Compositions and methods for treating and managing chronic inflammatory skin disorders

The present invention is directed to compositions and methods for treating and managing symptoms associated with inflammatory skin disorders by applying a synergistic blend of plant-based actives containing at least: (a) Calendula officinalis stem cell extract; (b) Symphytum officinale extract; and (c) a biotech-derived Haberlea rhodopensis extract.

FIELD OF THE INVENTION

The present invention generally relates to compositions and methods for treating and managing chronic inflammatory skin conditions. More particularly, the present invention provides ways for effectively treating and managing inflammatory skin disorders by addressing their symptoms which include a structurally impaired stratum corneum (SC), skin inflammation, and oxidative stress caused by excessive amounts of reactive oxygen species (ROS) present in the skin.

BACKGROUND OF THE INVENTION

Skin represents the body's first line of defense against external/environmental insults that would otherwise damage sensitive underlying tissue and organs. In addition, it also functions as a relatively water-impermeable barrier that keeps water/moisture from prematurely escaping from the body. This barrier is subject to numerous extrinsic and intrinsic negative factors that affect its appearance, physical properties, and physiological functions. Extrinsic factors include ultraviolet radiation, environmental pollution, heat, harsh surfactants, and mechanical stress caused by, for example, shaving and the like. Intrinsic factors include chronological aging, a person's genetic makeup, and other biological conditions such as atopic dermatitis (i.e., eczema), psoriasis, acne vulgaris, and Hidradenitis Suppurativa (HS).

The skin is divided into two main structural layers: the epidermis and the dermis. The vital barrier function of the skin resides primarily in the top stratum of the epidermis, the stratum corneum (SC), a.k.a. the skin barrier. The epidermis also has immunologic functions and provides some protection from UV light via its pigment system. The predominant cell type of the epidermis is the keratinocyte which represents approximately 90% of the cells comprising the epidermis. They originate in the deepest layer of the epidermis (stratum basal) and migrate up to the final barrier layer, the stratum corneum.

Keratinocytes proliferate in the basal layer and start differentiating on their way to the surface. During this process, they significantly change their morphology and start to produce keratin, cytokines, growth factors, and interleukins. They play an essential role in protection as they form a tight barrier that prevents foreign substances from entering the body, while minimizing the loss of moisture, heat, and other constituents. These cells have a structural role, forming tight bonds with other cells in the epidermis and maintaining them in their locations. In addition, keratinocytes function as immunomodulators following skin injuries. At each stage of differentiation, keratinocytes express not only specific keratins, but various biological markers as well such as involucrin, loricrin, transglutaminase, filaggrin, and caspase.

In addition to their structure-forming properties, keratinocytes also interact with other skin cells such as fibroblasts, melanocytes, lymphocytes, and Langerhans cells. Crosstalk between keratinocytes and fibroblasts is essential for maintaining skin homeostasis and for ensuring a balanced wound-healing process. These two cell types communicate via paracrine signaling mechanisms, the disruption of which can lead to the presence of chronic wounds. The interaction between keratinocytes and melanocytes is also critically important for skin barrier homeostasis. Melanocytes produce melanin which absorbs UV light and prevents DNA damage to the keratinocytes which, in turn, stimulate melanocyte functions such as proliferation, differentiation, and melanogenesis. Keratinocytes also possess immunomodulatory functions that interact with lymphocytes and Langerhans cells in the skin.

Keratinocytes also play a role in wound-healing via a process known as re-epithelialization, necessary for successful wound closure. When the skin is injured, keratinocytes are activated and migrate to the wound where they start proliferating to fill/close the wound. During wound healing, interactions between keratinocytes, fibroblasts, and immune cells are critically important to the healing process.

When the skin barrier/SC is compromised (i.e., structurally impaired) or when pathogens enter the skin, an inflammatory response is triggered. Keratinocytes actively participate in this process by expressing cytokines that transmit positive or negative signals to immune cells. They also play a role in chronic inflammatory skin conditions like eczema and psoriasis by recruiting and activating dendritic cells and leukocytes.

Corneocytes are essentially “dead” keratinocytes, i.e., they are devoid of nuclei and cytoplasmic organelles. They are primarily comprised of the protein keratin and are continually shed (desquamated) and renewed from lower epidermal layers, a process commonly referred to as cell turnover.

The SC is often analogized to a brick wall. The corneocytes with their resistant cell envelopes and keratin microfibrils are the bricks, whereas the layers of lipids found between the cells are the mortar. This lipid mortar is the main barrier to water prematurely escaping via the stratum corneum. Both the bricks and mortar of the stratum corneum are produced by keratinocytes at the stratum granulosum where keratinocytes release the lipids (mortar) into the spaces between the cells as they are being transformed into the corneocytes (bricks). Two proteins from the keratohyalin granules, filaggrin and loricrin, play key toles in the formation of the corneocytes. Filaggrin is an acronym for filament-aggregating protein.

The lamellar bodies that appear at the stratum granulosum contain lipids which are released into the intercellular spaces as the stratum corneum forms. The lipids are comprised of glucosyl ceramides, cholesterol esters, and long-chain fatty acids. In the intercellular space, the glucosyl ceramides are converted to ceramides. The lipids spontaneously organize into multiple layers between the stratum corneum cells.

This lipid mortar is critically important to skin barrier function, with ceramides being vital to the organization and functioning of the barrier. The lamellar bodies of the stratum corneum release other important molecules into the intercorneocyte spaces in addition to the lipids that form the permeability barrier. The proteolytic enzymes involved in desmosome hydrolysis as well as inhibitors of those enzymes to control rates of desquamation are released by the lamellar bodies, as are antimicrobial peptides called “defensins”, which play a role in protecting the skin from infection. Production of these peptides is increased in skin with psoriasis, but not in skin suffering from atopic dermatitis (eczema). This is believed to account for the increased susceptibility of eczema sufferers to skin infection and eczematous skin may be more likely to spread nosocomial infections.

The very specialized bricks and mortar of the stratum corneum work together to produce a covering for the skin that is both flexible and protective. When functioning properly, it defends the body against dehydration, external toxins, and bacterial assault, while protecting the more fragile keratinocytes below from mechanical disruption.

