Use of lotus extracts for treating and / or preventing skin ageing

Abstract

The invention is generally directed to active ingredients capable of countering dermal intrinsic ageing and its manifestations, compositions containing them and their use as cosmetic or medical (e.g. dermatological) dermal compositions. The agents inhibit or reduce the biosynthesis of secreted frizzled-related protein 4 (SFRP 4) and reduce expression of p21 in fibroblasts.

Description

USE OF ACTIVE INGREDIENTS FOR TREATING AND / OR PREVENTING SENESCENCE CLAIM OF PRIORITY

[0001] This patent application claims the benefit of priority to FR Application Serial No.2408591, filed August 2, 2024, and FR Application Serial No.2501793, filed February 21, 2025, which are incorporated by reference herein in their entireties. BACKGROUND

[0002] The invention is generally directed to active ingredients capable of countering dermal intrinsic ageing and its manifestations, compositions containing them and their use as cosmetic or medical (e.g. dermatological) dermal compositions.

[0003] Skin is the largest organ of the human body: it provides a barrier against harmful organisms and substances, protects against UV radiation and regulates water loss and body temperature. The skin is a complex organ, consisting of several compartments with different functions. The outermost epidermis is stratified into four sublayers (basal, spinous, granular and cornified layer) with keratinocytes being the predominant cell type. Pigment-producing melanocytes reside within the basal layer of the epidermis, determine skin color and possess photo-protecting properties. The dermal-epidermal junction (DEJ) connects the epidermis to the underlying dermis, which harbors dermal fibroblasts and appendages such as hair follicles, sebaceous glands, and sweat glands (1).

[0004] Skin aging is a multi-factorial process that affects nearly every aspect of its biology and function; it is driven by both intrinsic (e.g., time, genetic factors, hormones) and extrinsic (e.g., UV exposure, pollution) factors. The apparent signs of intrinsic ageing include the loss and descent of underlying fat, leading to hollowed cheeks and eye sockets, as well as loss of firmness and sagging skin as the bones shrink away from the skin due to bone loss. Other visible signs include thin and dry skin, fine wrinkles, decreased elasticity, aberrant pigmentation, hair graying and hair loss.

[0005] Intrinsic and extrinsic aging are partly due to the process of senescence and represent an inevitable change attributable to the passage of time alone and is representedprimarily by physiologic alterations with subtle but undoubtedly important consequences for both healthy and diseased skin and is largely genetically determined. Senescence, and in particular skin senescence, is a continuous process that usually begins in the mid-20s but may not become evident for decades. In a person’s 20s collagen production begins to slow and elastin has a bit less spring. Dead skin cells do not shed as quickly, and turnover of new skin cells decreases. The process of intrinsic skin aging resembles that seen in most internal organs and an explanation is thought to involve decreased proliferative capacity leading to cellular senescence, and altered biosynthetic activity of skin derived cells The intrinsic rate of skin aging in any individual can be dramatically influenced by personal and environmental factors, particularly the amount of exposure to ultraviolet light. For example, photodamage, which considerably accelerates the visible aging of skin, also greatly increases the risk of cutaneous neoplasms: the processes of intrinsic and extrinsic aging are superimposed. It is essential to stress however that the terms aging and cellular senescence cannot be used interchangeably: aging is a progressive decline with time whereas senescence occurs throughout the lifespan, including during embryogenesis. The number of senescent cells increases with age, but senescence also plays an important role during development.

[0006] Senescence does not occur only in the skin. Its role in all tissues is to prevent the replication of cells harboring damaged DNA, which serves an important anti-tumorigenic function. Senescence typically occurs in response to damaging stimuli, including telomere shortening (replicative senescence), DNA damage (DNA damage-induced senescence), and oncogenic signaling (oncogene-induced senescence).

[0007] Replicative senescence refers to the phenomenon whereby normal nonmalignant cells stop dividing in vitro, after approximately fifty divisions, which has been termed the Hayflick limit. Replicative senescence is induced by telomere shortening. With each round of DNA replication, telomeres are progressively shortened, eventually reaching a critical length which prevents further replication, thereby halting cell division. Critically short, uncapped telomeres initiate a DNA damage response which triggers senescence.

[0008] DNA damage triggers the DNA repair machinery, apoptosis, or senescence depending on the extent of damage and physiological context. Senescent cells are characterized by a persistent DNA damage response (DDR), including chronic ATM (Ataxia Telangiectasiamutated) and ATR (Ataxia Telangiectasia and Rad3 related) kinase signaling which ultimately invokes cell cycle arrest and senescence through activation of the p53 / p21 and p16 / pRb pathways. Persistent DNA damage and subsequent senescence can also be induced by ionizing radiation, chemotherapeutics, genotoxic stress, and oxidative stress.

[0009] Senescent cells are characterized by stable cell cycle arrest as well as morphological and metabolic changes, chromatin reorganization, altered gene expression, and acquisition of the senescence-associated secretory phenotype (SASP). One nefarious property of senescent cells is that they can also contaminate healthy cells, turning them into senescent cells too, and then accelerating their ageing. The new senescent cells adopt the same communication patterns with healthy cells, and the vicious cycle continues. Left unchecked, these cells can increase by up to three times after 65 years old.

[0010] Applied to the skin, this knowledge might reverse the concept of inside out beauty suggesting that treating senescence of the skin, our largest organ, could have an impact on how we age overall. It might be a whole new way to look at skin healthy longevity.

