphate, also inhibited the main inflammatory pathways in humans (pathway

R0138083 Α2 cyclooxygenase/ COX and the lipoxygenase/ LOX pathway), in addition, proved a stimulating activity on the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx); punctually, the aqueous fraction was the most effective in stimulating SOD activity, the chloroform fraction indicated the highest efficiency in inhibiting the pro-inflammatory COX pathway, the hexane fraction excelled in inhibiting the LOX pro-inflammatory pathway, at the same time it increased GPx activity (Saima Gul et al , 2014);

- dandelion (Taraxacum officinale) is intensively used in cosmetics for its anti-inflammatory properties by reducing the level of NO, PGE2 cytokines and by reducing the expression of iNOS and COX-2, respectively inactivating the MAP Kinase signaling pathway (Yoon-Jeoung Koh et al, 2010), also indicated antibacterial and antifungal effect; the data also indicate an anti-pigment effect and restoration of skin elasticity, particularly as a result of the proportionate content of Vitamins A, C and E;

- plantain (Plantago major) is another species widely used in the treatment of skin and mucous membranes, especially as a result of its calming, healing and cicatrizing effect, achieved through the synergism of mucilages, allantoin, minerals and active polyphenols that allow a cumulative anti-inflammatory, antioxidant effect , anticomplementary, antiallergic and antimicrobial (Anne Berit Samuelsen. 2000);

- pedicuta (Lycopodium clavatum) is shown to have an anti-inflammatory effect in the extraction and fatty, non-polar formulation, most likely induced by the alkaloid lycopodine (Ilkay Orhan et al, 2007);

- black pine cones (Pinus nigra) in non-polar extract indicated remarkable properties in speeding up wound healing (Ipek Suntar et al, 2012);

- Geranium (Geranium macrorrhizum) is active especially in oily formulation, and nanoemulsions have proven highly effective in the treatment of injuries related to severe burns; punctually, the data indicated a 4-fold decrease in the diameter of burned wounds compared to the untreated control, as well as a 3.81-fold decrease in the serum level of interleukin-6 (IL-6), and a 4-fold decrease in the zone of inhibition against Staphylococcus aureus, compared to the untreated control (Waleed Y. Rizg et al, 2022);

- small-flowered downy mildew (Epilobium parviflorium) is recognized for its marked antimicrobial and anti-inflammatory properties, which are the result of the high content in glycosylated derivatives of myricetin and myricetin-galloyl, it also proved the preference for formulations suitable for oily skin, which is particularly recommended the alcoholic extracts from the poufulita for the treatment of acne pustules, present in the case of people with oily skin (Lucia Pirvu at al, 2015, 2019);

- rostopasca (Chelidonium majus) in combination with microbial cellulose in an in vitro study proved that it reduces the level of secretion of the pro-inflammatory cytokines IL-1 β, IL-8 and INF-α, at the same time it also indicated a marked anti-inflammatory effect, in addition to activity antimicrobial and antibiofilm on Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans, which justifies its use in the treatment of inflamed and infected lesions, and as an occlusive surface dressing (Sylwia Zielihska et al, 2022);

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- acacia (Robinia pseudacacia) indicated an SOD activation effect (by 76.41%), as well as an inhibition effect on the Staphylococcus aureus strain, the formulation in liposomes bringing increased effectiveness in a point experiment of dermal permeability (Soo Hyeon Heo et al., 2022);

- willow bark (Salix alba) and salicin are the most used active ingredients in the cosmetic industry for formulas that address the problem of acne; compared to the separate active ingredient, salicin, the total extract from willow bark, rich in polyphenols, flavonoids and tannins, phenolic acids, salicin, beneficial vitamins and minerals, had the advantage of a synergistic composition (Adolf Nahrstedt et al, 2017), respectively indicated definite anti-inflammatory effect (Mohd Shara et al, 2015), in addition, a stripping effect on the superimposed and thickened layers of cells specific to the acne pustule;

- sage (Salvia pratense) is a plant species with a high content of phenolic acids and flavonoids with strong antioxidant activity, a fact that resides in an intense anti-inflammatory and calming effect, especially on tissues affected by oxidative stress, such as the skin of the face (Umut Kerem Kolac et al., 20170); salvia also indicated definite antimicrobial effects on Enterococcus faecalis, Bacillus cereus, B. subtilis, Staphylococcus epidermidis, S. aureus and S. typhimurium (Nikola Sreckovic et al, 2020);