FIG. 1 depicts how the skin barrier functions when healthy vs. when impaired. As can be seen in FIG. 1, a breakdown of the skin barrier results from a breakdown in the tight adhesion of the cells in the SC, and the loss of intercellular lipids (e.g.: ceramides) thereby allowing excess egress of moisture and pronounced ingress of pathogens. What is not so clear is whether the breakdown of the skin barrier is initially caused by external/environmental influences (e.g. UV rays, pollution) or by internal influences (e.g., cytokines, chemokines and ROS) residing below the epidermis.

Atopic dermatitis (eczema) and psoriasis are two common chronic immune-inflammatory diseases. While each of these diseases has its own unique etiology or set of etiologies, they all share a common trait. This trait is a compromised skin barrier, which is often accompanied by inflammation, and pruritus. This path can be reduced to a basic description which generally consists of three fundamental steps, though not necessarily in this precise order given the two competing theories for the genesis of these diseases. In the “outside-in” hypothesis, epidermal barrier dysfunction triggers immune activation; in contrast, based on the “inside-out” hypothesis, AD is primarily cytokine driven with secondary skin barrier dysfunction.

The modern approach to defining AD pathogenesis appears to now be centered on integrating these two mechanisms and is oriented toward characterizing their interplay in AD as it is not clear which individual model is implicated or whether a combination of the two models explains the disease process. (Facheris, P., Jeffery, J., Del Duca, E. et al. The translational revolution in atopic dermatitis: the paradigm shift from pathogenesis to treatment. Cell Mol Immunol 20, 448-474 (2023). doi.org/10.1038/s41423-023-00992-4)

Myriad central genetic mutations have been demonstrated in the pathogenesis of AD: mutations in structural epidermal barrier proteins, mutations in functional proteins that maintain the epidermal barrier, and mutations in factors that regulate the immune system. Skin barrier alterations, with dysfunction of barrier-related proteins such as filaggrin (FLG), loricrin (LOR), and involucrin (IVL), are the first steps that explain the subsequent sensitization against allergens and the so-called “atopic march” in the “extrinsic” form of AD, where the extrinsic form of AD has a signature increase in total serum IgE levels. Alteration of the skin barrier is also related to alteration of the intercorneocyte lipid composition (e.g. ceramides), with further trans-epidermal water loss (TEWL). This, in turn, contributes to the skin dryness typical of AD patients and increased penetration of allergens and pathogens creating a feedback loop.

Epidermal barrier dysfunction in AD is characterized by a lower expression of terminal differentiation markers, such as filaggrin (FLG) and loricrin (LOR), and by a higher permeability defect caused by skin lipid film impairment and higher trans-epidermal water loss. This makes AD skin more prone to the penetration of external agents (antigens, allergens, pollution, etc.) that are harmful to keratinocytes. Damaged keratinocytes produce epidermal alarmins such as IL-33, IL-25, and TSLP, which activate the dendritic cells (DCs) and type 2 innate lymphoid cells (ILC2s) that produce IL-5 and IL-13, which activate eosinophils and Th2 cells. Local Th2 polarization, in return, further diminishes barrier functions and sustains itching, causing skin barrier impairment and facilitating dysbiosis. AD skin shows a higher proliferation of the members of the genus Staphylococcus, especially S. aureus, which can further damage keratinocytes and sustain local inflammation.” (Facheris, P., Jeffery, J., Del Duca, E. et al. The translational revolution in atopic dermatitis: the paradigm shift from pathogenesis to treatment. Cell Mol Immunol 20, 448-474 (2023). doi.org/10.1038/s41423-023-00992-4)

Conversely, while sharing a similar Th2 signature, “intrinsic” AD is characterized by increased Th1 expression in the blood compared to extrinsic AD and lower CCL17 levels. Intrinsic AD also shows greater cellular infiltrates with T cells, Langerhans cells, and myeloid DCs. Th17- and Th22-driven expression of the antimicrobials S100A9 and S100A12 was found to be higher in intrinsic AD, with Th17 expression positively correlating with disease severity. (Facheris, P., Jeffery, J., Del Duca, E. et al. The translational revolution in atopic dermatitis: the paradigm shift from pathogenesis to treatment. Cell Mol Immunol 20, 448-474 (2023). doi.org/10.1038/s41423-023-00992-4)

The microenvironment of a compromised skin barrier contributes, especially in the acute phase, to the activation of the immune system with a predominantly, but not exclusively Th2-mediated reaction and the release of proinflammatory cytokines and chemokines. On the other hand, this process may occur in the opposite order with proinflammatory processes leading to a disturbed skin barrier. Either way, these cytokines and chemokines include alarmins such as tumor necrosis factor (TNF) and interleukins (IL-4, IL-9, IL-22). Epithelium-derived cytokines, such as thymic stromal lymphopoietin (TSLP), IL-25 and IL-33 also act as alarmins after appropriate stimulation, including oxidative stress (OS), and activate a Th2-mediated response in AD, contributing to the inflammatory state of the skin.

The persistence of Th2 inflammation and skin barrier disruption contributes to chronic inflammation and to the overproduction of reactive oxygen species (ROS), such as superoxide and hydrogen peroxide. In addition to this mechanism, the increase in ROS may also depend on other exogenous factors such as solar radiation, pollution, psychological stress, and infections. For example, Staphylococcus aureus, a pathogen that frequently causes skin infections and flares up in patients with AD because of a disruption in the skin barrier such as in the case of filaggrin deficiency, can sustain skin inflammation through ROS released by monocytes activated by the pathogen itself. Over time, the accumulation of ROS can eventually cause oxidative stress due to an imbalance between the generation of excess ROS and the body's ability to produce ROS-neutralizing antioxidants.

Inflammatory skin disorders characterized by itch as the primary symptom encompass a wide range of conditions, each with its unique underlying causes and manifestations. These disorders, collectively referred to as inflammatory itch disorders, often present with intense itching sensations, leadin to significant discomfort and distress for affected individuals. Conditions such as atopic dermatitis, psoriasis, and allergic contact dermatitis are prime examples of inflammatory skin disorders where itch plays a central role. The itching sensation associated with these conditions can be relentless, interfering with daily activities, disrupting sleep patterns, and negatively impacting quality of life. Despite varying etiologies, these disorders share common inflammatory pathways that contribute to the perception of itch, including activation of immune cells, release of proinflammatory mediators, and alteration in nerve signaling within the skin.

