WO2014093053A1 - Modulation de thymosine bêta-4 dans la peau - Google Patents

Modulation de thymosine bêta-4 dans la peau Download PDF

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Publication number
WO2014093053A1
WO2014093053A1 PCT/US2013/072731 US2013072731W WO2014093053A1 WO 2014093053 A1 WO2014093053 A1 WO 2014093053A1 US 2013072731 W US2013072731 W US 2013072731W WO 2014093053 A1 WO2014093053 A1 WO 2014093053A1
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Prior art keywords
skin
thymosin beta
active agent
human
inflammation
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PCT/US2013/072731
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English (en)
Inventor
Ying Chen
Qian Zheng
Russell J. WYBORSKI
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Avon Products, Inc.
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Publication date
Priority claimed from US13/710,585 external-priority patent/US8709507B2/en
Priority claimed from US13/735,659 external-priority patent/US20140193529A1/en
Application filed by Avon Products, Inc. filed Critical Avon Products, Inc.
Publication of WO2014093053A1 publication Critical patent/WO2014093053A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
    • A61K8/4926—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having six membered rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]

Definitions

  • the present invention relates generally to methods of improving the aesthetic appearance and health of human skin; to methods for identifying compounds useful for treating skin; to methods for modulating skin inflammatory responses; and also to methods for identifying compounds useful for modulating skin inflammatory responses.
  • the invention relates to compounds, other than retinoids, that stimulate thymosin beta-4 expression in skin and thereby modulate skin inflammatory responses.
  • Thymosins refer to a family of biochemically and functionally distinct proteins that were originally identified from the thymus, but which are now known to be present in many other tissues, and which have a variety of different physiological functions. More than 20 isoforms of beta-thymosin have been identified in different species, and among humans, three beta-thymosins have been identified, including thymosin beta-4, thymosin beta- 10, and thymosin beta- 15, all of which share significant amino acid sequence homology. Despite this homology, each of these thymosin beta proteins is a distinct gene product with different functions.
  • Thymosin beta-4 the most abundant of the beta-thymosins, is a highly conserved, water-soluble amino acid acidic polypeptide, which can be found in a variety of mammalian tissues and cell types.
  • the mammalian gene encoding thymosin beta-4 localizes to the X-chromosome.
  • Human thymosin beta-4 X-linked, has 44 amino acids (msdkpdmaei ekfdksklkk tetqeknplp sketieqekq ages, SEQ ID NO: l), and escapes X-inactivation by being processed into a 43 amino acid peptide, with a molecular weight of 4.9 kDa, by removal of the first methionyl residue (Girardi, M., et ah, Immunology, 2003, 109: 1-7). Thymosin beta- 4 is localized to both the cytoplasm and the nucleus of cells. Thymosin beta-4 is present in many tissues, and has multiple biological functions.
  • Inflammation is normally a highly complex, interdependent biochemical and physiological protective response to tissue injury (e.g., due to bacterial injury or trauma). This process serves to destroy, dilute, partition off, or remove the injurious agent and the injured tissues, thereby promoting tissue repair. When this crucial and normally beneficial response occurs in an uncontrolled or exaggerated manner, the result is inappropriate or excessive tissue damage that often results in chronic inflammation.
  • Acute inflammation is a normal process that protects and heals the body following physical injury or infection. Acute inflammation involves local dilation of blood vessels as well as increased vessel permeability to improve blood flow to the injured area.
  • mast cells, platelets, nerve endings, endothelial cells, and other resident cells release signaling molecules and chemoattractants that recruit leukocytes to the affected area.
  • Neutrophils a type of granulocyte, are the first leukocytes to appear at the injured site.
  • Neutrophils also release cytokines, including interleukin (IL)-l, IL-6, tumor necrosis factor (TNF)-alpha, gamma interferon (INF-gamma), and others.
  • IL interleukin
  • TNF tumor necrosis factor
  • INF-gamma gamma interferon
  • proinflammatory cytokines in turn induce the liver to synthesize various acute phase reactant proteins and also induce systemic inflammatory responses (e.g., fever and leukocytosis— a rise in the number of white blood cells).
  • Neutrophils are short-lived and are thus primarily involved in the early stages of inflammation.
  • inflammation can last days, months, and even years.
  • Chronic inflammation is primarily mediated by monocytes and long-lived macrophages; monocytes mature into macrophages once they leave the bloodstream and enter tissues. Macrophages engulf and digest microorganisms, foreign invaders, and senescent cells. Macrophages release several different chemical mediators, including IL-1, TNF-alpha, and prostaglandins, that perpetuate the pro-inflammatory response. At later stages, other cells, including lymphocytes, invade the affected tissues: T lymphocytes kill virus-infected cells and B lymphocytes produce antibodies that specifically target the invading microorganisms for destruction.
  • Inflammatory skin diseases are the most common problem in dermatology.
  • Skin inflammation can be characterized as acute or chronic. Acute inflammation can result from exposure to UV radiation (UVR), ionizing radiation, allergens, or to contact with chemical irritants (soaps, hair dyes, etc.). This type of inflammation is typically resolved within 1 to 2 weeks with little accompanying tissue destruction. In contrast, chronic inflammation results from a sustained immune cell mediated inflammatory response within the skin itself. This inflammation is long lasting and can cause significant and serious tissue destruction.
  • UV radiation UV radiation
  • ionizing radiation ionizing radiation
  • allergens or to contact with chemical irritants (soaps, hair dyes, etc.). This type of inflammation is typically resolved within 1 to 2 weeks with little accompanying tissue destruction.
  • chronic inflammation results from a sustained immune cell mediated inflammatory response within the skin itself. This inflammation is long lasting and can cause significant and serious tissue destruction.
  • Inflammatory skin conditions affect over 35 million Americans who annually spend over $2 billion to treat their symptoms.
  • Factors that contribute to skin inflammation include, but are not limited to, sun, hot water, environmental irritants (pollution, wind, smoke), components of products that may be topically applied to the skin, and possibly caffeine and alcohol.