[0011] As mentioned above, the hallmark of senescent cells is the appearance of the senescence-associated secretory phenotype (SASP). The SASP includes several families of soluble and insoluble factors that can be globally divided into the following major categories: soluble signaling factors (interleukins, chemokines, and growth factors), secreted proteases and secreted insoluble proteins / extracellular matrix (ECM) components.

[0012] Because of their potential involvement in many aging and disease processes, eliminating senescent cells and attenuating the senescence-associated secretory phenotype (SASP) have emerged as attractive therapeutic and cosmetic strategies; however, translation of these findings into relevant human applications is currently limited by the fragmentary understanding of both the basic molecular cell biology of in vivo senescent cells and the overall importance of senescence to age-related diseases.

[0013] Known SASP inhibitors belong to one of the following pharmacological categories: a. Glucocorticoids as potent suppressors of selected components of the SASP;b. Statins such as simvastatin, that can reduce the expression of pro- inflammatory cytokines (IL-6, IL-8, and MCP-1); c. JAK1 / 2 inhibitors such as ruxolitinib; d. NF-κB and p38 inhibitors; e. Mitochondrial depleters in the case of impaired mitophagy.

[0014] There is however the need to develop new SASP inhibitors to meet the desire to shift dermatological focus from ameliorating the cosmetic consequences of skin aging to decreasing the genuine morbidity associated with problems of the aging skin in the face of an ageing population.

[0015] There is also the need to develop new SASP inhibitors to selectively eliminate senescent cells in vivo, thereby reducing inflammation, enhancing immune system function, slowing the progression of age-related diseases, all of which would lead to increased health and lifespan.

[0016] Nelupure is a commercially available blended extract from two types of lotus flower, the Egyptian Lotus (Nymphaea caerulea) and the Sacred Lotus (Nelumbo nucifera). It is speculated that Nelupure acts via leptin-receptor inhibition to regulate the sebaceous glands, preventing the excess of sebum production and inflammatory processes that cause skin blemishes. SUMMARY OF THE INVENTION

[0017] The present invention is partially based on the observation that specific agents inhibit or at least significantly reduce the biosynthesis of secreted frizzled-related protein 4 (SFRP 4) and the expression of p21 in fibroblasts; and additionally inhibit expression of phosphor m-TOR by fibroblasts and increase Caspase-9 activity and Annexin-V binding on normal fibroblasts. The agents may also inhibit the senescence-associated secretory phenotype.

[0018] In one embodiment the active ingredient comprises a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera).

[0019] In another embodiment the active ingredient is one or more flavonoid of Formula (I)[Chem 1]Formula (I) wherein each is a single or double bond; X1= CH, C=O; X2-X10= H, OR; R=H, Alk, mono-, di- or trisaccharide; Alk=C1-C6, straight or branched, saturated or unsaturated, optionally substituted with OR’, NHR’, NR’2 or SR’; R’=H or Alk; and * = (+) if C(2)-O is a double bond

[0020] In one embodiment the flavonoid is one or more of malvidin, delphinidin, cyanidin, petunidin hexosides, and pentosides, kaempferol, quercetin, isorhamnetin, myricetin, apigenin, epicatechin, gallocatechin, apigenin, afzelin, hyperoside, prunin, naringenin, astragalin, trifolin, isorhamnetin, luteolin, rutin, miquelianin, quercetin or taxifolin, preferably wherein the active agent is one or more of kaempferol, miquelianin, myricitrin, myricetin, quercetin or taxifolin.

[0021] In one embodiment the flavonoid is in the form of an extract, preferably wherein the extract is one or more of an extract of mentha longifolia, an extract of camellia sinensis, anextract of disporopsis pernyi an extract of ginkgo biloba, an extract of hamamelis, an extract of salvia, and / or an extract of lavender.

[0022] Given this multiple biochemical activity, the agents can be used to treat and / or prevent senescence and therefore intrinsic and extrinsic aging like the apparent signs of intrinsic skin ageing like hollowed cheeks and eye sockets, loss of skin firmness, sagging, thin and dry skin, fine wrinkles and decreased skin elasticity.

[0023] Reduction in the sense of the present invention means preferably at least about minus 5%, minus 10%, minus 15%, minus 20%, about minus 25%, about minus 30%, about minus 35%, about minus 40%, about minus 45%, about minus 50%, about minus 55%, about minus 60%, about minus 65%, about minus 70%, about minus 75%, about minus 80%, about minus 85%, about minus 90%, about minus 95% compared to the level of the respective marker in non-treated senescent cells. All the percent reductions laying between any pair of the values above are hereby explicitly disclosed.

[0024] The present invention also provides the cosmetic use of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for treating and / or preventing skin senescence, particularly wherein skin senescence comprises the apparent signs of intrinsic skin ageing like hollowed cheeks and eye sockets; loss of skin firmness; sagging, thin and dry skin; fine wrinkles; decreased elasticity; aberrant pigmentation; and hair graying and loss.

[0025] The present invention in particular provides a therapeutically effective amount of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for use in treating and / or preventing senescence, in particular wherein senescence is replicative senescence, DNA-damage induced senescence or oncogene-induced senescence. More particularly senescence can be skin senescence.

[0026] The present invention also provides a therapeutically effective amount of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for use in treating and / or preventing age-associated diseases in humans. The age-associated disease can in particular be cancer, heart disease or dementia.