- the yellow gorse (Galium verum) has antibacterial and antifungal effects on pathogenic and opportunistic strains (e.g., Candida albicans, Trichophyton rubrum and Cryptococcus neoformans), it also has anti-inflammatory activity by inhibiting macrophage activation (Paulina Bolivar et al, 2011);

- the glitter (Potentilla erecta), rich in agrimonin, proved a dose-dependent inhibition effect against UVB-induced inflammation in vivo, also significantly reduced UVB-induced PGE2 production, respectively demonstrated anti-inflammatory properties in vitro and in vivo in the UVB-induced inflammation model, which is highly conclusive for skin lesions (Julia Hoffmann et al, 2016);

- the catfish (Euphrasia roskoviana) protected against UVB-induced photoaging, in a model of normal human fibroblasts, thus proving the benefit in dermal formulas (Ying Liu et al, 2018);

- elderflowers (Sambucus nigră) have an anti-inflammatory effect by modulating the functions of macrophages and neutrophils, through the effect on cell migration and promoting phagocytosis, at the same time they have indicated a relaxing effect on smooth, vascular and non-vascular muscle tissue, thus being an important tool for the management of acute inflammation in humans (Jose RobertoSantin et al, 2022);

- sulfina (Melilotus officinalis), punctual extracts in water, acetone, diethyl ether and ethanol indicated increased antioxidant effects, and the antimicrobial activity screening on 25 microorganisms indicated a greater effect on Gram-positive bacteria than on Gram bacteria -negative, the extract in acetone being the most competent in microbial inhibition (Katarina G. Mladenovic et al, 2016);

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- goose sole (Leonurus cardiacă), through phenols with a 3,4-dihydroxyphenyl type structure demonstrated the ability to restore the production of hypothiocyanite (-OSCN) by lactoperoxidase, an essential enzyme for the humoral immune response against pathogens (Jorg Flemmig et al, 2015);

- linden flowers (Tilia cordata) excel in terms of emollient, calming and anti-inflammatory effects, being an effective remedy in formulas for skin and mucous membrane injuries (Aleksandra Kruk et al, 2022);

- Shepherd's wort (Capsella bursa-pastoris), due to its content in isothiocyanate and sulforaphane, has marked anti-inflammatory and antibacterial properties, punctually it has been shown to decrease the level of nitric oxide (NO), the level of pro-inflammatory cytokines (IL-Ιβ, IL-6 and IL-10) and prostaglandin E2 (PGE2), also induced a decrease in the level of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), which results in a strong anti-inflammatory activity; to these, we add the demonstrated activity on vancomycin-resistant enterococci (VRE) and Bacillus anthracis, therefore it has a wide range of use in pharmaceutical chemistry (Woo Jin Choi et al, 2014);

- three spotted brothers (Viola tricolor), the tincture, significantly reduced the percentages of leukocytes and polymorphonuclear monocytes, it also induced the activation of circulating phagocytes, which resides in an anti-inflammatory effect in hard-to-reach places, such as fatty tissues in the bone marrow (Anca Toiu, 2007);

- the cod (Polygonum aviculare), in an alcoholic extract rich in luteolin derivatives, proved significant anti-inflammatory and analgesic effects, by reducing the release of PGE2 and suppressing the expression of genes mediated by the nuclear factor NF-κΒ, as well as the inhibition of COX-2 (Bing-BingShen et al., 2018);

- nettle (Urtica dioicd), in the form of tea, infusion, decoction or alcoholic extract, is intensively used in the treatment of erythema of the skin and scalp, especially in the context of association with fungal infections, therefore the benefit in skin conditions such as folliculitis is obvious ( Yasaman Taheri et al., 2022);

- raspberry (Rubus idaeus) is known for its antiseptic effect and its usefulness in the treatment of various skin diseases; punctually, it was shown that preparations in ethanol have definite bactericidal and fungicidal activity on the surface of the skin, also the fact that very hydrophilic preparations (logP < 0) feel more effective by being placed in the lipid bilayers in the aqueous core of the liposomes, and not in the lipid shell (Eukasz Kucharsk et al, 2022), which once again indicates the importance of the double hydrophilic and hydrophobic formulation of active plant extracts, as well as the necessity of alternating such complex preparations that sum up compounds with very different polarities, in the treatment of tissue injuries in which coexist an inflammatory process and an infectious process.

