A topical cosmetic formulation containing a novel peptide that improves skin appearance and regeneration.

Abstract

The present invention relates to novel naturally occurring and synthetic active peptides or peptide-derived agents designed for the cosmetic treatment of human skin, as well as cosmetic formulations and compositions containing them. The active agents are effective in restoring, promoting, and maintaining healthy skin. In particular, the present invention discloses a combination or set of skin active agents, including stem cell factors, that modulate the skin microenvironment and regulate skin stem cell behavior, thereby effectively healing, regenerating, and improving the condition of aged or damaged skin.

Description

[Technical Field]

[0001] This invention relates to novel naturally derived and synthetic active peptides or peptide-derived agents designed for cosmetic treatment of human skin, as well as cosmetic formulations and compositions containing them. The active agents are effective in restoring, promoting, and maintaining healthy skin.

[0002] In particular, the present invention discloses combinations or sets of the aforementioned skin-effective agents, including stem cell factors, that induce the repair and regeneration of skin cells, thereby effectively healing and improving the condition of aged or damaged skin through synergistic effects or mutual mitigation of undesirable effects. These novel sets of agents and factors are referred to as “trigger factor complexes” according to the present invention. [Background technology]

[0003] Skin homeostasis and wound healing Human skin is one of the body's primary barriers to the external environment and is constantly exposed to harmful damage. To prevent the continuous deterioration of skin condition, biology has developed various regenerative mechanisms. In maintaining homeostasis, a constant metabolic turnover of cells in the skin epithelium causes old cells to slough off the surface, and new cells are compensatoryly generated to replace the lost cells. In cases of more severe injury, such as trauma (wounds), radiation, chemical damage, or inflammation, damage cues accumulate in the skin, and a sophisticated natural repair program takes place. In both the homeostatic cellular turnover program and the dedicated repair program, stem cells and progenitor cells act as key executing factors in maintaining skin. In both cases, they ultimately provide new cells to replace old and lost cells. Typically, the dedicated repair program proceeds through the following four phases: (I) hemostasis (in the case of open wounds), (II) inflammation, (III) proliferation, and (IV) tissue remodeling. The processes of both the homeostatic cellular turnover program and the dedicated repair program (including their subphases) must be strictly regulated. For example, misregulation of cell proliferation due to the inability to limit the generation of new cells through both homeostasis and dedicated repair programs can lead to cancer development. Conversely, insufficient supply of new cells slows down the regeneration process and impairs the skin's barrier function. Similarly, the inability to limit or block inflammation can lead to chronic inflammatory diseases, local tissue degeneration, stem cell pool depletion, or autoimmune diseases. Conversely, a failure to produce a proper and significant inflammatory response to harmful injuries prevents the skin from exhibiting its full regenerative capacity, equally impairing the skin's barrier function and increasing the risk of opportunistic infections. At the tissue remodeling level, if this phase does not occur, the skin will be left with impaired temporary tissue, such as loss of skin appendages and suboptimal extracellular matrix composition. Conversely, if the tissue remodeling phase is not completed, an inaccurate extracellular matrix composition will occur, equally contributing to fibrosis. Therefore, the body has developed sophisticated mechanisms to ensure that the skin's maintenance and repair programs function in an orderly manner.However, tissue regeneration is not necessarily a linear path through a predefined series of steps, which contributes to further complexity. For example, when the natural repair mechanism is overwhelmed by very large wounds, aging, or constant injuries and the accompanying depletion of regenerative capacity, functional repair is impaired, and ultimately a "damage control" program with scarring occurs.

[0004] stem cells Stem cells and progenitor cells contribute to skin healing by providing new cells. Different stem cell populations give rise to functional skin cells according to skin compartments. The epidermis is the outermost layer of the skin, and the homeostasis during wound and cell turnover during re-epithelialization are mediated mostly by stem cells present in the epidermis. Most of the epidermal surface is covered by the interfollicular epidermis (IFE), and other epithelial structures of the skin epithelium include hair follicles and sweat glands. Different populations of stem cells and progenitor cells exist in their respective niches within the skin epithelium: stem cells in the IFE (Itg2α high , Itg1α high ), progenitor cells in the IFE (Inv + , Lgr6 + ), stem cells in the infundibulum (Lrig1 + ), stem cells in the sebaceous duct (Gata6 + ), stem cells in the isthmus and sebaceous gland (Lrig1 + , Lgr6 + , Blimp1 + , Plet1 + ), and stem cells in the bulge (K15 + , K19 + , Lgr5 + , CD34 + , Sox9 + , Tcf3 +(Dekoninck & Blanpain, 2019, Nature Cell Biology, 21(1), 18-24). All of these cells can contribute to the temporary replenishment of IFE epithelial cells at the wound site. Furthermore, both resident mesenchymal stem cells (MSCs) (Crigler et al., 2007, The FASEB Journal, 21(9), 2050-2063) and hematopoietic stem cells (HSCs) (Fan et al., 2006, Experimental Hematology, 34(5), 672-679) can contribute to epithelial regeneration to some extent. However, among all these sources, IFE stem cells and the aforementioned cells are the greatest epithelial cell contributors to both short-term and long-term wound repair (Blanpain & Fuchs, 2014, Science, 344(6189)). MSCs have been reported to reside in defined niches within the hair follicle's dermal papilla (DP) and connective tissue sheath (CTS) (Lau, Paus, Tiede, Day, & Bayat, 2009, Experimental Dermatology, 18(11), 921-933). Furthermore, the dermal and epidermal compartments are not isolated from each other, but rather communicate and cooperate. For example, MSCs are involved in paracrine secretion loops with keratinocytes and their precursor cells, thereby stimulating re-epithelialization (Lau et al., 2009).

[0005] Mesenchymal stem cells (MSCs) play a crucial role in skin homeostasis, cell turnover, ECM dynamics, and tissue regeneration. MSCs replenish the mesenchymal cell pool, participate in the deposition and degradation of ECM proteins, and regulate tissue dynamics through the secretion of growth factors and cytokines. Various subclasses of MSCs reside in different niches within the skin (Hu, Borrelli, Lorenz, Longaker, & Wan, 2018, Stem Cells International, 2018, 1-13). These include hair follicle (HF) resident cells such as dermal sheath cells and dermal papillary cells, interfollicular MSCs in the dermis, vascular-associated pericytes, and adipose-derived MSCs in the subcutaneous tissue. Furthermore, contributions from primarily bone marrow-derived MSCs infiltrating the skin from the vascular system have been reported. Stem cells are generally defined by their ability to regenerate and differentiate into functional cell types. Since normal fibroblasts are morphologically indistinguishable from MSCs and formally meet the definition criteria for (pluripotent) stem cells, the past distinction between MSCs and fibroblasts in terms of stem cell status is under debate (Soundararajan & Kannan, 2018, Journal of Cellular Physiology, Wiley-Liss Inc, December 1). Regardless of its formal classification, the role of fibroblasts in skin homeostasis, ECM dynamics, and wound healing is well established (Rognoni & Watt, 2018, Trends in Cell Biology, 28(9), 709-722). Nevertheless, the diversity of mesenchymal phenotypes is further detailed by lineages associated with the dermis, e.g., papillary (upper dermis) and reticular (lower dermis) fibroblasts (Driskell et al., 2013, Nature, 504(7479), 277-281). Papillary lineages have a "regeneration-promoting" phenotype and are necessary for hair follicle formation, while reticular lineages are necessary for rapid wound closure but also contribute to fibrosis-related ECM deposition in a "scarring-promoting" manner.Heterogeneity of mesenchymal cells has also been studied at the molecular level (Philippeos et al., 2018, Journal of Investigative Dermatology, 138(4), 811-825; Vaculik et al., 2012, Journal of Investigative Dermatology, 132(3 PART 1), 563-574), and more detailed phylogenetic relationships have recently been outlined (Lynch & Watt, 2018).

[0006] Furthermore, in addition to microenvironment (dermal layer)-related phenotypic diversity in mesenchymal cells, cell-specific heterogeneity also exists and is a dominant determinant of regenerative behavior. One major factor is the Engrailed-1 (En-1) state (Jiang et al., 2018, Nature Cell Biology, 20(4), 422-431; Rinkevich et al., 2015, Science, 348(6232)). En-1 negative fibroblasts (ENFs) mediate scarless wound healing during embryonic development. However, the number of ENFs decreases after embryonic development, and En-1 positive fibroblasts (EPFs) emerge as a dominant lineage, promoting scarring.

[0007] Attributes of aging skin In aging skin, cell replacement continues to decrease, impairing barrier function and mechanical protection, delaying wound healing and immune responses, impaired thermoregulation, and reduced sweat and sebum production (Farage, Miller, & Maibach, 2010, Textbook of Aging Skin, 1-1220). The slower rate of cell turnover leads to rough skin, delayed wound healing, and uneven pigmentation. Older adults often suffer from drier skin than younger, healthy individuals. This is based on the decreased function of the sebaceous glands in the production of natural moisturizing factors and lipids in the stratum corneum, resulting in a reduction of the lamellar bilayer and decreased water retention. Skin aging also involves large-scale remodeling of the extracellular matrix (ECM) in the dermis, aging of dermal fibroblasts, dramatic upregulation of matrix metalloproteinases (MMPs), and decreased collagen production. As a result, an overall deficiency and fragmentation of collagen, elastic fibers, and other ECM proteins occurs, leading to a loss of tensile strength and manifesting as wrinkles and sagging. Furthermore, flattening of the dermal papillae increases the risk of blister formation and, consequently, infection.

[0008] Skin regeneration and scarring in damaged skin correlate with signaling molecule patterns. Biological processes are regulated at various levels, including the cellular and molecular levels. Cells, including stem cells, integrate internal states with external cues to make biological decisions. Similarly, physiological skin homeostasis and regeneration are governed by a predetermined set of signaling molecules that act at predetermined locations and times.

[0009] Enhanced skin regeneration by trigger factors Despite its ability to maintain most functional skin, the body's homeostasis and self-repair mechanisms are not perfect. This can be exacerbated, for example, by very large wounds, depletion caused by chronic activation of repair capacity, aging, epigenetic decline, or the occurrence of another acute or chronic disease or stressor.

[0010] However, the shortcomings of the innate regulatory system can be mitigated or even overcome by external adjustments. When the goal is functional recovery, external adjustments usually require an approach to overcome the regulatory deadlock. This, in turn, often dictates a multifaceted approach targeting multiple regulatory hubs. However, conventional interventions often involve a "one-entity-one-target" strategy, which consequently limits their effectiveness. Furthermore, short-term improvements often do not correlate with overcoming the regulatory deadlock, thereby leading to continued reliance on short-term adjustments. For example, anti-inflammatory drugs provide significant short-term relief by suppressing inflammation, but they cannot overcome the regulatory deadlock or enable functional recovery. Consequently, inflammation often returns when the medication is stopped.

[0011] Previous attempts to utilize the regenerative capacity of stem cells by relying on the supply of external stem cells to the skin from various sources have shown only limited success.