  • Development of acne is also a contributing cause to subsequent skin inflammation. Using products such as moisturizers or topical cortisone preparations to temporarily hydrate the skin may actually make some conditions worse.
  • retinol is a potent upregulator of thymosin beta-4. This surprising result lead to the search for additional agents that upregulate thymosin beta-4, that could act as potential alternatives or replacements for retinol in the treatment of and in the improvement of the aesthetic appearance and health of human skin.
  • the present invention provides stimulators, other than retinoids, of thymosin beta-4 ("TB-4"), to improve one or more signs of dermatological aging.
  • the stimulators upregulate levels of thymosin beta-4 in the skin. It is believed that upregulators of thymosin beta-4 within dermal fibroblasts and epidermal keratinocytes lead to increased protein production within the cells, including, for example, the fibroblast specific proteins, collagen, elastin, and fibrillin, and the keratinocyte protein keratin.
  • upregulation of thymosin beta-4 will have a beneficial effect on reducing the appearance of aging on skin. It is also believed that upregulation of thymosin beta-4 within at least dermal fibroblasts yields inhibition of skin inflammatory response, including but not limited to suppression of IL-8 gene expression and /or pro-inflammatory cytokine and/or chemokine gene and /or protein expression.
  • a method for improving the aesthetic appearance of human skin comprising topically applying to an area of the skin in need thereof using an effective amount of an active agent, other than a retinoid, that stimulates thymosin beta-4 expression, for a time sufficient to improve the aesthetic appearance of said human skin.
  • an active agent other than a retinoid
  • the sufficiency of the time will be determined by individual response, in particular where wrinkles and/or fine lines are treated, but encompasses once daily administration for from one to four weeks.
  • a method is provided for using an effective amount of an active agent that stimulates TB-4 expression for a time sufficient to modulate the inflammatory response of human skin.
  • methods are provided for screening candidate substances to identify actives useful for improving the aesthetic appearance of skin and/or modulating (in one embodiment, inhibiting) the inflammatory response of skin.
  • the method comprises assaying candidate substances for ability to stimulate thymosin beta-4 expression in a dermal fibroblast or keratinocyte.
  • the method typically involves incubating human dermal fibroblasts or keratinocytes with a candidate substance and subsequently measuring the levels of mRNA encoding thymosin beta-4 by a quantitative technique such as quantitative polymerase chain reaction (qPCR).
  • the method may optionally additionally involve incubating human cells with TNF-alpha subsequent to incubation with a candidate substance and measuring relative inhibition of inflammatory response, including but not limited to suppression of IL-8 gene expression and /or pro-inflammatory cytokine and/or chemokine gene and /or protein expression.
  • the cell used in the assay may be any animal cell that expresses thymosin beta-4, but is preferably mammalian, and is more preferably a human or mouse cell.
  • Methods are also provided for inhibiting the inflammatory response of human skin comprising topically applying to an area of the skin in need thereof an effective amount of a substance that stimulates thymosin beta-4 levels in human skin cells, for example in dermal fibroblasts and epidermal keratinocytes.
  • the thymosin beta-4 stimulators are referred to herein as "actives" or upregulators, and typically will be formulated in a cosmetically acceptable vehicle and topically applied to a human integument, such as the skin of the face, neck, hands, chest, legs, etc., for a time sufficient to enhance the health or aesthetic appearance thereof.
  • a method for inhibiting the inflammatory responseof human skin comprising topically applying to an area of the skin in need thereof an effective amount of an active agent that stimulates (e.g., upregulates) cellular levels of thymosin beta-4, wherein the ability of the active agent to stimulate thymosin beta-4 has been determined by an assay which measures the expression level of thymosin beta-4 in a cell that has been contacted with the active agent.
  • the cell used in the assay may be any animal cell that expresses thymosin beta-4, but is preferably mammalian, and is more preferably a human or mouse cell.
  • the cell may be a skin cell, such as a fibroblast or keratinocyte.
  • the assay may measure the levels of any homolog, fragment or marker of thymosin beta-4, but typically what is measured are the expression levels of thymosin beta-4, which is expressed by the human gene TMSB4X or by the mammalian gene tmsb4x.
  • the expression levels of the human gene TMSB4X or the mouse gene tmsb4x may be determined, for example, by measuring the expression levels of the corresponding mRNA by any suitable technique, such as quantitative polymerase chain reaction (qPCR).
  • a method for inhibiting the inflammatory response of human skin comprising topically applying to an area of the skin in need thereof an effective amount of an active agent that inhibits skin inflammation, wherein the ability of the active agent to inhibit skin inflammation has been determined by an assay which measures the expression level ofIL-8 gene expression and /or pro-inflammatory cytokine and/or chemokine genes and /or proteins subsequent to incubation with a candidate substance and then incubation with TNF-alpha.
  • the cell used in the assay may be any animal cell that expresses thymosin beta-4 and is capable of mounting an inflammatory response to TNF-alpha challenge, but is preferably mammalian, and is more preferably a human or mouse cell.
  • the cell may be a skin cell, such as a fibroblast or keratinocyte.
  • the assay may measure the levels of any inflammatory biomarker, but typically what is measured are the expression levels of IL-8 and/or other pro-inflammatory chemokines and / or cytokines.
  • the expression levels of the IL-8 and/or other pro-inflammatory chemokines and / or cytokines may be determined, for example, by measuring the expression levels of the corresponding mRNA by any suitable technique, such as quantitative polymerase chain reaction (qPCR), or by any other sutiable genomic and/or proteomic methods of analysis.
  • qPCR quantitative polymerase chain reaction
  • thymosin beta-4 refers to a protein, such as the proteins encoded in humans by the TMSB4X gene and in mammals such as mice by the tmsb4x gene.
  • the thymosin beta-4 protein (and its associated RNA and/or DNA encoding thymosin beta-4) as described in the present invention may be from any animal, but is typically human or mouse thymosin beta-4, and preferably human thymosin beta-4.