One key player in the pathogensis of inflammatory itch disorders, particularly in children, is the gene interleukin-13 (IL-13). IL-13 is a cytokine, i.e., a small protein involved in cell signaling, that plays a crucial role in regulating immune responses and inflammation. In the contexgt of inflammatory skin conditions, IL-13 has been implicated in promoting the inflammatory cascade that triggers the itch sensation. Research has shown that elevated levels of IL-13 are present in the skin of individuals with conditions like atopic dermatitis, where itch is a predominant symptom.

Genetic studies have identified variation in the IL-13 gene that may predispose individuals, especially children, to develop inflammatory skin disorders characterized by severe itching. The overproduction of IL-13 triggers a cascade of events within the skin, including the recruitment of immune cells and the release of itch-inducing molecules, ultimately leading to heightened itch perception and exacerbation of the underlying skin inflammation.

When it comes to treatment options, topical corticosteroids are overwhelmingly the most frequently prescribed class of drugs for atopic dermatitis. However, long-term application of topical corticosteroids is not recommended because of the risk of skin atrophy, dyspigmentation, acneiform eruptions, and risks associated with systemic absorption such as Cushing's disease. Topical calcineurin inhibitors are generally effective and safe as short-term treatments, but concerns of skin malignancies and increased risk of lymphomas have prompted regulatory authorities to require a warning regarding their long-term safety.

First generation antihistamines are widely prescribed for acute symptomatic treatment of pruritis, although their effectiveness is limited and largely attributed to their sedating effect. Oral immunosuppressants and glucocorticoids are effective but are sometimes associated with severe toxicity and side effects, thus limiting their use to short courses and/or intermittent therapy. No systemic agents are approved in the treatment of atopic dermatitis in children. All systemic agents are used off label (cyclosporine, methotrexate, etc.) and have a broad immunosuppressive effect which predisposes the patients to serious infections and increased risk of malignancies if used for prolonged periods. Other reported significant side effects with these agents include gastritis, stunted growth, diabetes, weight gain, hypertension, osteoporosis, and adrenal suppression (corticosteroids), elevated liver enzymes and leukopenia. Moreover, a high proportion of patients in which the condition is initially controlled by systemic agents suffer from relapse once therapy is discontinued. As a result, there exists a significant unmet need for alternative compositions and methods for managing inflammatory skin conditions such as atopic dermatitis, psoriasis, acne vulgaris, and hidradenitis suppurativa.

Based on the foregoing, it is an object of the present invention to provide compositions capable of addressing the symptoms associated with inflammatory skin disorders which include (i) structural degradation of the skin barrier/SC, (ii) inflammation caused by the activation of the body's immune system and its release of proinflammatory cytokines, and (iii) oxidative stress caused by generation of excess amounts of reactive oxygen species (ROS), in order to effectively treat and manage such disorders.

It is also an object of the present invention to provide methods of treating and managing inflammatory skin disorders by effectively addressing the above-referenced symptoms.

SUMMARY OF THE INVENTION

According to certain aspects of the present invention, compositions and methods are provided for treating and managing symptoms associated with inflammatory skin disorders.

According to one embodiment, the present disclosure provides a skincare composition containing a synergistic blend of plant-based actives comprised of at least: (a) Calendula officinalis plant or stem cell extract; (b) Symphytum officinale extract; and (c) a Haberlea rhodopensis extract, e.g., a biotech-derived Haberlea rhodopensis extract, wherein the composition is capable of effectively treating and managing symptoms associated with inflammatory skin disorders.

According to certain embodiments, the present disclosure also includes methods for effectively treating and managing symptoms associated with inflammatory skin disorders involving the application of the above-disclosed composition onto skin.

According to another embodiment, the present disclosure includes a method of reducing dependence on topical corticosteroids in a person suffering from an inflammatory skin disorder involving consistently applying the above-disclosed composition onto skin.

According to yet another embodiment, the present disclosure is also directed to a natural preservative system that cooperates with the skincare composition and method for treating and managing inflammatory skin disorders. The preservative system may be a preservative system as discussed and/or claimed in U.S. Pat. No. 10,721,937 incorporated herein by reference in its entirety.

In some embodiments, the present disclosure provides a composition for application onto human skin including a synergistic blend of at least: (a) Calendula officinalis extract; (b) Symphytum officinale extract; and (c) Haberlea rhodopensis extract, wherein the composition is capable of effectively treating and managing symptoms associated with inflammatory skin disorders.

In some embodiments, (a) is present in an amount of from about 0.1 to about 5.0% by weight, (b) is present in an amount of from about 0.1 to about 5.0% by weight, and (c) is present in an amount of from about 0.1 to about 5.0% by weight, all weights based on total weight of the composition. In some embodiments, (a) is present in an amount of from about 0.15 to about 3.5% by weight, (b) is present in an amount of from about 0.15 to about 3.5% by weight, and (c) is present in an amount of from about 0.15 to about 3.5% by weight, all weights based on total weight of the composition. In some embodiments, (a) is present in an amount of from about 0.20 to about 2.0% by weight, (b) is present in an amount of from about 0.20 to about 2.0% by weight, and (c) is present in an amount of from about 0.25 to about 2.0% by weight, all weights based on total weight of the composition.

In some embodiments, the composition further includes at least one humectant. In some embodiments, the at least one humectant is present in an amount of from about 0.1 to about 10% by weight, based on total weight of the composition. In some embodiments, the at least one humectant includes hyaluronic acid.

In some embodiments, the composition further includes at least one ceramide. In some embodiments, the at least one ceramide is present in an amount of from about 0.01 to about 1.0% by weight, based on total weight of the composition. In some embodiments, the at least one ceramide is selected from ceramide 1, ceramide 2, ceramide 3, ceramide 4, ceramide 5, ceramide 1A, ceramide 6 II, ceramide AP, ceramide EOP, ceramide EOS, ceramide NP, ceramide NG, ceramide NS, ceramide AS, and ceramide NS dilaurate.

In some embodiments, the present disclosure provides a composition for application onto human skin including: (1) a synergistic blend of at least: (a) Calendula officinalis stem cell extract, (b) Symphytum officinale extract, and (c) a biotech-derived Haberlea rhodopensis extract; (2) at least one humectant; and (3) at least one ceramide, wherein the composition is capable of effectively treating and managing symptoms associated with inflammatory skin disorders.

