Compositions comprising peptide wkdeagkplvk

Abstract

To provide an alternative that helps muscle recovery, maintenance and / or muscle growth.SOLUTION: A composition comprises a peptide of SEQ ID NO: 1, or a glucose transport-promoting variant of SEQ ID NO: 1 with 1 to 3 amino acid alterations relative to SEQ ID NO: 1. The amino acid alterations occur in one or more positions of positions 1, 2, 3, 9, 10 and 11 of SEQ ID NO: 1. The amino acid alterations in the variant include insertion, addition, deletion and substitution of amino acids.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to cosmetic and pharmaceutical compositions. [Background technology]

[0002] Many compositions have been described and improvements are always desirable.

[0003] Currently, different approaches exist for improving muscle health or muscle glucose uptake, but it is desirable to find alternatives that aid in muscle recovery, maintenance, and / or muscle growth.

[0004] In particular, since people are now living longer and healthier lives, the increasing will to maintain a youthful appearance is actually leading to more and more research into new cosmetic and dermatological procedures to treat skin aging.In recent years, there has been an increasing interest in minimally invasive treatments and techniques designed to address issues such as wrinkles, weight loss, and other skin damage.The most common topical anti-aging solutions are creams and serums.

[0005] US 2004 / 0132667 discloses compositions comprising peptides, optionally in combination with one or more additional ingredients. The disclosed compositions provide relief from one or more skin conditions, including skin conditions caused by a variety of causes, such as stress, pollution, and general aging.

[0006] US Patent Application Publication No. 2014 / 0120141 discloses cosmetic and pharmaceutical compositions containing peptides for use in the treatment and / or care of skin and / or mucosal conditions, disorders, and / or diseases. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] US Patent Application Publication No. 2004 / 0132667 [Patent Document 2] US Patent Application Publication No. 2014 / 0120141 [Patent Document 3] European Patent Application Publication No. 2050437 [Patent Document 4] International Publication No. 2005 / 023290 Brochure [Patent Document 5] US Patent Application Publication No. 2010 / 098660 [Patent Document 6] US Patent Application Publication No. 2007 / 0053845 [Patent Document 7] US Patent Application Publication No. 2014 / 120131 [Patent Document 8] US Patent Application Publication No. 2004 / 132667 [Patent Document 9] U.S. Pat. No. 4,186,183 [Patent Document 10] U.S. Pat. No. 4,217,344 [Patent Document 11] U.S. Pat. No. 4,235,871 [Patent Document 12] U.S. Pat. No. 4,261,975 [Patent Document 13] U.S. Pat. No. 4,485,054 [Patent Document 14] U.S. Pat. No. 4,501,728 [Patent Document 15] U.S. Pat. No. 4,774,085 [Patent Document 16] U.S. Pat. No. 4,837,028 [Patent Document 17] U.S. Pat. No. 4,235,871 [Patent Document 18] U.S. Pat. No. 4,261,975 [Patent Document 19] U.S. Pat. No. 4,485,054 [Patent Document 20] U.S. Pat. No. 4,501,728 [Patent Document 21] U.S. Pat. No. 4,774,085 [Patent Document 22] U.S. Pat. No. 4,837,028 [Patent Document 23] U.S. Pat. No. 4,946,787 [Patent Document 24] PCT International Publication No. 91 / 17424 Pamphlet [Patent Document 25] US Patent Application Publication No. 2014120141 [Patent Document 26] U.S. Pat. No. 4,766,106 [Patent Document 27] U.S. Pat. No. 4,179,337 [Patent Document 28] U.S. Pat. No. 4,495,285 [Patent Document 29] U.S. Pat. No. 4,609,546 [Patent Document 30] U.S. Pat. No. 3,654,090 [Patent Document 31] U.S. Pat. No. 3,850,752 [Patent Document 32] U.S. Pat. No. 4,016,043 [Patent Document 33] U.S. Patent No. 6,165,779 [Patent Document 34] U.S. Patent No. 6,225,289 [Patent Document 35] U.S. Patent No. 5,708,025 [Patent Document 36] U.S. Pat. No. 5,994,106 [Patent Document 37] International Patent Application Publication No. 98 / 32859 [Patent Document 38] International Publication No. 00 / 55119 Brochure [Patent Document 39] U.S. Pat. No. 5,656,722 [Patent Document 40] U.S. Pat. No. 5,750,497 [Patent Document 41] U.S. Patent No. 6,251,856 [Patent Document 42] U.S. Patent No. 6,268,335 [Patent Document 43] U.S. Patent No. 6,451,970 [Patent Document 44] International Publication No. 01 / 62218 Brochure [Patent Document 45] International Publication No. 98 / 07744 Brochure [Patent Document 46] International Publication No. 99 / 18927 Brochure [Non-patent literature]

[0008] [Non-Patent Document 1] Topical drug delivery formulations edited by David Osborne and Antonio Aman, Taylor & Francis [Non-Patent Document 2] O'Riordan et al(Respir Care,2002,Nov.47) [Non-Patent Document 3] Handbook of Pharmaceutical Excipients, 2nd Edition, (1994), Edited by A Wade and PJ Weller [Non-Patent Document 4] Remington's Pharmaceutical Sciences, Mack Publishing Co. (ARGennaro edit. 1985) [Non-Patent Document 5] JMStewart and JDYoung, Solid Phase Peptide Synthesis, 2nd edition, Pierce Chemical Company, Rockford, Illinois (1984)

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[0009] It is an object of the present invention to provide a composition which overcomes at least one of the above mentioned problems. [Means for solving the problem]

[0010] A first aspect of the present invention provides a peptide comprising the amino acid sequence of SEQ ID NO:1 (hereinafter referred to as "the peptide of the present invention").

[0011] A second aspect of the present invention provides a composition (hereinafter "the composition of the present invention") comprising an effective amount of a peptide of the present invention. Preferably, the composition is a topical composition. Typically, the composition may be a cosmetic composition comprising a cosmetically effective amount of a peptide comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof. Suitably, the composition may be a pharmaceutical composition comprising a pharma- ceutical effective amount of a peptide comprising the amino acid sequence of SEQ ID NO:1 or a variant thereof. Preferably, the peptide is a bioactive peptide. Suitably the composition comprises a plurality of peptides. Typically, the composition further comprises at least one cosmetically or pharma- cetically acceptable excipient or additive. Typically, the composition further comprises at least one cosmetically or pharma- ceutical acceptable active ingredient. Suitably the peptide comprises the amino acid sequence consisting of SEQ ID NO:1. Typically, the peptide comprises a length of about 3 to about 50 amino acids, preferably about 5 to about 20 amino acids, preferably about 11 amino acids.

[0012] The present invention also relates to fragments of SEQ ID NO: 1 comprising a length of at least 3 amino acids. Typically, said fragments are biologically active. Preferably, the fragment is at least 4, 5, 6, 7, 8, 9, 10, or 11 amino acids in length. Preferably, the peptide or fragment has an activity selected from one or more of the following: anti-aging activity, glucose transport promoting activity, cell proliferation promoting activity, or anabolic activity. The activity can be a cosmetic activity (i.e., a non-therapeutic activity), a therapeutic activity, or both. Suitably the peptide or fragment has anti-ageing activity. Suitably the peptide or fragment has glucose transport enhancing activity. Suitably the peptide or fragment has cell proliferation promoting activity. Suitably, the peptide or fragment has anabolic activity. Typically, the anabolic activity is muscle growing activity. The peptide or fragment stimulates anabolic metabolism in a mammal.

[0013] Preferably, said peptide or fragment exhibits GLUT4 translocation promoting activity. Preferably, said peptide or fragment exhibits increased protein synthesis (stimulation of anabolic metabolic activity).

[0014] More preferably, the peptide comprises (or consists of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, in which case the peptide typically has anti-ageing activity. More preferably, the peptide comprises (or consists of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, in which case the peptide typically has glucose transport promoting activity. More preferably, the peptide comprises (or consists of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, in which case the peptide typically has cell proliferation promoting activity. More preferably, the peptide comprises (or consists of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, in which case the peptide typically has anabolic activity. More preferably, the peptide comprises (or consists essentially of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, where the peptide typically exhibits GLUT4 migration promoting activity. More preferably, the peptide comprises (or consists essentially of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, in which case the peptide typically exhibits increased protein synthesis (stimulation of anabolic metabolic activity). More preferably, the peptide comprises (or consists essentially of) the amino acid sequence of any one of SEQ ID NOs: 1 to 85, in which case the peptide typically exhibits cell proliferation promoting activity.

[0015] A further aspect of the invention provides the non-therapeutic use of a composition of the invention as a cosmetic.

[0016] A further aspect of the invention provides the composition of the invention as a medicament.

[0017] The following aspects can be cosmetic, i.e., non-therapeutic, or therapeutic.

[0018] An aspect of the present invention provides a composition for use in a method for anti-aging.

[0019] A further aspect of the invention provides a composition of the invention for use as a medicament.

[0020] The present invention also provides a peptide or composition of the present invention for use in a method for slowing or inhibiting or preventing aging of human skin.Typically, administration may be by plaster or patch or formulation suitable for topical application.

[0021] A further aspect of the invention provides a peptide or composition of the invention for use in a method of promoting tissue growth. The present invention also provides a peptide or composition of the invention for use in a method for promoting the proliferation of epithelial tissue. The present invention also provides a peptide or composition of the invention for use in a method for promoting skin proliferation. The present invention also provides a peptide or composition of the invention for use in a method for promoting organ growth. The present invention also provides a peptide or composition of the invention for use in a method for promoting the growth of an organism. Preferably, the cell, tissue, or organism has a normal pathology (e.g., aged skin). Typically, the cell, tissue, or skin has an abnormal pathology (e.g., tissue damaged due to trauma, drug use, or epithelial tissue in the digestive tract damaged due to inflammatory disorders). The use of promoting growth may be in vivo or in vitro. The use of promoting growth may involve administration to the outside (i.e., skin) or inside (i.e., gastrointestinal tract) of a mammal.

[0022] A further aspect of the invention provides a peptide or composition of the invention for use in a method for improving muscle condition in a mammal.

[0023] A further aspect of the invention provides a peptide or composition of the invention for use in a method for promoting muscle recovery, typically after physical exercise.

[0024] A further aspect of the invention provides a peptide or composition of the invention for use in a method for maintaining or restoring muscle health (eg lean tissue mass) in a mammal.

[0025] A further aspect of the invention provides a peptide or composition of the invention for use in a method for enhancing physical performance by local administration.

[0026] A further aspect of the invention provides a peptide or composition of the invention for use in a method for the treatment or prevention of a disease or condition characterised by lethargy or low energy levels.

[0027] A further aspect of the invention provides a peptide or composition of the invention for use in a method for muscle growth or muscle building. The composition of the invention can be used in a method for increasing lean muscle mass in beef cattle and other farm animals.

[0028] A further aspect of the invention provides a peptide or composition of the invention for use in a method for increasing protein synthesis.

[0029] A further aspect of the invention provides a peptide or composition of the invention for use in a method for treating muscle weakness.

[0030] Another aspect of the invention provides a peptide or composition of the invention for use in a method for treating a wound in a mammal.

[0031] A still further aspect of the invention provides a peptide or composition of the invention for use in a method for the treatment or prevention of pain in a mammal.

[0032] Another aspect of the invention provides a peptide or composition for use in a method for the treatment or prevention of a disease or condition characterised by epithelial cell or tissue damage and / or dermal or epithelial cell or tissue damage. Preferably, the disease or condition is characterised by dermal or epithelial cell or tissue damage and is selected from cancer, trauma.

[0033] A further aspect of the present invention relates to a method of treating, preventing or caring for any one of the diseases, conditions and / or disorders mentioned above and described herein, comprising administering a peptide or composition of the present invention. The composition can be administered topically.

[0034] The uses of the present invention may be therapeutic or cosmetic, ie non-therapeutic.

[0035] A further aspect of the present invention provides a peptide or composition of the present invention for use in a method for treating or preventing a metabolic disorder in a patient. In one embodiment, the metabolic disorder is a disease or syndrome characterized by a decrease in insulin response or production in the patient. In one embodiment, the metabolic disorder is selected from diabetes and prediabetes. The present invention also provides a peptide or composition of the present invention for use in treating or preventing a disease or condition selected from hyperlipidemia, obesity, hypertension, appetite-related syndromes, diabetes-related syndromes or diabetes-related conditions (e.g., reducing glucose levels and / or the rate of stroke, heart disease, kidney disease, blindness, and / or loss of sensation in the extremities associated with diabetes), and syndrome X.

[0036] A further aspect of the present invention provides the peptide or composition of the present invention for use in a method for treating a disease or condition characterized by muscle wasting, loss of lean muscle mass, reduced anabolic metabolism, catabolic wasting, or reduced protein synthesis.In one embodiment, the disease or condition is selected from sarcopenia or cachexia.In one embodiment, the patient to be treated is elderly (i.e., over 60 years old), and the disease is age-related sarcopenia or cachexia.In one embodiment, the patient has cancer.In one embodiment, the patient has lethargy.

[0037] A further aspect of the invention provides a peptide or composition of the invention for use in a method for treating or preventing a disease or condition characterised by epithelial cell or tissue damage and / or dermal or epithelial cell or tissue damage. Preferably, the disease or condition is characterised by epithelial cell or tissue damage and is selected from cancer, trauma, tissue damaged due to drug use or epithelial tissue in the gastrointestinal tract damaged due to an inflammatory disorder.

[0038] A further aspect of the invention provides a peptide or composition of the invention for use in a method of maintaining or restoring muscle health (eg lean tissue mass) in a mammal.

[0039] A further aspect of the invention provides a peptide or composition of the invention for use in a method of treatment or prevention of a disease or condition characterised by lethargy or low energy levels.

[0040] Another aspect of the invention provides a peptide or composition of the invention for use in a method of treating a wound in a mammal.

[0041] A still further aspect of the invention provides a peptide or composition of the invention for use in a method of treating or preventing pain in a mammal.

[0042] Another aspect relates to a topical composition of the invention for use in the treatment or prevention of a metabolic disorder in a mammal. The metabolic disorder may be diabetes. In one embodiment, the use of the present invention comprises the step of administering a therapeutically effective amount of a peptide of the present invention. In one embodiment, the compositions of the invention are administered systemically. In one embodiment, the compositions (especially pharmaceutical compositions) of the present invention are formulated for oral or parenteral administration, although other methods of administration are also described herein.

[0043] A further aspect of the invention relates to a composition comprising one or more of the peptides selected from SEQ ID NOs: 1-85. In one embodiment, a composition of the invention comprises substantially all of the peptides of SEQ ID NOs: 1-85.

[0044] A further aspect of the invention relates to an artificial treatment composition comprising the composition of the invention. Preferably, the treatment composition is an anti-aging composition. Preferably, the treatment is a muscle treatment composition. In an embodiment, the composition is a topical composition. In one embodiment, the composition comprises a cream, gel, lotion, rub, powder. Preferably, the treatment is a wound treatment composition.

[0045] The present invention also relates to a plaster, bandage or dressing suitable for application to keratinous tissue or a wound and comprising a peptide or composition of the invention. Preferably, the composition or product is man-made.

[0046] (definition) All publications, patents, patent applications, and other references mentioned in this specification are herein incorporated by reference in their entirety and for all purposes, to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference and its contents as fully set forth.

[0047] As used herein, and unless specifically indicated otherwise, the following terms are intended to have the following meanings, in addition to any broader (or narrower) meaning that the terms may have in the art.

[0048] Unless otherwise required by context, the use of the singular herein shall be read to include the plural and vice versa. The terms "a" and "an" when used in connection with entities shall be read to refer to one or more of those entities. Thus, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein.

[0049] Throughout this specification, the word "comprise" or variations thereof, such as "comprises" or "comprising," shall be read to indicate the inclusion of a recited integer (e.g., a feature, element, characteristic, property, method / process step, or limitation) or group of integers (e.g., features, elements, features, properties, method / process steps, or limitations) and not the exclusion of any other integer or group of integers. Thus, as used herein, the term "comprising" is inclusive or open-ended and does not exclude additional, unrecited integers or method / process steps.

[0050] As used herein, the term "disease" is used to define any abnormal condition that impairs physiological function and is associated with specific symptoms. The term is used broadly to include any disorder, disease, abnormality, condition, illness, state, or syndrome in which physiological function is impaired, regardless of the nature of the etiology (or, indeed, whether an etiological basis for the disease has been established). It thus includes conditions resulting from infection, trauma, injury, surgery, radiation ablation, poisoning, or malnutrition.

[0051] As used herein, the term "treatment" or "treating" refers to an intervention (e.g., administration of a drug to a subject) that cures, improves, or alleviates the symptoms of a disease, or eliminates its cause (or reduces its effects) (e.g., reducing the accumulation of pathological levels of lysosomal enzymes). In this context, the term is used synonymously with the term "therapy."

[0052] In addition, the term "treatment" or "treating" refers to an intervention (e.g., administration of a drug to a subject) that prevents or slows the onset or progression of a disease, or reduces its incidence (or eradicates) in a treated population. In this instance, the term treatment is used synonymously with the term "prophylaxis."

[0053] As used herein, an effective amount or therapeutically effective amount of a drug defines an amount that can be administered to a subject without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio, and that is sufficient to bring about the desired effect, e.g., treatment or prevention manifested by a permanent or temporary improvement of the subject's condition. The amount will vary from subject to subject, depending on the individual's age and general condition, the mode of administration, and other factors. Thus, it is not possible to specify an exact effective amount, but one skilled in the art will be able to determine the appropriate "effective" amount in any individual case, using routine experimentation and general background knowledge. Therapeutic results in this context include eradication or alleviation of symptoms, relief of pain or discomfort, prolonged survival, improved movement, and other markers of clinical improvement. Therapeutic results do not need to be a complete cure.

[0054] As used herein, the term "composition" shall be understood to mean something made by man and not including naturally occurring compositions.

[0055] The term "biological activity" as used herein refers to peptide or fragment having biological activity.For example, biological activity can be one or more of glucose transport promoting activity, growth promoting activity, anti-aging activity, anabolic activity, anabolic metabolism promoting activity, GLUT4 translocation promoting activity, and protein synthesis promoting activity.Activity can be anti-inflammatory activity or antibacterial activity.Activity can be antioxidant activity.

[0056] "Enhancing glucose transport" or "glucose transport enhancing activity" as applied to a peptide or fragment means a peptide, variant, or fragment that is capable of increasing glucose uptake into a cell in the glucose uptake assay described below.

[0057] "Promotion of GLUT4 translocation" or "GLUT4 translocation promoting activity" as applied to a peptide or fragment means a peptide fragment capable of increasing GLUT4 translocation into skeletal muscle compared to an untreated control in the in vitro assay described below.

[0058] "Anti-aging" refers to inhibiting or slowing down the appearance of aging in human skin and / or reversing the appearance of aging. "Slowing or inhibiting aging in skin" refers to slowing down or inhibiting the aging process in skin and / or reversing the appearance of aging. In one embodiment, "anti-aging" or "anti-aging activity" as applied to a peptide or fragment refers to a peptide or fragment that is capable of increasing collagen or elastin production in human dermal fibroblasts compared to untreated controls when examined in the in vitro assay described below and / or a peptide or fragment that is capable of increasing cell proliferation in the cell proliferation assay described below.

[0059] "Promoting cell proliferation" as applied to a peptide or fragment means a peptide, variant, or fragment capable of increasing elastin or collagen production or cell proliferation in the assays described below. Peptides that increase cell proliferation or have "cell proliferation promoting activity" are anti-aging peptides or anti-aging variants.

[0060] "Increasing protein synthesis" as applied to the peptides of the invention means a peptide capable of significantly increasing the degree of phosphorylation of MTOR in the phospho-MTOR assay described below, or significantly increasing the degree of protein synthesis in the puromycin assay described below. Protein synthesis increasing peptides and compositions can be utilized to stimulate anabolic metabolism in mammals and / or to treat diseases or conditions characterized by catabolic wasting.