  • thymosin beta-4 examples are that of the National Center for Biotechnology Information (“NCBI"), Accession Numbers NP_066932 (human, thymosin beta-4, protein), NM_021109 (human, thymosin beta-4, mRNA), NP_067253 (mouse, thymosin beta-4, protein), NM_021278 (mouse, thymosin beta-4, mRNA), which are hereby incorporated by reference and summarized in Table 1.
  • NCBI National Center for Biotechnology Information
  • the term "inflammatory response” means a tissue reaction to injury or an antigen that may include pain, swelling, itching, redness, heat, and loss of function.
  • the response may involve dilation of blood vessels and consequent leakage of fluid, causing edema; leukocytic exudation; and release of plasma proteases and vasoactive amines such as histamine.
  • Methods are also provided for treating a skin condition comprising topically applying to an area of the skin in need thereof an effective amount of an active agent that upregulates thymosin beta-4, wherein the ability of said active agent to stimulate the thymosin beta-4 expression has been determined by an assay which measures the expression level of thymosin beta-4 in a cell that has been contacted with said active agent.
  • a method of treating wrinkles and/or fines lines comprising topically applying to an area of the skin in need thereof an effective amount of an active agent that upregulates thymosin beta-4, wherein the ability of the active agent to upregulate thymosin beta-4 has been determined by an assay which measures the expression level of thymosin beta-4 in a cell that has been contacted with the active agent.
  • the assaying step most typically comprises incubating human dermal fibroblasts with a candidate substance and subsequently measuring the levels of mRNA encoding thymosin beta-4, wherein the candidate substance was selected for use as an active agent, in part, on the basis of its ability to upregulate thymosin beta-4 expression.
  • the cell used in the assay may be any animal cell that expresses thymosin beta-4, but is preferably mammalian, and is more preferably a human or mouse cell.
  • a cosmetic composition for improving the aesthetic appearance of human skin comprising a cosmetically acceptable vehicle in the form of a water-in-oil or oil-in-water emulsion, and an effective amount of an active agent, other than a retinoid, that upregulates thymosin beta-4.
  • a cosmetic composition for inhibiting the inflammatory response of human skin comprising a cosmetically acceptable vehicle in the form of a water-in-oil or oil-in-water emulsion, and an effective amount of an active agent, other than a retinoid, that upregulates thymosin beta-4.
  • the composition is intended for use as a non-therapeutic treatment.
  • the composition is an article intended to be rubbed, poured, sprinkled, or sprayed on, introduced into, or otherwise applied to the human body for cleansing, beautifying, promoting attractiveness, or altering the appearance, in accordance with the US FD&C Act, sec. 201(i).
  • a therapeutic composition for inhibiting the inflammatory response of human skin comprising a therapeutically acceptable vehicle in the form of a water-in-oil or oil-in-water emulsion, and an effective amount of an active agent, other than a retinoid, that upregulates thymosin beta-4 and that results in inhibition of subsequent inflammatory response in the skin comprising thymosin beta-4 - upregulated cells.
  • compositions comprising an amount of a botanical extract effective to upregulate thymosin beta-4 in a dermal fibroblast or keratinocyte, and a cosmetically acceptable vehicle, wherein said botanical extract is extracted from a plant selected from the group consisting of Maesa japonica, Celosia argentea, Berchemia Uneata, Ixora chinesis, Physalis minima, and combinations thereof.
  • thymosin beta-4 refers to a protein, such as the proteins encoded in humans by the TMSB4X gene and in mice by the tmsb4x gene.
  • the thymosin beta-4 protein (and its associated RNA and/or DNA encoding thymosin beta-4) as described in the present invention may be from any animal, but is typically human or mouse thymosin beta-4, and preferably human thymosin beta-4.
  • thymosin beta-4 examples are that of the National Center for Biotechnology Information (“NCBI"), Accession Numbers NP_066932 (human, thymosin beta-4, protein), NM_021109 (human, thymosin beta-4, mRNA), NP_067253 (mouse, thymosin beta-4, protein), NM_021278 (mouse, thymosin beta-4, mRNA), which are hereby incorporated by reference and summarized in Table 1 (see SEQ ID NOs: 1-4).
  • NCBI National Center for Biotechnology Information
  • Thymosin beta-4 as encompassed by the present invention includes any full- length, fragment or derivative of this protein having the biological functions of thymosin beta-4, as well as the corresponding RNA and/or DNA sequences encoding thymosin beta-4, as obtained from any animal.
  • thymosin beta-4 also refers to a genus of polypeptides that further encompasses proteins having the amino acid sequence of SEQ ID NO: l and SEQ ID NO:3, as well as those proteins and polypeptides having a high degree of similarity (at least 90% homology) with such amino acid sequences and which proteins and polypeptides have the biological functions of thymosin beta-4.
  • the oligonucleotide comprising a part of or an entire sequence of the nucleic acid sequence described in SEQ ID NO:2 or SEQ ID NO:4 for use in the present invention includes, for example, an oligonucleotide comprising a base sequence sharing about 70% or more, preferably about 80% or more, more preferably about 90% or more, and furthermore preferably about 95% or more of homology with the nucleic acid sequence described in SEQ ID NO:2 or SEQ ID NO:4, or an oligonucleotide comprising a part of or an entire sequence of the nucleic acid sequence described in SEQ ID NO:2 or SEQ ID NO:4 of the sequence listing.
  • the term “stimulator” encompasses any substance, including, without limitation, organic molecules; biomolecules (e.g., peptides, proteins, antibodies, nucleic acid oligomers, etc.); and combinations of substances, such as botanical extracts.
  • the stimulators may stimulate the cellular levels of thymosin beta-4, by which is meant that the cellular levels of thymosin beta-4 protein are increased by the active agent.
  • stimulation may refer to upregulation, induction, stimulation, and/or potentiation, or relief of inhibition.
  • the stimulators may also be, without limitation, activators, or agonists, which are compounds that, for example, bind to, stimulate, increase, open, activate, facilitate, enhance activation, sensitize, or upregulate expression levels of genes or thymosin beta-4 proteins or peptides.