Tamanu seed oil (Calophyllum inophyllum) is traditionally used in Indonesia to treat a wide range of skin injuries, from burns, scars and infected wounds to skin diseases such as dermatosis, urticaria and eczema. Tests on keratinocyte cells have proven, on the one hand, the safety in use and, on the other hand, the acceleration effect of wound healing up to 2.1 times higher than the control of untreated keratinocytes. In addition, it has been proven that tamanu oil has 8

R0138083 Α2 effect against Gram-positive bacteria by directly inhibiting mitotic growth, and against Gram-negative bacteria by increased release of Sheta-defensin 2 by activated macrophages, and the concentrations for inhibiting bacterial growth and promoting wound healing have been shown to be lower than those that induce cytotoxic effects. In addition, tamanu oil has been shown to be active on strains of Staphylococcus aureus, the components responsible for inhibiting bacterial growth being found to be in the more polar fraction of the oil, indicating good solubilization in heavy cream (O/W) formulas (Mathieu Cassien et al, 2021: Ravi Raj Pal et al, 2019; Win Min Oo, 2018).

In this very complex compositional context, the topical product, in the two formulation variants, cream and light hydrogel, subject of the invention, can be used as an adjuvant, or as a treatment for skin conditions that occur with inflammation and associated infection; according to the invention, it consists of 0.50% carbopol, 0.25...0.50% pemulen, 0.25...0.50% plurol, 0.5% triethanolamine, 1.5% vitamin E, 1.00...5.50% oil from tamanu fruits and 5.00% standardized extract from Hay Flowers, which gives a standardized continuum of total polyphenols expressed in gallic acid of 100 mg GAE/lOOg active product, and distilled water up to 100% (mass percentages).

The procedure for obtaining the topical product, according to the invention, consists in making the hydrogel base, respectively the cream, by combining at room temperature (18 - 20°C) with continuous stirring (300 - 500 r.p.m.) the carbopol and pemulen components with distilled water, followed by the incorporation into the hydrogel base, respectively the cream, first of the mixture of oils in combination with plurol, then of the standardized active extract from Flori defan so as to result in a standardized concentration of 100 mg GAE / lOOg active product, followed by pH adjustment of the composition at 5.3 - 5.5.

The topical product, in the light hydrogel and cream formulation variants, presents the following advantages: - due to the very high number of active compounds provided by the extract of Hayflowers, the topical product accumulates extensive anti-inflammatory properties, for a large range of pro-inflammatory cytokines, at the same time it has calming effects, speeding up tissue healing, and antimicrobial potential without allowing the installation of antibiotic resistance, infectious germs not being able to counteract such diverse antimicrobial information as that brought by 51 plant species;

- the extract of Hay flowers in combination with tamanu oil and Vitamin E, in the light hydrogel and cream version, provides the context of chemical composition necessary for a synergistic anti-inflammatory, calming and antimicrobial action, necessary for the healing of acne and folliculitis type lesions;

- through the different polarity of the formulas, the conditions for the manifestation of the biological activity of both polar and non-polar active components are created, but it requires the alternation of their application;

- through the proposed formulation, the topical product is homogeneous, stable and quickly penetrates the skin.

- due to the proposed formulation, the topical product does not contain perfume and chemical preservatives;

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- through the proposed formulation, the product has a pleasant smell of plants and self-preservation effect;

- through the standardized active extract from the raw material Flori de fen, the topical product capitalizes at a higher level knowledge from folk medicine, validated by hundreds of years of use.

Below are two examples of the invention, related to the two formulation options, and of the standardized active extract of Fennel flowers, which ensures the therapeutic properties of the topical product, in connection with the tables and figures showing the physico-chemical compositional characteristics, their qualitative and quantitative.