[0027] According to the present invention, the composition can be for topical application, in particular in the form of an oil-in-water emulsion, a water-in-oil emulsion, a multiple emulsion (Water / Oil / Water or Oil / Water / Oil), a microemulsion, a nanoemulsion, a solution, a suspension, a hydrodispersion, a gel, an ointment, a paste, an aerosol foam, a spray, an aqueous gel, a powder, a foundation, a transdermal patch, a cream or a mask.

[0028] The composition can also be suitable for enteral or parenteral administration, in particular intradermal or subcutaneous administration.

[0029] According to the present invention, each extract of the blend can be present in a cosmetically or pharmaceutically useful amount. For example, a cosmetic or therapeutic composition may comprise at least 0,000001875%by weight of each extract of the blend, preferably between 0.0000075% and 0.15%, more preferably between 0.0001% and 0.1%, even more preferably between 0.000025% and 0.05%, even more preferably between 0.00005% and 0.001%, most preferably 0.0001% of each extract of the blend by weight of the whole composition.

[0030] The ratio of extract of Egyptian Lotus (Nymphaea caerulea) to Sacred Lotus (Nelumbo nucifera) in the composition can be between 10:1 and 1:10, preferably 9, 8, 7, 6, 5, 4, 3, 2, 1:1 and 1:2, 3, 4, 5, 6, 7, 8, 9. BRIEF DESCRIPTION OF THE FIGURES

[0031] In the drawings, which are not necessarily drawn to scale, like numerals describe substantially similar components throughout the several views. The drawings illustrate generally, by way of example, but not by way of limitation, various aspects of the present invention.

[0032] [Fig 1] is a summary of Senescence-Associated SPRF4 (secreted frizzled- related protein 4) and p21 immunostaining of normal human fibroblasts in the presence or absence of an active ingredient comprising a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera).

[0033] [Fig 2] shows the inhibition of expression of phosphor m-TOR by a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) on normal human fibroblasts.

[0034] [Fig 3] shows the increased Caspase-9 activity of normal human fibroblasts treated with a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera).

[0035] [Fig 4] shows the increased Annexin-V binding of normal human fibroblasts treated with a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera). DETAILED DESCRIPTION OF THE INVENTION

[0036] Reference will now be made in detail to certain aspects of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated clauses, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0037] Throughout this document, values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0038] In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. The statement “at least one of A and B” or “at least one of A or B” has the same meaning as “A, B, or A and B.” In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting; information that is relevant to a section heading may occur within or outside of that particular section. Acomma can be used as a delimiter or digit group separator to the left or right of a decimal mark; for example, “0.000,1” is equivalent to “0.0001.” All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0039] The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range and includes the exact stated value or range. The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%. The term “substantially free of” as used herein can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that about 0 wt% to about 5 wt% of the composition is the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than or equal to about 4.5 wt%, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less, or about 0 wt%.

[0040] When percentages of ingredients in compositions are indicated, the standard error applies. For example, 3.0% is to be interpreted as 2.9% to 3.1%

[0041] The term “comprises” and variations thereof is to be interpreted as meaning both “includes”, “substantially consists of” and “consists of”, and variations thereof.

[0042] The present invention provides active agents having both senostatic and senolytic activity. Specifically, the agents according to the invention inhibit or at least significantly reduce the biosynthesis of secreted frizzled-related protein 4 (SFRP 4) and the expression of p21 in fibroblasts; and additionally inhibit expression of phosphor m-TOR by fibroblasts and increase Caspase-9 activity and Annexin-V binding on normal fibroblasts. The agents may also inhibit the senescence-associated secretory phenotype.

[0043] In one embodiment the active ingredient comprises a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera).

[0044] In another embodiment the active ingredient is one or more flavonoid of Formula (I) [Chem 1]Formula (I) wherein each is a single or double bond; X1= CH, C=O; X2-X10= H, OR; R=H, Alk, mono-, di- or trisaccharide; Alk=C1-C6, straight or branched, saturated or unsaturated, optionally substituted with OR’, NHR’, NR’2 or SR’; R’=H or Alk; and * = (+) if C(2)-O is a double bond

[0045] In one embodiment the flavonoid is one or more of malvidin, delphinidin, cyanidin, petunidin hexosides, and pentosides, kaempferol, quercetin, isorhamnetin, myricetin, apigenin, epicatechin, gallocatechin, apigenin, afzelin, hyperoside, prunin, naringenin, astragalin, trifolin, isorhamnetin, luteolin, rutin, miquelianin, quercetin or taxifolin, preferably wherein the active agent is one or more of kaempferol, miquelianin, myricitrin, myricetin, quercetin or taxifolin.

[0046] In one embodiment the flavonoid is in the form of an extract, preferably wherein the extract is one or more of an extract of mentha longifolia, an extract of camellia sinensis, an extract of disporopsis pernyi an extract of ginkgo biloba, an extract of hamamelis, an extract of salvia, and / or an extract of lavender.

[0047] In one embodiment the flavonoid is not myricetin or derivatives thereof

[0048] The present invention specifically provides the cosmetic use of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for treating and / or preventing skin senescence. The extract may be present in a cosmetically effective amount.

[0049] In particular, the present invention provides a cosmetic use of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for treating and / or preventing the apparent signs of intrinsic and extrinsic skin aging like hollowed cheeks, eye sockets, loss of skin firmness, sagging, thin and dry skin, fine wrinkles and decreased elasticity. The extract may be present in a cosmetically effective amount.