In some embodiments, (a) is present in an amount of from about 0.1 to about 5.0% by weight, (b) is present in an amount of from about 0.1 to about 5.0% by weight, (c) is present in an amount of from about 0.1 to about 5.0% by weight, (2) is present in an amount of from about 0.1 to about 10% by weight, and (3) is present in an amount of from about 0.01 to about 1.0% by weight, all weights based on total weight of the composition. In some embodiments, (a) is present in an amount of from about 0.20 to about 2.0% by weight, (b) is present in an amount of from about 0.20 to about 2.0% by weight, (c) is present in an amount of from about 0.20 to about 2.0% by weight, (2) is present in an amount of from about 0.1 to about 5.0% by weight, and (3) is present in an amount of from about 0.1 to about 0.3% by weight, all weights based on total weight of the composition.

In some embodiments, the composition is free of a skin-sensitizing amount of an essential oil and has a pH ranging from about 4.5 to about 5.5.

In some embodiments, the present disclosure provides a method of treating and managing symptoms associated with inflammatory skin disorders, comprising applying the composition described herein onto skin of a subject in need thereof. In some embodiments, the symptoms include at least one of a compromised skin barrier, inflammation, and oxidative stress.

In some embodiments, the present disclosure provides a method of reducing reliance on corticosteroids, comprising applying the composition described herein onto skin of a subject in need thereof. In some embodiments, the subject exhibits symptoms including at least one of a compromised skin barrier, inflammation, and oxidative stress.

Other embodiments of the present disclosure will become apparent from a review of the ensuing detailed disclosure.

DETAILED DESCRIPTION

For purposes of the present disclosure, the use of the term “inflammatory skin disorders” is intended to encompass those disorders whose symptoms include a structurally impaired skin barrier/SC (i.e., a compromised skin barrier), inflammation, and excessive amounts of reactive oxygen species/ROS (i.e., oxidative stress). Examples include atopic dermatitis/eczema, psoriasis, acne vulgaris, and hidradenitis suppurativa.

Compositions of the present disclosure can comprise, consist essentially of, or consist of, the components of the present disclosure as well as other ingredients described herein. The term “comprising” as used herein is meant to include various optional, compatible components that can be used in the preservative systems and cosmetic compositions of the present disclosure without limiting the inclusion, use of, or cooperation with other ingredients, excipients, uses, or otherwise. The term “consisting essentially of” as used herein means that the composition or component may include additional ingredients, but only if the additional ingredients do not materially alter the basic and novel characteristics of the compositions or methods.

As used herein, the words “preferred,” “preferably,” and variants thereof refer to embodiments of the disclosure that afford certain benefits under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the disclosure.

Numerical ranges as used herein are intended to include every number and subset of numbers contained within that range, whether specifically disclosed or not. Further, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers within that range.

All percentages, parts, proportions, and ratios as used herein are by weight of the total composition, unless otherwise specified. All such weights as they pertain to listed ingredients are based on the active level.

All references to singular characteristics or limitations of the present disclosure shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary by the context in which the reference is made.

All publications, articles, papers, patents, patent publications, and other references cited herein are hereby incorporated in their entireties for all purposes to the extent consistent with the disclosure herein.

The term “oxidative stress” as described herein refers to the disturbance in balance between reactive oxygen species (ROS) and antioxidants present in the skin caused by extrinsic and/or intrinsic factors. Extrinsic factors include, for example, exposure to UV radiation, high energy visible light, pollution, and products containing harsh chemicals. Intrinsic factors include, for example, chronological aging, a person's genetic makeup, and other biological changes that occur from within the skin such as atopic dermatitis, psoriasis, acne vulgaris, and hidradenitis suppurativa.

The term “consistently applying” as described herein refers to an individual suffering from chronic inflammatory skin disorders incorporating the composition of the present disclosure into their skincare regimen.

It has surprisingly and unexpectedly been discovered by the inventors that a combination of: (a) Calendula officinalis extract; (b) Symphytum officinale extract; and (c) Haberlea rhodopensis extract, results in a synergistic influence on certain genes that are known to help manage oxidative stress, restore skin barrier structure and integrity, reduce inflammation, restore skin barrier homeostasis, and support wound healing and tissue remodeling. The synergistic influence on these genes facilitate the effective treatment and management of certain symptoms associated with chronic inflammatory skin disorders, particularly atopic dermatitis, psoriasis, acne vulgaris, and hidradenitis suppurativa.

Calendula officinalis extract is derived from the cells of the marigold plant. Common bioactive compounds found therein include, for example, flavonoids, phenolic acids, carotenoids, triterpenic alcohols, polycarbohydrates, proteins, amino acids, saturated hydrocarbons, vitamin C, and mineral substances. The Calendula officinalis extract is commercially available from various suppliers including, for example, Innova BM, a company headquartered in Bulgaria, which sells the extract under the tradename InnovaStemCell Calendula W®.

In some embodiments, the Calendula officinalis extract is a stem cell extract. The Calendula officinalis stem cell extract of the present disclosure may be employed in an amount of from about 0.1 to about 10% by weight, including from about 0.1 to about 5.0% by weight, or from about 0.15 to about 3.5% by weight, or from about 0.20 to about 2.0% by weight, or from about 0.25 to about 2.0% by weight, all weights based on the total weight of the composition. The Calendula officinalis extract is employed in the form of an oil infusion derived from either a plant macerate or plant stem cells grown in a bioreactor. The biotech method employed by Innova BM uses an in-vitro cell culture of Calendula officinalis cells to create a sustainable, eco-friendly cell extract containing a higher concentration of beneficial phyto-chemicals.

Regarding the Symphytum officinale extract, the primary bioactive compounds found therein include allantoin, rosmarinic acid and ellagic acid.

The Symphytum officinale extract of the present disclosure may be employed in an amount of from about 0.1 to about 10% by weight, including from about 0.1 to about 5.0% by weight, or from about 0.15 to about 3.5% by weight, or from about 0.25 to about 2.0% by weight, or from about 0.20 to about 2.0% by weight, or from about 0.25 to about 2.0% by weight, all weights based on the total weight of the composition. The Symphytum officinale extract is employed in the form of an oil infusion derived from either a plant macerate or plant stem cells grown in a bioreactor. The biotech method employed by Mibelle uses an in-vitro cell culture of Symphytum officinale cells to create a sustainable, eco-friendly cell extract containing a higher concentration of beneficial phyto-chemicals.