[0061] "Anti-inflammatory" as applied to a peptide or fragment means a peptide or fragment capable of significantly reducing secretion of TNFα by LPS-stimulated J774.2 macrophages (compared to untreated, LPS-stimulated J774.2 macrophages) when the macrophages are treated with 100 μM of the peptide, variant, or fragment. J774.2 macrophages were treated with 100 μM of synthetic peptide for 24 hours and then stimulated with (A) LPS (10 ng / ml) for 5 hours or (B) LPS (10 ng / ml) for 5 hours followed by ATP (5 mM) for 1 hour. Supernatants were collected and TNFα levels were determined by ELISA.

[0062] "Antibacterial" or "antibacterial activity" as applied to a peptide or fragment means a peptide, variant, or fragment capable of visibly inhibiting bacterial growth in the following agar plate-based growth inhibition assay: peptide stock solution = 5 mg / mL dissolved in DMSO. Bacterial inocula were adjusted to a McFarland 0.5 standard and spread onto MHA plates. Blank discs were placed within the plates and 10 instances of each compound (at the maximum tested concentration of 64 μg / mL) were added. Plates were incubated at 37°C for 16-18 hours. Appropriate controls (DMSO; Mueller-Hinton medium alone; and two antibiotic discs: ciprofloxacin and tetracycline) were also set up.

[0063] "Antioxidant activity" as applied to the peptides of the invention means peptides that exhibit antioxidant activity as determined by the DPPH radical scavenging assay described below. The invention relates to the use of the peptides or compositions of the invention as antioxidants and in treating or preventing, or slowing the progression of, diseases or conditions characterized by oxidative stress (or an imbalance between pro-oxidants and antioxidants). Examples include cardiovascular disease (among others, atherosclerosis), cancer, neurodegenerative diseases (i.e., Parkinson's disease, Alzheimer's disease, and ALS), cataract formation, diabetes, and rheumatoid arthritis. Oxidation, the same chemical reaction that causes iron to rust, also plays a role in corrosion in our bodies. The process is called oxidative stress. Numerous studies have demonstrated that because these diseases are mediated by oxidative stress and a loss of balance between pro-oxidants and antioxidants, antioxidants may play a pivotal role in preventing or slowing the progression of these conditions. Some studies have shown an association between lower dietary antioxidant intake and an increased incidence of heart disease. On the other hand, people with higher blood levels of antioxidants have a lower risk of heart disease. For example, as seen in a study of nurses, people who regularly consumed high amounts of vitamin E had a 41% lower incidence of heart disease than those who consumed low amounts. Increasing antioxidant vitamins in the diet can reduce the risk of heart disease by 20-30%.

[0064] Cancer kills millions of people worldwide. Diet may be a contributing factor in up to 35% of all human cancers. Low dietary antioxidant intake may also be a contributing factor.

[0065] Pro-oxidants, substances that generate free radicals, stimulate cell division, initiating mutagenesis and tumor formation. When cells with damaged DNA strands divide, they result in clusters of distorted and distorted cells, which form cancer.

[0066] Antioxidants are Reduces oxidative damage to DNA -Reducing abnormal increases in cell division thereby exerting their protective effect.

[0067] In addition, smoking and chronic inflammation also lead to strong free radical generation, which is thought to be the reason for many cancers. Some studies indicate that smokers have lower antioxidant levels than non-smokers, which increases the risk of cancer for smokers.

[0068] The respiratory system is a well-known target of free radical attack, resulting from intrinsic factors as well as exposure to air pollutants and toxins, cigarette smoke, etc. Recent studies suggest that free radicals may play a role in the development of lung disorders, including asthma. Antioxidants are thought to reduce the development of asthmatic symptoms. Supplementation with vitamin C, vitamin E, and beta-carotene is associated with improved lung function.

[0069] Free radicals can also damage nerves and the brain. Nervous tissue is particularly susceptible to oxidative damage because the brain receives by far the most oxygen and has large amounts of polyunsaturated fatty acids, which are highly prone to oxidation and oxidative damage.

[0070] Diseases in which oxidative stress is implicated include: Alzheimer's disease Parkinson's disease Dementia, etc. Includes.

[0071] Cataract formation is thought to involve damage to lens proteins by free radicals. This leads to clouding of the lens. Cataract formation can be delayed by regular intake of supplemental antioxidants such as vitamin E, vitamin C, and carotenoids. Other diseases, such as diabetes and rheumatoid arthritis, are also associated with low antioxidant levels in the blood.

[0072] The term "topical composition" refers to a composition that is a formulation for topical administration. "Topical administration" refers to application to keratinous tissues, such as skin, hair, and nails. Topical delivery generally refers to delivery to the skin, but can also refer to delivery to epithelial cell-lined body cavities, such as the lungs or airways, the digestive tract, and the oral cavity. In particular, formulations for topical delivery are described in Topical drug delivery formulations edited by David Osborne and Antonio Aman, Taylor & Francis, the full contents of which are incorporated herein by reference. Compositions or formulations for delivery to the respiratory tract are described in O'Riordan et al (Respir Care, 2002, Nov. 47), EP2050437, WO2005023290, US2010 / 098660, and US2007 / 0053845. Compositions and formulations for delivery of active agents to the ileum, especially the proximal ileum, include microparticles and microcapsules in which the active agent is encapsulated in a protective matrix formed from polymers or dairy proteins that are resistant to acid but tend to dissolve in the more alkaline ileal environment. Examples of such delivery systems are described in EP1072600.2 and EP13171757.1. An alternative means of transdermal administration is by the use of a skin patch. For example, the active ingredient can be incorporated into a cream consisting of an aqueous emulsion of polyethylene glycol or liquid paraffin. Similarly, the active ingredient can be incorporated into an ointment consisting of a white wax or white soft paraffin base, together with stabilizers and preservatives that may be required, in a concentration between 1 and 10% by weight. Injectable forms may contain between 10 and 1000 mg, preferably between 10 and 250 mg, of active ingredient per dose.

[0073] Compositions may be formulated in unit dosage form, ie, in the form of discrete portions containing a unit dose, or a multiple or sub-unit of a unit dose.

[0074] The term "cosmetic composition" as used herein relates to a composition that can be used for cosmetic, personal care, and / or hygiene purposes. It will be understood that a composition may have more than one cosmetic purpose and can be used for more than one of these purposes at the same time. As used herein, "cosmetics" can include, but are not limited to, lipstick, mascara, lipstick, foundation, blusher, eyeliner, face and body powder, sunscreen, sunblock, nail polish, compact, solid, pencil.

[0075] "Pharmaceutical Composition": A further aspect of the present invention relates to a pharmaceutical composition comprising the peptide of the present invention or a composition of the peptide of the present invention mixed with one or more pharma- ceutically acceptable diluents, excipients, or carriers. The peptides and compositions of the present invention may be administered alone, but will generally be administered mixed with a pharmaceutical carrier, excipient, or diluent, particularly for human treatment. The pharmaceutical composition may be a composition for human or animal use, in human and veterinary medicine. Examples of such excipients suitable for the various different forms of the pharmaceutical compositions described herein can be found in Handbook of Pharmaceutical Excipients, 2nd Edition, (1994), Edited by A Wade and PJ Weller. In particular, formulations for topical delivery are described in Topical drug delivery formulations edited by David Osborne and Antonio Aman, Taylor & Francis, the full contents of which are incorporated herein by reference. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical arts and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (ARGennaro edit. 1985). Examples of suitable carriers include lactose, starch, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and the like. Examples of suitable diluents include ethanol, glycerol, and water. The choice of pharmaceutical carrier, excipient, or diluent can be selected in light of the intended route of administration and standard pharmaceutical practice. The pharmaceutical composition can include any suitable binder, lubricant, suspending agent, coating agent, solubilizing agent as, or in addition to, the carrier, excipient, or diluent.Examples of suitable binders include starch, gelatin, glucose, anhydrous lactose, liquid lactose, β-lactose, and other natural sugars, corn sweeteners, natural and synthetic gums, such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Preservatives, stabilizers, dyes, and even flavorings can be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid, and esters of hydroxybenzoic acid. Antioxidants and suspending agents can also be used.

[0076] The term "mammal" shall be understood to mean higher mammals, especially humans. However, the term also includes non-mammals, such as fish. The human may be an infant, toddler, child, juvenile, adult, or elderly human. In one embodiment of the invention, the human is, for example, 55 years of age or older. In one embodiment, the human is an elderly person experiencing a loss of lean tissue mass. In one embodiment, the human is an athlete. In one embodiment, the human is a pregnant woman. In one embodiment, the human suffers from lethargy or lack of energy.

[0077] As used herein, the term "dermatologically acceptable" means that a topical composition or components of the composition are suitable for use in contact with human skin or keratinous tissue without risk of toxicity, incompatibility, instability, and / or allergic response.

[0078] The term "sustained release" is used in its conventional sense in reference to a delivery system for a compound or active ingredient that provides for gradual release of the compound or active ingredient over a period of time, preferably, but not necessarily, at a relatively constant level of compound release over the period of time.

[0079] The term "peptide" as used herein typically refers to a polymer composed of monomers of up to 50 amino acids, e.g., 5 to 50 amino acids, linked via peptide bond linkages. The peptides of the present invention and peptides for use in the present invention (including fragments and variants thereof) can be produced completely or partially by chemical synthesis or by expression from nucleic acid. For example, the peptides of the present invention and peptides for use in the present invention can be readily prepared according to standard liquid phase peptide synthesis methods, or preferably solid phase peptide synthesis methods, well established in the art (see, e.g., JM Stewart and JD Young, Solid Phase Peptide Synthesis, 2nd edition, Pierce Chemical Company, Rockford, Illinois (1984), and M. Bodanzsky and A. Bodanzsky, The Practice of Peptide Synthesis, Springer Verlag, New York (1984)). If necessary, any of the peptides utilized in the present invention can be chemically modified to increase their stability. A chemically modified peptide or peptide analogue includes any functional chemical equivalent of a peptide that is characterized by increased stability and / or efficacy in vivo or in vitro with respect to the practice of the invention. The term peptide analogue also refers to any amino acid derivative of the peptides described herein. Peptide analogues can be produced by procedures including, but not limited to, side chain modifications, incorporation of non-natural amino acids and / or their derivatives during peptide synthesis, and the use of cross-linking agents and other methods that impart conformational constraints to the peptides or their analogues.Examples of side chain modifications include modifications of amino groups such as reductive alkylation by reaction with aldehydes followed by reduction with NaBH4; amidation with methylacetimidate; acetylation with acetic anhydride; carbamylation of amino groups with cyanate; trinitrobenzylation of amino groups with 2,4,6-trinitrobenzenesulfonic acid (TNBS); alkylation of amino groups with succinic anhydride and tetrahydrophthalic anhydride; and pyridoxylation of lysine by pyridoxa-5'-phosphate followed by reduction with NaBH4. The guanidino group of arginine residues can be modified by the formation of heterocyclic condensation products with reagents such as 2,3-butanedione, phenylglyoxal, and glyoxal. Carboxyl groups can be modified by carbodiimide activation with o-acylisourea formation followed by derivatization (e.g., to the corresponding amide). Sulfhydryl groups can be modified by methods such as carboxymethylation with iodoacetic acid or iodoacetamide; performic acid oxidation to cysteic acid; formation of mixed disulfides with other thiol compounds; reaction with maleimide, maleic anhydride, or other substituted maleimides; formation of mercuric derivatives using 4-chloromercuribenzoate, 4-chloromercuriphenylsulfonic acid, phenylmercuric chloride, 2-chloromerc-4-nitrophenol, and other mercuric; carbamylation of cyanates at alkaline pH. Tryptophan residues can be modified, for example, by oxidation with N-bromosuccinimide or alkylation of the indole ring with 2-hydroxy-5-nitrobenzyl bromide or sulfonyl halides. Tyrosine residues can be altered by nitration with tetranitromethane to form 3-nitrotyrosine derivatives. Modification of the imidazole ring of a histidine residue can be accomplished by alkylation with iodoacetic acid derivatives or N-carboethoxylation with diethylpyrocarbonate.Examples of incorporation of unnatural amino acids and derivatives during peptide synthesis include, but are not limited to, the use of norleucine, 4-aminobutyric acid, 4-amino-3-hydroxy-5-phenylpentanoic acid, 6-aminohexanoic acid, t-butylglycine, norvaline, phenylglycine, ornithine, sarcosine, 4-amino-3-hydroxy-6-methylheptanoic acid, 2-thienylalanine, and / or D-isomers of amino acids. Peptide structural modifications include the creation of retro-inverso peptides that contain reverse sequences encoded by D-amino acids. Changes can be those that reduce susceptibility to proteolysis, reduce susceptibility to oxidation, alter the binding affinity of the variant sequence (typically increasing affinity is desired), and / or confer or modify other physicochemical or functional properties to the variant / analog peptide of interest.

[0080] The term "peptide comprising the amino acid sequence of SEQ ID NO:1" as used herein typically refers to a polymer composed of up to 50 amino acids, e.g., 5 to 50 amino acids, monomers linked via peptide bond linkages, comprising SEQ ID NO:1 or an amino acid sequence essentially consisting of SEQ ID NO:1 that is substantially identical to SEQ ID NO:1 but modified with respect to the modification of one or more amino acid residues, e.g., one, two or three residues (hereinafter referred to as "variants" or "peptide variants"). Preferably, such modifications involve the insertion, addition, deletion and / or substitution of 11 or less amino acids, more preferably 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, preferably 5 or less, 4 or less, even more preferably 3 or less amino acids, most preferably only one or two amino acids. Insertions, additions and substitutions with natural and modified amino acids are envisaged. The peptides may have conservative amino acid changes, where the introduced amino acid is structurally, chemically or functionally similar to the amino acid being replaced. Generally, a peptide will have at least 70% amino acid sequence identity, preferably at least 80% sequence identity, more preferably at least 90% sequence identity, and ideally at least 95%, 96%, 97%, 98% or 99% sequence identity with the parent sequence.

[0081] The term "fragment" shall be understood to mean a segment of amino acid sequence SEQ ID NO: 1. Typically, fragments have a length between 3 and 10 consecutive amino acids. In general, fragments have a charge of -5 to +3. The charge of a peptide, fragment, or region is determined using the method of Cameselle, JC, Ribeiro, JM, and Sillero, A. (1986). Derivation and use of a formula to calculate the net charge of acid-base compounds. Its application to amino acids, proteins and nucleotides. Biochem. Educ. 14, 131-136.

[0082] As used herein, the term "sequence identity" includes both sequence identity and sequence similarity, i.e., a variant (or homologue) that shares 70% sequence identity with a reference sequence shall be understood as a variant (or homologue) in which any 70% of the aligned residues of the variant (or homologue) are identical to or are conservative substitutions of the corresponding residues in the reference sequence over the entire length of the sequence. Sequence identity is the amount of exact matched letters between two different sequences. As used herein, gaps are not counted and the measurement is relative to the shorter of the two sequences.

[0083] In the context of "sequence homology", the term shall be understood to mean that a variant (or homolog) shares a defined percentage of similarity or identity with a reference sequence when a defined percentage of aligned residues of the variant (or homolog) are identical to, or are conservative substitutions of, corresponding residues in the reference sequence, and the variant (or homolog) shares the same function as the reference sequence.

[0084] The alignment and percent homology or percent sequence identity can be determined using software programs known in the art, for example, one alignment program is BLAST using default parameters. Details of these programs can be found at the following internet address: http: / / www.ncbi.nlm.nih.gov / blast / Blast.cgi.

[0085] "Inflammatory disorder" refers to immune-mediated inflammatory conditions that affect humans and are generally characterized by dysregulation of the expression of one or more cytokines. Examples of inflammatory disorders include skin and joint inflammatory disorders, cardiovascular inflammatory disorders, certain autoimmune diseases, pulmonary and airway inflammatory disorders, and intestinal inflammatory disorders. Examples of skin inflammatory disorders include dermatitis, such as atopic dermatitis and contact dermatitis, acne vulgaris, and psoriasis. Examples of joint inflammatory disorders include rheumatoid arthritis. Examples of cardiovascular inflammatory disorders are cardiovascular disease and atherosclerosis. Examples of autoimmune diseases include type 1 diabetes, Graves' disease, Guillain-Barre disease, lupus, psoriatic arthritis, and ulcerative colitis. Examples of pulmonary and airway inflammatory disorders include asthma, cystic fibrosis, COPD, emphysema, and acute respiratory distress syndrome. Examples of inflammatory disorders of the intestine include colitis and inflammatory bowel disease. Other inflammatory disorders include cancer, hay fever, periodontal disease, allergies, hypersensitivity, ischemia, depression, systemic disease, post-infectious inflammation, and bronchitis. The peptides and compositions of the present invention can also be used in non-therapeutic treatment of inflammation. Examples of non-therapeutic treatment of inflammation include use to reduce normal, non-pathological inflammation, such as inflammation of muscles and joints after physical exercise.

[0086] "Disease or condition characterized by dermal or epidermal cell or tissue damage" refers to any condition or disease that results in dermal or epidermal tissue or cell or organ damage. One example is trauma, which often results in skin damage. Another example is inflammatory skin conditions, such as psoriasis or eczema, which often results in skin damage. Another example is inflammatory disorders of the lower intestine, which results in epithelial cell / tissue damage lining the lower intestine. Another example is epithelial cell / tissue damage lining the lower intestine caused by ingestion of toxic or damage-inducing substances (e.g., toxic chemicals or toxic drugs). Another example is cancer, for example, intestinal cancer, which results in epithelial tissue damage in the intestine. Another condition is peripheral inflammatory disorders, such as atopic dermatitis, which may result in skin damage in humans.

[0087] As used herein, the term "metabolic disorder" is to be understood to include prediabetes, diabetes; type 1 diabetes; type 2 diabetes; metabolic syndrome; obesity; diabetic dyslipidemia; hyperlipidemia; hypertension; hypertriglyceridemia; hyperfatty acidemia; hypercholesterolemia; hyperinsulinemia; and MODY.

[0088] "Bacterial infection characterized by a disease or condition" means any condition or disease characterized by having a pathology caused by bacterial proliferation or infection, including, for example, MRSA, Salmonella, Listerin pneumonia, Staphylococcus aureus food poisoning, bacterial meningitis. Specific examples are exemplified at https: / / en.wikipedia.org / wiki / List_of_infectious_diseases.

[0089] "Artificial" as applied to an edible product shall be understood to mean made by man and not occurring in nature.

[0090] "Improvement of muscle condition" means improvement of muscle health, e.g., promotion of skeletal muscle protein synthesis, skeletal glucose uptake, improvement of lean tissue mass in therapeutic or non-therapeutic situations, promotion of muscle recovery after active physical exercise in general, or improvement of muscle performance. The method or use may be therapeutic or non-therapeutic. The term "improvement of lean tissue mass condition" shall be understood to mean an increase in lean tissue mass, or inhibition or prevention of the rate of degradation of lean tissue mass.

[0091] By "promoting muscle recovery" is meant causing an increase in glucose uptake in skeletal muscle compared to untreated skeletal muscle.

[0092] "Disease or condition characterized by lethargy or low energy levels" means any symptom or disease characterized by fatigue or low energy. Examples include allergies, asthma, anemia, cancer and its treatment, chronic pain, heart disease, infection, depression, eating disorders, grief, sleep disorders, thyroid problems, medication side effects, alcohol use, or drug use.

[0093] The term "catabolic wasting" encompasses both sarcopenia and cachexia ("catabolism" refers to the breakdown of tissue and is the antonym of "anabolism," which refers to the building of tissue).