[0050] The present invention also provides a therapeutically effective amount of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for use in treating and / or preventing senescence, in particular wherein senescence is replicative senescence, DNA-damage induced senescence or oncogene-induced senescence. More particularly senescence can be skin senescence.

[0051] The present invention also provides a therapeutically effective amount of a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) or of a flavonoid as disclosed above for use in treating and / or preventing age-associated diseases in humans. The age-associated disease can in particular be cancer, heart disease or dementia.

[0052] According to the present invention, each extract of the blend can be present in a cosmetically or pharmaceutically useful amount. For example, a cosmetic or therapeutic composition comprises between 0.0000075% and 0.15%, more preferably between 0.0001%and 0.1%, even more preferably between 0.000025% and 0.05% of each extract of the blend by weight of the whole composition.

[0053] The ratio of extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) in the composition can be between 10:1 and 1:10, preferably 9, 8, 7, 6, 5, 4, 3, 2, 1:1 and 1:2, 3, 4, 5, 6, 7, 8, 9.

[0054] The compositions of the present invention may comprise other agents active to treat or prevent senescence.

[0055] The topical compositions according to the invention may comprise at least one further ingredient commonly found in topical cosmetic and / or medical (e.g. dermatological) compositions chosen from water, oils, which may be chosen in particular from volatile and / or non-volatile, linear or cyclic silicone oils, waxes (such as ozokerite, polyethylene wax, beeswax or carnauba wax), silicone elastomers, nonionic, anionic, cationic and / or amphoteric surfactants, co-surfactant (such as linear fatty alcohols), thickeners, gelling agents, humectants (such as polyols like glycerin), colorants, preservatives, fillers, tensors, sequestrants, perfumes, and mixtures hereof.

[0056] These compositions can be in particular in the form of oil-in-water emulsions, water-in-oil emulsions, multiple emulsions (Water / Oil / Water or Oil / Water / Oil) which can optionally be microemulsions or nanoemulsions, or in the form of solutions, suspensions, hydrodispersions, gels, ointments, pastes, aerosol foams or sprays, aqueous gels, powders, or foundations or transdermal patches. They can be more or less fluid and have the appearance of creams, emulsions, gels, masks or any other aspect of healthy skin care cosmetics. The person skilled in the art is aware of the procedures, ingredients and forms of common use in the preparation of dermal formulations.

[0057] A “gel” is a colloid in which the dispersed phase has combined with the continuous phase to produce a semisolid material, such as jelly.

[0058] An “oil” is a composition containing at least 95% wt of a lipophilic substance. Examples of lipophilic substances include but are not limited to naturally occurring and synthetic oils, fats, fatty acids, lecithins, triglycerides and combinations thereof.

[0059] A “continuous phase” refers to the liquid in which solids are suspended or droplets of another liquid are dispersed and is sometimes called the external phase. This also refers to the fluid phase of a colloid within which solid or fluid particles are distributed. If the continuous phase is water (or another hydrophilic solvent), water-soluble or hydrophilic drugs will dissolve in the continuous phase (as opposed to being dispersed). In a multiphase formulation (e.g., an emulsion), the discreet phase is suspended or dispersed in the continuous phase. Excipients for topical administration may include anti-microbial compounds, e.g. parabens, antioxidants, e.g. sodium ascorbyl acetate and alpha-tocopherol, stabilizers, e.g. sorbitol, and / or emulsifying agents to produce a stable emulsion with both a hydrophilic and a hydrophobic phase.

[0060] “Diluents” may be included in the formulations to dissolve, disperse or otherwise incorporate the carrier. Examples of diluents include, but are not limited to, water, buffered aqueous solutions, organic hydrophilic diluents, such as monovalent alcohols, and low molecular weight glycols and polyols (e.g. propylene glycol, polypropylene glycol, glycerol, butylene glycol).

[0061] Appropriate excipients are selected based on the type of formulation. Standard excipients include gelatin, casein, lecithin, gum acacia, cholesterol, tragacanth, stearic acid, benzalkonium chloride, calcium stearate, glyceryl monostearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyethylene glycols, polyoxyethylene stearates, colloidol silicon dioxide, phosphates, sodium dodecyl sulfate, carboxymethylcellulose calcium, carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethycellulose phthalate, noncrystalline cellulose, magnesium aluminum silicate, triethanolamine, polyvinyl alcohol, polyvinylpyrrolidone, sugars, and starches.

[0062] An emulsion is a preparation of one liquid distributed in small globules throughout the body of a second liquid. The dispersed liquid is the discontinuous phase, and the dispersion medium is the continuous phase. When oil is the dispersed liquid and an aqueous solution is the continuous phase, it is known as an oil-in-water emulsion, whereas when water or aqueous solution is the dispersed phase and oil or oleaginous substance is the continuousphase, it is known as a water-in-oil emulsion. The oil phase may consist at least in part of a propellant, such as an HFA propellant. Either or both of the oil phase and the aqueous phase may contain one or more surfactants, emulsifiers, emulsion stabilizers, buffers, and other excipients. Preferred excipients include surfactants, especially non-ionic surfactants; emulsifying agents, especially emulsifying waxes; and liquid non-volatile non-aqueous materials, particularly glycols such as propylene glycol. The oil phase may contain other oily pharmaceutically approved excipients. For example, materials such as hydroxylated castor oil or sesame oil may be used in the oil phase as surfactants or emulsifiers.