  • activators or agonists, which are compounds that, for example, bind to, stimulate, increase, open, activate, facilitate, enhance activation, sensitize, or upregulate expression levels of genes or thymosin beta-4 proteins or peptides.
  • the mechanism by which the protein level is modulated is not important.
  • inhibitor encompasses any substance, including, without limitation, organic molecules; biomolecules (e.g., peptides, proteins, antibodies, nucleic acid oligomers, etc.); and combinations of substances, such as botanical extracts.
  • the inhibitors may inhibit the cellular levels of thymosin beta-4, by which is meant that the cellular levels of thymosin beta-4 protein are increased by the active agent; and/or inhibit inflammatory response, by which is meant but is not limited to suppression of IL-8 gene expression and /or pro-inflammatory cytokine and/or chemokine gene and / or protein expression.
  • the term “inhibition” may refer to downregulation or relief of stimulation.
  • the stimulators may also be, without limitation, deactivators, or antagonists, which are compounds that, for example, bind to, decrease, close, block, limit activation, desensitize, or down regulate expression levels of genes or thymosin beta-4 proteins or peptides or IL-8 gene expression and /or proinflammatory cytokine and/or chemokine gene and /or protein expression.
  • deactivators or antagonists, which are compounds that, for example, bind to, decrease, close, block, limit activation, desensitize, or down regulate expression levels of genes or thymosin beta-4 proteins or peptides or IL-8 gene expression and /or proinflammatory cytokine and/or chemokine gene and /or protein expression.
  • the mechanism by which the protein level is modulated is not important.
  • the term "expression levels” refers to an amount of a gene and/or protein that is expressed in a cell.
  • a “gene” includes a polynucleotide containing at least one open reading frame that is capable of encoding a particular polypeptide.
  • polynucleotide is synonymous with “oligonucleotide” and includes polymeric forms of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof, including, without limitation, mRNA, DNA, cDNA, primers, probes, and the like.
  • an assay is provided for determining the expression levels of thymosin beta-4 after a cell has been treated, incubated, or otherwise contacted with a candidate substance.
  • candidate substance refers to any substance that is tested for activity as a stimulator of thymosin beta-4, whether or not the substance is suspected of possessing such activity, other than retinoids.
  • the cell can be any cell from any animal wherein the cell expresses thymosin beta-4.
  • the cell is a mammalian fibroblast.
  • the cell is a mammalian keratinocyte.
  • the cell is a human or mouse cell.
  • the cell After the cell has been incubated with a candidate substance for a sufficient length of time to provide a measurable change in expression levels, which will typically be at least one hour, and more typically from about 12 hours to about 72 hours, the cell is then lysed to release the cellular components, such as thymosin beta-4 and mRNA encoding thymosin beta-4.
  • the amount of thymosin beta-4 protein or mRNA may then be measured by any suitable technique for detection and quantitation of peptides and proteins and/or polynucleotides (e.g., mRNA).
  • an assay for determining the antiinflammatory effects of overexpressed thymosin beta-4 after a cell has been treated, incubated, or otherwise contacted with a candidate substance and subsequently challenged with TNF-alpha or another inflammation stimulator.
  • candidate substance refers to any substance that is tested for activity as a stimulator of thymosin beta-4 and as an inhibitor of inflammation, whether or not the substance is suspected of possessing such activity, other than retinoids.
  • the cell can be any cell from any animal wherein the cell expresses thymosin beta-4.
  • the cell is a mammalian fibroblast.
  • the cell is a mammalian keratinocyte.
  • the cell is a human or mouse cell.
  • a candidate substance for a sufficient length of time to provide a measurable change in expression levels, which will typically be at least one hour, and more typically from about 12 hours to about 72 hours
  • the cell is then challenged with TNF-alpha or another suitable inflammation stimulator and then lysed to release the cellular components, such as thymosin beta-4, mRNA encoding thymosin beta-4, IL-8, and/or mRNA encoding IL-8.
  • the amount of thymosin beta-4 or IL-8 protein or mRNA may then be measured by any suitable technique for detection and quantitation of peptides and proteins and/or polynucleotides (e.g., mRNA).
  • One method for measuring expression levels of thymosin beta-4 and/or proinflammatory chemokines or cytokines involves the quantitation of mRNA expression.
  • Suitable methods for determining mRNA expression include quantitative PCR (QPCR), realtime QPCR, reverse transcription PCR (RT-PCR), and quantitative reverse transcription PCR (QRT-PCR), as are well-known in the art. As described in detail in U.S. Patent Nos.
  • a quantitative reverse transcriptase polymerase chain reaction for detecting mRNA may include the steps of: (a) incubating an RNA sample from the cellular lysate with a reverse transcriptase and a high concentration of a target sequence-specific reverse transcriptase primer under conditions suitable to generate cDNA; (b) subsequently adding suitable polymerase chain reaction (PCR) reagents to the reverse transcriptase reaction, including a high concentration of a PCR primer set specific to the cDNA and a thermostable DNA polymerase to the reverse transcriptase reaction; and (c) cycling the PCR reaction for a desired number of cycles and under suitable conditions to generate PCR products ("amplicons") specific to the cDNA.
  • PCR polymerase chain reaction
  • the products of the QRT-PCR process may be compared after a fixed number of PCR cycles to determine the relative quantity of the RNA species as compared to a given reference gene, for example, GAPDH (glyceraldehyde-3- phosphate dehydrogenase). More typically, the progress of the PCR reaction is monitored by analyzing the relative rates of amplicon production for each PCR primer set, for example, by (1) non-specific fluorescent dyes that intercalate with any double-stranded DNA, and/or (2) sequence-specific DNA probes consisting of oligonucleotides that are labeled with a fluorescent reporter which permits detection only after hybridization of the probe with its complementary DNA target.