[0094] Loss of muscle and fat tissue due to chronic disease is called cachexia. The general loss of body weight and muscle mass that occurs with aging is called sarcopenia. In both cachexia and sarcopenia, muscle loss leads to frailty and has deleterious effects on a variety of clinical outcomes (Rolland 2011; Fearon 2013; Muscaritoli 2013). Individuals with cachexia and / or sarcopenia are at increased risk for death, infection, and falls; delayed wound healing; significantly reduced physical and respiratory capacity; and an overall decline in quality of life.

[0095] Cachexia typically causes more rapid and significant weight loss than sarcopenia and is generally characterized as a loss of muscle and adipose tissue totaling more than 5% of body weight, although losses of more than 20% of body weight are common. Often, individuals with cachexia continue to lose weight even when consuming sufficient calories. Severe chronic illnesses such as cancer, AIDS, and chronic obstructive pulmonary disease (COPD) are known causes of cachexia. It is estimated that 50% to 80% of all cancer patients experience cachexia, and cachexia is the leading cause of more than 20% of all cancer-related deaths. Cachexia in HIV / AIDS patients is common and occurred already before the advent of anti-HIV viral drugs. Sarcopenia (from Greek meaning "lack of flesh") generally refers to the loss of muscle mass and muscle function associated with aging. Approximately 50% of people over the age of 80 experience sarcopenia. Sarcopenia can also result from physical inactivity, malnutrition, or disease. Some researchers refer to muscle weakness associated with aging and not related to an underlying cause as "primary sarcopenia," and muscle weakness resulting from one or more other causes as "secondary sarcopenia." Sarcopenia is associated with increased risk of insulin resistance and type 2 diabetes in non-obese adults over 60 years of age. The conventional medical community has failed to provide early and aggressive intervention for cachexia, often resulting in poor clinical outcomes, including premature death and impaired physical function. Standard medical treatments for cachexia include promoting fluid and food intake, and the use of certain medications. However, many standard medical therapies to treat sarcopenia and cachexia carry the risk of adverse effects, such as nausea, edema, and fatigue, some of which have not been fully investigated in clinical trials. A number of nutritional, lifestyle, and innovative pharmacological interventions may be useful in preventing and treating catabolic wasting. Whey protein, creatine, and the amino acids glutamine, arginine, leucine, and methyl hydroxybutyrate or HMB (a leucine derivative) are, among others, important for building and maintaining lean muscle mass. Omega-3 fatty acids, conjugated linoleic acid, and vitamin D also combat lean tissue loss.There is a real and urgent need for new and emerging strategies to prevent muscle wasting. (Ribeiro S, Keheyias J. Sarcopenia and the analysis of body composition. Adv Nutr 2014:5:260-267. Muscaritoli A, Lucia S, Molfino A, Cederholm MT, Rossi Fanelli F, Muscle atrophy in aging and chronic diseases: is it sarcopenia or cachexia? Intern Emerg Med 2013;8: 553-560. Rolland Y, VanKan GA, Gillette-Guyonnet S, Vellas B. Cachexia versus sarcopenia. Curr Opin Clin Nutr Metab Care. 2010;14(1):15-21. Evans WJ, Morley JE, Argiles JM, et al. Cachexia: a new definition. Clin Nutr 2008;27:793-799. Fearon K, Arends J, Baracos V. Understanding the mechanisms and treatment options in cancer cachexia. Nat Rev Clin Oncol 2013;10(2):90-99. )

[0096] "Maintaining or restoring muscle health" means to maintain muscle health in a mammal or to help it recover from damage incurred during physical exercise. By promoting glucose transport in skeletal muscle, the peptides promote recovery from physical exercise and relieve muscle pain and damage associated with physical exercise. They can also be used to reduce and prevent muscle cramps and allow for faster recovery from muscle cramps. Cramps can result from physical stress, mental stress, and / or repetitive strain injury stress. By promoting glucose transport, the peptides reduce muscle myopathy, prevent sarcopenia in mammals, promote recovery from injury during physical exercise, and relieve muscle pain and damage associated with physical exercise. The present invention also relates to the peptide or composition of the present invention for use in maintaining or restoring muscle health in a mammal.

[0097] As used herein, the term "personal care product" shall be understood to mean a composition formulated for human use in cleaning or treating the human body, particularly the skin, teeth, nails, feet, and hair. Examples include shampoos, conditioners, skin creams and lotions, powders, dentifrices, shower gels or creams, bath gels or shower gels, hair dyes, soaps, body scrubs, exfoliants, anti-dandruff liquids, body lotions, shaving liquids, moisturizers, washing solutions, masks, oils, serums, as well as rinses, deodorants, and antiperspirants.

[0098] The term "skin aging" is used in the sense commonly and broadly used in the art of cosmetics and personal care products. Signs of skin aging include wrinkles, lines, fissures, bumps, red spots, enlarged pores, roughness, dullness, loss of elasticity, sagging, loss of firmness, discoloration, bumps, hyperpigmentation, age spots, keratosis, inflammation, collagen breakdown, and other histological changes within the skin layers including the underlying tissue.

[0099] The term "cosmetically or pharma- ceutically acceptable salt" refers to salts that are recognized for use in animals, and more particularly in humans, and includes salts used to form base addition salts or acid addition salts. Base addition salts are inorganic salts, such as, but not limited to, lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc, or aluminum, or organic salts, such as, but not limited to, ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine, or piperazine. Acid addition salts are organic salts, such as, but not limited to, acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, oxalic acid, pamoic acid, or gluconic acid, or inorganic salts, such as, but not limited to, chloride, sulfuric acid, boric acid, or carbonate. The nature of the salt is not critical, provided that it is cosmetically or pharma- ceutically acceptable. Cosmetically or pharma- ceutically acceptable salts of the peptides of the present invention can be obtained by conventional methods well known in the prior art [Berge SM et al., "Pharmaceutical Salts", J. Pharm. Sci., (1977), 66, 1-19].

[0100] "C-terminal domain" as applied to a fragment means the first three amino acids at the C-terminus of the fragment.

[0101] "N-terminal domain" as applied to a fragment means the last three amino acids at the N-terminus of the fragment.

[0102] A "homolog" of a reference protein shall be understood to mean a protein derived from a different species of plant having at least 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence homology with the reference protein.

[0103] As used herein, the terms "comprise, comprise, comprised, comprising" or any variation thereof and the terms "include, includes, included, including" or any variation thereof are considered to be fully interchangeable and are all to be given the broadest possible interpretation and vice versa.

[0104] The invention will be more clearly understood from the following description of embodiments thereof, given for illustrative purposes only, with reference to the accompanying drawings, in which: [Brief description of the drawings]

[0105] [Figure 1] FIG. 1 illustrates the results of a cell proliferation assay using untreated and peptide-treated human dermal fibroblasts (HDFs). [Diagram 2] FIG. 1 illustrates the results of a cell proliferation assay using untreated and peptide-treated HACAT cells. [Diagram 3] FIG. 1 illustrates the results of a collagen expression assay using untreated and peptide-treated human dermal fibroblasts (HDFs). [Figure 4] FIG. 1 illustrates the results of an elastin expression assay using untreated and peptide-treated human dermal fibroblasts (HDFs). [Diagram 5] FIG. 1 illustrates the results of a glucose uptake assay using untreated and peptide-treated human skeletal myoblasts. [Figure 6] FIG. 1 illustrates the results of a glucose uptake assay using untreated and peptide-treated human skeletal myoblasts. [Figure 7] FIG. 1 illustrates the results of a glucose uptake assay using untreated and peptide-treated human skeletal myoblasts. [Figure 8]FIG. 1 illustrates the results of a GLUT4 translocation assay using untreated and peptide-treated human skeletal myoblasts. [Figure 9] FIG. 1 illustrates the results of a muscle protein synthesis (mTOR) assay. [Figure 10] FIG. 1 illustrates the results of a muscle protein synthesis (puromycin) assay. [Figure 11] FIG. 1 illustrates the results of a cytotoxicity study in HSKMC cells using the peptide of SEQ ID NO:1. [Figure 12] FIG. 1 illustrates the results of a phosphoacetyl-CoA carboxylase (Ser79) ELISA (peptide of SEQ ID NO:1). [Figure 13] FIG. 1 illustrates the results of a phospho-Akt1 ELISA (peptide of SEQ ID NO:1). [Figure 14] FIG. 1 illustrates the results of a phospho-AMPKα ELISA (peptide of SEQ ID NO: 1). [Figure 15] FIG. 1 illustrates blood glucose levels in KKAY mice after 5 days of treatment with the peptide of SEQ ID NO:1. [Figure 16] FIG. 1 illustrates blood glucose levels in KKAY mice after 7 days of treatment with the peptide of SEQ ID NO:1. [Figure 17] FIG. 1 illustrates the body weight of KKAY mice after 13 days of treatment with the peptide of SEQ ID NO:1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0106] The invention will now be described with reference to specific examples, which are presented for illustrative purposes only and are not intended to limit in any manner the claimed exclusivity or scope of the invention described, which examples constitute the best modes currently contemplated for carrying out the invention.

[0107] In its broadest sense, a first aspect of the invention provides a peptide comprising the amino acid sequence of SEQ ID NO:1 (hereinafter referred to as "the peptide of the invention").

[0108] In a second aspect, the present invention provides a composition comprising an effective amount of a peptide of the present invention. The composition may be a topical composition. The composition may be a cosmetic composition comprising a cosmetically effective amount of a peptide comprising the amino acid sequence of SEQ ID NO:1 or a variant thereof. The composition may be a pharmaceutical composition comprising a pharma- ceutical effective amount of a peptide comprising the amino acid sequence of SEQ ID NO:1 or a variant thereof. It will be appreciated that the composition may provide a cosmetic benefit, i.e. a non-therapeutic benefit, or a therapeutic benefit. The composition may provide both a cosmetic benefit and a therapeutic benefit.

[0109] Examples of cosmetic applications include, but are not limited to, anti-aging, slowing or inhibiting or preventing aging of human skin, promoting tissue growth, promoting epithelial tissue growth, promoting skin growth, promoting organ growth, promoting organism growth, improving muscle condition in mammals, promoting muscle recovery, typically after physical exercise, maintaining or restoring muscle health in mammals, enhancing physical performance, muscle growth or muscle building, and treating muscle weakness.

[0110] The present invention also provides therapeutic uses / applications of the compositions or peptides of the invention, especially non-topical therapeutic applications.

[0111] Examples of therapeutic applications include, but are not limited to, promoting tissue growth, promoting epithelial tissue growth, promoting skin growth, promoting organ growth, promoting organism growth, improving muscle condition in a mammal, promoting muscle recovery, maintaining or restoring muscle health in a mammal, muscle growth or muscle building, treatment of muscle weakness, treatment or prevention of diseases or conditions characterized by muscle wasting or reduced anabolic activity or reduced protein synthesis (catabolic wasting), treatment or prevention of diseases or conditions characterized or mediated by oxidative stress, treatment or prevention of metabolic diseases (particularly diabetes or pre-diabetes), treatment or prevention of diseases or conditions characterized by lethargy or low energy levels, treatment of wounds in a mammal, treatment or prevention of pain in a mammal, treatment or prevention of diseases or conditions characterized by epithelial cell or tissue damage and / or dermal or epithelial cell or tissue damage, and treatment or prevention of metabolic disorders (such as diabetes) in a mammal.

[0112] SEQ ID NO:1 contains the following sequence: Has WKDEAGKPLVK.

[0113] In an embodiment of the invention, the peptide is bioactive.

[0114] In an embodiment, the peptide has glucose transport promoting activity. In an embodiment, the peptide has cell proliferation promoting activity. In an embodiment, the peptide has anti-aging activity. In an embodiment, the peptide has anabolic activity. It will be understood that the peptide may have two or three of glucose transport promoting activity, cell proliferation promoting activity, anti-aging activity, and anabolic activity. The peptide may have glucose transport promoting activity, cell proliferation promoting activity, anti-aging activity, and anabolic activity.

[0115] In an embodiment of the invention, the peptide comprises a length of about 3 to 50 amino acids, a length of about 14 to about 50 amino acids, preferably a length of about 15, 20, 25, 30, 35, 40, 45, or 49 amino acids, preferably a length of about 14, 15, 16, 17, 18, 19, or 20 amino acids.

[0116] In an embodiment of the invention, the peptide comprises a length of about 3 to about 11 amino acids, preferably about 4, 5, 6, 7, 8, 9, or 10 amino acids.

[0117] In one embodiment, the peptide has one to five amino acid changes compared to SEQ ID NO:1. In one embodiment, the peptide has one to four amino acid changes compared to SEQ ID NO:1. In one embodiment, the peptide has one to three amino acid changes compared to SEQ ID NO:1. In one embodiment, the peptide has one to two amino acid changes compared to SEQ ID NO:1. In one embodiment, the amino acid changes are conservative amino acid changes. In one embodiment, the amino acid changes are amino acid substitutions. In one embodiment, the amino acid substitutions are conservative substitutions. In one embodiment, the amino acid changes are amino acid additions. In one embodiment, the amino acid changes are amino acid deletions.

[0118] The peptides of the present invention include the following variant peptides:

[0119] Variants of SEQ ID NO:1 Variants of SEQ ID NO:1 (WKDEAGKPLVK) including variants with 1, 2 or 3 conservative amino acid substitutions, 1, 2 to 3 non-conservative amino acids, 1, 2 or 3 amino acid additions, 1, 2 or 3 amino acid deletions are provided below: One conservative amino acid substitution: WKEEAGKPLVK (SEQ ID NO:2);FKDEAGKPLVK (SEQ ID NO:3); WKDEAGKPMVK (SEQ ID NO: 4); WKDEAGRPLVK (SEQ ID NO: 5); WRDEAGKPLVK (SEQ ID NO: 6); WKDEAGKPLMK (SEQ ID NO: 7); WKDQAGKPLVK (SEQ ID NO: 8); WKDEATKPLVK (SEQ ID NO: 9) Two conservative amino acid substitutions: YKNEAGKPLVK (SEQ ID NO:10); WKNESGKPLVK (SEQ ID NO:11); WKDEAGKTLVR (SEQ ID NO:12); FKDEATKPLVK (SEQ ID NO:13); FKDEAGKPLIK (SEQ ID NO:14); WKDEAGKTLLK (SEQ ID NO:15); WKNEAGKPVVK (SEQ ID NO:16); WKDEAGRTLVK (SEQ ID NO:17) Three conservative amino acid substitutions: WEDESGKPLLK (SEQ ID NO: 18); WKEEAGKPIVQ (SEQ ID NO: 19); YKNEAGKPLVR (SEQ ID NO: 20); WKDQATRPLVK (SEQ ID NO: 21); WKDESGKPVLK (SEQ ID NO: 22); WQDDSGKPLVK (SEQ ID NO: 23); WKNEAGKTLLK (SEQ ID NO: 24); WKDKAGEPLVR (SEQ ID NO: 25) One non-conservative amino acid substitution: WKDEAGNPLVK (SEQ ID NO:26); CKDEAGKPLVK (SEQ ID NO:27); WKDEAGKPLGK (SEQ ID NO:28); WKDENGKPLVK (SEQ ID NO:29); WKDEARKPLVK (SEQ ID NO:30); WKDEAGKPLVT (SEQ ID NO:31); WKDEAGKRLVK (SEQ ID NO:32); WKWEAGKPLVK (SEQ ID NO:33) Two non-conservative amino acid substitutions: WKDEAGFPTVK (SEQ ID NO:34); WYDMAGKPLVK (SEQ ID NO:35); WKDYEGKPLVK (SEQ ID NO:36); WKREAGKPGVK (SEQ ID NO:37); WKLEKGKPLVK (SEQ ID NO:38); WKDEAGKPCVK (SEQ ID NO:39); WKKEAPKPLVK (SEQ ID NO:40); SKDEAGPPLVK (SEQ ID NO:41) Three non-conservative amino acid substitutions: WKHEPGKPLAK (SEQ ID NO: 42); WKDEREKPFVK (SEQ ID NO: 43); WKQEAGKPWRK (SEQ ID NO: 44); VKDEAKKPLVH (SEQ ID NO: 45); NWDEAGKMLVK (SEQ ID NO: 46); IKDEDGPPLVK (SEQ ID NO: 47); LKDEYGKPLVN (SEQ ID NO: 48); WKDRAGKELTK (SEQ ID NO: 49) Amino acid addition WKDEAGKPLPVK (SEQ ID NO:50); WKGDENYAGKPLVK (SEQ ID NO:51); LWKDEAGRKYPLVK (SEQ ID NO:52); WKDCEGAGKPLVK (SEQ ID NO:53); WKDEPAGKPLVVK (SEQ ID NO:54); WKDEAGPKPLVK (SEQ ID NO:55); WKDEAGWADKPLVK (SEQ ID NO:56); WKNDEAGKPLVK (SEQ ID NO:57) Amino acid deletion WKDAKPLVK (SEQ ID NO:58); WKEAGKPVK (SEQ ID NO:59); WKDEAKPLVK (SEQ ID NO: 60); WDEAGKPV (SEQ ID NO: 61); WKDEAGKPVK (SEQ ID NO: 62); WDAGKPLVK (SEQ ID NO: 63); WKDEAGKPLV (SEQ ID NO:64); WEAGKPLV (SEQ ID NO:65)

[0120] The variant peptide may be a bioactive variant.

[0121] The present invention also provides fragments of SEQ ID NO:1, and peptides comprising one or more of these fragments.

[0122] The fragment may be a bioactive fragment. In one embodiment, the fragment is an anti-aging fragment. In one embodiment, the fragment is a cell proliferation promoting fragment. In one embodiment, the fragment is a glucose transport promoting fragment. In one embodiment, the fragment is an anabolic fragment. It will be appreciated that in one embodiment, the bioactive fragment is two or more of an anti-aging fragment, a glucose transport promoting fragment, a cell proliferation promoting fragment, and an anabolic fragment. In one embodiment, the bioactive fragment is an anti-aging fragment, a glucose transport promoting fragment, a cell proliferation promoting fragment, and an anabolic fragment.

[0123] The fragment may be a bioactive fragment. In one embodiment, the fragment is a cell proliferation promoting fragment. In one embodiment, the fragment is a GLUT4 migration promoting fragment. In one embodiment, the fragment exhibits anabolic metabolism stimulating activity. In one embodiment, the fragment exhibits antioxidant activity or anti-inflammatory activity.

[0124] Examples of fragments of SEQ ID NO:1 are provided below. WKDEAG (SEQ ID NO: 66); WKDEA (SEQ ID NO: 67); KDEAGKPL (SEQ ID NO: 68); KDEAG (SEQ ID NO: 69); DEAGKPL (SEQ ID NO: 70); GKPLV (SEQ ID NO: 71); DEAGK (SEQ ID NO: 72); WKDEAGKPL (SEQ ID NO: 73); WKD (SEQ ID NO: 74); KDE (SEQ ID NO: 75); KPLVK (SEQ ID NO: 76); WKDE (SEQ ID NO: 77); AGKPL (SEQ ID NO: 78); EAG (SEQ ID NO: 79); AGK (SEQ ID NO: 80); KPL (SEQ ID NO: 81); LVK (SEQ ID NO: 82); GKP (SEQ ID NO: 83); DEA (SEQ ID NO: 84); PLV (SEQ ID NO: 85)

[0125] It will be appreciated that the composition may comprise more than one peptide or fragment. Preferably, the composition comprises at least two peptides of the invention.

[0126] Preferably, the composition comprises at least three peptides of the invention. Preferably, the composition comprises at least four peptides of the invention. Preferably, the composition comprises at least five peptides of the invention. Preferably, the composition comprises at least six peptides of the invention. Preferably, the composition comprises at least seven peptides of the invention. Preferably, the composition comprises at least eight peptides of the invention. Preferably, the composition comprises at least nine peptides of the invention. Preferably, the composition comprises at least ten peptides of the invention. In one embodiment, the composition comprises substantially all peptides. In one embodiment, the composition comprises substantially all variants. In one embodiment, the composition comprises substantially no other peptides.