[0063] “Emollients” are an externally applied agent that softens or soothes skin and are generally known in the art and listed in compendia, such as the “Handbook of Pharmaceutical Excipients”, 4thEd., Pharmaceutical Press, 2003. These include, without limitation, almond oil, castor oil, ceratonia extract, cetostearoyl alcohol, cetyl alcohol, cetyl esters wax, cholesterol, cottonseed oil, cyclomethicone, ethylene glycol palmitostearate, glycerin, glycerin monostearate, glyceryl monooleate, isopropyl myristate, isopropyl palmitate, lanolin, lecithin, light mineral oil, medium-chain triglycerides, mineral oil and lanolin alcohols, petrolatum, petrolatum and lanolin alcohols, soybean oil, starch, stearyl alcohol, sunflower oil, xylitol and combinations thereof. In one embodiment, the emollients are ethylhexylstearate and ethylhexyl palmitate.

[0064] “Surfactants” are surface-active agents that lower surface tension and thereby increase the emulsifying, foaming, dispersing, spreading and wetting properties of a product. Suitable non-ionic surfactants include emulsifying wax, glyceryl monooleate, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polysorbate, sorbitan esters, benzyl alcohol, benzyl benzoate, cyclodextrins, glycerin monostearate, poloxamer, povidone and combinations thereof. In one embodiment, the non-ionic surfactant is stearyl alcohol.

[0065] “Emulsifiers” are surface active substances which promote the suspension of one liquid in another and promote the formation of a stable mixture, or emulsion, of oil and water. Common emulsifiers are metallic soaps, certain animal and vegetable oils, and various polar compounds. Suitable emulsifiers include acacia, anionic emulsifying wax, calcium stearate, carbomers, cetostearyl alcohol, cetyl alcohol, cholesterol, diethanolamine, ethylene glycol palmitostearate, glycerin monostearate, glyceryl monooleate, hydroxpropyl cellulose,hypromellose, lanolin, hydrous, lanolin alcohols, lecithin, medium-chain triglycerides, methylcellulose, mineral oil and lanolin alcohols, monobasic sodium phosphate, monoethanolamine, nonionic emulsifying wax, oleic acid, poloxamer, poloxamers, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, propylene glycol alginate, self-emulsifying glyceryl monostearate, sodium citrate dehydrate, sodium lauryl sulfate, sorbitan esters, stearic acid, sunflower oil, tragacanth, triethanolamine, xanthan gum and combinations thereof. In one embodiment, the emulsifier is glycerol stearate.

[0066] A “lotion” is a low- to medium-viscosity liquid formulation. A lotion can contain finely powdered substances that are in soluble in the dispersion medium through the use of suspending agents and dispersing agents. Alternatively, lotions can have as the dispersed phase liquid substances that are immiscible with the vehicle and are usually dispersed by means of emulsifying agents or other suitable stabilizers. In one embodiment, the lotion is in the form of an emulsion having a viscosity of between 100 and 1000 centistokes. The fluidity of lotions permits rapid and uniform application over a wide surface area. Lotions are typically intended to dry on the skin leaving a thin coat of their medicinal components on the skin's surface. A “cream” is a viscous liquid or semi-solid emulsion of either the “oil-in-water” or “water-in-oil type”. Creams may contain emulsifying agents and / or other stabilizing agents. In one embodiment, the formulation is in the form of a cream having a viscosity of greater than 1000 centistokes, typically in the range of 20,000-50,000 centistokes. Creams are often time preferred over ointments as they are generally easier to spread and easier to remove.

[0068] An “ointment” is a semisolid preparation containing an ointment base and optionally one or more active agents. Examples of suitable ointment bases include hydrocarbon bases (e.g., petrolatum, white petrolatum, yellow ointment, and mineral oil); absorption bases (hydrophilic petrolatum, anhydrous lanolin, lanolin, and cold cream); water-removable bases (e.g., hydrophilic ointment), and water-soluble bases (e.g., polyethylene glycol ointments). Pastes typically differ from ointments in that they contain a larger percentage of solids. Pastes are typically more absorptive and less greasy that ointments prepared with the same components.

[0069] A “gel” is a semisolid system containing dispersions of small or large molecules in a liquid vehicle that is rendered semisolid by the action of a thickening agent or polymeric material dissolved or suspended in the liquid vehicle. The liquid may include a lipophilic component, an aqueous component or both. Some emulsions may be gels or otherwise include a gel component. Some gels, however, are not emulsions because they do not contain a homogenized blend of immiscible components. Suitable gelling agents include, but are not limited to, modified celluloses, such as hydroxypropyl cellulose and hydroxyethyl cellulose; Carbopol homopolymers and copolymers; and combinations thereof. Suitable solvents in the liquid vehicle include, but are not limited to, diglycol monoethyl ether; alkylene glycols, such as propylene glycol; dimethyl isosorbide; alcohols, such as isopropyl alcohol and ethanol. The solvents are typically selected for their ability to dissolve the drug. Other additives, which improve the skin feel and / or emollience of the formulation, may also be incorporated. Examples of such additives include, but are not limited, isopropyl myristate, ethyl acetate, C12- C15 alkyl benzoates, mineral oil, squalane, cyclomethicone, capric / caprylic triglycerides, and combinations thereof.

[0070] “Foams” consist of an emulsion in combination with a gaseous propellant. The gaseous propellant consists primarily of hydrofluoroalkanes (HFAs). Suitable propellants include HFAs such as 1,1,1,2-tetrafluoroethane (HFA 134a) and 1,1,1,2,3,3,3- heptafluoropropane (HFA 227), but mixtures and admixtures of these and other HFAs that are currently approved or may become approved for medical use are suitable. The propellants preferably are not hydrocarbon propellant gases which can produce flammable or explosive vapors during spraying. Furthermore, the compositions preferably contain no volatile alcohols, which can produce flammable or explosive vapors during use.