  • a given reference gene for example, GAPDH (glyceraldehyde-3- phosphate dehydrogenase). More typically, the progress of the PCR reaction is monitored by analyzing the relative rates of amplicon production for each PCR primer set, for example, by (1) non-specific fluorescent dyes that intercalate with any double-strand
  • the mRNA may be any mRNA that is associated with thymosin beta-4 or with pro-inflammatory chemokines and/or cytokines from any animal, including the mRNAs encoding thymosin beta-4, such as those identified in Table 1, or any polynucleotide, fragment or derivative thereof.
  • the mRNA encodes thymosin beta-4, among others.
  • the mRNA encodes human thymosin beta-4.
  • the level of mRNA expression may be compared to controls that are not treated with the candidate substance to determine the relative degree of stimulation of thymosin beta-4; and/or inhibition of stimulation of pro-inflammatory chemokines and/or cytokines.
  • the candidate substance will upregulate or downregulate mRNA expression by at least about 10%, more suitably at least about 20%, at least about 30%, at least about 40%, or at least about 50%.
  • the candidate substance will upregulate or downregulate mRNA expression by at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100%.
  • Candidate substances meeting these criteria, other than retinoids may be selected for use of for further evaluation.
  • Retinoids such as retinol A and retinoic acid
  • retinol A and retinoic acid are the most common active agents used in anti-wrinkle creams.
  • the use of retinoids on the skin has been associated with various side effects, e.g. skin irritation, redness and itching, skin peeling, dry skin, and sensitivity to sunlight.
  • thymosin beta-4 stimulators and inflammatory inhibitors of the invention may be used as active agents in cosmetic preparation and may be formulated with other cosmetically acceptable components, such as a vehicle, into a composition for topical application to the skin.
  • a cosmetically acceptable component such as a vehicle
  • the use of these thymosin beta-4 stimulators exhibiting inhibition of inflammation activity results in less skin irritation, redness and itching, skin peeling, dry skin, and sensitivity to sunlight compared to the use of retinoids.
  • the use of TB-4 stimulators also exhibiting inhibition of skin inflammation activity results in at least one of (a) the ability to use a greater quantity of thymosin beta-4 upregulator without excessive accompanying inflammation; and/or (b) the ability to use otherwise skin inflammatory amounts of skin active agents given the TB-4 upregulator's ability to decrease potential skin inflammation.
  • the compositions are topically applied to the skin in effective amounts, by which is meant an amount sufficient to achieve a measurable improvement in skin health or reduction in one or more dermatological signs of skin inflammation aging.
  • signs of skin aging include without limitation, the following:
  • compositions of the invention including agents that stimulate thymosin beta-4 expression levels and inhibit inflammation, alone, or in cosmetically acceptable vehicles, are applied to skin in need of treatment. That is, the compositions of the invention are applied to skin which suffers from a deficiency or loss in any of the foregoing attributes or which would otherwise benefit from improvement in any of the foregoing skin attributes.
  • the skin is typically treated once or twice daily.
  • the treatment may continue for a week, two weeks, four weeks, eight weeks, six months or longer.
  • the treatment is once daily administration for a period of at least four weeks.
  • the length of treatment is determined by response, in particular by the effective treatment of fine lines and/or wrinkles.
  • the active agents are topically applied, in a cosmetically acceptable vehicle, to skin suffering from fine lines and/or wrinkles to prevent, treat, and/or amelioration the appearance of the fine lines and/or wrinkles in the skin.
  • the compositions are applied to skin in need of treatment, by which is meant skin already having wrinkles and/or fine lines or skin that is at risk of developing fine lines and/or wrinkles.
  • the compositions are applied directly to the fine lines and/or wrinkles on the skin of the face, neck, chest, and/or hands.
  • Preferred agents according to this embodiment are upregulators of thymosin beta-4 expression.
  • the upregulators of thymosin beta-4 expression lead to an enhanced production of collagen, estastin and/or fibrillin in dermal fibroblasts.
  • the invention is directed to a method of improving the aesthetic appearance of skin by increasing the production of collagen, elastin, and/or fibrillin in the skin, the method comprising topically applying to an area of the skin in need thereof an effective amount of an agent that upregulates thymosin beta-4 expression, wherein said active agent has been identified for use by an assay which determines the ability of a substance to stimulate expression levels of thymosin beta-4, including the assay described herein.
  • the assay measures thymosin beta-4 mRNA expression levels in dermal fibroblasts.
  • the thymosin beta-4 stimulator and inflammation inhibitor comprises a natural plant material such as a botanical extract.
  • a natural plant material such as a botanical extract.
  • suitable extracts are extracts from plants selected from the following species and/or portions thereof: Maesa japonica (branches and leaves), Celosia argentea (flower), Berchemia lineata (whole plant), Ixora chinesis (flower), Physalis minima (whole plant), and combinations thereof.
  • the active agent which may be a botanical extract, upregulates thymosin beta-4 and inhibits inflammation in a dermal fibroblast or epidermal keratinocyte
  • the active agent is from Maesa japonica, and is combined with a topically acceptable vehicle.
  • the Maesa japonica active agent and topically acceptable vehicle are combined with a second active agent which upregulates thymosin beta-4 in a dermal fibroblast or epidermal keratinocyte.
  • thymosin beta-4 include the following compounds:
  • the invention is directed to a cosmetic composition comprising an effective amount of Compound 1 and at least one topically acceptable vehicle.
  • Another embodiment of the invention is directed to a method of improving the aesthetic appearance of human skin comprising topically applying to the skin in need thereof an effective amount of Compound 1 for a time sufficient to improve the aesthetic appearance of said human skin.
  • the plant materials may be in any form including, but not limited to, the whole plant, a dried plant, a ground plant, or parts thereof, including but not limited to, seeds, needles, leaves, roots, bark, cones, stems, rhizomes, callus cells, protoplasts, flowers, and meristems, or components and/or constituents found in, or isolated from, the natural plant material, and/or portions of the plant, or any combinations thereof.