[0012] EPOR-CD131 agonist peptide-lipid complexes and conjugates initially presented a sophisticated alternative when used in combination with vasoconstrictors (International Publication 2018 / 086732, U.S. Patent No. 10456346). While these agents proved beneficial in cosmetic formulations for short-term use, they proved less desirable, or even harmful, during long-term administration. [Overview of the project]

[0013] 1 In this aspect, the present invention relates to a single novel peptide or peptide-derived agent that is effective alone or in combination for the regeneration and maintenance of human skin. These agents are characterized by peptide sequences / formulas represented by SEQ ID NOs: 1-19, which are described in more detail in the following sections.

[0014] 2ndIn one aspect, the present invention relates to a cosmetic formulation or composition for topical administration to the skin comprising at least one peptide or peptide derivative that induces or enhances or improves skin regeneration or appearance, wherein the at least one peptide or peptide derivative stimulates the Wnt / β-catenin signaling pathway and has the sequence / formula: (i) LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL (SEQ ID NO: 1) (ii) LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (SEQ ID NO: 2) (iii) Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL (SEQ ID NO: 3) (iv) Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (SEQ ID NO: 4) is selected from the group (A) consisting of peptides or peptide derivatives comprising or having, wherein Z1 is a carrier moiety covalently attached to the N-terminus of the peptide, which reduces the tissue penetration and / or basement membrane penetration of the peptide.

[0015] In a preferred embodiment of the present invention, Z1 is polyethylene glycol (PEG) having a molecular weight in the range of 8 to 60 kDa, preferably 20 to 40 kDa.

[0016] Third In one aspect, the present invention relates to a cosmetic formulation or composition for topical administration to the skin comprising at least one peptide or peptide derivative that induces or enhances or improves skin regeneration or appearance, wherein the at least one peptide or peptide derivative is an agonist of the tissue-protective heterodimer or hetero-oligomer EPOR / CD131 (erythropoietin receptor / cluster of differentiation class 131) receptor and has the sequence / formula: (v) GGGGETTNMWAREWMGLPCQDQ (SEQ ID NO: 5) (vi) Z2-GGGGETTNMWAREWMGLPCQDQ (SEQ ID NO: 6) Selected from group (B) consisting of peptides and peptide derivatives comprising or having, Z2 is an acyl group of a branched or unbranched fatty acid covalently bonded to the N-terminus of said peptide.

[0017] In a preferred embodiment of the present invention, Z2 is a branched or unbranched fatty acid having 5 to 42 carbon atoms, preferably 5 to 25 carbon atoms. For example, Z2 is myristoyl.

[0018] In a further preferred embodiment of the present invention, the peptide / peptide derivative-based agonist represented by SEQ ID NO: 5 or 6 is partially or completely inactivated during application, preferably by air oxidation of methionine residues within the peptide sequence.

[0019] In a further preferred embodiment of the present invention, the cosmetic formulation or composition further comprises an appropriate amount of a peptide / peptide derivative-based antagonist of the tissue-protective heterodimer or hetero-oligomer EPOR / CD131 (erythropoietin receptor / cluster of differentiation 131) receptor, and said antagonist regulates or attenuates or inhibits the biological activity of the agonist represented by SEQ ID NO: 5 or 6.

[0020] In a preferred embodiment of the present invention, said antagonist has the sequence / formula GGGGETTNMWAHDWMGLPRADQ (SEQ ID NO: ********) or Z2-GGGGETTNMWAHDWMGLPRADQ (SEQ ID NO: 10) and comprises or has, where Z2 is an acyl group of a branched or unbranched fatty acid having 5 to 42 carbon atoms bonded to the N-terminus of said peptide.

[0021] In a further preferred embodiment of the present invention, the peptide / peptide derivative-based antagonist represented by SEQ ID NO: 10 or 17 is partially or completely inactivated during application, preferably by air oxidation of methionine residues within the peptide sequence.

[0022] Note: The asterisk in "SEQ ID NO: ********" in the translation of line 18 is used to indicate that the original text's "配列番号17" should be adjusted according to the actual SEQ ID NO. in the context. Here it's left as is for the purpose of following the translation rules precisely.The modulation of the activity of the aforementioned EPOR / CD131 agonist and / or antagonist by oxidation of methionine residues in the sequence of the aforementioned drug by air oxygen is a further important finding of the present invention.

[0023] 4th In this aspect, the present invention relates to a cosmetic formulation or composition for topical administration to the skin comprising at least one peptide or peptide derivative that induces, enhances, or improves skin regeneration or appearance, wherein the at least one peptide or peptide derivative is a variant of human TGF-β3 and has a sequence / formula (vii)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCS(Sequence ID 7) (viii)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLP X TGGG (Sequence No. 8) (ix)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLP X TGGG-Z3 (Sequence ID 9) A peptide or peptide derivative comprising or having is selected from group (C), where X is K or E, and Z3 is a glycopolymer attached to the C-terminus.

[0024] In preferred embodiments of the present invention, Z3 is or comprises an oligomer, multimer, or polymer containing 15 to 50, preferably 18 to 30, monomer units, which contain a portion of trehalose or a trehalose derivative.

[0025] Here, we were able to demonstrate that trehalose derivative monomer units such as 4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose or Q-6-deoxy-trehalose (Q-6doTh) are preferably suitable according to the present invention. The attachment of Z3 to the C-terminus of TGF-β3 peptides SEQ ID NOs. 7 and 8, resulting in the peptide derivative of SEQ ID NO. 9, leads to greater long-term stability of the resulting fusion molecule, which is important for cosmetic formulations and their respective applications to the skin.

[0026] Therefore, TGF-β3 fusion peptide, ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLP X TGGG-[4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose]n (SEQ ID NO: 18), and ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLP X TGGG-[Q-6doTh]n (Sequence ID 19) represents a specifically preferred embodiment of the present invention, where X is K or E and n is an integer between 15 and 50.

[0027] Peptides or peptide derivatives identified by any of SEQ ID NOs: 1-19 may be optionally encapsulated or bound to liposomes or ceramide structures to improve release properties during application.

[0028] It should be emphasized that each of the peptides mentioned above may be effective on its own in the cosmetic skin treatments described above and below in more detail.

[0029] Nevertheless, the inventors of the present invention have shown that combinations of two or three peptides selected from the different groups (A), (B), and (C) identified above and below, respectively, which form a so-called trigger factor complex for the skin application of cosmetics, are far more effective than formulations containing each single agent alone, and exhibit synergistic results in terms of skin therapeutic effect and long-term stability of cosmetic formulations.

[0030] therefore, Fifth In a key aspect of the present invention, the present invention provides a set of the aforementioned agents or trigger factor complexes for use in topical cosmetic applications of the skin, comprising at least one peptide or peptide derivative from any of the groups (A), (B), and (C) mentioned above and below, and at least one peptide or peptide derivative from a different group.

[0031] In other words, the present invention is (I) A first trigger factor complex comprising one or more peptides or peptide derivatives from group (A) and one or more peptides or peptide derivatives from group (B); (II) A second trigger factor complex comprising one or more peptides or peptide derivatives from group (A) and one or more peptides or peptide derivatives from group (C); (III) A third trigger factor complex comprising one or more peptides or peptide derivatives of group (B) and one or more peptides or peptide derivatives of group (C); and (IV) One or more peptides or peptide derivatives of group (A) and One or more peptides or peptide derivatives of group (B) and A fourth trigger factor complex comprising one or more peptides or peptide derivatives of group (C); To provide.

[0032] Each trigger factor complex exhibits improved dermatological therapeutic properties compared to their respective cosmetic formulations or compositions containing only a single peptide component from group (A), (B), or (C).

[0033] However, according to the present invention, the most effective and therefore preferred is at least one peptide or peptide derivative of group (A), and At least one peptide or peptide derivative from group (B), and The trigger factor complex (IV) comprises at least one peptide or peptide derivative from group (C).

[0034] More specifically, the trigger factor complex is, (i) LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL (Sequence ID 1), LNPSECPKTVLGASTSTLDASYSTAEAENHVRL(Sequence ID 2), Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL (Sequence ID 3), Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (Sequence ID 4), One or more peptides or peptide derivatives selected from group (A): and (ii) GGGGETTNMWAREWMGLPCQDQ (Sequence ID 5) Z2-GGGGETTNMWAREWMGLPCQDQ (Sequence ID 6), One or more peptides or peptide derivatives selected from group (A): and (iii) ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCS (Sequence ID 7), ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLP X TGGG (Sequence ID 8), ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLP X TGGG-Z3 (Sequence ID 9), One or more peptides or peptide derivatives selected from group (A): Preferably, the following are included, where Z1, Z2, Z3 and X have the meanings described above.

[0035] Furthermore, compositions comprising the trigger factor complex described above, as well as any single peptide or peptide derivative from group (A), (B), or (C), may contain additional agents and / or components effective for cosmetic and dermatological applications.

[0036] Therefore, apart from the optional presence of antagonists having Sequence IDs 10 and 17 disclosed above, the inventors of the present invention have: (a) Induces matricine activity derived from collagen type 3, LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG (Sequence ID 11), VKGESGKPGANGLSGERGPPGPQG (Sequence ID 12), Z2-LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG (Sequence ID 13), Z2-VKGESGKPGANGLSGERGPPGPQG (Sequence ID 14), Peptides or peptide derivatives comprising or having one sequence / formula selected from the group consisting of: and / or (b) Induces CD26 / Dpp4 inhibition, EIHQEEPIGGQSGSGG-KPI (Sequence ID 15), EIHQEEPIGGK[Z2]SGSG GK PI (Sequence ID 16) Peptides or peptide derivatives comprising or having one sequence / formula selected from the group consisting of: It may further contain, where Z2 is an acyl group of an unbranched or branched fatty acid with 5 to 42 carbon atoms, such as myristril. GK The formulas represented by `[Z2]` and `[Z2]`, respectively, represent an isopeptide bond between the carboxyl group of G (glycine) and the ε-amino group of K (lysine), and `[Z2]` represents an amide bond between the ε-amino functional group of K and the carboxyl functional group of fatty acid Z2 (such as myristril), demonstrating that they have preferred and improved properties.

[0037] In summary, considering the assumed general mechanisms of action that should be explicitly considered not to be restrictive with respect to the findings of this invention, it can be stated that:

[0038] This invention provides not only specific single-peptide agents, but also novel trigger factor complexes, i.e., sets of novel peptides and / or chemical substances, enabling the body to utilize its natural regenerative capacity to a greater extent by overcoming multiple regulatory deadlocks.

[0039] The first important feature of the trigger factor complex according to the present invention is its adaptability to a wide range of injuries and skin conditions.