[0127] In an embodiment, the peptide or composition of the present invention has an activity selected from one or more of antibacterial activity, anti-inflammatory activity, and antioxidant activity. The activity can be cosmetic activity (i.e., non-therapeutic activity), therapeutic activity, or both. The present invention also provides the composition of the present invention for use in a method for maintaining or restoring gastrointestinal health in a mammal. The present invention also provides the composition of the present invention for use in a method for treating bacterial infection. A further aspect of the present invention provides the composition of the present invention for use in a method for treating or preventing inflammatory disorders and / or inflammation in a mammal. Preferably, the inflammation is symptomatic inflammation. This use can be in addition to or alternative to the use of the present invention discussed above.

[0128] In an embodiment, the peptide or fragment or composition of the invention has an activity selected from one or more of antibacterial activity, anti-inflammatory activity, and antioxidant activity.

[0129] The composition may be a topical composition. The topical composition may be provided as a formulation selected from the group including creams, multiple emulsions, anhydrous compositions, aqueous dispersions, oils, milks, balsams, foams, lotions, gels, cream gels, hydroalcoholic solutions, hydroglycolic solutions, cosmetics, personal care products, hydrogels, liniments, serums, soaps, dusting powders, pastes, semi-solid formulations, liniments, serums, shampoos, conditioners, ointments, optional rinse formulations, talcs, mousses, powders, sprays, aerosols, solutions, suspensions, emulsions, syrups, elixirs, polysaccharide films, patches, gel patches, bandages, adhesive systems, water-in-oil emulsions, oil-in-water emulsions, and silicone emulsions.

[0130] In an embodiment of the present invention, the emulsion contains lipids or oils. The emulsion can be, but is not limited to, oil-in-water, water-in-oil, water-in-oil-in-water, and oil-in-water-in-silicone emulsions. The emulsion can contain a humectant. The emulsion can contain an antifoaming agent, such as silicone. The emulsion can have any suitable viscosity. The emulsion can further contain an emulsifier and / or an antifoaming agent. Methods for preparing emulsions are known to those skilled in the art.

[0131] The compositions of the present invention can be provided, prepared, and / or administered in various suitable forms. Such forms include, but are not limited to, liquid, semi-solid, and solid dosage forms, such as, for example, solutions (e.g., injection and infusion solutions), dispersions or suspensions, emulsions, microemulsions, tablets, pills, powders, liposomes, dendrimers and other nanoparticles, microparticles, and suppositories. It will be understood that the form may depend on the intended mode of administration, the nature of the composition or combination, and the therapeutic application or other intended use. The formulations may also include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, vesicles containing lipids (cationic or anionic), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions, carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax.

[0132] It should be understood that what is considered to be an "active" ingredient in one product may be a "functional" or "excipient" ingredient in another product, and vice versa. It will also be understood that some ingredients play a dual role as both an active ingredient and a functional or excipient ingredient.

[0133] In a particularly preferred embodiment, the methods and uses of the present invention involve the administration of the peptides or compositions of the present invention in combination with one or more other active agents (e.g., commercially available, existing growth-promoting drugs or pharmacological enhancers). In such cases, the compounds of the present invention can be administered sequentially, simultaneously, or sequentially with one or more other active agents.

[0134] In a preferred embodiment, repeated use of the composition is provided.

[0135] The composition of the present invention can be incorporated into a medical device for administration. Such devices include, but are not limited to, fabrics, patches, bandages, gauze, socks, tights, underwear, dressings, gloves, masks, adhesive patches, non-adhesive patches, occlusive patches, and microcurrent patches, or suitable adhesive systems. In such embodiments, the device is in direct contact with a keratin layer, such as the skin, thereby releasing the peptide of the present invention. It will be understood that the topical composition can be incorporated in any suitable form, as detailed herein. For example, the topical composition or topical peptide of the present invention can be incorporated into the device or present on the surface of the device, or present in a cream, gel, or wax formulation, or any suitable formulation as defined herein, and incorporated into or on the surface of the device.

[0136] The device can be adapted to be attached or bonded to the skin. In one embodiment, the device is adapted to release a certain amount of the composition or peptide of the present invention. It will be understood that the amount of composition contained in the sustained release system depends, for example, on where the composition is administered, the kinetics and duration of release of the composition of the present invention, as well as the nature of the condition, disorder, and / or disease being treated and / or cared for. The device can be such that the composition is released due to the biodegradation of the device or due to friction between the device and the body, due to body moisture, skin pH, or body temperature.

[0137] In an embodiment of the present invention, the composition may further comprise at least one cosmetically or pharma- ceutically acceptable excipient. Excipients may be used interchangeably with functional ingredients or additives. It will be understood that the topical composition of the present invention may be administered alone, but will generally be administered in admixture with cosmetic or pharmaceutical excipients. Cosmetically or pharma-ceutically acceptable excipients are well known in the art, and any known excipient may be used, provided that it is suitable for topical administration and dermatologically acceptable without undue toxicity, incompatibility, and / or allergic reactions.

[0138] Preferably, any excipient incorporated is present in a trace amount. The amount of excipient incorporated depends on a number of factors, including the type of excipient used, the nature of the excipient, the ingredients of the topical composition, the amount of active ingredient or peptide in the topical composition, and / or the intended use of the topical composition. The nature and amount of any excipient should not unacceptably alter the usefulness of the peptide of the present invention.

[0139] In embodiments of the present invention, the excipient may be a suitable diluent, carrier, binder, lubricant, suspending agent, coating agent, preservative, stabilizer, dye, vehicle, solubilizer, base, emollient, emulsifier, fragrance, moisturizer, and / or surfactant.

[0140] Examples of suitable diluents include, but are not limited to, any diluent disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667. Examples include ethanol, glycerol, and water. Examples of suitable carriers include, but are not limited to, lactose, starch, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and any suitable carrier disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667.

[0141] Examples of suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose, anhydrous lactose, fluid lactose, beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, and polyethylene glycol, as well as any suitable binders disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0142] Examples of suitable lubricants include, but are not limited to, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride, as well as any suitable lubricant disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0143] The carrier can be any suitable carrier known in the art or disclosed in US Patent Publication No. 2014 / 0120131 or US Patent Publication No. 2004 / 0132667. In some embodiments, the carrier can include, but is not limited to, liquids such as water, oil, or surfactants, including surfactants of petroleum, animal, vegetable, or synthetic origin, polymers, oils such as peanut oil, mineral oil, castor oil, soybean oil, alcohols, polysorbates, sorbitan esters, ether sulfates, sulfates, betaines, glycosides, maltosides, fatty alcohols, nonoxynol, poloxamers, polyoxyethylene, polyethylene glycols, dextrose, glycerol, or digitonin. It will be understood that the carrier is dermatologically acceptable. Preferred carriers include emulsions such as oil-in-water, water-in-oil, water-in-oil-in-water, and oil-in-water-in-silicone emulsions. The emulsions can further include emulsifiers and / or antifoaming agents.

[0144] In the embodiment of the present invention, the composition may further comprise one or more additional components.The composition of the present invention may be administered sequentially, simultaneously, or sequentially with one or more other additional agents.Such additional components may be beneficial to be incorporated into topical composition or may be beneficial components depending on the intended use of topical composition.The additional components may be active or functional components, or both.

[0145] Examples of such additional ingredients include, but are not limited to, one or more of cleansing agents, conditioning agents, sunscreens, pigments, moisturizers, thickening agents, gelling agents, essential oils, astringents, pigments, anti-caking agents, anti-foaming agents, binders, additives, buffering agents, chelating agents, topical analgesics, film formers or materials, bulking agents, polymers, opacifying agents, pH adjusting agents, propellants, reducing agents, sequestering agents, skin depigmenting and skin lightening agents, skin conditioning agents, aloe, healing agents, soothing agents, smoothing agents, pantothenic acid, treatment agents, thickening agents, vitamins, coloring agents, medicinal agents, preservatives, anti-foaming agents, buffering agents, astringents, polymers, pH adjusting agents, anti-odor agents, or any other dermatologically acceptable carrier or surfactant.

[0146] It is understood that the additional ingredients listed may provide more than one benefit, and the classifications made herein are for clarity and convenience only and are not intended to limit the additional ingredients to the particular application or type listed.

[0147] Any additional ingredients should be suitable for application to the skin without undue toxicity, incompatibility, and / or allergic reactions.

[0148] In some embodiments, the additional ingredient has or contributes to glucose transport activity. In some embodiments, the additional ingredient has anabolic activity. In some embodiments, the additional ingredient has or contributes to anti-inflammatory activity. In some embodiments, the additional ingredient has or contributes to anti-aging activity. In some embodiments, the additional ingredient is an ingredient for keratin layer health and / or development, skin health and / or development, and / or muscle health, recovery, and / or development. The active agent may be a pharmacological enhancer. Such active agents are known and commercially available. In such cases, the topical composition of the present invention may be administered sequentially, simultaneously, or sequentially with one or more other active agents.

[0149] In some embodiments, the additional ingredients can be farnesol ([2E,6E]-3,7,11-trimethyl-2,6,10-dodecatrien-1-ol), phytantriol (3,7,11,15-tetramethylhexadecane-1,2,3-triol), desquamating actives, enzymes, enzyme inhibitors, enzyme activators, plant extracts, and marine extracts, anti-acne actives, anti-wrinkle actives or anti-atrophy actives, antioxidants / radical scavengers, chelating agents, flavonoids, anti-inflammatory agents, anti-cellulite agents, topical anesthetics, tanning actives, skin lightening agents, skin healing agents, bisabolol, antimicrobial or antifungal actives, sunscreen actives, particulate materials, conditioning agents, structuring agents, thickening agents. The desquamating active can be any suitable agent that enhances skin appearance or texture, as disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0150] Examples of anti-acne actives are disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667 and include resorcinol, salicylic acid, erythromycin, zinc, sulfur, benzoyl peroxide.

[0151] Examples of thickening agents are disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667 and include carboxylic acid polymers, crosslinked polyacrylate polymers, polyacrylamide polymers, polysaccharides.

[0152] Examples of conditioning agents are disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667 and include humectants, moisturizers, or skin conditioners.

[0153] Examples of structuring agents are disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667, and include any agent that provides rheological characteristics to the composition and contributes to the stability of the composition.

[0154] Any suitable antimicrobial or antifungal active ingredient can be used, examples of which are disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667. Such active ingredients can destroy microorganisms and prevent their growth or action. Examples include, but are not limited to, β-lactam drugs, quinolones, tetracyclines, erythromycin, streptomycin sulfate, salicylic acid, and benzoyl peroxide.

[0155] Examples of particulate materials include metal oxides.

[0156] Examples of anti-cellulite agents include xanthine agents.

[0157] Examples of tanning actives include 1,3-dihydroxy-2-propanone and the tanning actives disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667.

[0158] Examples of local anesthetics include benzocaine, lidocaine, and bupivacaine, as well as the local anesthetics disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667.

[0159] Examples of skin lightening agents include any agent known in the art, such as kojic acid, ascorbic acid, and the skin lightening agents disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0160] Examples of sunscreen actives include any suitable organic or inorganic sunscreen actives. Examples include metal oxides, 2-ethylhexyl p-methoxycinnamate, and the sunscreen actives disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0161] Examples of skin healing agents include pantheonic acid, as disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667.

[0162] Examples of anti-inflammatory agents include any agent that enhances the appearance, tone, or color of the skin, including, but not limited to, corticosteroids, hydrocortisone, nonsteroidal agents such as ibuprofen and aspirin, and the anti-inflammatory agents disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0163] Examples of flavonoids include flavanones, methoxyflavonones, unsubstituted chalcones, and mixtures thereof, as well as the flavonoids disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667.

[0164] Examples of enzymes include lipase, protease, catalase, superoxide dismutase, amylase, peroxidase, glucuronidase, ceramidase, hyaluronidase. Examples of enzyme inhibitors include trypsin inhibitor, Bowman-Birk inhibitor, chymotrypsin inhibitor, plant extract, flavonoid, quercetin chalcone, and enzymes disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667, and mixtures thereof. Examples of enzyme activators include coenzyme A, coenzyme Q10 (ubiquinone), glycyrrhizin, berberine, chrysin, and enzyme activators disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667, and mixtures thereof.

[0165] Examples of anti-wrinkle or anti-atrophy actives include sulfur-containing D- and L-amino acids, particularly N-acyl derivatives such as N-acetyl-L-cysteine, hydroxyl acids, phytic acid, lipoic acid, lysophosphatidic acid, skin exfoliants, vitamin B3, retinoids, and the anti-wrinkle or anti-atrophy actives disclosed in U.S. Patent Application Publication No. 2014 / 0120131 or U.S. Patent Application Publication No. 2004 / 0132667, and mixtures thereof.

[0166] The antioxidant / radical scavenger can be any agent useful for providing protection against UV radiation or other environmental agents that may cause skin damage, such as the antioxidants / radical scavengers disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667. Examples of antioxidants / radical scavengers include ascorbic acid, its salts and derivatives (vitamin C), tocopherol, its salts and derivatives (vitamin E), butylated hydroxylbenzoic acid and its salts, peroxides, gallic acid and alkyl esters, sorbic acid, lipoic acid, amines, lysine pidolate, arginine pyrrolate, nordihydroguaiaretic acid, bioflavonoids, curcumin, lysine, methionine, proline, superoxide dismutase, sirmarin, tea extract, and mixtures thereof.

[0167] Examples of chelating agents include EDTA, NTA, hydroxamic acid, phytic acid, lactoferrin, and chelating agents disclosed in US Patent Application Publication No. 2014 / 0120131 or US Patent Application Publication No. 2004 / 0132667, and mixtures thereof. Chelating agents refer to agents that can remove metal ions by forming complexes so that the metal ions cannot participate in or catalyze chemical reactions. Chelating agents are useful for protection against UV radiation or other environmental agents that can cause skin damage.

[0168] It will be understood that multiple additional components may be added. The amount of the additional component may be about 0.001% to about 50% by weight of the composition, preferably about 0.01% to about 20% by weight of the composition, preferably about 0.1% to about 10%, about 0.5% to about 10%, about 1% to about 5%, preferably 2%. The amount of the additional component depends on a number of factors, including the type of additional component used, the nature of the additional component, the components of the topical composition, the amount of the active ingredient or peptide in the topical composition, and / or the intended use of the topical composition. The nature and amount of any additional component should not unacceptably alter the benefits of the peptide of the present invention.

[0169] The topical composition can be alcohol-free.

[0170] In some embodiments of the present invention, the composition further comprises one or more additional active agents in addition to the peptide of the present invention (also known as the active ingredient of the composition). In addition, or alternatively, the composition can be administered with one or more other additional active agents. Typically, the additional active agents are present only in trace amounts. In some embodiments, there may be no additional active agents present in the composition. The amount of additional active agents incorporated depends on a number of factors, including the type of additional active agent used, the nature of the additional active agent, the ingredients of the topical composition, the amount of active ingredient or peptide in the topical composition, and / or the intended use of the topical composition. The nature and amount of any additional active agent should not unacceptably alter the benefits of the peptide of the present invention.

[0171] It should be understood that what is considered to be an "active" ingredient in one product may be a "functional" or "excipient" ingredient in another product, and vice versa. It will also be understood that some ingredients play a dual role as both an active ingredient and a functional or excipient ingredient.

[0172] Examples of additional active agents include glucose transport promoters, skin supplements, agents for skin treatment and / or care, anti-inflammatory agents, anti-aging agents, anabolic agents, cell growth promoters, and pharmacological enhancers.It will be understood that such agents are well known in the art, and any suitable additional active agent can be used.Additional active agents for skin treatment and / or care can include collagen synthesis agents, retinoids, exfoliants, anti-cellulite agents, elastase inhibitors, melanin synthesis stimulators or inhibitors, self-tanning agents, anti-aging agents, antimicrobial agents, antifungal agents, fungistatic agents, bactericidal agents, and healing agents.Active agents also include anti-inflammatory agents.

[0173] Any additional active agent should be suitable for application to the skin without undue toxicity, incompatibility, and / or allergic reaction. It will be understood that the classifications made herein are for clarity and convenience only and are not intended to limit the additional ingredients, excipients, or active ingredients to the particular applications or types listed.

[0174] In a particularly preferred embodiment, the method and use of the present invention involves administration of the peptide or composition of the present invention in combination with one or more other active agents (e.g., commercially available, existing growth-promoting or pharmacological enhancers). In such cases, the compound of the present invention can be administered sequentially, simultaneously, or sequentially with one or more other active agents.

[0175] In an embodiment, the effect of the present invention is achieved by topical application or topical administration of the topical composition of the present invention described herein to a person, animal, or patient in need of treatment or care.Topical delivery preferably refers to delivery to keratin layers such as skin, hair, and / or nails, but can also refer to delivery to body cavities covered with epithelial cells (e.g., lungs or airways, digestive tract, oral cavity).The effect can be limited to the surface of the skin or can be intradermal, or a combination of both.

[0176] The topical composition of the present invention is administered in a cosmetically or pharmacologic effective amount. In other words, it is administered in a non-toxic but sufficient amount to produce the desired effect. It will be understood that a person skilled in the art can determine the appropriate dose for administering the topical composition of the present invention without undue experimentation. Alternatively, a physician will determine the actual dose that is most suitable for a patient depending on the specific condition, disease, or disorder being treated or cared for, and the age, weight, and / or health of the person. It will depend on a variety of factors, including the activity, metabolic stability, and length of action of the specific compound utilized, the age, weight, general health, sex, diet, mode and frequency of administration, excretion rate, drug combination, the severity of the specific condition, and the individual being treated. Of course, there may be individual cases where higher or lower dosage ranges are appropriate, and such cases are also within the scope of the present invention. For example, the composition can be administered at a dose of 0.01 to 50 mg / kg body weight, more preferably 0.1 to 20 mg / kg body weight, more preferably 0.1 to 10 mg / kg body weight, preferably 0.1 to 5 mg / kg body weight. In an exemplary embodiment, one or more doses of 10 to 300 mg / day, or more preferably 10 to 150 mg / day, are administered to the patient. The amount and frequency are as best suited for the purpose. The frequency of application or administration can vary widely according to the needs of each subject, with a recommended application or administration range of once a month to 10 times a day, preferably once a week to 4 times a day, more preferably 3 times a week to 3 times a day, and even more preferably once or twice a day.

[0177] In a preferred embodiment, repeated applications of the composition are provided.

[0178] The topical composition can be applied to the keratinous tissue, skin, or body area to be treated or cared for by, but is not limited to, rubbing or rubbing. In some embodiments, the composition is left on the body area or is not removed from the body area. In other embodiments, the composition is removed after a time period, such as, but not limited to, about 2 minutes to about 60 minutes, about 5 minutes to about 30 minutes, preferably about 10 minutes to about 20 minutes. The composition can be removed immediately after application. In some embodiments of the present invention, the composition of the present invention can be applied to the area to be treated by, but is not limited to, any means to achieve greater penetration of the composition and / or peptide of the present invention, such as, but not limited to, iontophoresis, sonophoresis, electroporation, microcurrent patch, mechanical pressure, osmotic gradient, occlusion healing, microinjection, or needleless injection by pressure (such as injection by oxygen pressure), or any combination thereof.