Example 1. Topical product in the form of hydrogel easily usable as an adjuvant or treatment in dermatological conditions that associate an inflammatory process with an infectious process, and which, according to the invention, consists of the following components: carbopol, pemulen, plurol, Vitamin E, triethanolamine, tamanu fruit oil (Calophyllum inophyllum), standardized extract of Hayflower (with a content of 51 plant species). Obtaining procedure: Weigh 0.5 g carbopol, 0.25 g pemulen, 0.25 g plurol, 1.5 g Vitamin E, 1 g tamanu fruit oil (Calophyllum inophyllum), 0.5 g triethanolamine and 5 g standardized extract from Hay Flowers and 91 g distilled water. The ingredients carbopol and pemulen are mixed with distilled water and the hydrogel base is prepared. On top of the hydrogel base, drop by drop, at room temperature, add the mixture of Vitamin E, tamanu fruit oil and plurol. Next, add the standardized extract from Hay Flowers and triethanolamine. It results in 100 g of composition in the form of a light, stable, white-yellow hydrogel with a pleasant herbal smell.

Example 2. Topical product in the form of a cream usable as an adjuvant or treatment in dermatological conditions that associate an inflammatory process with an infectious process, and which, according to the invention, consists of the following components: carbopol, pemulen, plurol, Vitamin E, triethanolamine, oil from tamanu fruits (Calophyllum inophyllum), standardized extract of Hay flowers (with a continuum of 51 plant species). Production process: Weigh 0.5 g carbopol, 0.5 g pemulen, 0.5 g plurol, 1.5 g Vitamin E, 5.5 g tamanu fruit oil (Calophyllum inophyllum), 0.5 g triethanolamine and 5 g of standardized extract from Hay Flowers and 86 g of distilled water. The carbopol and pemulen ingredients are mixed with distilled water and the hydrogel base is prepared. On top of the hydrogel base, drop by drop, at room temperature, add the mixture of Vitamin E, tamanu fruit oil and plurol. Next, add the standardized extract from Hay Flowers and triethanolamine. The result is 100 g of composition in the form of cream, stable, white-yellow in color, with a pleasant herbal smell.

The standardized extract of Hayflowers is obtained from the mixture of dried and ground plants known as *Hayflower Tea, produced and marketed by specialized units (e.g., Hypericum Impex SRL). Periodically, ethanol 70% (v/v), 1:10 g/v, is added over the dry and ground plant, and it is left to macerate for 10-12 days, in the dark, with occasional stirring. The hydroalcoholic extract io

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32 the result is analyzed according to the content in polyphenols, by qualitative methods (e.g., thin layer chromatography, HPTLC) and appropriate quantitative methods (cf. the Pharmacopoeia in force), it is concentrated to the residue, and the residue is resuspended in 40% ethanol (v/ v) so as to result in a standardized continuum in total polyphenols (eg, 2% GAE, w/w). The standardized extract is used as such in the preparation of the topical product.

Note: The inherent variability of plant extracts and secondary metabolites present in them, depending on the annual growth conditions, is compensated quantitatively by using a standardized extract, and from a qualitative point of view it is an advantage because it avoids the installation of resistance.

Analytical and pharmacological testing methods used

Analytical tests on the topical product, in the two formulation variants, and on the standardized active extract of Fennel flowers were performed in accordance with the provisions of the Romanian Pharmacopoeia, Xth Edition, FRX, chapter 2.

For the pharmacological testing, an in vivo model was used, precisely the rat paw edema model induced with a phlogistic agent (carrageenan), and it was carried out in the context of using the patent to create a finished product of the medical device type. The rat paw edema model induced with a phlogistic agent is a common, unanimously accepted model for demonstrating the anti-inflammatory effect of a medicinal product in humans, the size of the edema/inflammation being evaluated by the computerized plethysmometry method. We mention the fact that the pharmacological tests were carried out in accordance with the provisions and regulations of FELASA (Federation of European Laboratory Animal Science Associations), taken over by ARSAL (Romanian Association for Laboratory Animal Science), in force. In order to limit the number of animals tested, the tests were performed only on the topical hydrogel formula.

Thus, according to the qualitative and quantitative analytical tests carried out, the two active formulas present the following physico-chemical and organoleptic characteristics:

No. crt. Characteristics Results and observations per formulation option Light hydrogel formulation Formulation Cream 1. appearance Homogeneous product without separate phases Homogeneous product without separate phases 2. Color White - yellowish White - yellowish 3. Smell Characteristic plants, pleasant Characteristic plants, pleasant 4. pH composition (at 25°C) 5.3-5.5 5.3-5.5 5. Product stability tested at 35°C, minimum 8h Stable Stable

R0138083 Α2

The standardized active extract of Hay flowers in ethanol 40% (v/v) presents the following physico-chemical, quantitative and qualitative compositional characteristics:

No. crt. Quantitative physical-chemical characteristics Hay flower extract in ethanol 40% (v/v) Conditions of admissibility result 1. Density, d ²⁰ 20: 0.970 ± 10% (0.873 - 1.067) 0.98 2. Dosage of total polyphenols, expressed in gallic acid, GAE (g/100 mL): 2.0 ±7.5% (1.85-2.15) 2.05 Qualitative physico-chemical characteristics Thin layer chromatography (HPTLC) is performed for each batch of topical product, necessary to evaluate and compare annual compositional variability 1. It is analyzed: - The appearance of the characteristic bands in the migration and development system for vegetable polyphenols, - 7?/-ul of the bands compared to the reference substances > Routine (Rf-~Q.35 - 0.44) > Ciorogenic acid (Λ/Μ),45 -0.52) > Caffeic acid (Λ/&0.95- 0.98) Below are 2 examples of chromatograms for raw materials, Fan Flower Tea, purchased in two consecutive years (2018 and 2019) ' (iiit iiii w w* j (111 QO '-'W Ibi

The topical product based on the extract of Flori de fam light hydrogel, presented the results of the in vivo testing of the anti-inflammatory effect in the carrageenan-induced rat paw edema model, in comparison with a reference topical product based on diclofenac gel:

Test batches Inflammatory indices (%) at The variation in the volume of edema/inflammation (%) is the end of the experiment 3 hours 5 hours Untreated irritated control lot 29,26 18.56 37% - decrease in volume Irritated test batch treated with a topical product based on diclofenac gel (reference substance) 17.33 27,23 57% - volume increase Irritated test batch treated with the hydrogel base - subject of the invention 46.62 51,52 11% - volume increase Irritated test batch treated with the topical product based on the extract of Fennel flowers in a hydrogel formulation - subject of the invention 58.77 44.85 24% - decrease in volume

CONCLUSION - From the analysis of the volume variation (%) of the edema/inflammation of the test groups compared to the untreated irritated control group, the net superiority of the topical product based on the extract of Hayflowers is found, which supports the synergistic, extended anti-inflammatory effect of the topical product which is the subject of the invention.

Claims (3)

R0138083 Α2 demand 1. Composition for a topical product, formulated as a light hydrogel or cream, which can be used as an adjuvant, or as a treatment for skin conditions that combine an inflammatory process with an infectious process, such as folliculitis and acne, characterized by the fact that it is consisting of 0.50% carbopol, 0.25...0.50% pemulen, 0.25...0.50% plurol, 0.5% triethanolamine, 1.5% vitamin E, 1.00.. ..5.50% tamanu fruit oil and 5.00% standardized extract from Ceai Flori de fen, and distilled water up to 100% (mass percentages). 2. The procedure for obtaining a topical product in the form of a light hydrogel that can be used as an adjuvant or treatment of skin conditions that occur with inflammation and infection, characterized by the fact that a hydrogel base is made from 0.5 g carbopol, 0 .25 g of pemulen and 91 g of distilled water to which is added, first the oily component consisting of the mixture of 1.5 g of Vitamin E, 1 g of tamanu fruit oil (Calophyllum inophylluiri) and 0.25 g of plurol, followed by the hydrophilic consisting of 5 g of active extract standardized in 40% ethanol from Fenugreek flowers which ensures a content of 100 mg GAE /100 g of hydrogel, and the pH is adjusted with triethanolamine. It results in 100 g of composition in the form of a light, stable, yellow-white hydrogel with a pleasant herbal smell. 3. The procedure for obtaining a topical product in the form of a cream that can be used as an adjuvant or treatment of skin conditions that occur with inflammation and infection, characterized by the fact that a cream base is made from 0.5 g carbopol, 0, 5 g of pemulen and 86 g of distilled water to which is added, first the oily component consisting of the mixture of 1.5 g of Vitamin E, 5.5 g of tamanu fruit oil (Calophyllum inophylluiri) and 0.50 g of plurol, followed by the hydrophilic component consisting of 5 g of active extract standardized in 40% ethanol from Fan flowers which ensures a consistency of 100 mg GAE / 100 g cream, and finally the pH is adjusted with triethanolamine. The result is 100 g of composition in the form of cream, stable, white-yellow in color, with a pleasant herbal smell.