Haberlea rhodopensis is a tertiary relict plant endemic to the Balkan region, especially the Rhodope mountains in Bulgaria. The Haberlea rhodopensis extract is employed in the form of an oil infusion generated from either a plant macerate or plant stem cells grown in a bioreactor. A Haberlea rhodopensis extract produced from plant macerate or plant stem cells grown in a bioreactor is referred to herein as a “biotech-derived Haberlea rhodopensis extract.” The Haberlea rhodopensis extract of the present disclosure may be employed in an amount of from about 0.1 to about 10% by weight, including from about 0.1 to about 5.0% by weight, or from about 0.15 to about 3.5% by weight, or from about 0.25 to about 2.0% by weight, or from about 0.20 to about 2.0% by weight, or from about 0.25 to about 2.0% by weight, all weights based on the total weight of the composition. The Haberlea rhodopensis extract of the present invention is commercially available from various suppliers including, for example, from Innova BM, a company headquartered in Bulgaria, under the tradename InnovaBio Tech Haberlea®. The biotech method employed by Innova BM uses an in-vitro cell culture of Haberlea rhodopensis cells cultivated from the plant to create a sustainable, eco-friendly cell extract containing a higher concentration of beneficial phyto-chemicals.

Though not required, the extracts of the present disclosure are typically employed in the form of an oil infusion, i.e., the extracts are infused in a plant-based oil. The oil component of each infusion is employed in an amount of from about 0.2% to about 3% by weight, including about 1% and 2% by weight, based on the total weight of the composition. While any plant-based oil may be used as the oil component in the above-described oil infusion, a preferred oil component is sunflower seed oil. Without being bound by theory, sunflower seed oil may be particularly preferred because of the high content of linoleic acid present therein. Linoleic acid provides additional anti-inflammatory properties to the formulation, is an excellent moisturizer, and helps fortify and protect the skin barrier against external irritants.

According to one embodiment, the composition of the present disclosure contains: (a) from about 0.1 to about 10% by weight, including from about 0.1 to about 5.0% by weight, or from about 0.15 to about 3.5% by weight, or about 0.20 to about 2.0% by weight, or about 0.25 to about 2.0% by weight of Calendula officinalis stem cell extract; (b) from about 0.1 to about 10% by weight, including from about 0.1 to about 5.0% by weight, or from about 0.15 to about 3.5% by weight, or about 0.20 to about 2.0% by weight, or about 0.25 to about 2.0% by weight Symphytum officinale extract; and (c) 0.1 to about 5.0% by weight, or from about 0.15 to about 3.5% by weight, or about 0.20 to about 2.0% by weight, or about 0.25 to about 2.0% by weight of biotech-derived Haberlea rhodopensis extract, all weights based on the total weight of the composition.

According to another embodiment of the present disclosure, the composition further includes at least one humectant for providing enhanced hydration and moisturization to de-hydrated skin effected by a chronic inflammatory skin disorder. The humectant may be employed in an amount of from about 0.1 to about 10% by weight, including from about 0.1 to about 8.0% by weight, or from about 0.1 to about 5.0% by weight, all weights based on the total weight of the composition. Examples of suitable humectants include, but are not limited to, hyaluronic acid and its derivatives such as sodium hyaluronate and hydrolyzed hyaluronic acid, lecithin, aloe vera, panthenol, glycerin, and seaweed. A particularly preferred humectant for use in embodiments of the present disclosure is hyaluronic acid.

Ceramides are a group of natural waxy, fatty substances in the skin, composed of sphingosine and lipids (fatty acids) bonded together. Ceramides make up about 50% of all skin lipids and are manufactured in the lower, living cells of the epidermis. As the cells mature and move to the surface, ceramides are released to the topmost layer, the stratum corneum. In the stratum corneum layer, ceramides combine with cholesterol (another important lipid found in the skin) and fatty acids to form an ordered, tightly packed, layered, sheet-like arrangement between the dead cells. Ceramides and cholesterol protect against moisture loss to keep skin youthful and supple and support the structure of the stratum corneum. Exemplary ceramides include ceramide 1, ceramide 2, ceramide 3, ceramide 4, ceramide 5, ceramide 1A, ceramide 6 II, ceramide AP, ceramide EOP, ceramide EOS, ceramide NP, ceramide NG, ceramide NS, ceramide AS, and ceramide NS dilaurate.

According to another embodiment of the present disclosure, the composition further includes at least one ceramide for structurally enhancing the stratum corneum. The ceramide may be employed in an amount of from about 0.01 about 1.0% by weight, including from about 0.05% to about 0.5%% by weight, and from about 0.1% to about 0.3% by weight, all weights based on the total weight of the composition, it being understood that the ceramide component of the composition may be comprised of a mixture of two or more different ceramides. In some embodiments, the at least one ceramide is selected from ceramide 1, ceramide 2, ceramide 3, ceramide 4, ceramide 5, ceramide 1A, ceramide 6 II, ceramide AP, ceramide EOP, ceramide EOS, ceramide NP, ceramide NG, ceramide NS, ceramide AS, ceramide NS dilaurate, or a combination thereof.

In yet another embodiment of the present disclosure, the compositions are free of a skin-sensitizing amount of an essential oil. In some embodiments, the compositions have a pH ranging from about 4.5 to about 5.5, and preferably from about 4.8 to about 5.3. In some embodiments, a composition “free of” a particular component means that the composition includes less than about 1% by weight, or less than about 0.8% by weight, or less than about 0.5% by weight, or less than about 0.2% by weight, or less than about 0.1% by weight, or less than about 0.05% by weight, or less than 0.01% by weight of the particular component.

In another embodiment, the present disclosure provides a composition containing: (1) a therapeutic blend of plant-based actives comprising at least: (a) Calendula officinalis extract; (b) Symphytum officinale extract; and (c) Haberlea rhodopensis extract; (2) at least one humectant; (3) at least one ceramide; (4) an emulsifier; and (5) a dermatologically acceptable carrier, wherein the composition is capable of effectively treating and managing symptoms associated with inflammatory skin disorders.