[0179] The peptides or fragments of the present invention are used in the topical cosmetic or pharmaceutical compositions of the present invention at a cosmetically, pharma- ceutical or therapeutically effective concentration to achieve the desired effect; in a preferred embodiment, the concentration is between 0.00000001% (by weight) and 20% (by weight) based on the total weight of the composition; preferably, between 0.000001% (by weight) and 15% (by weight), more preferably, between 0.0001% (by weight) and 10% (by weight), and even more preferably, between 0.0001% (by weight) and 5% (by weight). Ideally, the peptides of the present invention are used in an amount of preferably about 0.00001% w / w to about 0.5% w / w, more preferably, 0.00005% w / w to about 0.05% w / w, and most preferably, about 0.0001% w / w to about 0.01% w / w of the composition. Ideally, the peptides of the present invention are used in an amount of from about 0.0001% w / w to about 0.004% w / w of the composition.

[0180] The compositions of the present invention can be administered individually or in combination with other pharmacologically active agents.It will be understood that such combination therapy includes different treatment regimes, including administration of multiple agents together in a single dosage form, or administration of multiple agents in separate, independent dosage forms, without any limitation.When the agents are present in different dosage forms, administration can be simultaneous or near simultaneous, or can follow any predetermined regimen, including administration of different agents.Suitable active agents can be as described herein.

[0181] In some embodiments of the invention, the compositions may be delivered via any one of liposomes, mixed liposomes, oleosomes, niosomes, ethosomes, millicapsules, capsules, macrocapsules, nanocapsules, nanostructured lipid carriers, sponges, cyclodextrins, vesicles, micelles, mixed surfactant micelles, mixed surfactant-phospholipid micelles, millispheres, spheres, lipospheres, particles, nanospheres, nanoparticles, milliparticles, solid nanoparticles, as well as microemulsions including water-in-oil microemulsions with reverse micelle internal structure, and nanoemulsion microspheres, microparticles.

[0182] A variety of methods are available for preparing liposomes, see, e.g., Szoka et al, Ann. Rev. Biophys. Bioeng. 9:467 (1980), U.S. Patent Nos. 4,186,183, 4,217,344, 4,235,871, 4,261,975, 4,485,054, 4,501,728, 4,774,085, 4,837,028, 4,235,871, 4,261,975, 4,485,054, 4,501,728, 4,774,085, 4,837,028, 4,946,787, PCT International Publication No. 91 / 17424, Deamer & Bangham, Biochim. Biophys. Acta 443:629-634 (1976);Fraley, et al, PNAS 76:3348-3352 (1979);Hope et al, Biochim. Biophys. Acta 812:55-65 (1985);Mayer et al, Biochim. Biophys. Acta 858:161-168 (1986);Williams et al, PNAS 85:242-246 (1988);Liposomes (Ostro (ed.), 1983, Chapter 1);Hope et al, Chem. Phys. Lip. 40:89 (1986);Gregoriadis, Liposome Technology (1984);and Lasic, Liposomes: from Physics to Applications (1993). Suitable methods include, for example, sonication, extrusion, high pressure / homogenization, microfluidization, detergent dialysis, calcium-induced small liposome vesicle fusion, and ether fusion methods, all of which are well known in the art.

[0183] These delivery systems can be adapted to achieve greater penetration of the compounds and / or peptides of the present invention. This can improve pharmacokinetic and pharmacodynamic properties. The delivery system can be a sustained release system that gradually releases the compounds or peptides of the present invention during a certain period of time, preferably at a certain release rate over a certain period of time. The delivery system is prepared by methods known in the art. The amount of peptide contained in the sustained release system depends on the location where the composition is delivered and the duration of release as well as the type of symptoms, diseases, and / or disorders that are treated or cared for.

[0184] The compound of the present invention can be administered by oral administration.The compound (and other ingredients, if desired) can also be enclosed in hard or soft gelatin capsules, compressed into tablets, or directly incorporated into the diet of a subject.For therapeutic oral administration, the compound can be incorporated with excipients and used in the form of ingestible tablets, lozenges, troches, capsules, elixirs, suspensions, syrups, wafers, etc.The compound can be coated with a material that prevents its inactivation, or the compound can be co-administered with it.

[0185] The compositions of the present invention may be a food or beverage product. In one embodiment, the artificial edible product is a sports nutrition product, such as a beverage, snack, or supplement. In one embodiment, the artificial edible product is a beverage. In one embodiment, the artificial edible product is a bakery product. In one embodiment, the artificial edible product is a dairy product. In one embodiment, the artificial edible product is a snack product. In one embodiment, the artificial edible product is a baked extruded product. In one embodiment, the artificial edible product is a milk powder. In one embodiment, the artificial edible product is an artificial nutritional dairy product. In one embodiment, the artificial edible product is a confectionery product. In one embodiment, the artificial edible product is a yogurt. In one embodiment, the artificial edible product is a yogurt drink. In one embodiment, the artificial edible product is an ice cream product. In one embodiment, the artificial edible product is a frozen food product. In one embodiment, the artificial edible product is a breakfast cereal. In one embodiment, the artificial edible product is a bread. In one embodiment, the artificial edible product is a flavored milk drink. In one embodiment, the artificial edible material is a candy bar. In one embodiment, the artificial edible material is tea or a tea product. In one embodiment, the artificial edible material is a based extruded snack product. In one embodiment, the artificial edible material is a fried snack product. In one embodiment, the artificial edible material is a dietary supplement. In one embodiment, the artificial edible material is a sports dietary supplement. In one embodiment, the artificial edible material is a baby food. In one embodiment, the artificial edible material is a specialized food for immune-compromised individuals. In one embodiment, the artificial edible material is a food for elderly patients.

[0186] In one embodiment, the composition is a plant food. In one embodiment, the composition is a cell culture medium. In one embodiment, the composition is an animal feed. In one embodiment, the composition is an animal feed supplement. In one embodiment, the composition is a medical food.

[0187] In embodiments, the composition of the invention can be administered parenterally (e.g., intravenously, subcutaneously, intraperitoneally, and / or intramuscularly). For example, it can be administered by intravenous infusion or injection, or by intramuscular or subcutaneous injection.

[0188] The compositions of the invention may be for human or animal use, in human and veterinary medicine.

[0189] The compositions of the present invention can be used for pharmaceutical, personal care, and / or cosmetic uses.

[0190] The compositions can be used to treat or care for any disease, disorder, or condition of the skin, including, but not limited to, psoriasis, dermatitis, allergic dermatitis, eczema, spongiosis, edema, skin cancer, ulcers, acne, scars, cellulitis, elastosis, keratosis, rosacea, esophageal varices, and inflammatory disorders.

[0191] The compositions can be used for the non-topical treatment of any skin disease, disorder, or condition, including, but not limited to, psoriasis, dermatitis, allergic dermatitis, eczema, spongiosis, edema, skin cancer, ulcers, acne, scars, cellulitis, elastosis, keratosis, rosacea, esophageal varices, inflammatory disorders.

[0192] The composition can be used for non-topical treatment of wounds in mammals.In another embodiment, the composition is for use in non-topical treatment or prevention of epidermal cell or tissue damage and / or disease or condition characterized by dermal or epidermal cell or tissue damage.The disease can be, but is not limited to, cancer and trauma.

[0193] The composition can be used to treat or care for visible signs of aging, including but not limited to wrinkles, stretch marks and dark circles under the eyes, dryness, fine lines, age spots, red spots, sagging skin, and conditions caused by sun exposure, stress, pollution, and diet, including sunburn.Topical compositions can also be used to delay, delay, or inhibit the onset of skin aging.The composition can be administered by medical devices such as plasters or patches as described herein.

[0194] The composition can be used to treat or care for wounds in mammals.In another embodiment, the topical composition is a topical composition for use in treating or preventing epidermal cell or tissue damage and / or disease or condition characterized by dermal or epidermal cell or tissue damage.Diseases can be, but are not limited to, cancer and trauma.

[0195] The compositions can be used to treat or care for any muscle condition, such as to improve muscle condition in a mammal, promote muscle recovery, typically after physical exercise, maintain or restore muscle health (e.g., lean tissue mass) in a mammal, and enhance physical performance, in the treatment or prevention of diseases or conditions characterized by lethargy or low energy levels.

[0196] The compositions can be used to increase or stimulate muscle growth activity.

[0197] The composition can be used to promote tissue growth, promote epithelial tissue growth, promote skin growth, promote organ growth, promote organism growth, etc. The skin can have a normal pathology and / or an abnormal pathology.

[0198] The composition can also be used to treat or care for any inflammatory disorder. In one embodiment, the inflammatory disorder is an inflammatory disorder of the joints. In one embodiment, the inflammatory disorder is an inflammatory disorder of the cardiovascular system. In one embodiment, the inflammatory disorder is an autoimmune disease. In one embodiment, the inflammatory disorder is a pulmonary inflammatory disorder and an airway inflammatory disorder. In one embodiment, the inflammatory disorder is an intestinal inflammatory disorder. In one embodiment, the inflammatory disorder is dermatitis. In one embodiment, the inflammatory disorder is acne vulgaris. In one embodiment, the inflammatory disorder is psoriasis. In one embodiment, the inflammatory disorder is rheumatoid arthritis. In one embodiment, the inflammatory disorder is a cardiovascular disease. In one embodiment, the inflammatory disorder is atherosclerosis. In one embodiment, the inflammatory disorder is type I diabetes.

[0199] In one embodiment, the inflammatory disorder is Graves' disease. In one embodiment, the inflammatory disorder is Guillain-Barre disease. In one embodiment, the inflammatory disorder is lupus. In one embodiment, the inflammatory disorder is psoriatic arthritis. In one embodiment, the inflammatory disorder is ulcerative colitis. In one embodiment, the inflammatory disorder is asthma. In one embodiment, the inflammatory disorder is cystic fibrosis. In one embodiment, the inflammatory disorder is COPD. In one embodiment, the inflammatory disorder is emphysema. In one embodiment, the inflammatory disorder is acute respiratory distress syndrome. In one embodiment, the inflammatory disorder is colitis. In one embodiment, the inflammatory disorder is inflammatory bowel disease.

[0200] The compositions can also be used to treat or care for metabolic disorders. In one embodiment, the metabolic disorder is prediabetes. In one embodiment, the metabolic disorder is diabetes. In one embodiment, the metabolic disorder is type 1 diabetes. In one embodiment, the metabolic disorder is type 2 diabetes. In one embodiment, the metabolic disorder is metabolic syndrome. In one embodiment, the metabolic disorder is obesity. In one embodiment, the metabolic disorder is diabetic dyslipidemia. In one embodiment, the metabolic disorder is hyperlipidemia. In one embodiment, the metabolic disorder is hypertension. In one embodiment, the metabolic disorder is hypertriglyceridemia. In one embodiment, the metabolic disorder is hyperfatty acidemia. In one embodiment, the metabolic disorder is hypercholesterolemia. In one embodiment, the metabolic disorder is hyperinsulinemia. In one embodiment, the metabolic disorder is MODY. The compositions of the present invention can also be used in methods for treating patients suffering from disease symptoms associated with or caused by hyperinsulinemia, hypoglycemia, hypokalemia, and / or hypophosphatemia. The present invention further relates to a method for treating blood glucose-related disease or disorder, comprising administering insulin derivative or insulin conjugate. Blood glucose-related disease or disorder includes type I and type II diabetes, and gestational diabetes. Cystic fibrosis, polycystic ovarian syndrome, pancreatitis, and other pancreas-related diseases can also be treated.

[0201] In an embodiment of the invention, the composition is for use in maintaining or restoring gastrointestinal health.

[0202] In an embodiment of the invention, the composition is for use in a method for the treatment or prevention of localized pain.

[0203] The compositions have uses as personal care products, supplements, cosmetics and pharmaceuticals.

[0204] Also provided is a method of treating, preventing or caring for any one of the diseases, disorders or symptoms described herein, comprising administering the composition or topical composition of the present invention.The composition can be administered to skin, hair, nails.The composition can be administered at any dose or frequency, by any method of administration or any method of topical application disclosed herein.

[0205] In one embodiment, the composition is a cosmetic composition. In one embodiment, the composition is a pharmaceutical composition. It will be appreciated that a composition may have dual functionality and may be both a cosmetic composition and a pharmaceutical composition.

[0206] It will be appreciated that the composition may be a therapeutic composition or a non-therapeutic composition. The composition may have a dual role.

[0207] Compositions may be formulated in unit dosage form, ie, in the form of discrete portions containing a unit dose, or a multiple or sub-unit of a unit dose.

[0208] In a particularly preferred embodiment, the method and use of the present invention involves the administration of the peptide or composition of the present invention in combination with one or more other active agents that are commercially available. In such cases, the compound of the present invention can be administered sequentially, simultaneously, or sequentially with one or more other active agents.

[0209] Modified Peptides In one embodiment, the peptides of the present invention (including peptide fragments and peptide variants) may be modified peptides. The term "modified peptide" is used interchangeably with the term peptide derivative. In one embodiment, the term "modified peptide" refers to a peptide that has been modified to exhibit one or more of the following properties compared to the unmodified peptide: increased plasma half-life; increased lipophilicity of the peptide; increased renal clearance of the modified peptide; increased activity of the modified peptide, and increased resistance of the modified peptide to proteolysis (i.e., by mammalian, particularly human, gastrointestinal proteases). Disclosed herein are various methods of modifying the peptides of the present invention to exhibit these properties, including conjugating the peptide to a binding partner (e.g., albumin that binds small molecules, large polymers, long-lived plasma proteins, or antibodies or antibody fragments), cyclization, addition of N-terminal or C-terminal or side chain protecting groups, replacement of one or more L-amino acids with D-isomers, amino acid modification, increased plasma protein binding, increased albumin binding. Modified peptides include, but are not limited to, peptides that are substituted with one or more groups, or conjugated with binding partners, or cyclized, as defined herein.Generally, peptides are modified to extend their half-life in animals in vivo.Various modification methods are presented below.

[0210] In one embodiment, the modification may be any modification that provides the peptide and / or composition of the present invention with an increased ability to penetrate cell membranes. In one embodiment, the modification may be any modification that extends the half-life of the composition or peptide of the present invention. In one embodiment, the modification may be any modification that increases the activity of the composition or peptide of the present invention. In one embodiment, the modification may be any modification that increases the selectivity of the composition or peptide of the present invention.

[0211] In one embodiment, the group is a protecting group. The protecting group can be an N-terminal protecting group, a C-terminal protecting group, or a side chain protecting group. The peptide can have one or more of these protecting groups.

[0212] Those skilled in the art are aware of suitable techniques for reacting amino acids with these protecting groups. These groups can be added by preparation methods known in the art, for example, the methods outlined in US Patent Publication No. 2014 / 0120141, paragraphs

[0104] to

[0107] . The groups can remain on the peptide or can be removed. The protecting groups can be added during synthesis.

[0213] In an embodiment of the invention, the peptides may be substituted with one or more groups selected from linear or branched, long or short chain, saturated or unsaturated, having 1 to 29 carbon atoms, substituted with hydroxyl, amino, aminoacyl, sulfate, or sulfide groups, or unsubstituted groups. N-acyl derivatives include acyl groups derived from acetic acid, capric acid, lauric acid, myristic acid, octanoic acid, palmitic acid, stearic acid, behenic acid, linoleic acid, linolenic acid, lipoic acid, oleic acid, isostearic acid, elaidic acid, 2-ethylhexanoic acid, coconut oil fatty acid, tallow fatty acid, hardened tallow fatty acid, palm kernel fatty acid, lanolin fatty acid, or similar acids. These may be substituted or unsubstituted. If substituted, they are preferably substituted with hydroxyl, or sulfur-containing groups such as, but not limited to, SO3H, SH, or SS.

[0214] In an embodiment of the invention, the peptide is R1-X-R2.

[0215] The R1 and / or R2 groups are attached to the amino-terminus (N-terminus) and carboxyl-terminus (C-terminus) of the peptide sequence, respectively.

[0216] In one embodiment, the peptide is R1-X. Alternatively, the peptide is X-R2.

[0217] Preferably, R1 is H, C 1~4 is alkyl, acetyl, benzoyl, or trifluoroacetyl; X is a peptide of the present invention; and R2 is OH or NH2.

[0218] In an embodiment, R1 is selected from H, acyclic substituted or unsubstituted aliphatic groups, substituted or unsubstituted cycloaliphatic groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, tert-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (Fmoc), and the group formed by R5-CO-, wherein R5 is selected from the group formed by H, acyclic substituted or unsubstituted aliphatic groups, substituted or unsubstituted cycloaliphatic groups, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, substituted or unsubstituted heterocyclic groups, and substituted or unsubstituted heteroarylalkyl; R2 is selected from the group formed by -NR3R4, -OR3, and -SR3, where R3 and R4 are independently selected from the group formed by H, an acyclic substituted or unsubstituted aliphatic group, a substituted or unsubstituted cycloaliphatic group, a substituted or unsubstituted heterocyclic group, a substituted or unsubstituted heteroarylalkyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted aralkyl; and provided that R1 and R2 are not an α-amino acid.

[0219] According to another preferred embodiment, R2 is -NR3R4, -OR3 or -SR3, wherein R3 and R4 are independently H, substituted or unsubstituted C1-C 24 Alkyl, substituted or unsubstituted C2-C 24 Alkenyl, tert-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (Fmoc), substituted or unsubstituted C2-C 24 Alkynyl, substituted or unsubstituted C3-C 24 Cycloalkyl, substituted or unsubstituted C5-C 24 Cycloalkenyl, substituted or unsubstituted C5-C 24Cycloalkynyl, substituted or unsubstituted C6-C 30 Aryl, substituted or unsubstituted C7-C 24 aralkyl, 3-10 membered substituted or unsubstituted heterocycle, and substituted or unsubstituted heteroarylalkyl having 2-24 carbon atoms and 1-3 atoms other than carbon, the alkyl chain being formed by heteroarylalkyl having 1-6 carbon atoms. Optionally, R3 and R4 can be linked by a saturated or unsaturated carbon-carbon bond to form a ring with the nitrogen atom. More preferably, R2 is -NR3R4 or -OR3, where R3 and R4 are independently H, substituted or unsubstituted C1-C 24 Alkyl, substituted or unsubstituted C2-C 24 Alkenyl, substituted or unsubstituted C2-C 24 Alkynyl, substituted or unsubstituted C3-C 10 Cycloalkyl, substituted or unsubstituted C6-C 15 aryl, a substituted or unsubstituted heterocyclic group having 3 to 10 members, and a substituted or unsubstituted heteroarylalkyl group with a 3 to 10 membered ring and an alkyl chain having 1 to 6 carbon atoms. More preferably, R3 and R4 are selected from the group formed by H, methyl, ethyl, hexyl, dodecyl, or hexadecyl. Even more preferably, R3 is H and R4 is selected from the group formed by H, methyl, ethyl, hexyl, dodecyl, or hexadecyl. According to an even more preferred embodiment, R2 is selected from -OH and -NH2.

[0220] According to another embodiment of the present invention, R1 is selected from the group formed by H, acetyl, lauroyl, myristoyl, or palmitoyl, and R2 is -NR3R4 or -OR3, where R3 and R4 are independently selected from H, methyl, ethyl, hexyl, dodecyl, and hexadecyl, and preferably R2 is -OH or -NH2. More preferably, R1 is acetyl or palmitoyl, and R2 is -NH2.

[0221] In a preferred embodiment, an acyl (or acetyl) group is attached to the N-terminus of at least one amino acid of the peptide.