[0071] Buffers are used to control pH of a composition. Preferably, the buffers buffer the composition from a pH of about 4 to a pH of about 7.5, more preferably from a pH of about 4 to a pH of about 7, and most preferably from a pH of about 5 to a pH of about 7. In a preferred embodiment, the buffer is based on triethanolamine, citric acid, sodium hydroxyde, disodium phosphate and potassium phosphate.

[0072] Preservatives can be used to prevent the growth of fungi and microorganisms. Suitable antifungal and antimicrobial agents include, but are not limited to, benzoic acid,butylparaben, ethyl paraben, methyl paraben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, potassium sorbate, caprylyl alcohol, 1,2- hexandiol, and thimerosal.

[0073] Antioxidants can be used in the compositions of the present invention. Examples of antioxidants are tocopherol, hydroxyacetophenone, dimethylmethoxy chromanol and phenethyl alcohol.

[0074] Viscosity modifiers can additionally be present, such as potassium chloride and natural gums such as xanthan gum, gum arabic, gum tragacanth, carrageenan, chitosan, guar gum, konjac gum, sclerotium gum, dextrins and starch.

[0075] Chelating agents may additionally be present, such as sodium glucuronate.

[0076] Fragrances may be added to the compositions of the invention.

[0077] Penetration enhancers are frequently used to promote transdermal delivery of drugs across the skin, in particular across the stratum corneum. The more commonly used enhancers include urea, (carbonyldiamide), imidurea, N, N-diethylformamide, N-methyl-2- pyrrolidine, 1-dodecal-azacyclopheptane-2-one, calcium thioglycate, 2-pyyrolidine, N,N- diethyl-m-toluamide, oleic acid and its ester derivatives, such as methyl, ethyl, propyl, isopropyl, butyl, vinyl and glycerylmonooleate, sorbitan esters, such as sorbitan monolaurate and sorbitan monooleate, other fatty acid esters such as isopropyl laurate, isopropyl myristate, isopropyl palmitate, diisopropyl adipate, propylene glycol monolaurate, propylene glycol monooleatea and non-ionic detergents such as BRIJ® 76 (stearyl poly(10 oxyethylene ether), BRIJ® 78 (stearyl poly(20)oxyethylene ether), BRIJ® 96 (oleyl poly(10)oxyethylene ether), and BRIJ® 721 (stearyl poly (21) oxyethylene ether) (ICI Americas Inc. Corp.).

[0078] Emulsion stabilizers, surfactants and emulsifying agents may additionally be present, such as sorbitan isostearate, hydroxyethyl acrylate / sodium acryloxydiethyl taurate copolymer and sorbitan isostearate.

[0079] Humectants are crucial in skincare by providing essential skin hydration and maintaining the skin's moisture balance. Exemplary humectant are Glycerin, Hyaluronic acid, Aloe vera, Honey, Hydrolyzed proteins, Panthenol, Allantoin, Seaweed & algae, Sodium PCA(sodium pyrrolidone carboxylic acid), Saccharide Isomerate, Sugar Alcohols (e.g. sorbitol), Hydroxy acids (e.g.. lactic acid and gluconolactone), Agave nectar, Betaine, Chitosan, Trehalose, Beta-glucan, Topical collagen, Galactoarabinan and propanediol.

[0080] Solvents used in the compositions of the present invention may include water, alcohols such as ethanol and isopropyl alcohol, alkanes such as C15-19 Alkane, silicone oils and mineral oils.

[0081] Skin conditioning agents may be added to the compositions of the present invention, for example caprylic / capric triglyceride, alpha-glucan triglycerides, palmitoyl tripeptide-5, glycerine, diglucosyl gallic acid, panthenol, pantolactone, sodium stearoyl glutamate, lactic acid, hydrolysed hyaluronic acid, sodium hyaluronate and maltodextrin.

[0082] The compositions may additionally contain SPF and UVA protective agents.