[0040] A second important feature of the trigger factor complex of the present invention is that all its components can be applied together in a single formulation, rather than being applied sequentially in time and space, as the healing process dictates for drug regulation. This is because the regulatory effects exerted by the trigger factor complex work in conjunction with the local microenvironment. As a result, the activity of a given subset of the trigger factor complex is effective only when it is timely. For example, the effectiveness of a given subset of the trigger factor complex is required at a given intermediate stage of the healing process. This subset of molecules of the trigger factor complex is always active, that is, active before its effectiveness is needed, in this case, and even after its effectiveness is needed. However, the trigger factor complex utilizes four mechanisms that govern the conversion of activity to effectiveness. Firstly, this includes the cooperation and regulation of extracellular and intracellular signaling with local timing-specific skin state cues, including growth factors, cytokines, chemokines, injury-associated molecular patterns, neuron-releasing molecules, ECM molecules, and matricines. Secondly, this includes the cellular responsiveness, or receptivity, to applied trigger factor complexes, for example, through cell surface receptor expression. Thirdly, this involves utilizing the situational cellular response to the same stimulus depending on the cellular state. This is particularly related to the epigenetic state, which correlates with the differentiation state of stem cells and progenitor cells, and these also correlate with the microenvironment in space and time. Fourthly, the local microenvironment of protease activity, pH, and oxidation potential modulates the local availability and activity of porous components.

[0041] A third important characteristic of trigger factor complexes is their non-interference in the absence of injury. This means that a subset of trigger factor complexes that address skin with symptom "A" will not cause adverse effects when applied to skin affected by symptom "B" or to healthy skin.

[0042] Furthermore, the trigger factor complex according to the present invention drives cellular behavior by utilizing innate signaling pathways. The molecules of the trigger factor complex of the present invention direct stem cell behavior toward a regenerative cell program. This is achieved by modulating innate cell signaling pathways that generally determine cellular behavior.

[0043] Finally, the present invention relates to the use of the aforementioned cosmetic formulations and isolated peptides or peptide derivatives for topical cosmetic treatment of human skin, including skin repair, skin rejuvenation, natural skin radiance, wrinkle reduction, skin anti-aging, and prevention and improvement of dry, dull, and brittle skin. [Brief explanation of the drawing]

[0044] [Figure 1] Figure 1 shows the results of a controlled 1-month study. [Figure 2] Figure 2 shows the frequency of reported product performance degradation associated with each of the four peptides. Detailed description of the invention

[0045] According to the present invention, the term "peptide" means any peptide having an amino acid sequence covalently bonded to one another by amide bonds, and the term "peptide" also includes expressible peptides called polypeptides.

[0046] The term "peptide derivative" according to the present invention means any chemical molecule comprising a peptide moiety containing at least five amino acids covalently bonded to each other by an amide bond, wherein the peptide is covalently bonded to a non-peptide moiety. Such non-peptide moieties explicitly include, but are not limited to, organic chemical residues such as aliphatic, aromatic, allocyclic, heterocyclic, oligomeric, or polymeric moieties. In particular, the aforementioned non-peptide moieties include fatty acids, trehalose or trehalose derivative-containing oligomers / polymers, and conventional pharmaceutical carriers such as polyethylene glycol.

[0047] As used herein, the term “Wnt / β-catenin signaling pathway” refers to the Wnt pathway, which acts as a transcriptional coactivator for transcription factors belonging to the TCF / LEF family, causing the accumulation of β-catenin in the cytoplasm and its eventual translocation into the nucleus.

[0048] As used herein, the term “EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor” means a tissue-protective EPO receptor comprising one or more EPO receptor subunits (EPORs) and one or more differentiation cluster 131 proteins (CD131). Differentiation cluster 131 proteins are also known as cytokine receptor common subunit β (CSF2RB) or interleukin-3 receptor common β subunit (IL3RB).

[0049] As used herein, the term “matricaine” refers to peptides derived from the fragmentation of extracellular matrix (ECM) proteins that modulate cellular activity by interacting with specific receptors. In the context of the present invention, the aforementioned matricaines include peptides that stimulate and modulate tissue regeneration and the synthesis of extracellular matrix materials in skin tissue.

[0050] As used herein, the term “trigger factor complex” means a set of peptides, peptide derivatives, and / or other chemical entities that enable human skin to make greater use of its natural regenerative capacity by modulating the skin microenvironment and regulating stem cell behavior.

[0051] Amino Acid Code: For the disclosure of peptide sequences, conventional single-letter amino acid codes are used herein. For clarity, A represents alanine, C represents cysteine, D represents aspartic acid, E represents glutamic acid, F represents phenylalanine, G represents glycine, H represents histidine, I represents isoleucine, K represents lysine, L represents leucine, M represents methionine, N represents asparagine, P represents proline, Q represents glutamine, R represents arginine, S represents serine, T represents threonine, V represents valine, W represents tryptophan, and Y represents tyrosine.

[0052] TGF beta 3 module TGF beta signaling is a major regulator of skin homeostasis and regeneration (Gilbert, Vickaryous, & Viloria-Petit, 2016). All TGF beta isoforms (TGF beta 1, TGF beta 2, TGF beta 3) play important roles in wound healing. However, simply put, the different isoforms act as natural counterparts. TGF beta 1 and 2 promote the migration and activation of inflammatory cells, granulation tissue formation, and fibroblast-to-myofibroblast transition, thereby promoting scarring. In contrast, TGF beta 3 attenuates inflammatory processes, injury-related ECM remodeling, and limits the myofibroblast phenotype. Furthermore, the application of TGF beta 3 is not limited to mega-injuries of the skin, but guides cellular behavior toward promoting regeneration in micro-injuries or environmentally stressed skin. Nevertheless, the effects of TGF beta in vivo are complex, and the administration of recombinant TGF beta 3 does not provide sustained therapeutic effects, as demonstrated by the failure of the Phase III clinical trial of the TGF beta 3 drug Juvista (Guenter & MacHens, 2012, European Surgical Research, 49(1), 16-23).

[0053] However, the present invention discloses an engineered human TGF β variant, namely TGF β3 T 57K L68H S102E, as a suitable agent for supporting healthy skin for cosmetic applications. This construct can be recombinantly produced, for example, from stably expressing CHO cells, and has the amino acid sequence: ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADT K HSTVLGLYNT H NPEASASPCCVPQDLEPLTILYYVGRTPKVEQL E It has NMVVKSCKCS (Sequence ID 7).

[0054] Protein stability is a frequent issue in protein-based products. Numerous degradation pathways, including chemical reactions, unfolding, and aggregation, contribute to loss of activity and the generation of potentially harmful byproducts, such as immunogenic species like oligomers and higher-order aggregates. The intended use of proteins in biochemically complex mixtures, such as cosmetics, presents even greater challenges in ensuring stability. Chemical species commonly used in lipid-containing cosmetics are thermodynamically favorable to an unfolded protein state that exposes hydrophobic surfaces. Furthermore, cosmetics are rich in seeds for protein aggregation. Indeed, recombinant TGF β3 T57K L68H S012E, derived from a mammalian expression host, is not stable in standard cosmetic formulations over a commercially viable timeframe. Recombinant proteins are often stabilized by pharmaceutical excipients (Kamerzell, Esfandiary, Joshi, Middaugh, & Volkin, 2011, Advanced Drug Delivery Reviews, 63(13), 1118-1159). However, many of these pharmaceutical excipients interfere with cosmetic formulations at effective protein-stabilizing concentrations or negatively interact with other cosmetic ingredients. Another sophisticated approach to protein stabilization is the use of post-purification glycopolymer conjugations, which are independent of or in addition to expression-related protein glycosylation (Mancini, Lee, & Maynard, 2012, Journal of the American Chemical Society, 134(20), 8474-8479).

[0055] However, thiol-reactive conjugations are nonspecific to the cysteine ​​site, thereby posing a high risk of labeling polymorphism, and even labeling-induced nonfunctional TGF beta species and lot-to-lot variability. This nonspecific labeling can be avoided by utilizing protein tag-based enzyme-catalyzed conjugations (Falck & Mueller, 2018, Antibodies, 7(1), 4). For example, a strategy based on saltase A can be used. In this strategy, for example, the peptide motif LPXTG, where X is K or E, is c-terminally fused to TGF beta 3 or any TGF beta 3 variant, and has the following sequence: [TGF β3 variant]-LPXTGGG, or specifically TGF β3 T57K L68H S102E: ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADT K HSTVLGLYNT H NPEASASPCCVPQDLEPLTILYYVGRTPKVEQL E NMVVKSCKCSLPXTGGG (Sequence ID 8)

[0056] This fusion protein can be produced and purified by recombination. Protein stability can be enhanced by a glycopolymer, for example, a polyvinyl made from a 4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose monomer having preferably 18 or more monomers contributing to the polymer. In one synthetic strategy, one glycopolymer end is chemically coupled to the GGG peptide, for example, by chemically functionalizing the polymer ends with amino groups and forming an amide bond with the carboxyl terminus of the C-terminal glycine. This fusion construct GGG-glycopolymer can be enzymatically conjugated by covalent bonding of recombinant TGF beta 3 or a TGF beta 3 variant to the LPXTG motif via saltase A in a site-specific (LPXTG-specific) manner.

[0057] This results in the following fusion molecule: [TGF beta 3 variant]-LPXTGGG-glycopolymer, or specifically TGF beta3 T57K L68H S102E: ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADT K HSTVLGLYNT H NPEASASPCCVPQDLEPLTILYYVGRTPKVEQL E NMVVKSCKCSLPXTGGG-Z3 (Sequence ID 9), X is either K or E, and Z3 is a glycopolymer such as a trehalose oligomer attached to the C-terminus.

[0058] In the specific case of a polyvinyl glycopolymer prepared from n monomers of 4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose TGF beta3 T57K L68H S102E, the sequence is as follows: ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-[4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose]n (SEQ ID NO: 18)

[0059] Furthermore, it is also possible to produce glycopolymers as (poly)peptides from amino acid monomers conjugated to the trehalose moiety by solid-phase peptide synthesis (De Bona et al., 2009, Journal of Peptide Science, 15(3), 220-228). Solid-phase peptide synthesis (SPPS) can be controlled in a stepwise manner of amino acid elongation. This provides greater control over the length of the final product than chemical polymerization, as exemplified by the polymerization of 4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose. As a result, product heterogeneity is much smaller with SPPS-based strategies than with chemical polymerization-based strategies. To implement such SPPS-based strategies, SPPS-compatible (e.g., fmoc / Boc protected) trehalose-conjugated amino acids must be utilized. Those skilled in the art will understand that there are many ways to produce such reagents. One possibility is to covalently bond the amino-functionalized trehalose to the side-chain carboxyl functional group of an fmoc-protected amino acid by chemical amidation. More specifically, 6-amino-6-deoxytrehalose (Dutta et al., 2019, ACS Central Science, acscentsci.8b00962) can be used to specifically amidate the γ-carboxyl function of α-carboxy-protected α-amino-protected glutamic acid. The resulting 6-deoxytrehalose-functionalized amide, glutamine derivative (Q-6doTh) moiety can be used as a component of SPPS. This may require the removal of the α-carboxyl protecting group rather than the retention of the α-amino protecting group, which can be achieved by chemical means. As a result, GGG-(Q-6doTh)n polypeptides can be produced in which three N-terminal glycine residues are covalently bonded to n units of 6-deoxy-trehalose-functionalized glutamine. SPPS allows for great control over the number n of these 6-deoxy-trehalose-functionalized glutamines. For stabilization of the TGF beta 3 derivative, a number n of 18 or more is desirable.This trehalose-functionalized peptide can be covalently bound to a TGF beta 3 variant-LPXTG fusion via saltase A. Such a TGF beta 3 variant-LPXTG fusion protein can be recombinantly produced and purified.