Any ingredient capable of emulsifying the composition may be employed as an emulsifier without departing from the spirit of the invention, so long as it is natural and dermatologically acceptable. Examples thereof include, but are not limited to, glyceryl stearate, cetyl alcohol, sodium stearoyl lactylate, sorbitan olivate, cetearyl olivate, cetearyl alcohol, cetearyl glucoside, sodium cetearyl sulfate, and the like. It is also particularly preferred that the emulsifier be free of palm oil.

The compositions of the present disclosure may be in a wide variety of product forms that include, but are not limited to, solutions, suspensions, lotions, creams, gels, sprays, foams, ointments, and serums.

According to embodiments of the present disclosure, the compositions can also additionally comprise suitable optional ingredients as desired. For example, the composition can optionally include other active or inactive ingredients, provided they do not unacceptably alter the benefits of the composition and do not promote skin sensitization. The precise amount of optional ingredients chosen will be determined by those skilled in the art.

The dermatologically acceptable carrier can encompass a wide variety of forms. In some cases, the solubility or dispersibility of the components in the composition may dictate the form and character of the carrier. Non-limiting examples include simple solutions (e.g., aqueous or anhydrous), dispersions, emulsions and solid forms. In certain embodiments, the dermatologically acceptable carrier is in the form of an emulsion. An emulsion can be generally classified as having a continuous aqueous phase (e.g., oil-in-water and water-in-oil-in-water) or a continuous oil phase (e.g., water-in-oil or oil-in-water). While the oil phase may comprise any vegetable oil, so long as it does not cause skin sensitization, a particularly preferred oil component is sunflower oil. Further exemplary vegetable oils are provided herein.

The inventors have surprisingly discovered that the use of sunflower oil enables bioactive compounds present in the botanical extract to effectively penetrate into the skin, without the need of having to use skin-sensitizing essential oils, while still facilitating the desired degree of efficacy. The high concentration of linoleic acid (at least 60%) helps maintain the skin barrier and serve as an emollient to lock in moisture and reduce trans-epidermal water loss (TEWL). Sunflower oil is also rich in vitamin E, an antioxidant that helps protect the skin barrier from external stressors like sun exposure and air pollution, vitamins C and D as well as beta-carotene (pre-cursor to vitamin A) that can combine with fatty acids to promote new skin cell growth and prevent of bacterial infection and omega-6 fatty acids that help reduce redness and smooth roughness. In addition, sunflower oil is safe for people with tree nut allergies. Other vegetable oils that may also be used include, but are not limited to, almond oil, olive oil, jojoba oil, babassu oil, castor oil, coconut oil, corn oil, cotton seed oil, linseed oil, mustard oil, safflower oil, sesame oil, soybean oil, sunflower-seed oil, wheat germ oil, argan oil and marula oil.

According to another embodiment, there is provided a method of treating and managing symptoms associated with inflammatory skin disorders by applying one of the above-disclosed compositions onto skin of a subject in need thereof. In some embodiments, the symptoms include at least one of a compromised skin barrier, inflammation, and oxidative stress.

According to yet another embodiment, the present disclosure is also directed to a method of reducing reliance on corticosteroids for treating inflammatory skin disorders by consistently applying one of the above-disclosed compositions onto skin of a subject in need thereof. In some embodiments, the subject exhibits symptoms including at least one of a compromised skin barrier, inflammation, and oxidative stress.

EXAMPLES

The following examples as set forth herein are intended for illustrative purposes only and are not intended to limit the scope of the disclosure in any way, as many variations thereof are possible without departing from the spirit and scope of the disclosure. In the examples, all concentrations are listed as weight percent, unless otherwise specified.

Calendula officinalis stem cell extract in an amount of 2%, an oil infusion of Symphytum officinale extract in an amount of 1%, and a biotech-derived Haberlea rhodopensis extract in an amount of 1% were evaluated, both individually and in combination, to determine what, if any, gene expression effects they may indicate, per the below-indicated protocol.

Reconstructed Human Epidermis (RHE) tissues were obtained from ZenBio (Research Triangle Park, NC; lot #RHE051820) and were used immediately. Tissues were transferred to 6 well plates and were equilibrated for an hour in 1 ml of pre-warmed medium/well ZenSkin provided by the ZenBio. Samples of the above ingredients were then added non-diluted, in triplicates at 3 mg/cm2 with the positive displacement pipette and were spread evenly on top of the RHE tissues. Sterile distilled water was the negative control. Treated tissues were allowed to incubate overnight. The results of the gene expression testing fell into three distinct categories.

At the end of the incubation RNA was extracted and purified with RNeasy Mini Kit cat. #74104 from Qiagen (Germantown, MD), using a QiaCube Connect robotic station (Qiagen). Purified total RNA was assessed at 260 nm and 280 nm with a Thermo Fisher Scientific (Waltham, MA) NanoDrop™ Lite Spectrophotometer.

cDNA was prepared using a High-Capacity RNA-to-cDNA™ Kit (Applied Biosystems, Thermo Fisher) and the expression of the genes of interest was measured by real-time quantitative PCR with a BioRad iCycler iQ Detection System using PCR primers from Realtimeprimers (Elkins Park, PA) and AzuraView GreenFast qPCR Blue Mix LR available from Azura Genomics (Raynham, MA). Efficiency ΔΔCt method was used for quantification of results, after the normalization of gene expression to HPRT1 and GAPDH (housekeeping genes).

Genes were considered differentially expressed if the p value, as determined by the two-tailed t-test, was ≤0.10 and the modulation was ≥1.8.

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with anti-oxidative function, as compared to each individual ingredient, is shown in Table 1, below.

In the presence of the mixture, antioxidant genes SOD1 and GPX1 were downregulated and HMOX1/NQO1 were activated to provide a more comprehensive ROS reduction mechanism that not only scavenges ROS but protects skin tissue from damage. (See, e.g., Wagener et al., Int. J Mol. Sci. 14(5):9126-9167 (2013)) Moreover, HMOX1 counteracts the cytotoxic effects caused by high concentrations of free heme in the eczematic lesion and enables the NRF2/HMOX1 axis to mediate anti-inflammatory activity, whereas NQO1 provides NRF2 a target to help detoxify xenobiotic compounds as well as ROS.

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with ceramide production, as compared to each individual ingredient, is shown in Table 2, below.