[0083] By way of illustration of organic UV filters and in a non-limiting manner, mention may be made of: - anthranilates, in particular menthyl anthranilate; - benzophenones, in particular 1-benzophenone, 3-benzophenone or oxybenzone, 5- benzophenone, 6-benzophenone, 8-benzophenone, 9-benzophenone, 12- benzophenone, and preferably 2-benzophenone (Oxybenzone), or 4-benzophenone (Uvinul MS40® available from B.A.S.F.); - benzylidenecamphor, in particular 3-benzylidenecamphor, benzylidenecamphorsulphonic acid, benzalkoniumcamphor methosulfate, polyacrylamidomethylbenzylidenecamphor, terephthalylidenecamphorsulphonic acid, and preferably 4-methylbenzylidenecamphor (Eusolex 6300® available from Merck); - benzimidazoles, in particular benzimidazilate (Neo Heliopan AP® available from Haarmann and Reimer), or phenylbenzimidazole sulfonic acid (PARSOL HS® available from DSM); - benzotriazoles, in particular drometrizole trisiloxane, or methylene bis- benzotriazolyltetramethylbutylphenol (Tinosorb M® available from Ciba); - cinnamates, in particular cinoxate, DEA methoxycinnamate, diisopropyl methylcinnamate, dimethoxycinnamate glyceryl ethylhexanoate, isopropyl methoxycinnamate, isoamyl cinnamate, Kaempferia galanga root extract (TEGOGALANGA from EVONIK containing 98% ethyl-p-methoxycinnamate) and preferably octylmethoxycinnamate (Parsol MCX® available from Hoffmann La Roche); - diphenylacrylates, in particular ethocrylene (Uvinul N35® available from B.A.S.F.), or octocrylene (Uvinul 539® available from B.A.S.F.) or Ethylhexyl methoxycrylene (SOLASTAY available from HALLSTAR); - dibenzoylmethanes, in particular butyl methoxydibenzoylmethane (Parsol 1789®); imidazolines, in particular ethylhexyl dimethoxybenzylidenedioxoimidazoline; - PABA, in particular ethyl dihydroxypropyl PABA, ethylhexyldimethyl PABA, glyceryl PABA, PABA, PEG-25 PABA, or ethyl PABA (benzocaine); - triazines, in particular anisotriazine (Tinosorb S® available from Ciba) or diethylhexylbutamido-triazone (Uvasorb HEB® available from 3V Sigma), ethylhexyltriazone (llvinul T150® available from B. A. S. F.), Tris-Biphenyl Triazine (Tinosorb 2AB available from BASF), benzoates, in particular N-hexyl 2-(4- diethylamino-2-hydroxybenzoyl)benzoate (llvinul A+ available from BASF) or as a mixture with octyl methoxycinnamate (Uvinul A+B available from BASF); - benzalmalonates, in particular Polysilicone-15 (Parsol SLX available from DSM); - benzoxazoles, in particular 2,4-bis[4-[5-(1 .1 -dimethylpropyl)benzoxazol-2- yl]phenylimino]-6-[(2-ethylhexyl)imino]-1.3,5-triazine, (Uvasorb K2A available from Sigma 3 V); - salicylates, in particular dipropylene glycol salicylate, ethylhexyl salicylate, homosalate, butyloctyl salicylate (HALLBRITE BHB available from HALLSTAR) or TEA salicylate; - Phenylbenzimidazole sulfonic acid (eg Parsol HS), Octocrylene (eg Parsol 340), Ethylhexyl Methoxycinnamate (eg Parsol MCX), Methylene Bis-Benzotriazolyl Tetramethylbutylphenol (eg Tinosorb M), Bis-Ethylhexyloxyphenol Methoxyphenyl Triazine (eg Tinosorb S), Tris-Biphenyl Triazine (eg Tinosorb 2AB), Ethylhexyl Triazone (eg UVINUL T150) and Diethylhexyl Butamido Triazone (eg UVASORB HEB), Diethylamino Hydroxybenzoyl Hexyl Benzoate (eg Uvinul A PLUS), Butyl Methoxydibenzoylmethane (eg Parsol 1789), Polysilicone-15 (eg Parsol SLX), Benzophenone-4 (eg Uvinul MS 40), Benzophenone-3, Ethylhexyl Salicylate (eg Parsol EHS), Homosalate;- and mixtures thereof.

[0084] Inorganic UV screening agents can be used, for example metal oxide particles having an average elementary particle size of less than or equal to 100 nm. They may be chosen in particular from titanium oxide, zinc oxide, iron oxide, zirconium oxide, cerium oxide, and mixtures thereof.

[0085] Sun Protection Factor (SPF) indicates a sunscreen’s, cosmetic or pharmaceutical preparation’s or homecare product’s ability to protect against UVB rays but does not adequately designate to what extent (if any) a sunscreen composition protects against UVA radiation. SPF is a measure of how much solar energy (UV radiation) is required to produce sunburn on protected skin, i.e., in the presence of sunscreen, relative to the amount of solar energy required to produce sunburn on unprotected skin. As the SPF value increases, sunburn protection increases.

[0086] After formulation, the product is filled into an appropriate dispenser and shipped to the end user. Examples of final container may include a pump bottle, squeeze bottle, jar, tube or vial.

[0087] The invention will now be illustrated by non-limiting examples of compositions according to the invention and by test results. EXEMPLES Example 1

[0088] Normal human dermal fibroblasts were cultured with DMEM medium (Dulbecco’s Modified Eagle Medium) containing FBS (fetal bovine serum) at 5%. Cellular senescence was induced by a chronic oxidant stress generated with repeated exposure of hydrogen peroxide (H2O2 solution). During the cellular senescence induction, cells were treated with the composition. Non treated cells were cultivated in the same conditions. Cellular senescence was studied by the quantification of SFRP4 (secreted frizzled-related protein 4) and p21 through immunostaining.Results

[0089] Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract tested at 0.0015% each induced a significant reduction of respectively 55% and 26% of SFRP4 and p21 expression. Example 2

[0090] Normal human dermal fibroblasts were cultured with DMEM medium (Dulbecco’s Modified Eagle Medium) containing FBS (fetal bovine serum) at 10%. Cellular senescence was induced by a chronic oxidant stress generated with repeated exposure of hydrogen peroxide (H2O2 solution). After stress induced senescence, cells were treated with the composition comprising Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract for 6 days. Non treated cells were cultivated in the same conditions. Phospho m-TOR was quantified using Elisa essay and normalized with the total proteins content. Results

[0091] Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract tested at 0.0015% each induced a significant decrease of phosphor m-TOR by 26%. Example 3