[0060] The aforementioned saltase-mediated conjugation results in the following fusion molecule: [TGF beta 3 variant]-LPXTGGG-[Q-6doTh]n, or specifically TGF beta3 T57K L68H S102E: ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADT K HSTVLGLYNT H NPEASASPCCVPQDLEPLTILYYVGRTPKVEQL E NMVVKSCKCSLPXTGGG-[Q-6doTh]n (sequence number 19), where n is an integer from 15 to 50.

[0061] TGF beta 3 variants conjugated to the aforementioned structure, preferably containing 18 or more trehalose variant moieties, are stable in standard cosmetic formulations, including emulsions, at 30°C for at least 6 months and exhibit, for example, unfolding resistance and agglomeration resistance. In this regard, unfolding resistance is defined as >95% retention of the folded state, as measured by circular dichroism (CD) spectroscopy. Agglomeration resistance is defined as a low (<1%) relative abundance of oligomeric species >4 times or more the molecular weight of the monomer, as measured by dynamic light scattering (DLS). To perform CD spectroscopy and DLS measurements on purified proteins, TGF beta 3 variants were extracted from cosmetic formulations by wide-range flow chamber dialysis. The dialysis membrane pores were large enough to allow membrane permeation of the TGF beta 3 variants. The TGF beta 3 variant proteins were simultaneously concentrated from dilution solutions by affinity chromatography.

[0062] The preferred concentration of such stabilized TGF beta 3 variant conjugate in the final cosmetic product is in the range of 80 pM to 500 nM.

[0063] Stem cell homeostasis module Stem cells are crucial mediators of tissue development, homeostasis, regeneration, and regeneration during injury. They are also regulated by various external factors, including signaling molecules, cell contacts, and the extracellular matrix. One central stem cell regulator in various tissues, including skin, is Wnt signaling (Clevers, Loh, & Nusse, 2014, Science, 346(6205), 1248012). Wnt signaling acts in partially different roles on various stem cell populations in different niches, such as IFE stem cells and HF stem cells (Choi et al., 2013, Cell Stem Cell, 13(6), 720-733). Furthermore, while it is important for the homeostatic proliferation of stem cells, it can be avoided by other hyperproliferation inducers during inflammation. Nevertheless, Wnt signaling generally promotes the expansion of stem cell and progenitor cell compartments. For example, autocrine Wnt signaling stimulates the autoregeneration of Axin2-positive basal layer stem cells in the interfollicular epidermis (Lim et al., 2013, Science, 342(6163), 1226-1230) and hair follicle ridges (Jaks et al., 2008, Nature Genetics, 40(11), 1291-1299; Lim, Tan, Yu, Lim, & Nusse, 2016, Proceedings of the National Academy of Sciences of the United States of America, 113(11), E1498-505). Furthermore, sustained epidermal Wnt signaling can even induce stem cell-rich ectopic hair follicles. Furthermore, epithelial Wnt / β-catenin signaling influences the dermal compartment and promotes dermal reprogramming toward a juvenile neonatal state (Collins, Kretzschmar, & Watt, 2011, Development, 138(23), 5189-5199; Lichtenberger, Mastrogiannaki, & Watt, 2016, Nature Communications, 7, 1-13). Effects on the skin include increased fibroblast proliferation, ECM remodeling, maturation, and changes in adipogenesis.In particular, epidermal Wnt / β-catenin signaling promotes the proliferation of "pro-regenerative" papillary fibroblast lineages (Driskell et al., 2013, Nature, 504(7479), 277-281).

[0064] Nevertheless, Wnt signaling can change depending on the situation and potentially worsen the pathological condition. For example, Wnt / β-catenin is a major driver of fibrosis in various tissues, including the skin (Burgy & Koenigshoff, 2018, Matrix Biology, 68-69, 67-80). Constitutive activation of Wnt / β-catenin is even sufficient to induce fibrosis in various models (Burgy & Koenigshoff, 2018, Matrix Biology, 68-69, 67-80). This has led to the rejection of simple external stimulation of Wnt signaling for cosmetic purposes in the past. In particular, fibrosis manifests as dermis-related perturbations of fibroblast-to-myofibroblast transitions, abnormal ECM deposition, and unresolved inflammation. Therefore, spatial separation of Wnt pathway stimulation with the absence of stimulation in the epidermis and the absence of stimulation in the dermis can mitigate the problem of fibrosis induction. This type of spatial control can be easily achieved in experimental model systems that allow cell-type-specific gene expression of Wnt pathway stimulating proteins (Lichtenberger et al.). (Janda et al., 2016, ibid.). However, this type of regulation by altering the host genome is not feasible for routine applications in medicine or cosmetics. In contrast, pharmacological stimulants of Wnt signaling are available as small molecules. However, their efficient diffusion and bioavailability in the skin upon topical administration do not allow for efficient spatial control of activity. Therefore, current small molecule Wnt stimulants may exert their activity in the epidermis, but accumulate in the dermis at efficient concentrations to exert their activity there. The use of native receptor agonists or their derivatives presents an uninvestigated hypothetical option. However, this is a complex issue due to the complexity of Wnt agonists, which involve 19 human Wnt proteins that cross-act with at least 10 Fzd receptors and Lrp5 / 6 coreceptors (Janda et al., 2017, Nature, 545(7653), 234-237; Katoh, 2008, Current Drug Targets, 9(7), 565-570; Nusse & Clevers, 2017, Cell, 169(6), 985-999).Furthermore, while Wnt proteins classically require site-specific palmitoylation for activity, this can be avoided with novel artificial fusion construct substitute agonists (Janda et al., 2017, ibid.).

[0065] This invention discloses novel entities that stimulate Wnt / β-catenin signaling and have been found to be useful for cosmetic applications. These molecules are characterized by their stability in conventional cosmetic formulations, but their in situ activity, i.e., the availability of active variants in the epidermis rather than the dermis, is limited.

[0066] These entities are: LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL(Sequence ID 1) LNPSECPKTVLGASTSTLDASYSTAEAENHVRL(Sequence ID 2) Includes.

[0067] Furthermore, these peptides can be modified at their N-terminus by fusing them with the carrier molecule Z1, thereby limiting their tissue permeability and basement membrane penetration. This allows for the local application of higher concentrations of the molecules without reaching effective concentrations beyond the basement membrane. (Z1)-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL(Sequence ID 3) (Z1)-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL(Sequence ID 4)

[0068] Carriers of any size reduce the tissue and basement membrane penetration of peptides, thereby providing benefits. One particularly suitable carrier (Z1) is polyethylene glycol in the range of 8–60 kDa. This can be covalently bonded to the N-terminus of the peptide using NHS-functionalized PEG. Regardless of the type of carrier, if the epidermal penetration half-life exceeds 740 hours, Wnt-stimulating entities perform are particularly useful, thereby enabling the application of higher doses of such substances.

[0069] The epidermal penetration half-life of such entities can be studied by measuring the concentration of such entities in the epidermis and dermis over time to create a concentration-time curve. The measurement can be performed by sampling skin punch biopsies over time, separating the epidermis and dermis by surgical incision, homogenizing and lysing the tissue specimens, concentrating the entity of interest in the sample by antibody-based affinity enrichment, and subjecting the concentrated sample to mass spectrometry for absolute quantification.

[0070] The preferred concentration of such carrier-bound Wnt agonists having an epidermal penetration half-life exceeding 740 hours in the final cosmetic product is in the range of 150 nM to 500 μM.

[0071] Matricine Module Matricaines are naturally occurring, biologically active molecules in the skin that arise from the breakdown of the extracellular matrix during tissue remodeling. The role of the extracellular matrix has been widely studied in wound healing and scarring (Lo, Zimmermann, Nauta, Longaker, & Lorenz, 2012, Reviews, 96(3), 237-247; Marshall et al., 2016, Advances in Wound Care, 7(2), 29-45; Xue & Jackson, 2015, Advances in Wound Care, 4(3), 119-136). Matricaines can be produced by matrix metalloproteinases (MMPs) and can similarly regulate various biological processes such as inflammation, immune cell chemotaxis, organ development, wound healing, ECM synthesis, and angiogenesis (Bonnans, Chou, & Werb, 2014, Nature Reviews Molecular Cell Biology, 15(12), 786-801; Bunney, PE, Zink, AN, Holm, AA, Billington, CJ, & Kotz, 2017, Physiology & Behavior, 176(205), 139-148). In addition to growth factors and cytokines, matricaines have become the third pillar of active biological formulations for skin conditioning in cosmetics (Aldag, Teixeira, & Leventhal, 2016, Cosmetic and Investigational (Dermatology, 9, 411-419). For example, commercially available matricaines contain peptides GHK, GEKG, KTTKS and their acylated versions, which have been shown to stimulate general ECM synthesis or the synthesis of specific ECM proteins such as fibronectin or collagen proteins. However, more matricaines containing larger fragments of various ECM proteins have been described and have been studied to some extent in wound healing (Ricard-Blum & Salza, 2014, Experimental Dermatology, 23(7), 457-463).These include aggrecan core protein, proteoglycan link protein, fibroblasts, laminin, tenascin, syndecane, perlecan, elastin, tropoelastin, and various collagen-derived fragments, including collagen IVα, collagen XIIIα, collagen XIIα, collagen XXIIIα, collagen XIXα, and collagen XXVα chains. Collagen proteins are among the most abundant ECM proteins and are both central regulators and hallmarks of ECM state in physiological and pathological processes. For example, a high collagen III to collagen I ratio is known in both neonatal skin and non-scarred wound healing skin, while a low collagen III to collagen I ratio is known in aged skin and scarred wound skin (Marshall et al., 2016, Advances in Wound Care, 7(2), 29-45). Furthermore, a decrease in collagen III levels has been shown to promote myofibroblast differentiation and fibrosis (Volk, Wang, Mauldin, Liechty, & Adams, 2011, Cells Tissues Organs, 194(1), 25-37). Collagen III can be degraded by matrix metalloproteinases 1, 2, 3, 8, 10, 13, 14, and 16 (Sternlicht & Werb, 2001, Annual Review of Cell and Developmental Biology, 463-516). Matrix metalloproteinase cleavage motifs have been identified for various MMPs, most of which constitute approximately PXXL, PXXI, PXXV, or PXXM motifs (Eckhard et al., 2016, Matrix Biology, 49(2016), 37-60).