  • the natural plant material is in the form of an extract derived from the whole plant or from a select portion of the plant, such as the leaves of the plant. It is to be understood that "natural plant material" also includes an ingredient, component, constituent, or extract derived from the natural plant material.
  • the plant extract as used herein also includes "synthetic" extracts, i.e., various combinations of known plant components and/or constituents that are combined to substantially mimic the composition and/or activity of a plant extract of natural origin.
  • synthetic extracts are included in the term "plant extract.”
  • the synthetic extracts will have two or more, three or more, or four or more active ingredients in common with a plant. Most preferably, the synthetic extracts will have substantially the same number of active ingredients as a natural extract.
  • the correspondence of the numerical incidence of active ingredients between the synthetic extracts and the plant or a natural extract may also be described in terms of "percent commonality.”
  • the synthetic extract has about 50 percent or more commonality to the chemical composition of a plant or natural extract.
  • the synthetic extract has about 50 percent or more of the active ingredients found in the plant or a natural extract. More preferably, the chemical composition of the synthetic extract has about 70 percent or more commonality to the chemical composition of a plant or a natural extract. Optimally, a synthetic extract has about 90 percent or more commonality to the chemical composition of a plant or a natural extract.
  • the plant extract or components and/or active constituents are preferably derived directly from the plant.
  • the components may be in a pure form, a semi-pure form, or unpurified form.
  • the components are in the form of an extract obtained by aqueous or organic solvent extraction.
  • organic solvents include acetic acid, diethyl ether, ethyl acetate, lower alcohols (e.g., methanol, ethanol, isopropanol, or butanol), dichloromethane, chloroform, hexane, benzene, toluene, xylene, petroleum ether, and combinations thereof.
  • the solvent may be either polar or non-polar, protic or aprotic, water-miscible or water- immiscible.
  • the pH may be acidic, neutral, or alkaline.
  • Well-known methods in the art may be used for aqueous or organic solvent extraction. An extraction time between about 1-8 hours at a temperature between about 30°C to about 90 °C is typically suitable.
  • the collected extract is then fine-filtered to remove debris, and may be used directly, or is concentrated, for example by distilling the solvent or by other conventional processing, and the extract can also be provided in powder form.
  • compositions according to the invention can be formulated in a variety of forms for topical application and will comprise an effective amount of an active agent that modulates or upregulates thymosin beta-4 and inhibits inflammation, wherein the active agent or agents comprises from about 0.00001% to about 90% by weight of the composition, and preferably will comprise from about 0.001% to about 25% by weight, and more preferably from about 0.01% to about 10% by weight.
  • the active agent may also be present from about 0.5% to about 5% by weight.
  • compositions can include a cosmetically acceptable vehicle.
  • vehicles may take the form of any known in the art suitable for application to skin and may include, but are not limited to, water; vegetable oils; mineral oils; esters such as octal palmitate, isopropyl myristate and isopropyl palmitate; ethers such as dicapryl ether and dimethyl isosorbide; alcohols such as ethanol and isopropanol; fatty alcohols such as cetyl alcohol, cetearyl alcohol, stearyl alcohol and biphenyl alcohol; isoparaffins such as isooctane, isododecane and is hexadecane; silicone oils such as cyclomethicone, hydrocarbon oils such as mineral oil, petrolatum, isoeicosane and polyisobutene; polyols such as propylene glycol, glycerin, butylene glycol, pentylene glycol and hexylene glycol; liposomes;
  • the vehicle may comprise an aqueous phase, an oil phase, an alcohol, a silicone phase or mixtures thereof and may be in the form of an emulsion.
  • suitable emulsions include water-in-oil emulsions, oil-in-water emulsions, silicone-in-water emulsions, water-in-silicone emulsions, glycerin-in-oil emulsions, wax-in- water emulsions, water-oil-water triple emulsions or the like.
  • the emulsion may include an emulsifier, such as a nonionic, anionic or amphoteric surfactant, or a gelling agent.
  • the topical composition will typically have a pH range from 1 to 8, with a pH in the range of from 2 to 7 being preferred. In some embodiment, the composition will have a pH in the range of from 3.5 to 5.5. Suitable pH adjusters such as citric acid and triethanolamine may be added to bring the pH within the desired range.
  • compositions may include additional skin actives, including but not limited to, botanicals, keratolytic agents, desquamating agents, keratinocyte proliferation enhancers, collagenase inhibitors, elastase inhibitors, depigmenting agents, anti-inflammatory agents, steroids, anti-acne agents, antioxidants, and advanced glycation end-product (AGE) inhibitors.
  • additional skin actives including but not limited to, botanicals, keratolytic agents, desquamating agents, keratinocyte proliferation enhancers, collagenase inhibitors, elastase inhibitors, depigmenting agents, anti-inflammatory agents, steroids, anti-acne agents, antioxidants, and advanced glycation end-product (AGE) inhibitors.
  • botanicals including but not limited to, botanicals, keratolytic agents, desquamating agents, keratinocyte proliferation enhancers, collagenase inhibitors, elastase inhibitors, depigmenting agents, anti-inflammatory agents,
  • composition may comprise additional active ingredients having anti-aging benefits, as it is contemplated that synergistic improvements may be obtained with such combinations.
  • anti-aging components include, without limitation, botanicals (e.g., Butea frondosa extract); phytol; thiodipropionic acid (TDPA) and esters thereof; retinoids (e.g., 9-cis retinoic acid, 13-cis retinoic acid, all-trans retinoic acid and derivatives thereof, phytanic acid, retinol (Vitamin A) and esters thereof, such as retinol palmitate, retinol acetate and retinol propionate, and salts thereof and others); hydroxy acids (including alpha-hydroxy acids and beta-hydroxy acids), salicylic acid and alkyl salicylates; exfoliating agents (e.g., glycolic acid, 3,6,9-trioxaundecanedioic acid, etc.), estrogen synthet
  • retinoids include, without limitation, retinoic acid (e.g., all-trans or 13-cis) and derivatives thereof, retinol (Vitamin A) and esters thereof, such as retinol palmitate, retinol acetate and retinol propionate, and salts thereof.