[0222] In an embodiment of the invention, the peptide is modified to include a side chain protecting group. The side chain protecting group may be one or more of the following: benzyl or benzyl-based groups, t-butyl-based groups, benzyloxycarbonyl (Z) groups, and allyloxycarbonyl (alloc) protecting groups. The side chain protecting group may be derived from an achiral amino acid, such as achiral glycine. The use of an achiral amino acid helps to stabilize the resulting peptide and also facilitates the facile synthetic route of the invention. Preferably, the peptide further includes a modified C-terminus, preferably an amidated C-terminus. The achiral residue may be α-aminoisobutyric acid (methylalanine). It will be understood that the specific side chain protecting group used will depend on the sequence of the peptide and the type of N-terminal protecting group used. In one embodiment of the invention, the peptide is conjugated, linked, or fused to one or more polyethylene glycol polymers or other compounds, such as compounds that increase molecular weight. The molecular weight increasing compound is any compound that increases the molecular weight, typically by 10% to 90%, or 20% to 50%, of the resulting conjugate and may have a molecular weight between 200 and 20,000, preferably between 500 and 10,000. The molecular weight increasing compound may be PEG, any water-soluble (amphiphilic or hydrophilic) polymeric moiety, homopolymer or copolymer of PEG, monomethyl-substituted polymer of PEG (mPEG), and polyoxyethylene glycerol (POG), polyamino acids such as polylysine, polyglutamic acid, polyaspartic acid (especially those with L-configuration), pharmacologically inactive proteins such as albumin, gelatin, fatty acids, polysaccharides, lipids, amino acids, and dextran. The polymeric moiety may be linear or branched and may have a molecular weight of 500 to 40000 Da, 5000 to 10000 Da, 10000 to 5000 Da. The compound can be any suitable cell membrane permeable compound, such as tat peptide, penetratin, pep-1, etc. The compound can be an antibody molecule. The compound can be a lipophilic moiety or a polymeric moiety.

[0223] Lipophilic and polymeric substituents are known in the art. Lipophilic substituents include acyl groups, sulfonyl groups, N-, O-, or S-atoms, which form part of esters, sulfonyl esters, thioesters, amides, or sulfonamides. Lipophilic moieties can include a hydrocarbon chain with 4 to 30 C-atoms, preferably 8 to 12 C-atoms. It can be linear or branched, saturated or unsaturated. The hydrocarbon chain can be further substituted. It can be a cycloalkane or heterocycloalkane. The peptide can be modified at the N-terminus, C-terminus, or both. The polymer or compound is preferably linked to an amino, carboxyl, or thio group, and can be linked by the N-terminus or C-terminus of the side chain of any amino acid residue. The polymer or compound can be conjugated to the side chain of any suitable residue.

[0224] The polymer or compound can be conjugated via a spacer, which can be a natural or unnatural amino acid, succinic acid, lysyl, glutamyl, asparagyl, glycyl, β-alanyl, γ-aminobutanoyl.

[0225] The polymer or compound can be conjugated via an ester, sulfonyl ester, thioester, amide, carbamate, urea, sulfonamide.

[0226] Those skilled in the art are aware of suitable means for preparing the described conjugates.

[0227] Peptides can be chemically modified by covalent conjugation to polymers, for example, to extend their circulatory half-life.The exemplary polymers and methods for conjugating such polymers to peptides are illustrated in, for example, U.S. Patent Nos. 4,766,106; 4,179,337; 4,495,285; and 4,609,546.Further exemplary polymers include polyoxyethylated polyols and polyethylene glycol (PEG) moieties.

[0228] The peptides of the present invention can be modified at one or more sites to manipulate the storage stability, pharmacokinetics, and / or any aspect of the peptide's bioactivity (e.g., potency, selectivity, drug interactions, etc.). Chemical modifications of the peptides include, but are not limited to, conjugation of the peptide to one or more of polyethylene glycol (PEG), monomethoxypolyethylene glycol, dextran, poly(N-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymer, poly(propylene oxide / ethylene oxide) copolymer, polypropylene glycol, polyoxyethylated polyols (e.g., glycerol) and polyvinyl alcohol, colominic acid, or other carbohydrate-based polymers, polymers of amino acids, and biotin derivatives. PEG conjugation of proteins at Cys residues is disclosed, for example, in Goodson, RJ & Katre, NV (1990) Bio / Technology 8, 343; and Kogan, TP (1992) Synthetic Comm. 22, 2417.

[0229] Modified peptides may also include sequences in which one or more residues have been modified (i.e., by phosphorylation, sulfation, acylation, amidation, PEGylation, etc.), and mutants that contain one or more modified residues relative to the parent sequence. Amino acid sequences may also be modified with a label capable of directly or indirectly producing a detectable signal, including, but not limited to, radioisotope labels, fluorescent labels, and enzyme labels. Fluorescent labels include, for example, Cy3, Cy5, Alexa, BODIPY, fluorescein (e.g., FluorX, DTAF, and FITC), rhodamine (e.g., TRITC), auramine, Texas Red, AMCA blue, and Lucifer Yellow. Preferred isotopic labels include: 3 H, 14 C. 32 P, 35 S, 36 Cl, 51 Cr, 57 Co, 58 Co, 59 Fe, 90 Y, 125 I, 131 I, and 286Re. Preferred enzyme labels include peroxidase, β-glucuronidase, β-D-glucosidase, β-D-galactosidase, urease, glucose oxidase + peroxidase, and alkaline phosphatase (see, e.g., U.S. Pat. Nos. 3,654,090; 3,850,752; and 4,016,043). Enzymes can be conjugated by reactions involving molecules such as carbodiimides, diisocyanates, glutaraldehyde, and the like. Enzyme labels can be detected visually or can be measured by calorimetry, spectrophotometry, fluorescence spectroscopy, amperometry, or gasometry. Other labeling systems, such as avidin / biotin, Tyramide Signal Amplification (TSA™), etc., are known in the art and are commercially available (see, e.g., ABC kit (Vector Laboratories, Inc., Burlingame, Calif.); NEN® (Life Science Products, Inc., Boston, Mass.)).

[0230] In embodiments, the peptide, variant, and / or composition is modified to increase drug efficacy. In embodiments, the peptide, variant, and / or composition is modified to increase stability, permeability, maintain efficacy, avoid toxicity, and / or extend half-life. Modification can be as described above. For example, modification can be N-terminus and C-terminus protection, which can be amino acid modification, cyclization, amino acid replacement, and / or conjugation with macromolecular or large polymer or long-lived plasma protein. Strategies to extend half-life are as described by Strohl, et al (BioDrugs, 2015), Schlapschy, et al (Protein Eng Des Sel. 2013), Podust, VN, et al (Protein Eng Des Sel. 2013), Zhang, L et al (Curr Med Chem. 2012), Gaberc-Porekar, V, et al (Curr Opin Drug Discov Devel. 2008). Examples include the use of PEGylation, lipidation (covalent attachment of fatty acids to peptide side chains), fusion to Fc domains and human serum albumin, fusion with hydrophilic amino acid polymers (e.g., XTEN or PAS), and / or fusion with half-life extending proteins.

[0231] Peptides or proteins may contain fragile sites within their sequences that are susceptible to proteolysis when in an environment of enhanced proteolysis, e.g., in the blood or in the gastrointestinal tract. In embodiments, the peptides, variants, and / or compositions contain modifications of one or more fragile sites such that the peptides, variants, and / or compositions are not subject to proteolysis / cleavage or are subject to a reduced amount of proteolysis / cleavage compared to the unmodified peptide or protein. Thus, peptides can be modified to increase the resistance of the modified peptide to proteolysis by mammalian gastrointestinal proteases. Suitable modifications are described in Diao et al (Clinical pharmacokinetics 52. 10 (2013): 855-868).

[0232] Peptide modifications that extend the in vivo half-life of peptides are described in the literature, for example, Strategies to improve plasma half life time of peptide and protein drugs. Werle M, Bernkop-Schnurch A. Amino Acids. 2006 Jun; 30(4): 351-67.

[0233] Due to the obvious advantages of long-acting peptide and protein drugs, there is a great demand for strategies to extend the plasma half-life of such compounds. Short plasma half-life times are generally due to rapid renal clearance as well as enzymatic degradation occurring during systemic circulation. Modifications of peptides / proteins can result in an extension of the plasma half-life time. By shortening the total amino acid content of somatostatin and replacing L-analogue amino acids with D-amino acids, the plasma half-life time of the derivative octreotide was 1.5 hours compared to only a few minutes for somatostatin. The PEG (2.40K) conjugate of INF-α-2b exhibited a 330-fold extended plasma half-life compared to the native protein. It was the aim of this review to present an overview of possible strategies to extend plasma half-life such as N- and C-terminal modifications or PEGylation, as well as methods to assess the efficacy of drug modifications. Furthermore, basic data on the most important proteolytic enzymes in human blood, liver, and kidney, as well as their cleavage specificities and inhibitors against them, are presented to predict the enzymatic cleavage of peptide and protein drugs during systemic circulation. Strategic Approaches to Optimizing Peptide ADME Properties. Li Di AAPS J. 2015 Jan; 17(1): 134-143

[0234] Strategies to stabilize peptides against proteolysis Many methods are available for enhancing peptide stability through structural modification.Some methods not only improve stability, but also enhance other ADME properties, for example, cyclization can increase stability and permeability, and conjugation with polymer can improve stability and reduce renal clearance.It is important to improve stability and ADME properties while maintaining peptide efficacy and avoiding toxicity.

[0235] ·N- and C-terminal protection Many proteolytic enzymes in blood / plasma, liver or kidney are exopeptidases, aminopeptidases and carboxypeptidases, which degrade peptide sequences from the N-terminus and C-terminus.Modification of N-terminus or / and C-terminus can often improve peptide stability.Many examples have reported that N-acetylation and C-amidation increase resistance to proteolysis.

[0236] Substitution of L-amino acids with D-amino acids Substitution of natural L-amino acids with natural D-amino acids reduces substrate recognition and binding affinity and increases stability of proteolytic enzymes. One example is vasopressin, which contains L-Arg and has a half-life of 10-35 minutes in humans. The D-Arg analog desmopressin has a half-life of 3.7 hours in healthy human volunteers. In a study of bicyclic peptide inhibitors of urokinase-type plasminogen activator (uPA), a cancer-associated protease, substitution of certain glycines with D-serines not only improved potency by 1.8-fold, but also increased stability in mouse plasma by 4-fold.

[0237] Amino acid modification Modifications of natural amino acids can improve peptide stability by introducing steric hindrance or disrupting enzyme recognition. For example, gonadotropin releasing hormone has an extremely short half-life (minutes), but buserelin, in which one Gly is replaced by t-butyl-D-Ser and another by an ethylamide, has a much longer half-life in humans.

[0238] ·Cyclization Cyclization introduces conformational constraints, reduces peptide flexibility, and increases stability and permeability. Depending on the functional groups, peptides can be cyclized head-to-tail, head / tail-to-side chain, or side chain-to-side chain. Cyclization is generally achieved by lactamization, lactonization, and sulfide-based bridging. Disulfide bridges create folding and conformational constraints that can improve potency, selectivity, and stability. Many disulfide bond-rich peptides, such as linaclotide, lepirudin, and ziconotide, are commercially available or in preclinical or clinical development.

[0239] Conjugation with polymers Conjugation with macromolecules (eg, polyethylene glycol (PEG), albumin) is an effective strategy to improve peptide stability and reduce renal clearance.

[0240] Renal clearance Many peptides exhibit promising in vitro pharmacological activity but fail to demonstrate efficacy in vivo due to extremely short in vivo half-lives (minutes). The rapid clearance and short half-life of peptides hinder their successful development into drugs. The major causes of rapid clearance of peptides from the systemic circulation are enzymatic proteolysis or / and renal clearance. Glomeruli have a pore size of approximately 8 nm, and hydrophilic peptides with MW < 2-25 kDa are subject to rapid filtration through the renal glomerulus. Because peptides are not easily reabsorbed by the renal tubules, peptides often have large renal clearance and short half-lives. Other minor pathways of peptide clearance are endocytosis, and degradation by the proteasome and liver. Comparison of systemic and renal clearance in animal models provides useful information on whether renal clearance is the major elimination route.

[0241] Inappropriate dosing in patients with renal insufficiency may cause toxicity or therapeutic failure, so dose adjustments may be required in patients with renal impairment to avoid accumulation of peptide drugs and high drug exposure. Several strategies have been developed to reduce peptide renal clearance and extend half-life. These are reviewed below.

[0242] Increases plasma protein binding Renal clearance of peptides is reduced when they are bound to membrane or serum proteins. An example is the cyclic peptide drug octreotide, a treatment for endocrine tumors, which has a half-life of about 100 minutes in humans (unbound fraction: 0.65 minutes) due to binding to lipoproteins.

[0243] Covalent linkage to albumin-binding small molecules Covalent conjugation of albumin-binding small molecules to peptides can reduce glomerular filtration, improve proteolytic stability, and extend half-life by indirectly interacting with albumin through the highly conjugated small molecule.

[0244] Conjugation with large polymers Conjugation of peptides to large synthetic or natural polymers or carbohydrates can increase their molecular weight and hydrodynamic capacity, thereby reducing their renal clearance. Common polymers used for peptide conjugation are PEG, polysialic acid (PSA), and hydroxyethyl starch (HES).

[0245] Fusion with long-lived plasma proteins Plasma proteins such as albumin and immunoglobulin (IgG) fragments have a long half-life of 19-21 days in humans. Due to their high MW (67-150 kDa), these proteins have little renal clearance, and their binding to the neonatal Fc receptor (FcRn) reduces excretion via pinocytosis by the vascular epithelium. Covalent linkage of peptides to albumin or IgG fragments can reduce renal clearance and extend half-life. Fusion Proteins for Half-Life Extension of Biologies as a Strategy to Make Biobetters William R. Strohl BioDrugs. 2015; 29(4): 215-239 Schlapschy, M, Binder, U, Borger, C et al. PASYlation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins. Protein Eng Des Sel. 2013; 26(8): 489-501 Podust, VN, Sim, BC, Kothari, D et al. Extension of in vivo half-life of biologically active peptides via chemical conjugation to XTEN protein polymer. Protein Eng Des Sel. 2013; 26(11): 743-53 Zhang, L, Bulaj, G. Converting Peptides into Drug Leads by Lipidation. Curr Med Chem. 2012; 19(11): 1602-18 Gaberc-Porekar, V, Zore, I, Podobnik, B et al. Obstacles and pitfalls in the PEGylation of therapeutic proteins. Curr Opin Drug Discov Devel. 2008; 11(2): 242-50 By Dr Ronald V. Swanson-Long live peptides evolution of peptide half-life extension technologies and emerging hybrid approaches. From Drug Discovery World on line. Spring 2014

[0246] PEGylation PEGylation (the attachment of long chains of the hydrophilic polymer polyethylene glycol to molecules of interest) was originally thought to be a modification that would prevent recognition of foreign proteins by the immune system, thereby enabling their usefulness as therapeutic agents. Once formed, antibodies against the unmodified drug can rapidly neutralize and eliminate protein drugs. Unexpectedly, PEGylation improved the pharmacokinetics of proteins even in the absence of anti-drug antibodies. Simply by making the drug molecule larger, PEGylation resulted in drugs that were filtered more slowly by the kidney. The empirical observation that increasing the size or hydrodynamic radius then resulted in reduced renal clearance and extended half-life became the primary rationale for PEGylation of protein and peptide drugs. PEGylation can have a variety of effects on molecules, including increasing the water solubility of proteins or peptides and protecting them from degradation by proteolytic enzymes. PEGylation also affects the binding of therapeutic proteins to their cognate cellular receptors, usually reducing affinity. Varying the size, structure, and conjugation mode of the PEG polymer can affect the biological activity of the conjugated drug.

[0247] First generation PEGylation methods were fraught with challenges. However, the chemistry of PEGylation is quite simple. The process involves the covalent attachment of polyethylene glycol chains to reactive side chains of proteins or peptides. For example, PEG is easily attached to the amino groups of lysines on the surface of proteins or peptides. The reaction is pH dependent. At high pH (above 8.0), N-hydroxysuccinimide covalently attaches the lysine side chain amino groups to PEG. This method typically results in a family of products containing different numbers of PEG chains attached to different sites on the protein, rather than a single individual product. The first approved PEGylated pharmaceutical agents were bovine pegademase (PEGylated bovine adenosine deamidase), an enzyme replacement therapy for severe combined immunodeficiency, and pegaspargase (PEGylated asperginase) for the treatment of acute lymphoblastic leukemia. These drugs were complex mixtures of diverse PEGylated species, but with improved therapeutic properties over the native enzymes, including increased serum half-life of the proteins and reduced immunogenicity. Due to the inherent polydispersity of PEG, quality and batch-to-batch reproducibility was difficult. Despite this limitation, two PEGylated interferons (PegInterferon Alpha 2b and PegInterferon Alpha 2a), which are heterogeneous populations of multiple monoPEGylated positional isomers, have been approved by the FDA for the treatment of Hepatitis C. These drugs were marketed in 2001 and 2002, respectively.

[0248] Various enhancements and modifications have been made to the basic PEGylation technique. Second generation PEGylation processes have introduced the use of branched structures as well as alternative chemistries for PEG conjugation. In particular, PEG with cysteine ​​reactive groups, such as maleimide or iodoacetamide, allows for targeting of PEGylation to single residues within a peptide or protein, reducing the heterogeneity of the final product without eliminating heterogeneity due to the polydispersity of the PEG itself.

[0249] Although the original rationale for PEGylation was to reduce immunogenicity, there are also a few examples of immunogenic PEGylated proteins. One example is PEGylated urate oxidase, an enzyme that reduces plasma uric acid levels in gout patients. In clinical trials, a relatively high percentage of gout patients did not respond to treatment and developed antibodies specific to PEG but not to the uricase protein. Some studies have found that PEGylated liposomes, which are also generally considered to be non-immunogenic, are immunogenic. PEGylated liposomes induce a strong anti-PEG immunoglobulin M (IgM) response. In addition, multiple injections of PEG-glucuronidase have been shown to induce the development of specific anti-PEG IgM antibodies, thus accelerating the clearance of PEG-modified proteins from the body.

[0250] The major potential drawback of using PEG as a modifying agent is that it is non-biodegradable. The U.S. Food and Drug Administration (FDA) has approved PEG for use as a vehicle in pharmaceutical agents, including injectable, topical, rectal, and nasal formulations. PEG exhibits little toxicity and is excreted intact from the body by the kidneys (for PEGs <30 kDa) or feces (for PEGs >20 kDa). Repeated administration of some PEGylated proteins to animals has resulted in the observation of cellular vacuolation in the renal tubules. Choroid plexus epithelial cells have also been seen recently in toxicity studies of proteins conjugated with large (>40 kDa) PEG. Choroid plexus epithelial cells produce cerebrospinal fluid and form the blood-CSF barrier. The long-term negative consequences of cellular vacuolation are unknown, but represent an undesirable consequence for some potential therapeutic agents. One possible alternative would be the substitution of a biodegradable polymer for PEG. Polymers such as hydroxyethyl starch (HES) are possible alternatives. HES is non-toxic, biodegradable, and is used as a blood expander. The HES process functions similarly to PEGylation in reducing renal clearance by increasing the hydrodynamic radius of the peptide, but because of its biodegradability, it confers less of a tendency for accumulation. However, like PEG, HES and other proposed biodegradable polymers that are PEG alternatives are polydisperse, making characterization of the end products and metabolites difficult. One emerging solution that alleviates both concerns is the use of chemically defined polypeptides as polymer components, an approach that is discussed later in this paper.

[0251] Lipidization The second major chemical modification method to extend peptide half-life is lipidation, which involves the covalent attachment of fatty acids to the peptide side chains. Originally conceived and developed as a way to extend the half-life of insulin, lipidation shares the same basic mechanism of half-life extension as PEGylation, namely, increasing the hydrodynamic radius and reducing renal filtration. However, the lipid moiety itself is relatively small, and the effect is indirect, via the non-covalent attachment of the lipid moiety to circulating albumin. Albumin, a protein with a large size (67 KDa) and abundance in human serum (35-50 g / L), naturally functions to transport molecules (including lipids) throughout the body. As also seen with PEGylation, binding to plasma proteins protects the peptide from attack by peptidases through steric hindrance. One consequence of lipidation is that it reduces the water solubility of the peptide, but this effect can be tuned by manipulating the linker between the peptide and the fatty acid, for example, by the use of glutamic acid or mini-PEGs in the linker. Manipulation of the linker and modification of the lipid moiety can affect self-aggregation, which may contribute to increased half-life by slowing biodistribution independent of albumin.