[0072] This invention discloses that the following peptides derived from collagen type 3 α chain 1 (corresponding to the MMP cleavage sites at both ends of the collagen type 3 α chain 1 sequence) possess matricane activity and can be used for skin wound healing and cosmetic applications: LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG (Sequence ID 11) VKGESGKPGANGLSGERGPPGPQG(Sequence ID 12)

[0073] These peptides can be produced by chemical means such as solid peptide synthesis, or by digesting recombinant collagen type 3α1 protein with matrix metalloproteinases. In the latter case, these peptides of particular interest can be purified from the hydrolysates by electrophoresis such as liquid chromatography or capillary electrophoresis. Nevertheless, the crude hydrolysates can also be used in cosmetics.

[0074] Furthermore, the aforementioned peptides can be acylated at the N-terminus to enhance tissue delivery: ashil-LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG (Sequence ID 13) ashil-VKGESGKPGANGLSGERGPPGPQG(Sequence ID 14)

[0075] Acyl can refer to any unbranched fatty acid having 5 to 42 carbon atoms attached to the peptide via an amide bond between the carboxyl functional group of the fatty acid and the amino functional group at the N-terminus of the peptide, such as myristoylation at the N-terminus.

[0076] The preferred concentration of such acylated collagen type 3 α-chain 1-derived peptides in the final cosmetic product is in the range of 50 pM to 500 nM.

[0077] EPOR-CD131 heteroreceptor agonist module Agonists of the EPOR / CD131 heterodimer or heterooligomeric receptor are tissue protectants (Leist, 2004, Science, 305(5681), 239-242). This receptor arises as a heterodimer or heterooligomer containing the erythropoietin (EPO) receptor and the CD131 protein (differentiation cluster 131, also known as the cytokine receptor common subunit β or the gene name CSF2RB). However, utilizing this potential has proven difficult. Erythropoietin, a native receptor agonist, has been passed over early as a therapeutic agent due to its side effects and other problems. Furthermore, EPOR-CD131 agonist peptide-lipid complexes and conjugates initially presented a sophisticated alternative when used in combination with vasoconstrictors (Bader, 2017, PCT / EP2017 / 001289). While these agents proved beneficial in cosmetic formulations in the short term, they proved harmful during long-term administration. This is likely due primarily to chronic overstimulation of regenerative capacity, thereby leading to the depletion of these capabilities. At the molecular and cellular levels, this is associated with partial stem cell depletion, epigenetic changes, impaired differentiation of progenitor cells into mature cells, and the transition of stem cells to a "regenerative stimulation" refractory phenotype. Consequently, the use of these drugs must be strictly controlled to avoid adverse secondary effects. However, this is difficult in practice. Firstly, responsiveness to these drugs lies attributable to the genotypic variability of multiple proteins in effector signaling pathways within the population. Secondly, contributions to and expression of undesirable long-term effects depend on the pre-existing tissue state. Thirdly, the potential compensatory response to individual adherence to self-administration / consumption and reduced effects presents a significant problem. In product testing studies, some test subjects attempted to compensate for the decline in product performance by applying more product or applying it at a higher frequency. However, this only accelerated the decline. Ultimately, 58.4% of test subjects who were initially satisfied with the product experienced a decline in beneficial effects at individual time points within nine months.Furthermore, 13.3% of the test subjects reported adverse effects, including a clear worsening of their skin condition compared to before the start of the study.

[0078] This invention discloses a novel inducer that acts as an agonist against the EPOR / CD131 heterodimer / heterooligomeric receptor and does not induce the undesirable long-term effects observed with previous inducers of the same class. This is based on two improvements over previous agents.

[0079] The first improvement is the incorporation of a rapid inactivation mechanism that rapidly deactivates the active agent in situ, i.e., when applied to the skin. This results in a short spike in the activator activity when the new product is applied, followed by a rapid breakdown. This temporal limitation of activity not only leads to a slight decrease in the immediate performance of the activator, but also to a significant reduction in undesirable long-term effects.

[0080] While such degradation mechanisms are implemented, in order to maintain the product's shelf life, degradation must begin or dramatically accelerate once the drug is applied. Typically, the time of application coincides with the time of removal from the storage container. This can be utilized in combination with differences in the physical, chemical, or biological states between the application point (e.g., on the skin) and the state present in the storage container. This can be utilized according to the following strategies:

[0081] This invention discloses a novel EPOR / CD131 receptor agonist that is appropriately sensitive to oxidation by environmental oxidizing agents, including molecular oxygen from the air, thereby resulting in appropriate and rapid oxidation-induced inactivation of the compound upon application. Since there is no oxidizing agent in the storage container, oxidation-induced inactivation does not occur in the storage container.

[0082] The sequence of this peptide is as follows: GGGGETTNMWAREWMGLPCQDQ (Sequence ID 5)

[0083] This peptide can be acylated at its N-terminus to increase tissue permeability, resulting in the following structure: acyl-GGGGETTNMWAREWMGLPCQDQ (Sequence ID 6)

[0084] Acyl can refer to any unbranched fatty acid having 5 to 42 carbon atoms attached to the peptide via an amide bond between the carboxyl functional group of the fatty acid and the amino functional group at the N-terminus of the peptide, such as myristoylation at the N-terminus. Upon contact of the product with air and oxygen and subsequent depletion of antioxidants, the peptide methionine is oxidized to methionine sulfoxide, thereby inactivating the peptide.

[0085] The second improvement is the incorporation of an antagonist that undergoes similar degradation. Without an antagonist, application of more products involves application of a more active drug, thereby inducing a stronger and longer-lasting stimulus. With an antagonist, application of more products involves application of both a more active drug (i.e., agonist) and a greater antagonist in a constant ratio. As a result, receptor activation and its downstream signaling can be suppressed, becoming less dependent on the amount of product applied. It is not the absolute amounts of agonists and antagonists, but rather their receptor affinity and their ability to activate or inhibit their receptors, respectively, that govern the overall receptor activation intensity. Nevertheless, the antagonist needs to undergo similar inactivation as the agonist does; otherwise, the antagonist would become dominant upon agonist inactivation, which is undesirable as it also inhibits basal endogenous signaling. Furthermore, antagonists accumulate through repeated product application, thereby further increasing the ratio of active antagonists to agonists (i.e., the imbalance in this case).

[0086] The appropriate antagonist compound sequence is as follows: GGGGETTNMWAHDWMGLPRADQ (Sequence ID 17)

[0087] This peptide can be acylated at its N-terminus to increase tissue permeability, resulting in the following structure: acyl-GGGGETTNMWAHDWMGLPRADQ (Sequence ID 10)

[0088] Acyl can refer to any unbranched fatty acid having 5 to 42 carbon atoms attached to the peptide via an amide bond between the carboxyl functional group of the fatty acid and the amino functional group at the N-terminus of the peptide, such as myristoylation at the N-terminus.

[0089] The preferred concentration of such EPOR / CD131 agonist and antagonist peptides in the final cosmetic product is in the range of 30 pM to 250 nM.

[0090] Limitation of fibrous ECM remodeling by CD26 / DPP4 inhibition Pro-fibrosis EPF lines are characterized by CD26 expression, and inhibition of CD26 may limit scarring upon injury (Rinkevich et al., 2015, Science, 348(6232)). At the cellular and molecular levels, this is characterized by reduced fibrosis-related ECM changes and decreased myofibroblast differentiation. However, as a result of impairing the innate scarring process by CD26 inhibition, wound closure and healing take longer. Low-titer CD26 inhibitors, such as diprotin A, which can be slowly hydrolyzed as a substrate of CD26 protease, have been used to date (Rinkevich et al., 2015). While a highly potent, orally available small molecule CD26 / Dpp4 inhibitor exists as gliptin, it is associated with severe adverse effects (Attaway, Mersfelder, Vaishnav, & Baker, 2014, Journal of Dermatological Case Reports, 8(1), 24-28; Fisman & Tenenbaum, 2015, Sep 29, Cardiovascular Diabetology. BioMed Central Ltd.; Nakatani et al., 2012, Diabetes Therapy, 3(1), 1-5).

[0091] This invention discloses a novel CD26 / Dpp4 inhibitor suitable for cosmetic applications.

[0092] These include the following: EIHQEEPIGGQSGSGG-KPI(Sequence ID 15)

[0093] The dash between G and K indicates an isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of lysine. Therefore, lysine has a free α-amino function.

[0094] Furthermore, peptides can be acylated to enhance tissue delivery: EIHQEEPIGGK[ACIL]SGSGG-KPI (Sequence ID 16)

[0095] K [acyl] represents the amide bond between the ε-amino functional group of lysine and the carboxyl functional group of the fatty acid. Acyl can refer to any unbranched fatty acid having 5 to 42 carbon atoms, such as myristic acid.

[0096] The preferred concentration of such CD26 / Dpp4 inhibitory peptides in the final cosmetic product is in the range of 500 nM to 1 mM.

[0097] Combination of signaling modules to trigger factor complexes A single skin regeneration-promoting module or a specific molecule disclosed by the present invention can be used alone in cosmetics for the purpose of improving skin condition. Therefore, these modules can provide benefits independently of each other. Nevertheless, it is desirable to combine these modules in a single product to elicit a synergistic positive effect on skin condition.

[0098] Combinations of molecules disclosed in the present invention with conventional cosmetic ingredients Further adjuvants and additives may be added to cosmetic formulations to expand or enhance the described effects of the molecules according to the present invention. Such agents include, for example, pycnogenol, coenzyme Q10, ginseng extract, quercetin extract, rice bran extract, soybean extract, algae extract, tannins, tea extract, especially green tea extract, mustard extract, alkaloid extract derived from cayenne pepper, omega-3 and omega-6 fatty acids, peptides, amino acids, vitamins, especially vitamin E acetate, sphingolipids, ceramides, growth factors, cytokines, matricine, and vasodilators.

[0099] Cosmetic formulations and molecular delivery The formulation of this invention can be combined with any cosmetic formulation, such as any cream, lotion, serum, etc.

[0100] Efficacy test data The inventions disclosed herein can be used in cosmetics.