  • retinoic acid e.g., all-trans or 13-cis
  • retinol Vitamin A
  • esters thereof such as retinol palmitate, retinol acetate and retinol propionate, and salts thereof.
  • the topical compositions of the present invention may also include one or more of the following: a skin penetration enhancer; an emollient, such as isopropyl myristate, petrolatum, silicones (e.g., methicone, dimethicone), oils, mineral oils, and fatty acid esters; a humectant, such as glycerin or caprylyl glycol; a skin plumper, such as palmitoyl oligopeptide, collagen, or collagen and/or glycosaminoglycan (GAG) enhancing agents; a sunscreen, such as avobenzone; an exfoliating agent; and an antioxidant.
  • a skin penetration enhancer such as isopropyl myristate, petrolatum, silicones (e.g., methicone, dimethicone), oils, mineral oils, and fatty acid esters
  • a humectant such as glycerin or caprylyl glycol
  • a skin plumper such
  • Suitable exfoliating agents include, for example, alpha-hydroxy acids, beta- hydroxy acids, oxa-acids, oxadiacids, and their derivatives such as esters, anhydrides and salts thereof.
  • Suitable hydroxy acids include, for example, glycolic acid, lactic acid, malic acid, tartaric acid, citric acid, 2-hydroxyalkanoic acid, mandelic acid, salicylic acid and derivatives thereof.
  • a preferred exfoliating agent is glycolic acid.
  • the exfoliating agent may comprise from about 0.1% by weight to about 80 % by weight of the composition.
  • antioxidants examples include compounds having phenolic hydroxy functions, such as ascorbic acid and its derivatives/esters; beta-carotene; catechins; curcumin; ferulic acid derivatives (e.g., ethyl ferulate, sodium ferulate); gallic acid derivatives (e.g., propyl gallate); lycopene; reductic acid; rosmarinic acid; tannic acid; tetrahydrocurcumin; tocopherol and its derivatives; uric acid; or any mixtures thereof.
  • ferulic acid derivatives e.g., ethyl ferulate, sodium ferulate
  • gallic acid derivatives e.g., propyl gallate
  • lycopene reductic acid
  • rosmarinic acid tannic acid
  • tetrahydrocurcumin tocopherol and its derivatives
  • uric acid or any mixtures thereof.
  • antioxidants are those that have one or more thiol functions (-SH), in either reduced or non-reduced form, such as glutathione, lipoic acid, thioglycolic acid, and other sulfhydryl compounds.
  • the antioxidant may be inorganic, such as bisulfites, metabisulfites, sulfites, or other inorganic salts and acids containing sulfur.
  • Compositions of the present invention may comprise an antioxidant preferably from about 0.001 wt % to about 10 wt %, and more preferably from about 0.01 wt % to about 5 wt %, of the total weight of the composition.
  • compositions of the invention may contain any other compound for the treatment of skin disorders.
  • the composition may be formulated in a variety of product forms, such as, for example, an emulsion, lotion, cream, serum, spray, aerosol, cake, ointment, essence, gel, paste, patch, pencil, towelette, mask, stick, foam, elixir, concentrate, and the like, particularly for topical administration.
  • the composition is formulated as an emulsion, lotion, cream, ointment, serum or gel.
  • the invention provides a method for treating aging skin by topically applying a composition comprising an active agent that stimulates thymosin beta-4, preferably in a cosmetically acceptable vehicle, over the affected area for a period of time sufficient to reduce, ameliorate, reverse or prevent dermatological signs of aging.
  • the invention provides a method for treating, preventing, and/ or ameliorating skin subject to inflammatory response by topically applying a composition comprising an active agent that stimulates thymosin beta-4 and inhibits inflammatory response, preferably in a cosmetically acceptable vehicle, over the affected area for a period of time sufficient to reduce, ameliorate, reverse or prevent dermatological signs of inflammation.
  • the compositions of the invention may be applied topically to skin that is inflamed as a result of injury such as wounds or burns, introduction of a toxin, e.g. by an insect bite or other noxious chemical agent, or the presence of an agent generally used to provide a benefit, e.g. an active agent, but that may occasion an inflammatory response in certain individuals at specific levels.
  • the presence of the thymosin beta-4 upregulator that provides an antiinflammatory response may permit compositions having higher active agent concentrations, but which would be in the aggregate non-inflammatory.
  • the improvement in the condition and/or aesthetic appearance is selected from the group consisting of: reducing dermatological signs of chronological aging, photo-aging, hormonal aging, and/or actinic aging; preventing and/or reducing the appearance of lines and/or wrinkles; reducing the noticeability of facial lines and wrinkles, facial wrinkles on the cheeks, forehead, perpendicular wrinkles between the eyes, horizontal wrinkles above the eyes, and around the mouth, marionette lines, and particularly deep wrinkles or creases; improving the appearance of suborbital lines and/or periorbital lines; reducing the appearance of crow's feet; rejuvenating and/or revitalizing skin, particularly aging skin; reducing skin fragility; preventing and/or reversing of loss of glycosaminoglycans and/or collagen; ameliorating the effects of estrogen imbalance; preventing skin atrophy; preventing, reducing, and/or treating hyperpigmentation or hypopigmentation; minimizing skin discoloration; improving skin tone, radiance,
  • the compositions will comprise from about 0.00001% to about 90%, more typically from about 0.001% to about 25%, including from about 0.01% to about 10% by weight of a stimulator of thymosin beta-4.
  • the stimulator may be an upregulator of thymosin beta-4 in fibroblasts.
  • the stimulator may be an upregulator of thymosin beta-4 in keratinocytes.
  • the stimulator is an upregulator of thymosin beta-4 in both fibroblasts and keratinocytes. Combinations of stimulators are also contemplated.
  • the stimulator is a combination of two or more substances that are upregulators of thymosin beta-4 in fibroblasts and/or keratinocytes.