Ceramides play a key role in reducing permeability of the skin barrier. In eczema, skin lipids may contain less ceramide and ceramide composition may be altered.

SMPD1 was surprisingly upregulated by the mixture to increase ceramide production. The protein encoded by this gene is a lysosomal acid (sphingomyelinase) that converts sphingomyelin to ceramide. (See, e.g., Kitatani et al., Cells 10(9):2386 (2021)).

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with skin barrier repair/restructuring, as compared to each individual ingredient, is shown in Table 3, below.

Skin barrier structure and integrity genes of Table 3, associated with a properly functioning stratum corneum, were significantly upregulated in the presence of the mixture as compared to the individual components, especially FLG which is typically under-expressed in skin suffering from atopic dermatitis. Note that this includes both structural protein genes and binding protein genes.

CDSN codes for corneodesmosin and plays a vital role in the structural and functional integrity of the epidermis and hair follicle integrity by preventing the rupture of corneodesmosome, i.e., the main intercellular adhesive structures in the stratum corneum that are transformed from desmosomes at the most superficial layer of the stratum granulosum of the epidermis. The major compositional distinction as compared to desmosomes is the presence of corneodesmosin in the extracellular portion. CDSN was upregulated by the mixture, thereby enhancing the structural integrity of the stratum corneum through improved adhesion, which is helpful for improving lesions in individuals suffering from AD and psoriasis. (See, e.g., Ishida-Yamamoto et al., Cell Tissue Res. 360(3):477-482 (2015) and Gordon et al., Pediatr. Dermatol. 39(2):268-272 (2022)) FLG was significantly upregulated (more than the sum of the individual components, indicating synergistic amplification) which is known to increase the production of pro-filaggrin, a critically important protein for skin barrier structure, and under produced in those with atopic dermatitis.

LOR was upregulated to produce the skin barrier protein loricrin, which is a major component of the skin's cornified envelope. Links between loricrin and other components of the envelope, like filaggrin, hold the corneocytes together and help give the stratum corneum its structural strength.

GBA, an enzyme involved in ceramide production, was also significantly upregulated (compared to downregulated in comfrey alone) evidencing the ability of the mixture to enhance the structural integrity of the stratum corneum.

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with inflammation reduction, as compared to each individual ingredient, is shown in Table 4, below.

Inflammation modulation of atopic dermatitis: AHR/NRF2 also play critical roles in the maintenance of skin homeostasis. Specific disruption of AHR function in the skin has been found to be associated with the pathogenesis of atopic dermatitis. (See, e.g., Edamitsu et al., Antioxidants (Basel) 11(2):227 (2022)).

AGER was downregulated in the presence of the mixture, evidencing a reduction in the production of cell surface receptors for advanced glycation end-products (RAGE) that can cause an unfavorable proinflammatory state. (See, e.g., Serveaux-Dancer et al., Dis. Markers 2019:6067353 (2019) and Budu-Aggrey et al., Nature Comm. 14:6172 (2023)).

AHR was downregulated in the presence of the mixture. Without intending to be bound by theory, this downregulation is believed to evidence a reduction in immune mediated skin responses based on AHR's ability to inhibit: (i) autophagy, (ii) extracellular matrix degeneration, and (iii) oxidative stress. (See, e.g., Salminen, Cell. Mol. Life Sci. 79:489 (2022)).

EDN1 was downregulated in the presence of the mixture, which evidences a reduction in itch response as this gene is associated with the production of endogenous pruritogens [endothelin (ET)-1] in the skin. Elevated plasma ET-1 were significantly correlated with AD clinical severity, itch intensity, and serum IgE levels. (See, e.g., Tsybikov et al., Allergy Asthma Proc. 36(4):320-324 (2015) and Nakahara et al., Curr Treat. Options Allergy 6:156-163 (2019)).

ICAM1 gene was upregulated in the presence of the mixture, evidencing an increase in endothelial cell activation by acting as a leukocyte adhesion molecule thereby helping the affected area fight infection and/or inflammation caused by mechanical/frictional damage such as by scratching. (See, e.g., Marinovid Kulisid et al., Life 13:933 (2023) and Werfel, J. Invest. Dermatol. 129(8):1878-1891 (2009)).

IRF4 gene was downregulated in the presence of the mixture, evidencing a reduction in the inflammatory cascade of NLRP3-mediated IL-33 cytokines such as those triggered in AD lesions, as IRF4 interacts with NLRP3 to control IL-33 expression in epithelial cells. (See, e.g., Zheng et al., Cell Disease Death 12:871 (2021)).

IL8 gene was upregulated in the presence of the mixture, evidencing production of the IL-8 cytokine that is an attractant for neutrophils and T-cells and stimulates the migration of keratinocytes for wound healing. IL-8 is typically suppressed in patients with AD because it is inhibited by TH2 cytokines. (See, e.g., Nomura et al., J. Immunol. 171(6):3262-3269 (2003) and Fania et al., Int. J. Mol. Sci. 23(5):2684 (2022)).

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with skin barrier homeostasis, as compared to each individual ingredient, is shown in Table 5, below.

PPARD was significantly upregulated in the presence of the mixture, evidencing an acceleration in wound healing of a structurally impaired (wounded) skin barrier. PPARD has been found to inhibit keratinocyte apoptosis and promote the re-epithelialization of the skin by enhancing keratinocyte adhesion and migration. (See, e.g., Blunder et al., Int. J. Mol. Sci. 22(14):7354 (2021)).

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with cell death (apoptosis), as compared to each individual ingredient, is shown in Table 6, below.

CSF2 (Colony-stimulating factor 2) was significantly upregulated in the presence of the mixture to produce an endogenous damage signal that promotes the therapeutic effects of mesenchymal stem cells by enhancing their multi-lineage differentiation and migratory capacities. CSF2 also stimulates the proliferation and differentiation of granulocytes and macrophages for addressing infection. As a result, its stimulation is believed to enhance the skin barrier's response to atopic dermatitis flareups. (See, e.g., Park et al., Mol Therapy 27(6):1087-1100 (2019)).

The synergistic influence exhibited by the mixture of the present disclosure on certain genes associated with tissue remodeling/wound healing, as compared to each individual ingredient, is shown in Table 7, below.