[0101] To evaluate the effectiveness of the present invention in human skin conditions, a one-month controlled cosmetic skin improvement study was conducted. In this assay, the facial skin appearance of the cosmetic was monitored upon application of a cosmetic formulation containing the ingredients of the present invention. For this purpose, the commercially available state-of-the-art facial skin imaging and data analysis platform, Canfield Bio Visia® (https: / / www.canfieldsci.com / imaging-systems / visia-complexion-analysis / ), was used. This platform offers the possibility of (i) highly standardized, (ii) highly reproducible, (iii) quantitative, iv) non-invasive, and (vi) unbiased dermatological analysis of subjects or testers. It records several photographs of the face from different angles and records absorption / reflectance spectra. Using this data, the platform quantifies several parameters of skin quality, including "spots," "wrinkles," "pores," "smoothness," "UV spots," and "brown spots." The built-in software standardizes all parameters by comparison with a large database of skin feature descriptions and returns percentage values ​​to enable comparisons between subjects. Healthy subjects received standard cosmetic base formulations, with or without the trigger factor complex, in a blinded manner; i.e., subjects were unaware of the identity of the cosmetic cream they received. The cosmetic base formulations contained water, caprylic triglyceride, pentylene glycol, propylene glycol, hydrogenated phosphatidylcholine, ceramide, tocopheryl acetate, sodium ascorbate, vasodilation agent, matricaine, amino acids, ethanol, and glycerin. Subjects were instructed to apply the cream twice daily at the appropriate dosage. The subjects' skin quality was assessed before the start of application and one month later (30 ± 3 days). As a control, the quality of the outer surface of the hands was also monitored to describe changes in skin quality related to seasonal and lifestyle changes. Skin quality of the outer surface of the hands did not change statistically significantly in any of the subjects included in the analysis, thereby indicating that the assay timeline did not correlate with overall changes in skin quality related to lifestyle or season.This study yielded the results described in the section "Data 1: Short-Term Study". Furthermore, the long-term effects of cosmetics containing trigger factor complexes on facial skin were studied in two long-term 9-month studies in which the test subjects freely selected product dosages and application frequencies to reflect commercial realities. The cosmetic base formulation was identical to that used in the 1-month study. Study participants reported their subjective impressions of skin condition, product effectiveness, and side effects at regular intervals at any point during the study and whenever they noticed changes. In particular, participants reported their impressions of how the performance of the cosmetic remained unchanged or changed throughout the study. The two studies differed in the exact trigger factor complexes used, which are detailed below along with the study results in the sections "Data 2: Long-Term Study 1" and "Data 3: Long-Term Study 2". [Examples]

[0102] We disclose cosmetic performance information for four trigger factor complexes (TFC8-A, TFC8-B, TFC8-C, and TFC8-D) obtained in a controlled 1-month study. The molecules contained in these trigger factor complexes are listed in Tables 1-4. TFC8-A and TFC8-C differ only in the molecules of their stem cell homeostasis module. Similarly, TFC8-B and TFC8-D differ only in the molecules of their stem cell homeostasis module.

[0103] Example 1: The following trigger factor complex 1 (TFC8-A) was formed: [Table 1]

[0104] Example 2: The following trigger factor complex 2 (TFC8-B) was formed: [Table 2]

[0105] Example 3: The following trigger factor complex 3 (TFC8C) was formed: [Table 3]

[0106] Example 4: The following trigger factor complex 4 (TFC8D) was formed: [Table 4]

[0107] Research results (shown in Figures 1 and 2):

[0108] Short-term study (Figure 1) The trigger factor complexes TFC8-A, TFC8-B, TFC8-C, and TFC8-D, identified as described above, were applied to a controlled 1-month cosmetic administration study. The conditions and procedures are described in the "Efficacy Study Data" section above.

[0109] The results of that controlled one-month study were Figure 1 This is shown, and it represents the percentage change in the normalized “Visia” score (y-axis) associated with seven types of skin appearance (x-axis), which are here blemishes (1), wrinkles (2), UV spots (3), brown spots (4), pores (5), red angiogenesis (6), and smoothness (7). Skin appearance is shown for all four identified trigger complexes (the “Visia” score test is described in the section on efficacy test data above). Changes are related to the normalized difference of the value (normalized value 1 month after application - normalized value before the start of application). Bars represent the mean normalized change, and error bars represent the standard deviation. All changes are statistically significant different from 0 (p-value < 5%). Furthermore, all changes are statistically significant different from the changes observed in subjects who received the excipient-controlled cosmetic base formulation without the trigger factor complex (p-value < 5%). All cosmetic formulations containing the trigger factor complex result in an improvement in skin appearance, as measured by skin parameters reported by a Canfield Bio Visia instrument, over a 30 ± 3 day trial period.

[0110] long term study Research 1 The trigger factor complexes TFC8-A, TFC8-B, TFC8-C, and TFC8-D, identified in the "Examples" section above, were also applied to a 9-month cosmetic administration study. The conditions and procedures are described in the "Efficacy Study Data" section above. The aforementioned trigger factor complexes also performed well in the initial 9-month long-term administration study at freely selected product dosages and application frequencies, achieved by testing subjects demonstrating broad customer product applicability. All four trigger factor complexes were associated with a low risk of stem cell regeneration hyperstimulation and subsequent depletion. Overall, only 7.2% of subjects reported a decrease in product performance, and only 1.6% of subjects reported a noticeable deterioration in subjectively perceived skin condition during the study period. No serious adverse effects were reported.

[0111] Study 2 (Figure 2): To further investigate the role of the stem cell homeostasis module molecules of the trigger factor complex in the effects of long-term product administration on skin condition, another 9-month long-term study was conducted. The conditions and procedures are described in the "Efficacy Test Data" section above. To ensure comparability and to specifically examine the four peptides of the stem cell homeostasis module (SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4), these four peptides were tested together with the same set of molecules of the other modules identified by the trigger factor complex TFC8-D in the "Examples" section above. As before, the carrier for the peptide derivatives of SEQ ID NO: 3 and SEQ ID NO: 4 was polyethylene glycol with a molecular weight of 35 kDa. This second long-term study revealed that the stem cell homeostasis module molecules had different associations with product performance degradation throughout the study. The reported frequency of product performance degradation associated with each of the four peptides is shown in Figure 2. Error bars represent the 95% confidence interval of the observed frequency. Overall, the amino acid sequence of SEQ ID NO: 2, which is also included in SEQ ID NO: 4, was more strongly associated with a decline in product performance during the study period than the sequence of SEQ ID NO: 1, which is also included in SEQ ID NO: 3. These differences were statistically significant (p-value < 5%). Secondly, the polyethylene glycol carrier moderately reduced the frequency of product performance decline in both amino acid sequences. Therefore, this frequency was lower for SEQ ID NO: 3 than for SEQ ID NO: 1, and lower for SEQ ID NO: 4 than for SEQ ID NO: 2. The invention described in the original claims of this application is listed below. [1] A cosmetic formulation or composition for topical administration to the skin, comprising at least one peptide or peptide derivative that induces, enhances, or improves skin regeneration or appearance, wherein the at least one peptide or peptide derivative is from three groups: (A) (i)LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRLL (SEQ ID NO: 1) (ii) LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (Sequence ID 2) (iii)Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL(Sequence ID 3) (iv) A peptide and peptide derivative that stimulates the Wnt / β-catenin signaling pathway, comprising or having the sequence / formula Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (SEQ ID NO: 4), wherein Z1 is a carrier portion covalently bonded to the N-terminus of the peptide, which reduces tissue permeability and / or basement membrane penetration of the peptide; (B) A peptide and peptide derivative that is an agonist of the tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, wherein the peptide or peptide derivative has the sequence / formula: (v) GGGGETTNMWAREWMGLPCQDQ (Sequence ID 5) (vi) comprising or having Z2-GGGGETTNMWAREWMGLPCQDQ (SEQ ID NO: 6), where Z2 is an acyl group of a branched or unbranched fatty acid covalently bonded to the N-terminus of the peptide; (C) Sequence / Formula: (vii)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCS(Sequence ID 7) (viii)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG (Sequence ID 8) (ix) A peptide and peptide derivative that is a human TGF-β3 variant comprising or having TGGG-Z3 (SEQ ID NO: 9), where X is K or E, and Z3 is a glycopolymer attached to the C-terminus; a cosmetic formulation or composition selected from at least one of these. [2] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (A). [3] A cosmetic formulation or composition of [2] in which at least one peptide or peptide derivative comprises or has the sequence / formula of SEQ ID NO: 1, 2, 3, or 4, and Z1 is polyethylene glycol having a molecular weight in the range of 8 to 60 kDa. [4] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (B). [5] The cosmetic formulation or composition of [4] wherein the at least one peptide or peptide derivative comprises or has the sequence / formula of SEQ ID NO: 5 or 6, and Z2 is a branched or unbranched fatty acid with 5 to 42 carbon atoms. [6] A cosmetic formulation or composition of [4] or [5] further comprising an appropriate amount of a tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor peptide / peptide derivative-based antagonist, wherein the peptide or peptide derivative modulates, attenuates, or inhibits the biological activity of the agonist indicated by SEQ ID NO: 5 or 6. [7] The peptide / peptide derivative-based antagonist has the following sequence / formula: GGGGETTNMWAHDWMGLPRADQ (Sequence ID 17) or A cosmetic formulation or composition comprising or having Z2-GGGGETTNMWAHDWMGLPRADQ (SEQ ID NO: 10), wherein Z2 is an acyl group of a branched or unbranched fatty acid with 5 to 42 carbon atoms bonded to the N-terminus of the peptide [6]. [8] The peptide / peptide derivative-based agonist, as indicated by Sequence ID No. 5 or 6, is partially or completely inactivated during application in any one of the cosmetic formulations or compositions from [4] to [7]. [9] The peptide / peptide derivative-based antagonist represented by SEQ ID NO: 10 or 17 is partially or completely inactivated during application, in a cosmetic formulation or composition of [6] or [7].

[10] The inactivation of the peptide / peptide derivative-based agonist or antagonist is induced by air oxidation of a methionine residue in the sequence of the peptide / peptide derivative agonist or antagonist, as described in [8] or [9], cosmetic formulations or compositions.

[11] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (C).

[12] A cosmetic formulation or composition of

[11] in which at least one peptide or peptide derivative comprises or has the sequence / formula of SEQ ID NO: 7, 8, or 9, and Z3 is or comprises an oligomer, multimer, or polymer comprising at least 15 monomer units containing a portion of trehalose or a trehalose derivative.

[13] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (A) and at least one peptide or peptide derivative selected from group (B).

[14] (a) At least one peptide or peptide derivative of group (A) comprises or has the sequence / formula of SEQ ID NO: 1, 2, 3 or 4, where Z1 is polyethylene glycol having a molecular weight in the range of 8 to 60 kDa, and (b) At least one peptide or peptide derivative of group (B) comprises or has the sequence / formula of SEQ ID NO: 5 or 6, where Z2 is a branched or unbranched fatty acid having 5 to 42 carbon atoms,

[13] a cosmetic formulation or composition.

[15] A cosmetic formulation or composition of

[13] or

[14] further comprising an appropriate amount of a tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor peptide / peptide derivative-based antagonist, wherein the peptide / peptide derivative modulates, attenuates, or inhibits the biological activity of the agonist indicated by SEQ ID NO: 5 or 6.

[16] The aforementioned antagonist is the sequence / formula: GGGGETTNMWAHDWMGLPRADQ (Sequence ID 17) or A cosmetic formulation or composition

[15] comprising or having Z2-GGGGETTNMWAHDWMGLPRADQ (SEQ ID NO: 10), wherein Z2 is an acyl group of an unbranched or branched fatty acid with 5 to 42 carbon atoms bonded to the N-terminus of the peptide.