[0252] After the pioneering work on insulin, lipidation of various peptides has been explored, especially in the field of diabetes, including human glucagon-like peptide 1 (GLP-1) analogs, glucose-dependent insulinotropic polypeptides, and GLP-1R / glucagon receptor coagonists. Currently, two lipidated peptide drugs are approved by the FDA for use in humans. These are the GLP-1 analog liraglutide and insulin detemir, both of which are long-acting antidiabetic agents.

[0253] A potentially pharmacologically significant difference between PEGylation and lipidation is that therapeutically active peptides are covalently linked to the much larger PEG, whereas small fatty acyl-peptide conjugates are non-covalently associated with the larger albumin, resulting in the existence of bound and unbound forms in equilibrium. This can lead to differences in biodistribution that can result in different pharmacology, because access to receptors localized in different tissues can induce differential effects. In some cases, a more restricted biodistribution may be desired, while in other cases, greater tissue penetration may be important. An interesting variant of the PEG method that addresses this issue, using PEG conjugates that can be released with predictable cleavage rates, has been developed by Santi et al.

[0254] Both PEGylation and lipidation directly or indirectly provide protection against proteases and peptidases by shielding through steric hindrance, and extend circulation half-life by directly or indirectly increasing hydrodynamic radius. Both methods are flexible in that they use chemical conjugation and are tolerant of the means used to generate the modified peptide, whether biologically or synthetically produced. The advantage of using synthetic peptides is that they can incorporate unnatural amino acids designed to address a number of unique problems, including instability due to known susceptibility to proteolytic cleavage. They can also be more flexible in terms of the choice of conjugation site, which is crucial when activity or potency is highly dependent on modified residues, such as free or C-terminal amides.

[0255] Classical gene fusions: Fc and HSA Classical genetic fusions with long-lived serum proteins provide an alternative method of half-life extension that differs from chemical conjugation with PEG or lipids. Two major proteins have been used as fusion partners: antibody Fc domains and human serum albumin (HAS) (Figure 2). Fc fusions involve the fusion of peptides, proteins, or receptor extracellular domains to the Fc portion of an antibody. Both Fc and albumin fusions achieve half-life extension not only by increasing the size of the peptide drug, but also by taking advantage of the body's natural circulatory mechanism: the neonatal Fc receptor, FcRn. The pH-dependent binding of these proteins to FcRn prevents degradation of the fusion protein in endosomes. Fusions based on these proteins can have half-lives ranging from 3 to 16 days, much longer than typical PEGylated or lipidated peptides. Fusion with antibody Fc can improve the solubility and stability of peptide or protein drugs. An example of a peptide Fc fusion is dulaglutide, a GLP-1 receptor agonist currently in late-stage clinical trials. Human serum albumin, the same protein utilized by fatty acid acylated peptides, is another common fusion partner. Albiglutide is a GLP-1 receptor agonist based on this platform. The main difference between Fc and albumin is the dimeric nature of Fc versus the monomeric structure of HAS, resulting in the expression of the fusion peptide as a dimer or monomer, depending on the choice of fusion partner. If the target receptors are spaced close enough together or are themselves dimeric, the dimeric nature of the peptide-Fc fusion may result in an avidity effect. This may or may not be desirable, depending on the target.

[0256] Designed Polypeptide Fusions: XTEN and PAS An interesting variation of the recombinant fusion concept has been the development of low-complexity sequences designed as fusion partners (Figure 1). The sequences are essentially unstructured, hydrophilic amino acid polymers that are functional analogues of PEG. The inherent biodegradability of the polypeptide platform makes it an attractive, potentially better alternative to PEG. Another advantage is the precise molecular structure of the recombinant molecule, in contrast to the polydispersity of PEG. Unlike HSA and Fc peptide fusions, which require maintaining the three-dimensional folding of the fusion partner, recombinant fusions with unstructured partners can often be subjected to harsh conditions such as high temperature or HPLC purification.

[0257] The most advanced of this class of polypeptides, termed XTEN (Amunix), is 864 amino acids long and contains six amino acids (A, E, G, P, S, and T) (Figure 1). Made possible by the biodegradable nature of the polymer, it is much larger than the typically used 40 KDa PEG, conferring a concomitant large half-life extension. Fusion of XTEN to peptides results in a 60- to 130-fold half-life extension over the native molecule. Two fully recombinantly produced XTENylated products, VRS-859 (exenatide-XTEN) and VRS-317 (human growth hormone-XTEN), are undergoing clinical trials. In a Phase Ia study, VRS-859 was found to be well tolerated and effective in patients with type 2 diabetes. VRS-317 has been reported to have superior pharmacokinetic and pharmacodynamic properties compared to previously studied rhGH products, with the potential for once-monthly dosing. A second polymer based on a similar conceptual study is PAS (XL-Protein GmbH). The random coil polymer was constructed from a more constrained set of only three small uncharged amino acids, proline, alanine, and serine (Figure 1). It is not yet known whether the differences in the biophysical properties of PAS and the highly negatively charged XTEN contribute to differences in biodistribution and / or in vivo activity. However, this will become clear as these polypeptides are incorporated into more therapeutic agents and the behavior of the fusions is characterized.

[0258] All peptide protein fusions, whether the partner is Fc, HSA, XTEN, or PAS, are genetically encoded and, as a result, suffer from similar limitations. One limitation is that only natural amino acids are incorporated, unlike methods that utilize chemical conjugation, which allow the use of synthetic peptides that incorporate unnatural amino acids. Methods that overcome this by expanding the genetic code are being developed by companies such as Ambrx or Sutro, but these are not widely used. A second limitation is that the N- or C-terminus of the peptide must be fused to the partner. Often, the peptide termini are involved in receptor interactions, and fusion to one or both termini can significantly impair activity. The site of PEG or lipid conjugation can be anywhere on the peptide, and can be optimized to maximize the biological activity of the resulting therapeutic.

[0259] A hybrid method for integrating synthetic peptides with half-life extending proteins Although gene fusions have historically offered the potential for greater half-life extension, they lack the advantages offered by methods using chemical conjugation (PEGylation and lipidation) in terms of the flexibility of attachment sites and incorporation of non-natural amino acids or modifications to the peptide backbone. One of the first efforts to integrate the advantages of gene fusions with chemical conjugation for half-life extension was carried out by researchers at the Scripps Research Institute in La Jolla, a technology that later formed the platform for the biotech company CovX. Using a catalytic aldolase antibody, these researchers developed a platform in which the active site lysine of the antibody forms a reversible covalent enamine bond with a β-diketone incorporated into a peptide or small molecule. The resulting conjugate is called CovXBody™. This approach combines the functional qualities of peptide drugs or small molecules with the long serum half-life of antibodies through chemical linkage rather than through gene fusion. After initial technology demonstration, researchers expanded to use the CovX-Body™ prototype, based on an integrin-targeting peptidomimetic pharmacophore. At least three molecules based on this architecture have entered clinical development: CVX-096, a Glp-1R agonist; CVX-060, an angiopoietin 2-binding peptide; and CVX-045, a thrombospondin mimetic.

[0260] Similarly, XTEN polypeptide has been used in recent years in a chemical conjugation method that makes it more directly similar to PEG.The first example of XTEN peptide created using this method is GLP2-2G-XTEN, which uses maleimide-thiol chemical reaction to chemically conjugate peptide to XTEN protein polymer.The chemically conjugated GLP2-2GXTEN molecule exhibits in vitro activity, in vitro plasma stability and pharmacokinetics comparable to recombinant fusion GLP2-2G-XTEN in rats.

[0261] The number and spacing of reactive groups such as lysine or cysteine ​​side chains in the fully designed sequence of XTEN or PAS polypeptide can be precisely controlled through site-directed alteration due to the constrained amino acid set that composes them.This allows for more flexibility than the methods that can be used for Fc or albumin, whose sequences naturally contain many reactive groups, and contrasts with CovX technology, which relies on reactive residues in highly specialized active sites.In addition, the lack of tertiary structure of XTEN or PAS should allow for greater flexibility over the conditions and chemistry used in coupling and purification of conjugates.

[0262] In summary, hybrid peptide half-life extension methods are emerging that combine the advantages of chemical conjugation and gene fusion methods and overcome the limitations of each. These methods confer long half-life and free the therapeutic peptide moiety from the limitation of being composed only of natural L-amino acids or as linear unidirectional polypeptides fused at the N-terminus or C-terminus, allowing the creation of molecules based on recombinant polypeptide-based partners. This opens the door to a wide range of long-acting peptide-based drugs.

[0263] Pharmaceutical Compositions In the context of preparing a pharma- ceutically acceptable composition comprising one or more peptides of the invention, the peptides of the invention (including variants and modified peptides) can be combined with one or more carriers (diluents, excipients, etc.) appropriate for the intended route or routes of administration.

[0264] The peptides of the present invention can be mixed with, for example, lactose, sucrose, powder (e.g., starch powder), cellulose esters of alkanoic acids, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphate and sulfate, acacia, gelatin, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol, and can be optionally further tableted or encapsulated for conventional administration. Alternatively, the peptides of the present invention can be dissolved in saline, water, polyethylene glycol, propylene glycol, carboxymethylcellulose colloidal solution, ethanol, corn oil, peanut oil, cottonseed oil, sesame oil, tragacanth gum, and / or various buffers. Other carriers, adjuvants, and modes of administration are also well known in the pharmaceutical arts. Carriers or diluents can include time-delay materials, such as glyceryl monostearate or glyceryl distearate, alone or with wax or other functionally similar materials.

[0265] Pharmaceutically acceptable carriers generally also include any and all suitable solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc., that are physiologically compatible with the insulin analogue. Examples of pharmaceutically acceptable carriers include water, saline, phosphate buffered saline (PBS), dextrose, glycerol, ethanol, etc., as well as any combinations thereof. In many cases, it may be desirable to include an isotonic agent, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride. Pharmaceutically acceptable substances, such as wetting agents, or minor amounts of auxiliary substances (such as moisturizing or emulsifying agents, preservatives or buffers), which may enhance the shelf life or effectiveness of the insulin analogue, related compositions, or combinations thereof, may be used. The suitability of carriers, and other components of pharmaceutical compositions, can be determined based on the lack of significant negative impact on a desired biological property of the insulin analog, related composition, or combination (e.g., less than about a 20%, 15%, 10%, 5%, or 1% reduction in insulin receptor (IR) binding and / or activation; or the ability to reduce blood glucose in a target host).

[0266] The compositions according to the invention, related compositions, and combinations can be presented, prepared, and / or administered in a variety of suitable forms. Such forms include liquid, semi-solid, and solid dosage forms, such as, for example, liquid solutions (e.g., injection and infusion solutions), dispersions or suspensions, emulsions, microemulsions, tablets, pills, powders, liposomes, dendrimers and other nanoparticles (see, for example, Baek et al, Methods Enzymol. 2003; 362: 240-9; Nigavekar et al, Pharm Res. 2004 March; 21(3): 476-83), microparticles, and suppositories. The optimal form for any peptide of the compositions related to the invention depends on the intended mode of administration, the nature of the composition or combination, and the therapeutic application or other intended use. Formulations may also include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, vesicles containing lipids (cationic or anionic), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions, carbowax (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. Any of the above mixtures may be suitable for treatment and therapy according to the present invention, provided that the binding of the peptide to its cognate IR is not significantly inhibited by the formulation, and the formulation is physiologically compatible and tolerated with the route of administration. For further information related to excipients and carriers well known to medicinal chemists, see, for example, Powell et al. "Compendium of excipients for parenteral formulations" PDA J Pharm Sci Technol. 52: 238-311 (1998), and references cited therein.

[0267] In certain embodiments, the peptides of the present invention are administered via liposomes. In other embodiments, the peptides of the present invention are administered via liposomes along with one or more ancillary agents, such as one or more antidiabetic agents.

[0268] The composition of the present invention also includes compositions that include any suitable combination of the peptide of the present invention and a suitable salt thereof. Any suitable salt can be used in stabilizing the peptide of the present invention, such as any suitable form of alkaline earth metal salt (e.g., buffer salt) (preferably, the amount of salt is such that the oxidation and / or precipitation of the peptide is avoided). Suitable salts typically include sodium chloride, sodium succinate, sodium sulfate, potassium chloride, magnesium chloride, magnesium sulfate, and calcium chloride. Compositions that include a base and one or more peptides of the present invention are also provided.

[0269] The typical mode for delivery of the composition of the present invention is by parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, and / or intramuscular administration). In one embodiment, the composition or peptide of the present invention is administered to a human patient by intravenous infusion or intravenous injection. In another embodiment, the composition or peptide of the present invention is administered by intramuscular or subcutaneous injection. As indicated above, in certain treatment regimens, intratumoral administration may also be useful.

[0270] Thus, the peptides of the present invention can be formulated, for example, in solid preparations (including, for example, granules, powders, projectiles, or suppositories), semi-solid forms (such as gels, creams, etc.), or liquid forms (such as solutions, suspensions, or emulsions). The compositions or peptides of the present invention can be applied by various solutions. Solutions suitable for use according to the present invention are typically sterile, capable of dissolving a sufficient amount of the peptides of the present invention and other components of the composition, stable under the conditions of manufacture and storage, and not harmful to the subject for the proposed application. The peptides of the present invention may be subjected to conventional pharmaceutical processes, such as sterilization, and / or may contain conventional adjuvants (preservatives, stabilizers, humectants, emulsifiers, buffers, etc.). The compositions can also be formulated as solutions, microemulsions, dispersions, powders, macroemulsions, liposomes, or other ordered structures suitable for high drug concentration. For example, the desired fluidity of the solution can be maintained by the use of a coating, such as lecithin, to maintain the required particle size in the case of dispersions, and by the use of surfactants. Prolonged absorption of the injectable compositions can be brought about by incorporating into the composition an agent that delays absorption (e.g., monostearate salts and gelatin). These and other components of the pharma- ceutically acceptable compositions of the invention can impart advantageous properties, such as improved entry, delivery, and tolerability.

[0271] Compositions for pharmaceutical use may contain various diluents, fillers, salts, buffers, detergents (e.g., non-ionic detergents such as Tween-80), stabilizers (e.g., sugars or non-protein containing amino acids), preservatives, tissue fixatives, solubilizers, and / or other materials suitable for incorporation into compositions for pharmaceutical use. Examples of suitable ingredients are also described in, for example, Berge et al, J. Pharm. Sci., 6661), 1-19 (1977); Wang and Hanson, J. Parenteral. Sci. Tech: 42, S4-S6 (1988); U.S. Patent Nos. 6,165,779 and 6,225,289; and other documents cited herein. Such pharmaceutical compositions may also contain preservatives, antioxidants, or other additives known to those skilled in the art. Additional pharma- ceutically acceptable carriers are known in the art and are described, for example, in Urquhart et al, Lancet, 16, 367 (1980); Lieberman et al, Pharmaceutical Dosage Forms-Disperse Systems (2nd ed.,vol.3,1998); Ansel et al, Pharmaceutical Dosage Forms & Drug Delivery Systems (7th ed.2000); Martindale, The Extra Pharmacopeia (31st edition); Remington's Pharmaceutical Sciences (16th-20th editions); The Pharmacological Basis Of Therapeutics, Goodman and Gilman, Eds. (9th ed.-1996); Wilson and Gisvolds' TEXTBOOK OF ORGANIC MEDICINAL AND PHARMACEUTICAL CHEMISTRY, Delgado and Remers, Eds. (10th ed.-1998), and in U.S. Pat. Nos. 5,708,025 and 5,994,106.The principles of formulating pharma- ceutically acceptable compositions are also described, for example, in Piatt, Clin. Lab Med., 7: 289-99 (1987); Aulton, Pharmaceutics: The Science Of Dosage Form Design, Churchill Livingstone (New York) (1988); EXTEMPORANEOUS ORAL LIQUID DOSAGE PREPARATIONS, CSHP (1998); and "Drug Dosage," J. Kans. Med. Soc, 70(I), 30-32(1969). Additional pharma- ceutically acceptable carriers that are particularly suitable for administration of the peptides or compositions of the invention and related compositions (e.g., compositions comprising a nucleic acid encoding the peptide of the invention or a vector comprising a nucleic acid encoding the peptide of the invention) are described, for example, in WO 98 / 32859.

[0272] The peptide or composition of the present invention can be prepared with a carrier that protects compound against rapid release, such as controlled release formulations, including implants, transdermal patches, and microencapsulated delivery systems.To produce such compositions, biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid, and any combination thereof.The method for preparing such compositions is known.See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0273] In another embodiment, the composition of the present invention is orally administered, for example, with an inert diluent or an assimilable edible carrier (specific oral administration formulations and methods are also described separately elsewhere in this specification).The compound (and other ingredients, if desired) can also be enclosed in hard or soft gelatin capsules, compressed into tablets, or directly incorporated into the subject's diet.For therapeutic oral administration, the compound can be incorporated with excipients and used in the form of ingestible tablets, lozenges, troches, capsules, elixirs, suspensions, syrups, wafers, etc. In order to administer the compound of the present invention by other than parenteral administration, it may be necessary to coat the compound with or co-administer it with a material that prevents its inactivation.

[0274] The preparation of pharmaceutical compositions containing peptides as active ingredients is well understood in the art. Typically, such compositions are prepared as injectables as liquid solutions or suspensions, but can also be prepared in solid forms suitable for dissolving or suspending in liquid before injection. Preparations can also be emulsified. Therapeutic active ingredients are often mixed with excipients that are pharma- ceutically (i.e., physiologically) acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, and the like, and combinations thereof. In addition, if desired, the composition can contain minor amounts of auxiliary substances, such as humectants or emulsifiers, pH buffering agents, and the like, which enhance the effectiveness of the active ingredient.

[0275] The peptides of the present invention can be formulated into pharmaceutical compositions as physiologically acceptable neutral salt forms.Suitable salts include acid addition salts (i.e., formed with free amino groups of peptide molecules) formed with inorganic acids such as hydrochloric acid or phosphoric acid, or organic acids such as acetic acid, oxalic acid, tartaric acid, mandelic acid, etc. Salts formed from free carboxyl groups can also be derived from inorganic bases such as sodium, potassium, ammonium, calcium, or ferric hydroxide, and organic bases such as isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

[0276] For combination compositions (discussed further herein), the peptides of the invention can be co-formulated and / or co-administered with one or more additional therapeutic agents (e.g., antidiabetic agents such as insulin, insulin analogs, metformin, or other antidiabetic biguanides, glucagon receptor antagonists, sulfonylureas, thiazolidinediones, α-glucosidase inhibitors, meglitinides, glucagon-like peptide-1 (GLP-1), GLP-1 analogs, etc.). Such combination therapies may require lower dosages of the peptides of the invention and / or co-administered agents so as to avoid possible toxicities or complications associated with various monotherapies.

[0277] therapeutic application As indicated above, the peptide or composition of the present invention can be administered individually or in combination with other pharmacologically active agents.It will be understood that such combination therapy includes different treatment regimes.The treatment regimes include, but are not limited to, administration of multiple agents together in a single dosage form, or administration of multiple agents in separate, independent dosage forms.When agents are present in different dosage forms, administration can be simultaneous or near simultaneous, or can follow any predetermined regimen that includes administration of different agents.