Various tissue remodeling and wound healing genes that help mitigate alterations in skin barrier structure in regions suffering from AD or psoriatic inflammation were influenced by the mixture, as compared to each individual ingredient.

BMP4 was upregulated by the mixture. This gene has been observed as an upstream regulator in skin inflammatory disease lesions like AD and psoriasis which activates the RHO family of GTPases, important regulators for diverse cellular processes, including cellular polarization and morphogenesis as well as rearrangement of the actin skeleton in cells to facilitate their removal by phagocytosis. This upregulation enables the clearance of dying keratinocytes for wound healing. (See, e.g., Coleman and Olson, Cell Death Differ. 9:493-504 (2002) and Bang et al., Sci Report 12:5889 (2022)).

GRHL3—transcription factor, which activates gene expression programs required for cell adhesion, lipid production, cornified envelope formation and protein crosslinking, was significantly upregulated by the mixture (almost three times more than in Calendula alone showing amplificatory synergy). It is essential for skin barrier formation and adult epidermal repair. This upregulation helps to reinforce and restore the stratum corneum. (See, e.g., Klein et al., PLOS Genet. e1006745 (2017)).

PAX3 was upregulated by the mixture. The protein made by the PAX3 gene directs the activity of other genes that signal neural crest cells to form specialized tissues or cell types including nerve cells and melanocytes. Upregulation of this gene therefore serves to support melanocyte differentiation, survival, proliferation, and migration for healing inflamed regions of skin and local hair follicles. (See, e.g., Boudjadi et al., Gene 666:145-157 (2018) and Medic et al., Biochem. Biophys. Res. Comm. 411(4):832-837 (2011)).

SERPINH1 which encodes for Hsp47 (heat shock protein), a collagen-specific molecular chaperone that localizes in the endoplasmic reticulum (ER), is indispensable for molecular maturation of collagen by ensuring correct folding of procollagen. This gene was downregulated in the presence of the mixture to decrease procollagen folding in the early stages of wound healing to allow for the fibroblasts to accumulate more components such as hyaluronan and fibronectin in the extracellular matrix and not form scar tissue within the inflammatory lesions. (See, e.g., Tyavambiza et al., Bioengineering 9(11):712 (2022) and Ito and Nagata, Seminar Cell Dev. Biol. 62:142-151 (2017)).

TIMP2 inhibits several MMPs that cleave a range of substrates that impact structural and cellular aspects of tissue architecture. This gene was downregulated in the presence of the mixture, thereby enabling it to maximize the effectiveness of those MMP genes involved in tissue remodeling/wound healing. (See, e.g., Costanzo et al., Pulm. Med. 2022:3632764 (2022)).

In summary, the gene expression data of Tables 1-7 establishes the synergy realized by combining the ingredients of the present disclosure when it comes to treating and managing oxidative stress, inflammation, and skin barrier/SC restructuring and repair to help mitigate symptoms associated with inflammatory skin disorders.

mixed with Lactobacillus

The composition of Example 2 was clinically tested on a first cohort of test subjects with very dry, rough skin (to simulate a damaged skin barrier), in order to determine its efficacy in enhancing skin health and appearance. Twenty-two female individuals, aged 40-65, were asked to apply the composition twice a day at home, once in the morning and once in the evening, under normal use conditions, as a replacement for their normal skin treatment routine. Skin health and appearance data was collected on day 0, day 28 and day 56.

Moisturization/hydration data was obtained using a corneometer, wherein an increase in coreometer value corresponds to an increase in moisturization/hydration. The data showed that 95% of test subjects experienced an increase of more than 32% on day 28; and 71% of test subjects experienced an increase of more than 16% on day 56.

The skin smoothing and anti-roughness effect of the composition was determined in-vivo using the fringe projection system DermaTOP® and its attendant acquisition software. This system measures the height of projections emanating from the skin used to assess skin roughness, to simulate skin disorders such as eczema or psoriasis. Rough skin patches are often the result of dryness or a buildup of dead skin cells. A decrease in projection height evidences smoother skin. The data showed that approximately 72% of test subjects experienced a significant decrease of approximately 5% in peak surface roughness at day 28. The data clearly evidences the smoothing effect of the composition, implying improved skin barrier structure and correlating to improved hydration.

The composition of Example 2 was also clinically tested on a second cohort of test subjects with dry skin, in order to determine its efficacy in enhancing skin health and appearance. Thirty-two female individuals, aged 40-65, were asked to apply the composition twice a day at home, once in the morning and once in the evening, under normal use conditions, as a replacement for their normal skin treatment routine. Skin health and appearance data was collected on day 0 and day 28.

Moisturization/hydration data was obtained using a corneometer, wherein an increase in corneometer value corresponds to an increase in moisturization/hydration. The data showed that 75% of test subjects experienced an increase of 20% on day 28.

Transepidermal water loss (TEWL) was evaluated using an Aquaflux AF200 (BIOX) system. This device measures skin's ability to retain water. When water passes through the skin, which is a natural biological process, and evaporates from the skin's surface, this phenomenon is referred to as TEWL and is a measure of skin barrier's ability to retain moisture present in the dermis. The data showed that approximately 52% of test subjects experienced a significant decrease of approximately 10% in TEWL on day 28. The data clearly evidences the skin barrier integrity and reinforcement properties facilitated by the composition.

Regarding user experience, all participants in these two studies agreed with and further supported the measured data: 97% of subjects reported their skin appearing smoother and softer; 93% observed that the product helped smooth uneven texture and transformed their skin; 90% agreed that their skin felt less itchy and their skin looked less red/inflamed.

The above-discussed in-vivo clinical testing clearly establishes, quantitatively, the significant impact on skin health and appearance achieved by the use the inventive composition of Example 1.

The above-referenced examples establish the effectiveness of the disclosed embodiments to treat and manage symptoms associated with chronic inflammatory skin disorders. Specifically, the synergistic influence exerted on certain genes associated with (i) skin barrier repair/restructuring, (ii) inflammation reduction/management, and (iii) reduction/management of oxidative stress caused by ROS in the presence of the disclosed embodiments facilitates the treatment and management of symptoms experienced by those suffering from chronic inflammatory disorders. Moreover, consistent application of the disclosed embodiments on the skin of those suffering from chronic inflammatory skin disorders can reduce their reliance on corticosteroids for relief from their symptoms.