[17] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (A), at least one peptide or peptide derivative selected from group (B), and at least one peptide or peptide derivative selected from group (C).

[18] (a) The at least one peptide or peptide derivative of group (A) comprises or has the sequence / formula of SEQ ID NO: 1, 2, 3 or 4, and Z1 is polyethylene glycol having a molecular weight in the range of 8 to 60 kDa; (b) The at least one peptide or peptide derivative of group (B) comprises or has the sequence / formula of SEQ ID NO: 5 or 6, and Z2 is a branched or unbranched fatty acid having 5 to 42 carbon atoms; (c) The at least one peptide or peptide derivative of group (C) comprises or has SEQ ID NO: 7, 8 or 9, and Z3 is or has an oligomer or multimer or polymer comprising at least 15 monomer units containing a portion of trehalose or a trehalose derivative

[17] .

[19] A cosmetic formulation or composition of

[17] or

[18] further comprising an appropriate amount of a tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor peptide / peptide derivative-based antagonist, wherein the peptide or peptide derivative modulates, attenuates, or inhibits the biological activity of the agonist indicated by SEQ ID NO: 5 or 6.

[20] The aforementioned antagonist is the sequence / formula: GGGGETTNMWAHDWMGLPRADQ (Sequence ID 17) or A cosmetic formulation or composition comprising or having Z2-GGGGETTNMWAHDWMGLPRADQ (SEQ ID NO: 10), wherein Z2 is an acyl group of an unbranched or branched fatty acid with 5 to 42 carbon atoms bonded to the N-terminus

[19] .

[21] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (A) and at least one peptide or peptide derivative selected from group (C).

[22] (a) at least one peptide derivative of group (A) comprises or has the sequence / formula of SEQ ID NO: 3 or 4, and Z1 is polyethylene glycol having a molecular size in the range of 8 to 60 kDa; (b) at least one peptide derivative of group (C) comprises or has the sequence / formula of SEQ ID NO: 7, 8 or 9, and Z3 is or has an oligomer, multimer or polymer comprising at least 15 monomer units containing a portion of trehalose or a trehalose derivative

[21] .

[23] A cosmetic formulation or composition of [1] comprising at least one peptide or peptide derivative selected from group (B) and at least one peptide or peptide derivative selected from group (C).

[24] (a) at least one peptide derivative of group (B) comprises or has the sequence / formula of SEQ ID NO: 5 or 6, and Z2 is a branched or unbranched fatty acid with 5 to 42 carbon atoms; (b) the at least one peptide derivative of group (C) comprises or has the sequence / formula of SEQ ID NO: 7, 8 or 9, and Z3 is or has an oligomer, multimer or polymer comprising at least 15 monomer units containing a portion of trehalose or a trehalose derivative

[23] .

[25] A cosmetic formulation or composition of

[23] or

[24] further comprising an appropriate amount of a tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor peptide / peptide derivative-based antagonist, wherein the peptide or peptide derivative modulates, attenuates, or inhibits the biological activity of the agonist indicated by SEQ ID NO: 5 or 6.

[26] The aforementioned antagonist is the sequence / formula: GGGGETTNMWAHDWMGLPRADQ (Sequence ID 17) or A cosmetic formulation or composition comprising or having Z2-GGGGETTNMWAHDWMGLPRADQ (SEQ ID NO: 10), wherein Z2 is an acyl group of an unbranched or branched fatty acid with 5 to 42 carbon atoms bonded to the N-terminus of the peptide

[25] .

[27] The formulation further comprises at least one peptide or peptide derivative that induces matricine activity derived from collagen type 3, LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG (Sequence ID 11) VKGESGKPGANGLSGERGPPGPQG(Sequence ID 12) Z2-LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG(Sequence ID 13) A cosmetic formulation or composition from any one of [1] to

[26] comprising or having one sequence / formula selected from the group consisting of Z2-VKGESGKPGANGLSGERGPPGPQG (SEQ ID NO: 14), wherein Z2 is an acyl group of an unbranched or branched fatty acid with 5 to 42 carbon atoms bonded to the N-terminus of the peptide.

[28] It induces CD26 / Dpp4 inhibition, EIHQEEPIGGQSGSGG-KPI (Sequence ID 15) and A cosmetic formulation or composition from any one of [1] to

[27] further comprising at least one peptide or peptide derivative comprising or having one of the sequences / formulas selected from the group consisting of EIHQEEPIGGK[Z2]SGSGG-KPI (SEQ ID NO: 16), wherein GK represents an isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of K, Z2 represents an acyl group of an unbranched or branched fatty acid having 5 to 42 carbon atoms, and K[Z2] represents an amide bond between the ε-amino functional group of K and the carboxyl functional group of the fatty acid Z2.

[29] The peptide or peptide derivative is encapsulated or bound to a liposome or ceramide structure to improve or enhance tissue delivery, in any one of the cosmetic formulations or compositions from [1] to

[28] .

[30] (i)LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL(Sequence ID 1) (ii) LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (Sequence ID 2) (iii)Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL(Sequence ID 3) (iv) An isolated peptide or peptide derivative having or containing a sequence / formula selected from the group consisting of Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL (SEQ ID NO: 4): stimulates the Wnt / β-catenin signaling pathway, wherein Z1 is a carrier moiety covalently bonded to the N-terminus of the peptide, which reduces tissue permeability and / or basement membrane penetration of the peptide.

[31] Z1 is a polyethylene glycol having a molecular size in the range of 8-60 kDa, an isolated peptide or peptide derivative of

[30] .

[32] An isolated peptide or peptide derivative that acts as an agonist for the tissue-protective heterodimer or heterooligomeric EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, (i) GGGGETTNMWAREWMGLPCQDQ (Sequence ID 5) (ii) Having or containing an sequence / formula selected from the group consisting of Z2-GGGGETTNMWAREWMGLPCQDQ (sequence number 6), An isolated peptide or peptide derivative, wherein Z2 is an acyl group of a branched or unbranched chain fatty acid with 5 to 42 carbon atoms bonded to the N-terminus of the peptide.

[33] An isolated peptide or peptide derivative that acts as an antagonist of the tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, (i) Z2-GGGGETTNMWAHDWMGLPRADQ (Sequence ID 10), (ii) Having or containing an sequence / formula selected from the group consisting of GGGGETTNMWAHDWMGLPRADQ (Sequence ID 17), Z2 is an isolated peptide or peptide derivative, wherein Z2 is an acyl group of an unbranched or branched fatty acid with 5 to 42 carbon atoms bonded to the N-terminus of the peptide.

[34] (i)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCS(Sequence ID 7) (ii)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG (Sequence ID 8) (iii)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-Z3 (Sequence ID 9) (iv)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-[4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose]n (SEQ ID NO: 18) (v)ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-[Q-6-deoxy-trehalose]n (SEQ ID NO: 19): Induces the biological activity of human TGF-β3 having a sequence / formula selected from the group consisting of these, An isolated peptide or peptide derivative in which X is K or E, Z3 is a glycopolymer bonded to the C-terminus, and n is an integer from 15 to 50, preferably from 15 to 30.

[35] One isolated peptide or peptide derivative from

[30] to

[34] , optionally encapsulated or conjugated in a liposome or ceramide structure, for use in topical cosmetic treatments of the skin, including skin repair, skin rejuvenation, natural skin radiance, wrinkle reduction, anti-aging of the skin, and prevention and improvement of dry, dull, and brittle skin.

Claims

1. A cosmetic formulation for topical application to the skin in the form of a cream, lotion, or serum, which regenerates or enhances or improves the appearance of the skin, The aforementioned formulation (i) Among the first biologically effective group (A) which is contained in the cosmetic formulation to stimulate the Wnt / β-catenin signaling pathway, exerts a local effect in the epidermis but does not act in the dermis, Sequence ID 2: LNPSECPKTVLGASTSTLDASYSTAEAENHVRL, or Sequence ID 4: Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL, or Sequence ID 1: LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRLL, or Sequence ID 3: Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL, A peptide or peptide fusion molecule having a sequence or formula defined by, The peptide or peptide fusion molecule wherein Z1 is polyethylene glycol (PEG) having a molecular weight in the range of 20 to 40 kDa, which is covalently bonded to the N-terminus of the peptide and reduces tissue permeability and / or basement membrane penetration of the peptide; (ii) Of the second biologically effective group (B) contained in the cosmetic formulation to act as an agonist for tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, Sequence ID 6: Z2-GGGGGETTNMWAREWMGLPCQDQ, A first peptide fusion molecule having the sequence or formula defined by, and To act as an antagonist of the receptor in the cosmetic formulation, among the second biologically effective group (B) contained in the cosmetic formulation, Sequence ID 10: Z2-GGGGGETTNMWAHDWMGLPRADQ A second peptide fusion molecule having the sequence or formula defined by, The first peptide fusion molecule and the second peptide fusion molecule, wherein Z2 is myristoyl bonded to the N-terminus of the peptide; (iii) Human transform growth factor beta-3 (TGF-β3) variant, which is included in the cosmetic formulation to support healthy skin, and is a third biologically effective group (C), Sequence ID 9: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASAASPPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-Z3 A peptide fusion molecule having a sequence or formula defined by, The peptide fusion molecule wherein X is E, and Z3 is an oligomer consisting of 18 to 30 trehalose or trehalose derivative units, and the amino acid sequence includes three amino acid substitutions, T57K, L68H, and S102E, compared to natural human TGF-β3; (iv) CD26 / Dpp4 inhibition is induced in the cosmetic formulation, Sequence ID 15: EIHQEEPIGGQSGSGGG-KPI, or Sequence ID 16: EIHQEEPIGGK[Z2]SGSGG-KPI A peptide having a sequence or formula defined by, G-K represents an isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of K, Z2 is myristoyl bonded to the N-terminus of the peptide, and K[Z2] represents an amide bond between the ε-amino functional group of K and the carboxyl functional group of Z2, the peptide; and (V) Contained in the cosmetic formulation to induce matricine activity derived from collagen type 3, Sequence ID 11: LQGLPGTGPGPGENGKPGEGPGKGDAGAAPGAAPGGKGDAGAAPGERGPPG, and Sequence ID 12: VKGESGKPGANGLSGERGPPGPQG, or Sequence ID 13: Z2-LQGLPGTGPPPGENGKPGEPPGKGDAGAAPGAAPGGKGDAGAAPGERGPPG, and Sequence ID 14: Z2-VKGESGKPGANGLSGERGPPGPQG Two peptides having a sequence or formula defined by, The Z2 is myristoyl bound to the N-terminus of the peptide, the peptide: Cosmetic formulations, including those mentioned above.

2. In Sequence ID No. 9, Z3 is [4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose] 18 (Sequence ID No. 18), or [Q-6-deoxy-trehalose] 18 (Sequence ID 19) A cosmetic formulation according to claim 1.