  • the composition will typically be applied to the skin one, two, or three times daily for as long as is necessary to achieve desired results.
  • the treatment regiment may comprise daily application for at least one week, at least two weeks, at least four weeks, at least eight weeks, or at least twelve weeks or more.
  • Chronic treatment regimens are also contemplated.
  • the effect of a composition on the formation or appearance of fine lines and wrinkles can be evaluated qualitatively, e.g., by visual inspection, or quantitatively, e.g., by microscopic or computer assisted measurements of wrinkle morphology (e.g., the number, depth, length, area, volume and/or width of wrinkles per unit area of skin).
  • the composition of the invention will be applied to the skin in an amount from about 0.001 to about 100 mg/cm 2 , more typically from about 0.01 to about 20 mg/cm 2 , and preferably about 0.1 to about 10 mg/cm 2 .
  • compositions of the invention will be useful for treating thin skin by topically applying the composition to thin skin of an individual in need thereof.
  • Thiin skin is intended to include skin that is thinned due to chronological aging, menopause, or photo-damage and skin that is thinning prematurely.
  • the treatment is for thin skin in men, whereas other embodiments treat thin skin in women, pre-menopausal or post-menopausal, as it is believed that skin thins differently with age in men and women, and in particular in women at different stages of life.
  • the method of the invention may be employed prophylactically to forestall aging including in individuals that have not manifested signs of skin aging, most commonly in individuals under 25 years of age.
  • the method may also reverse or treat signs of aging once manifested as is common in individuals over 25 years of age, or to slow the progression of dermatological aging in such individuals.
  • TMSB4X thymosin beta- 4
  • TMSB4X Percent increase in mRNA for thymosin beta- 4
  • Thymosin beta-4 according to the method of Example 1.
  • the results for these active agents are provided in Table 3.
  • the concentrations of each extract are provided based on the dry weight of the given plant extract, by which is meant the weight of the extract after volatile extraction solvents have been removed.
  • the cells tested were primary human dermal fibroblasts.
  • Synthetic compounds 1-4 were also tested as active agents for the ability to upregulate thymosin beta-4 according to the method of Example 1. The results are provided in Table 4.
  • TB-4 human dermal fibroblasts were treated with tumor necrosis factor alpha for 6 hours to induce inflammation.
  • Cells were either pre-treated with recombinant TB-4 or control media to assess its anti-inflammatory effect.
  • Cell lysates and media were collected and qPCR and ELISA were performed to assess IL-8 gene and protein expression.
  • Human cytokines and chemokines PCR array was also performed to investigate the impact of TB-4 on the expression profile of key secreted proteins in the inflammation pathway.
  • TB-4 also significantly regulates a variety of key cytokines and chemikines genes to play important roles in skin cell inflammation response. Therefore, modulating endogenous TB-4 could be adopted as an avenue to battle inflammatory skin damage resulting from skin chronological aging and photoaging.
  • human dermal fibroblasts will be treated with potential anti-inflammatory agents that upregulate TB-4 according to Example 1 above. After incubation, the human dermal fibroblasts may be treated with TNF-alpha, with control cells (those that were not treated with TB-4 upregulators) also treated with TNF-alpha. After treatment, cells will be lysed and the techniques of Example 3 will be applied to assess the impact of treatment with TB-4 upregulators on the expression profile of key secreted proteins in the inflammation pathway. It is expected that cells treated with TB-4 upregulators will demonstrate a significant suppression of IL-8 gene expression and protein secretion caused by TNF-alpha stimulation. [0095] EXAMPLE 5
  • Example of a skincare regimen utilizing anti-inflammatory TB-4 stimulators [0096] Example of a skincare regimen utilizing anti-inflammatory TB-4 stimulators.
  • a cosmetic or therapeutic composition comprising one or more TB-4 stimulators will be applied to skin once or multiple times daily for a period of days or weeks. It is expected that such a treatment regimen will yield skin that demonstrates prevented, reversed, and/or reduced inflammation.

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Abstract

L'invention concerne des méthodes pour prévenir, soulager, ou réduire les signes dermatologiques d'inflammation, qui font appel à des principes actifs, autres qu'un rétinoïde, qui stimulent l'expression de la thymosine bêta-4 dans la peau et inhibent la production de chimiokines et/ou de cytokines pro-inflammatoires. L'invention concerne également des procédés de criblage de substances qui stimulent les niveaux d'expression de la thymosine bêta-4 et les procédés d'utilisation des principes actifs identifiés par le protocole de criblage pour le traitement de la peau.
PCT/US2013/072731 2012-12-11 2013-12-03 Modulation de thymosine bêta-4 dans la peau WO2014093053A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US13/710,585 US8709507B2 (en) 2011-12-13 2012-12-11 Maesa japonica extracts and methods of use
US13/710,585 2012-12-11
US13/735,659 2013-01-07
US13/735,659 US20140193529A1 (en) 2013-01-07 2013-01-07 Modulation of Thymosin Beta-4 in Skin

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WO2002091969A1 (fr) * 2001-05-17 2002-11-21 Regenerx Biopharmaceuticals, Inc. Traitement de l'epidermolyse bulleuse par la thymosine beta 4
US20060292572A1 (en) * 2004-01-09 2006-12-28 Stuart Robert O Cell-type-specific patterns of gene expression
US20050154046A1 (en) * 2004-01-12 2005-07-14 Longgui Wang Methods of treating an inflammatory-related disease
US20110111010A1 (en) * 2004-03-04 2011-05-12 Yissum, Research Development Company Of The Hebrew University Of Jerusalem Ltd. Method and Composition for Burned Skin
US20110177970A1 (en) * 2008-10-02 2011-07-21 Inserm (Institut National De La Sante Et De La Recherche Medicale) Methods for predicting or monitoring whether a patient affected by a cancer is responsive to a treatment with a molecule of the taxoid family
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EP4103215A4 (fr) * 2020-02-13 2024-03-06 Hlb Therapeutics Co Ltd Compositions et méthodes de traitement ou de prévention du prurit

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