[0092] Normal human dermal fibroblasts were cultured with DMEM medium (Dulbecco’s Modified Eagle Medium) containing FBS (fetal bovine serum) at 10%. Cellular senescence was induced by a chronic oxidant stress generated with repeated exposure of hydrogen peroxide (H2O2 solution). After stress induced senescence, cells were treated with the composition comprising Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract for 4 days. Non treated cells were cultivated in the same conditions. Caspase-9 activity was quantified using Caspase-Glo 9 essay and normalized with the total proteins content. Results

[0093] Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract tested at 0.0015% each induced a significant increase of Caspase-9 activity by 135%.Example 4

[0094] Normal human dermal fibroblasts were cultured with DMEM medium (Dulbecco’s Modified Eagle Medium) containing FBS (fetal bovine serum) at 10%. Cellular senescence was induced by a chronic oxidant stress generated with repeated exposure of hydrogen peroxide (H2O2 solution). After stress induced senescence, cells were treated with the composition comprising Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract for 4 days. Non treated cells were cultivated in the same conditions. Annexin- V activity was quantified using bioluminescence essay and normalized with the total proteins content. Results

[0095] Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract tested at 0.0015% each induced a significant increase of Annexin-V binding by 61% . Discussion and conclusions

[0096] These studies demonstrate that a blend of Nymphaea Caerulea Flower Extract and Nelumbo Nucifera Flower Extract inhibit the biosynthesis of secreted frizzled-related protein 4 (SFRP 4) and the expression of p21 in fibroblasts, which are known markers of cellular senescence, and additionally inhibit expression of phosphor m-TOR by fibroblasts and increase Caspase-9 activity and Annexin-V binding on normal fibroblasts, which are markers of senolytic activity.

[0097] By minimizing cellular senescence, these actives can be beneficial for improving skin texture and decelerating the skin aging process.

[0098] This anti senescence approach can be implemented in antiaging skincare strategies to help increase healthy cells, aged or not, in their metabolic functions and in particular in collagen synthesis.

Claims

CLAIMS What is claimed is:

1. Cosmetic use of an active agent having senolytic and senostatic activity for treating and / or preventing skin senescence.

2. Cosmetic use according to Claim 1, wherein the agent has senostatic activity by inhibiting or reducing the biosynthesis of secreted frizzled-related protein 4 (SFRP 4) and reducing expression of p21 in fibroblasts.

3. Cosmetic use according to Claim 1, wherein the agent has senolytic activity and is capable of increasing Caspase-9 activity and Annexin-V binding on normal fibroblasts.

4. Cosmetic use according to any of Claims 1-3, wherein the active agent is a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera).

5. Cosmetic use according to any of the preceding Claims, wherein skin senescence comprises the apparent signs of intrinsic and / or extrinsic skin ageing.

6. Cosmetic use according to any of the preceding Claims, wherein the apparent signs of intrinsic skin ageing comprise hollowed cheeks, eye sockets, loss of skin firmness, sagging, thin and dry skin, fine wrinkles and decreased elasticity.

7. Cosmetic use according to any of the preceding Claims, wherein the agent supports healthy cells, aged or not, in their metabolic functions, in particular in collagen synthesis.

8. Cosmetic use according to any of the preceding Claims, wherein the extracts are present in a cosmetically useful amount.

9. Therapeutically effective amount of an active agent having senolytic and senostatic activity for use in treating and / or preventing skin senescence or age-associated diseases in humans.

10. Therapeutically effective amount of an active agent for use according to Claim 9 wherein the agent has senostatic activity by inhibiting or reducing the biosynthesis of secreted frizzled-related protein 4 (SFRP 4) and reducing expression of p21 in fibroblasts.

11. Therapeutically effective amount of an active agent for use according to Claim 9, wherein the active agent has senolytic activity and is capable of increasing Caspase-9 activity and Annexin-V binding on normal fibroblasts.

12. Therapeutically effective amount of an active agent for use according to any of Claims 9-11, wherein the active agent is a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera).

13. Therapeutically effective amount for use according to any of Claims 9-12, wherein senescence is replicative senescence, DNA-damage induced senescence or oncogene-induced senescence.

14. Therapeutically effective amount for use according to any of claims 8 to 13, wherein the age-associated disease is cancer, heart disease or dementia.

15. Cosmetic or pharmaceutic composition comprising a blended extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) and further comprising an SPF or UVA protective agent.

16. Cosmetic or pharmaceutic composition according to claim 15, wherein each extract of the blend is present in an amount between 0.0000075% and 0.15%, more preferably between 0.0001% and 0.1%, even more preferably between 0.000025% and 0.05% by weight of the whole composition.

17. Cosmetic or pharmaceutic composition according to claim 16, wherein the ratio of extract of Egyptian Lotus (Nymphaea caerulea) and Sacred Lotus (Nelumbo nucifera) in the composition can be between 10:1 and 1:

10.

18. Cosmetic or pharmaceutic composition according to any of claims 15 to 17, wherein the composition is in the form of an oil-in-water emulsion, a water-in-oil emulsion, a multiple emulsion (Water / Oil / Water or Oil / Water / Oil), a microemulsion, a nanoemulsion, a solution, a suspension, a hydrodispersion, a gel, an ointment, a paste, an aerosol foam, a spray, an aqueous gel, a powder, a foundation, a transdermal patch, a cream or a mask.

19. Cosmetic or pharmaceutic composition according to any of claims 15 to 18, wherein the composition is for enteral or parenteral administration, in particular intradermal or subcutaneous administration.

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