3. The aforementioned formulation is named TFC8-A, and, (i) contained in the cosmetic formulation to stimulate the Wnt / β-catenin signaling pathway, Sequence ID 2: LNPSECPKTVLGASTSTLDASYSTAEAENHVRL A peptide having the sequence or formula defined by; (ii) A tissue-protective heterodimer or heterooligomer contained in the cosmetic formulation to act as an agonist of the EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, Sequence ID 6: Z2-GGGGGETTNMWAREWMGLPCQDQ A first peptide fusion molecule having the sequence or formula defined by, and A substance contained in the cosmetic formulation is used to act as an antagonist of the receptor of the cosmetic formulation, and Sequence ID 10: Z2-GGGGGETTNMWAHDWMGLPRADQ A second peptide fusion molecule having the sequence or formula defined by, The first peptide fusion molecule and the second peptide fusion molecule, wherein Z2 is myristoyl bonded to the N-terminus of the peptide; (iii) Human transforming growth factor beta-3 (TGF-β3) mutant, Sequence ID 18: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANF-CSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLE-PLTILYYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-[4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose]18, A peptide fusion molecule having a sequence or formula defined by, X is E, the peptide fusion molecule; (iv) CD26 / Dpp4 inhibition is induced in the cosmetic formulation, Sequence ID 15: EIHQEEPIGQSGSGG-KPI A peptide having a sequence or formula defined by, G-K represents the isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of K, the peptide; and (V) Contained in the cosmetic formulation to induce matricine activity derived from collagen type 3, Sequence ID 11: LQGLPGTGPGPGENGKPGEGPGKGDAGAAPGAAPGGKGDAGAAPGERGPPG, and Sequence ID 12: VKGESGKPGANGLSGERGPPGPQG, Two peptides having the sequence or formula defined by; A cosmetic formulation according to claim 1, comprising:

4. The aforementioned formulation is named TFC8-B, and, (i) contained in the cosmetic formulation to stimulate the Wnt / β-catenin signaling pathway, Sequence ID 4: Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL A peptide having a sequence or formula defined by, The peptide is wherein Z1 is a polyethylene glycol (PEG) bonded to the N-terminus of the peptide and having a molecular weight of 35 kDa; (ii) A tissue-protective heterodimer or heterooligomer contained in the cosmetic formulation to act as an agonist of the EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, Sequence ID 6: Z2-GGGGGETTNMWAREWMGLPCQDQ A first peptide fusion molecule having the sequence or formula defined by, and A substance contained in the cosmetic formulation is used to act as an antagonist of the receptor of the cosmetic formulation, and Sequence ID 10: Z2-GGGGGETTNMWAHDWMGLPRADQ A second peptide fusion molecule having the sequence or formula defined by, The first peptide fusion molecule and the second peptide fusion molecule, wherein Z2 is myristoyl bonded to the N-terminus of the peptide; (iii) A human transforming growth factor beta-3 (TGF-β3) mutant, and Sequence ID 19: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANF-CSGPCPPYLRSAADTKHSTVLGLYNTHNPEASASPCCVPQDLE-PLTILYYYVGRTPPKVEQLENMVVKSCKCSLPXTGGG-[Q-6-deoxy-trehalose]18, A peptide fusion molecule having a sequence or formula defined by, The peptide fusion molecule in which X is E; (iv) CD26 / Dpp4 inhibition is induced in the cosmetic formulation, Sequence ID 16: EIHQEEPIGGK[Z2]SGSGG-KPI A peptide having a sequence or formula defined by, G-K represents an isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of K, and K[Z2] represents an amide bond between the ε-amino functional group of K and the carboxyl functional group of Z2, the peptide; and (V) Contained in the cosmetic formulation to induce matricine activity derived from collagen type 3, Sequence ID 13: Z2-LQGLPGTGPGPGENGKPGEGPKGDAGAPGAPGGKGDA-GAPGERGPPG, and Sequence ID 14: Z2-VKGESGKPGANGLSGERGPPGPQG, Two peptides having a sequence or formula defined by, The peptide is such that Z2 is myristoyl bonded to the N-terminus of the peptide; A cosmetic formulation according to claim 1, comprising:

5. The aforementioned formulation is named TFC8-C, and, (i) contained in the cosmetic formulation to stimulate the Wnt / β-catenin signaling pathway, Sequence ID 1: LNPSECPKTVLGAYGKTLDASYSTAEAENHVRL Peptides having a sequence or formula defined by; (ii) A tissue-protective heterodimer or heterooligomer contained in the cosmetic formulation to act as an agonist of the EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, Sequence ID 6: Z2-GGGGGETTNMWAREWMGLPCQDQ A first peptide fusion molecule having the sequence or formula defined by, and A substance contained in the cosmetic formulation is used to act as an antagonist of the receptor of the cosmetic formulation, and Sequence ID 10: Z2-GGGGGETTNMWAHDWMGLPRADQ A second peptide fusion molecule having the sequence or formula defined by, The first peptide fusion molecule and the second peptide fusion molecule, wherein Z2 is myristoyl bonded to the N-terminus of the peptide; (iii) Human transforming growth factor beta-3 (TGF-β3) mutant, Sequence ID 18: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANF-CSGPCPYLRSADTKHSTVLGLYNTHNPEASASPCCVPQDLE-PLTILYYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-[4,6-O-(4-vinylbenzylidene)-α,α-D-trehalose]18, A peptide fusion molecule having a sequence or formula defined by, The peptide fusion molecule in which X is E; (iv) CD26 / Dpp4 inhibition is induced in the cosmetic formulation, Sequence ID 15: EIHQEEPIGQSGSGG-KPI A peptide having a sequence or formula defined by, G-K represents the isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of K, the peptide; and (V) Contained in the cosmetic formulation to induce matricine activity derived from collagen type 3, Sequence ID 11: LQGLPGTGPGPGENGKPGEGPGKGDAGAAPGAAPGGKGDAGAAPGERGPPG, and Sequence ID 12: VKGESGKPGANGLSGERGPPGPQG, Two peptides having the sequence or formula defined by; A cosmetic formulation according to claim 1, comprising:

6. The aforementioned formulation is named TFC8-D, and, (i) contained in the cosmetic formulation to stimulate the Wnt / β-catenin signaling pathway, Sequence ID 3: Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL A peptide having a sequence or formula defined by, The peptide is wherein Z1 is a polyethylene glycol (PEG) bonded to the N-terminus of the peptide and having a molecular weight of 35 kDa; (ii) A tissue-protective heterodimer or heterooligomer contained in the cosmetic formulation to act as an agonist of the EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, Sequence ID 6: Z2-GGGGGETTNMWAREWMGLPCQDQ A first peptide fusion molecule having the sequence or formula defined by, and The cosmetic formulation contains the following to act as an antagonist to the receptor of the cosmetic formulation: Sequence ID 10: Z2-GGGGGETTNMWAHDWMGLPRADQ A second peptide fusion molecule having the sequence or formula defined by, The second peptide fusion molecule wherein Z2 is myristoyl bonded to the N-terminus of the peptide; (iii) Human transforming growth factor beta-3 (TGF-β3) mutant, Sequence ID 19: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANF-CSGPCPYLRSADTKHSTVLGLYNTHNPEASAASPCCVPQDLEPLTILYYVGRTPPKVEQLENMVVKSCKCSLPXTGGG-[Q-6-deoxy-trehalose]18, A peptide fusion molecule having a sequence or formula defined by, The peptide fusion molecule in which X is E; (iv) CD26 / Dpp4 inhibition is induced in the cosmetic formulation, Sequence ID 16: EIHQEEPIGGK[Z2]SGSGG-KPI A peptide having a sequence or formula defined by, The peptide wherein G-K represents an isopeptide bond between the carboxyl functional group of G and the ε-amino functional group of K, and K[Z2] represents an amide bond between the ε-amino functional group of K and the carboxyl functional group of Z2; and (V) Contained in the cosmetic formulation to induce matricine activity derived from collagen type 3, Sequence ID 13: Z2-LQGLPGTGPPPGENGKPGEPPGKGDAGAAPGAAPGGKGDAGAAPGERGPPG, and Sequence ID 14: Z2-VKGESGKPGANGLSGERGPPGPQG, Two peptides having a sequence or formula defined by, The peptide wherein Z2 is myristoyl bound to the N-terminus of the peptide; A cosmetic formulation according to claim 1, comprising:

7. Contains in order to stimulate the Wnt / β-catenin signaling pathway, In cosmetic formulations for topical administration to the skin, the effects are exerted through combination with selected peptides having different biological functions. Sequence ID 1: LNPSECPKTVLGAYGKTLDASYSTAEAENHVRL, or Sequence ID 3: Z1-LNPSECPKTVLGAEYGKTLDASYSTAEAENHVRL, or Sequence ID 2: LNPSECPKTVLGASTSTLDASYSTAEAENHVRL, or Sequence ID 4: Z1-LNPSECPKTVLGASTSTLDASYSTAEAENHVRL Having an arrangement or expression defined by, The Z1 is an isolated peptide or peptide fusion molecule, wherein Z1 is polyethylene glycol (PEG) having a molecular weight in the range of 20 to 40 kDa, covalently bonded to the N-terminus of the peptide, thereby reducing tissue permeability and / or basement membrane penetration of the peptide.

8. Acting as an agonist or antagonist of a tissue-protective heterodimer or heterooligomer EPOR / CD131 (erythropoietin receptor / differentiation cluster 131) receptor, In cosmetic formulations for topical administration to the skin, the effects are exerted through combination with selected peptides having different biological functions. The aforementioned agonist, Sequence ID 5: GGGGETTNMWAREWMGLPCQDQ, or Sequence ID 6: Z2-GGGGGETTNMWAREWMGLPCQDQ Having an arrangement or formula defined by, The aforementioned antagonist, Sequence ID 10: Z2-GGGGGETTNMWAHDWMGLPRADQ, or Sequence ID 17: GGGGETTNMWAHDWMGLPRADQ Having an arrangement or expression defined by, The Z2 is an isolated peptide or peptide fusion molecule, wherein Z2 is myristoyl bound to the N-terminus of the peptide.

9. A human transform growth factor beta-3 (TGF-β3) mutant, In cosmetic formulations for topical administration to the skin, the effects are exerted through combination with selected peptides having different biological functions. Sequence ID 7: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPPYLRSADTKHSTVLGLYNTHNPEASAASPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCS, Having an arrangement or expression defined by, The isolated peptide's amino acid sequence contains three amino acid substitutions—T57K, L68H, and S102E—compared to that of natural human TGF-β3.

10. The peptide is fused with trehalose or a trehalose derivative at the C-terminus via a peptide motif, Sequence ID 9: ALDTNYCFRNLEENCCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTKHSTVLGLYNTHNPEASAASPPCCVPQDLEPLTILYYVGRTPKVEQLENMVVKSCKCSLPXTGGG-Z3, Form a peptide fusion molecule having the sequence defined by, The isolated peptide according to claim 9, wherein LPXTGGG is the peptide motif, X is E, and Z3 is an oligomer consisting of 18 to 30 trehalose or trehalose derivative units.

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