[0278] For example, when used to treat diabetes or other diseases or syndromes associated with reduced insulin response or production, hyperlipidemia, obesity, appetite-related syndromes, etc., the peptides of the present invention may be advantageously administered in a combination treatment regimen with one or more agents, including but not limited to insulin, insulin analogs, insulin derivatives, glucagon-like peptides 1 or 2 (GLP-1, GLP-2), derivatives or analogs of GLP-1 or GLP-2 (such as those disclosed in WO 00 / 55119). It will be understood that an "analog" of insulin, GLP-1, or GLP-2 as used herein refers to a peptide that contains one or more amino acid substitutions compared to the native sequence of insulin, GLP-1, or GLP-2, as applicable; a "derivative" of insulin, GLP-1, or GLP-2 as used herein refers in particular to a native or analog insulin, GLP-1, or GLP-2 peptide that has undergone one or more additional chemical modifications of the amino acid sequence compared to the native sequence. Insulin derivatives and insulin analogues are disclosed, for example, in U.S. Patent Nos. 5,656,722, 5,750,497, 6,251,856 and 6,268,335.In some embodiments, the combination agent is one of LysB29(ε-myristoyl)des(B30) human insulin, LysB29(ε-tetradecanoyl)des(B30) human insulin, and B29-Nε-(N-lithocholyl-γ-glutamyl)-des(B30) human insulin.Non-peptide hypoglycemic agents, antidyslipidemic agents, etc., such as those known in the art, are also suitable for combination therapy.

[0279] In one embodiment, the invention encompasses a method of treating diabetes, or a related syndrome or associated condition (e.g., reducing the rate of stroke, heart disease, kidney disease, blindness, and / or loss of sensation in the extremities associated with glucose levels and / or associated with diabetes), comprising the step of delivering an effective amount of at least one peptide of the invention (by gene expression, by homogenous peptide delivery, or by administration of a pharma- ceutically acceptable composition, typically comprising one or more peptides and one or more pharma- ceutically acceptable carriers, as described above). In another aspect, the invention provides the use of a peptide or composition (such as a combination composition) of the invention in the manufacture of a medicament for use in the treatment of type 1 or type 2 diabetes.

[0280] The peptides of the present invention can be used generally in the treatment of type 1 or type 2 diabetes, ie, both insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM).

[0281] In an exemplary combination therapy aspect, the present invention provides a method of treating diabetes (e.g., reducing one or more symptoms associated therewith in a host and / or administering to a host an amount of a composition that has been demonstrated to be therapeutically effective in at least a substantial proportion of a similar host population), comprising delivering a first amount of a peptide or composition of the present invention and a second amount of a long-acting insulin analog (e.g., LysB29(ε-myristoyl)des(B30) human insulin, LysB29(ε-tetradecanoyl)des(B30) human insulin, or B29-Nε-(N-lithocholyl-γ-glutamyl)-des(B30) human insulin, etc.), wherein the first amount and the second amount together are effective to treat the syndrome. As used herein, a long-acting insulin analog is an insulin analog that exhibits a prolonged profile of action compared to native human insulin, as disclosed, for example, in U.S. Pat. No. 6,451,970. In another aspect, the present invention provides the use of a combination composition comprising a therapeutically effective combination of at least one peptide or composition of the present invention and at least one insulin or insulin analogue in the manufacture of a medicament for use in the treatment of a disease, such as the treatment of diabetes mellitus type 1 or type 2. Similar compositions comprising a combination of one or more peptides or compositions of the present invention with one or more long-acting insulin analogues and / or short-acting insulin analogues may also be suitable for therapeutic methods, such as the treatment of diabetes.

[0282] In one aspect, the present invention provides a method of treating symptoms and / or underlying conditions associated with type 2 diabetes in a patient in need of treatment (due to diagnosis of the disease and / or substantial risk of its development), comprising the step of delivering to the patient a therapeutically or prophylactically effective amount of a peptide or composition of the present invention to treat such symptoms and / or conditions. In a particular aspect, the present invention provides a method of treating a patient with type 2 diabetes and high insulin blood levels (hyperinsulinemia). In one such aspect, the patient is obese. In another aspect, the patient also comprises, concomitantly or alternatively, an insulin resistance genotype / mutation.

[0283] In another aspect, the present invention provides a method of lowering blood pressure in a patient with insulin / IR-associated hypertension, comprising administering or otherwise delivering a therapeutically effective amount of a peptide or composition of the present invention to lower the patient's blood pressure.

[0284] In another aspect, the invention provides a method of treating symptoms and / or the underlying condition of Syndrome X, or aspects thereof (e.g., hyperlipidemia, hypertension, and / or obesity) in a patient, comprising administering or otherwise delivering to the patient a therapeutically and / or prophylactically effective amount of a peptide or composition of the invention to treat Syndrome X or a Syndrome X condition.

[0285] In yet another aspect, the present invention provides a method for treating a non-diabetic IR-mediated condition, disorder, or disease in a patient, such as IR-associated neurodegenerative disease; IR-associated non-diabetic autoimmune disease, comprising administering to the patient a therapeutically or prophylactically effective amount of a peptide or composition of the present invention to treat such condition / condition.

[0286] In another aspect, the present invention provides a method for preventing weight gain in a patient in need thereof, comprising administering to the patient a therapeutically or prophylactically effective amount of a peptide or composition of the present invention to prevent IR-associated weight gain.

[0287] In a related aspect, the present invention provides a method for treating obesity, comprising administering to a patient a therapeutically or prophylactically effective amount of a peptide or composition of the present invention to treat obesity or a condition associated therewith (by stabilizing and / or reducing the patient's body weight).

[0288] In another aspect, the present invention provides a method of treating a patient suffering from a disease condition associated with or caused by hyperinsulinemia, hypoglycemia, hypokalemia, and / or hypophosphatemia, comprising administering (or otherwise delivering: in any case) to the patient a therapeutically or prophylactically effective amount of a peptide or composition of the present invention to treat such condition / condition. The present invention further relates to a method for treating a blood glucose-related disease or disorder, comprising administration of an insulin derivative or insulin conjugate. Blood glucose-related diseases or disorders include diabetes mellitus type I and type II, and gestational diabetes. Cystic fibrosis, polypustular ovarian syndrome, pancreatitis, and other pancreas-related diseases can also be treated by administration of the insulin derivative or insulin conjugate of the present invention. Insulin is also known as a growth factor, and therefore the insulin derivative or insulin conjugate of the present invention may also be useful in topical administration for wound healing and other related indications.

[0289] In another aspect, the invention provides the use of a peptide or composition (such as a combination composition) of the invention in the manufacture of a medicament for use in the treatment of any of the foregoing conditions.

[0290] In one general aspect, the invention provides a method of modulating glucose levels in an individual, comprising administering a physiologically effective amount of a peptide or composition of the invention to detectably modulate glucose levels in the patient. In another aspect, the invention provides the use of a peptide or composition (such as a combination composition) of the invention in the manufacture of a medicament for use in reducing blood glucose levels.

[0291] In another general aspect, the present invention provides a method of mediating IR activity comprising administering a physiologically effective amount of a peptide or composition of the present invention such that responsive IR on an IR-presenting cell binds in an amount and under conditions sufficient to induce, promote, enhance, and / or otherwise modulate IR-mediated activity or response. For example, a peptide of the present invention can be delivered to a host by binding to site 1 or site 2 to direct insulin or insulin analog molecules to other sites and modify the profile of insulin or insulin analog treatment.

[0292] In still further aspects, the present invention provides methods of delivering or otherwise administering to a patient a therapeutically or prophylactically effective amount of a peptide or composition of the invention to induce, promote, and / or enhance such physiological responses, thereby providing methods of: modulating nitric oxide production levels in a patient, such as within a patient's endothelial cells; mediating RAS, RAF, MEK, and / or mitogen-activated protein (MAP) kinase pathways; modulating vascular tissue growth and / or smooth muscle cell, monocyte, macrophage, and / or endothelial cell proliferation and / or migration; stimulating the production of plasminogen activator inhibitor type 1 (PAI-1); modulating endothelin production; modulating IR-associated pro-atherosclerotic pathway biological events; modulating IR-associated inflammation; treating and / or reducing the risk of arterial damage; treating and / or preventing atherosclerosis; and / or reducing IR-associated inflammatory molecules, such as LDL cholesterol, within a patient's vascular wall.

[0293] The peptides and compositions of the present invention can also be used to treat catabolic wasting (e.g., cachexia and sarcopenia) and for non-therapeutic use, for example, in increasing muscle mass in athletes, sportsmen, or bodybuilders. The peptides are also used for their anabolic effects on muscle mass in athletes (GHRP stands for growth hormone releasing hexapeptide, a type of growth hormone releasing hormone).

[0294] This can be useful in several ways. Obviously, athletes need to heal and be productive quickly immediately after an injury. Peptides will aid the muscle or soft tissue in this reconstructive healing process. Supplements that provide an anabolic effect can also be used during preseason and other periods when building muscle mass is important. Muscle mass can be built rapidly because athletes create small tears in the muscle, which are then healed on a rapid schedule, and the process is repeated: the end effect is an increase in muscle mass and a reduction in body fat within a short time frame. The bodybuilding community uses the most effective peptides in this second format, since the new peptides do not carry the side effects of anabolic steroids.

[0295] Exemplary Dosages and Administration Strategies As described above, the compositions of the present invention may comprise a "therapeutically effective amount" or a "prophylactically effective amount" of the peptides of the present invention (or a first amount and a second amount in the case of a combination composition comprising a peptide of the present invention and a second component; a first, second, and third amount in the case of a combination composition comprising two peptides of the present invention and a secondary agent, or a peptide of the present invention and two secondary agents, etc.). In order to better illustrate certain embodiments, a detailed discussion of dosage principles is now further provided.

[0296] The amount or dosage range of the peptide of the invention utilized in practicing the invention is typically a range that effectively induces, promotes, or enhances a physiological response associated with the binding of the peptide of the invention to the cognate IR. In one embodiment, the dosage range is selected such that the peptide of the invention utilized induces, promotes, or enhances a medially significant effect, for example, in patients suffering from or at substantial risk of developing a related condition (such as a form of diabetes) that is at least partially regulated by IR activity. The medially significant effect is associated with activation, signal transduction, and / or biological modification (e.g., phosphorylation) of the cognate IR.

[0297] In yet another embodiment, a dosage of about 0.01 to 100 milligrams of active ingredient (e.g., a peptide of the invention) per kilogram of body weight per day is provided to the patient. Typically, about 1 to about 5 or about 1 to about 10 milligrams per kilogram per day given in divided doses from about 1 to about 6 times daily, or in sustained release form, can be effective to obtain the desired results.

[0298] As non-limiting examples, treatment of IR-associated conditions in humans or animals can be achieved using a single or divided dose every about 24, 12, 8, 6, 4, or 2 hours, or any combination thereof, for at least one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 days, or alternatively, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 days. In one embodiment, the method comprises administering a daily dosage of a peptide of the invention in an amount of about 0.1 to 100 mg / kg (such as 0.5, 0.9, 1.0, 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90, or 100 mg / kg) per day during at least one of weeks 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or any combination thereof.

[0299] In one aspect, the method of the invention comprises administering or otherwise delivering two different peptides of the invention over a period of one month, either at the initiation of treatment with the second peptide of the invention or at any time when a significant immune response against the first peptide of the invention is generated in the host such that continued use of the first peptide of the invention becomes detrimental to the patient. EXAMPLES

[0300] The peptide referred to in the examples is the peptide having the sequence SEQ ID NO:1.

[0301] Example 1 Cell proliferation assay Study description BrDu is incorporated into newly synthesized DNA strands of actively proliferating cells and can be detected immunochemically after partial denaturation of double-stranded DNA, allowing assessment of cell populations that are synthesizing DNA.

[0302] method Human dermal fibroblasts (HDF-Sigma 10605a) were seeded at 10,000 cells per well in 96-well plates in DMEM containing 10% fetal calf serum (FCS), 1% Pen / strep, 1% L-glutamine and allowed to adhere for 24 hours.

[0303] After an initial 24 hour incubation, cells were incubated with 5 μg / ml, 0.5 μg / ml, or 0.05 μg / ml of each of the synthetic peptides for 24 hours.

[0304] After 18 h incubation with synthetic peptides, 20 μL of BrDu reagent was added to each well. After 24 h incubation, cells were fixed and the amount of 2-DG6P was measured using the BrdU cell proliferation assay, all steps were performed according to the manufacturer's instructions.

[0305] This experiment was then repeated using HACAT cells.

[0306] result Results were calculated as a percentage of untreated controls. As illustrated in Figure 1 (HDF cells) and Figure 2 (HACAT cells), an increase in optical density reading indicates an increase in BrDu incorporation and an increase in cell proliferation. As illustrated in Figure 1 and Figure 2, samples stimulated with the peptides of the present invention showed an increase in cell proliferation compared to untreated controls.

[0307] conclusion These results support the effectiveness of the peptide in facilitating cell proliferation.

[0308] Example 2 Collagen production assay Study description Hydroxyproline in tissue preparations is a direct measure of the amount of collagen present. The FIRELISA Human Hydroxyproline ELISA Kit assay is designed to measure hydroxyproline in tissue or peptide compositions. method

[0309] Human dermal fibroblasts (HDF Sigma 10605a) were seeded at 50,000 cells per well in 24-well plates in DMEM containing 10% fetal calf serum (FCS), 1% Pen / strep, 1% L-glutamine and allowed to adhere for 24 hours.

[0310] After an initial 24 hour incubation, cells were incubated with 5 μg / ml, 1 μg / ml, or 0.1 μg / ml of each of the synthetic peptides for 96 hours.

[0311] After treatment, cell supernatants were removed and centrifuged. 50 μL of each supernatant was assayed using the FIRELISA human hydroxyproline ELISA kit. All steps were performed according to the manufacturer's instructions.

[0312] result Results were calculated as a percentage of the untreated control. An increase in optical density reading indicates an increase in collagen content. As illustrated by Figure 3, an increase in collagen production was observed compared to the untreated control.

[0313] conclusion These results support the effectiveness of the peptide in facilitating collagen production.

[0314] Example 3 Elastin production assay Study description Elastin is a highly elastic protein in connective tissue that allows many tissues in the body to regain their shape after stretching. The FIRELISA Human Elastin ELISA Kit assay is designed to measure elastin in tissue compositions or protein / peptide compositions. method

[0315] Human dermal fibroblasts (HDFs) were seeded at 50,000 cells per well in 24-well plates in DMEM containing 10% fetal calf serum (FCS), 1% Pen / strep, 1% L-glutamine and allowed to adhere for 24 hours.

[0316] After an initial 24 hour incubation, cells were incubated with 5 μg / ml, 1 μg / ml, or 0.1 μg / ml of each of the synthetic peptides for 96 hours.

[0317] After treatment, cells were lysed using cell lysis buffer and sonicated for 10 seconds. Lysed cells were centrifuged and the supernatant was removed from each sample. Total protein concentration was determined using a BCA assay. Each sample was diluted with assay buffer to contain 10 μg total protein. 50 μL of each sample was then assayed using the FIRELISA human elastin ELISA kit. All steps were performed according to the manufacturer's instructions.

[0318] result Results were calculated as a percentage of the untreated control. An increase in optical density reading indicates an increase in elastin content. As illustrated by Figure 4, samples stimulated with the peptides of the present invention showed increased elastin production compared to the untreated control.

[0319] conclusion These results support the effectiveness of the peptide in facilitating elastin production.

[0320] Example 4 Glucose uptake Study description Glucose uptake in skeletal muscle cells was measured using 2-deoxyglucose (2-DG). 2-DG is taken up by glucose transporters and metabolized to 2-DG-6-phosphate (2-DG6P). The accumulation of non-metabolized 2-DG6P is proportional to the glucose uptake by the cells. method

[0321] Human skeletal myoblasts (Sigma 150-05a) were seeded at 10,000 cells per well in 96-well plates in skeletal muscle differentiation medium and allowed to differentiate for 72 hours prior to experimentation.

[0322] Differentiated cells were serum starved for 24 hours before stimulation with insulin or synthetic peptides. After starvation, serum-free medium was removed, cells were rinsed with phosphate-buffered saline (PBS), and medium was replaced with 100 μL Krebs-Ringer-phosphate-HEPES (KRPH) and incubated for 1 hour.

[0323] Cells were then stimulated with 100 nM insulin for 30 min or with 5 μg / ml, 0.5 μg / ml, or 0.05 μg / ml of each of the synthetic peptides for 3 h.

[0324] After stimulation, cells were incubated with 10 μL of 2-DG solution per well for 40 min, and glucose uptake was measured using the "PrismColor Glucose Uptake Assay Kit" (Molecutools), all steps were performed according to the manufacturer's instructions. The experiment was repeated using HACAT cells.

[0325] result Results were calculated as a percentage of untreated controls. An increase in optical density reading indicates increased incorporation of 2-DG6P and increased glucose uptake. All experiments were performed in duplicate (6 wells per condition) on triplicate plates. Student's t-test ( * p<0.05 compared with control;** p<0.01 compared with control; *** Significance was determined using p<0.001 compared to control. Figure 6 illustrates the results for 0.5 μg / ml of synthetic peptide, and Figure 7 illustrates the results for 5 μg / ml of synthetic peptide. Figure 5 displays the results for HACAT cells. As illustrated by these figures, samples stimulated with the peptides of the invention showed increased glucose uptake compared to untreated controls.

[0326] conclusion These results support the efficacy of the peptide in facilitating glucose uptake in skeletal muscle.

[0327] Example 5 GLUT-4 migration method HSKMC cells were cultured in 6-well plates and then treated with peptides / hydrolysates at concentrations of 0.5 μg / ml and 5 μg / ml for 2 hours. Upon completion of treatment, cell membrane proteins were harvested. Briefly, cells were harvested from wells with a cell scraper and centrifuged at 300×g for 5 minutes. The cell pellet was washed with 300 μl of cell wash solution and then centrifuged again at 300×g for 5 minutes. The supernatant was discarded and cells were resuspended in 1.5 ml of cell wash solution and transferred to a 2 ml centrifuge tube. The cells were then centrifuged at 300×g for 5 minutes and the supernatant was discarded. 150 μl of permeabilization buffer was added to the cell pellet and vortexed briefly to obtain a homogenous cell suspension. The suspension was then incubated at 4° C. for 10 minutes with constant mixing. The permeabilized cells were then centrifuged at 16000×g for 15 min, and the supernatant containing the cytosolic proteins was transferred to a fresh tube. 100 μl of solubilization buffer was added to the pellet and resuspended by gentle pipetting. The sample was then incubated at 4° C. for 10 min with constant mixing. The sample was then centrifuged at 16000×g for 15 min and at 4° C. for 10 min. The supernatant containing the solubilized membrane and membrane-associated protein fraction was transferred to a fresh tube and stored at −80° C. until ready for evaluation.

[0328] GLUT-4 was assessed in the cell membrane fraction using a sandwich ELISA from LSBio. 10 μg of protein from each sample was loaded into individual wells on a 96-well plate to a final volume of 100 μl. GLUT-4 standards (0.3 ng / ml to 20 ng / ml) were also added to the plate in the same volume. The plate was then sealed and incubated at 37° C. for 1 hour. Liquid was aspirated from each well and then 100 μl of detection reagent A was added to each well, at which point the plate was sealed, gently swirled to ensure mixing, and then incubated at 37° C. for 1 hour. Liquid was aspirated from each well and the wells were washed 3 times using 350 μl of wash buffer. 100 μl of detection reagent B was added to each well, at which point the plate was sealed and incubated at 37° C. for 30 minutes. The wells were washed five times as outlined above, and 90 μl of TMB substrate was added to each well. The plate was sealed, protected from light, and incubated at 37° C. for 10 minutes to ensure optimal color development. 50 μl of stop solution was added to each well, and the optical density of the samples was determined using a microplate reader set to 450 nm. GLUT-4 concentrations in the samples were calculated from the standard curve after generating a four-parameter logistic (4-PL) curve fit. The sample concentration readings of the curve were then corrected by multiplying the results by the dilution factor.

[0329] result As illustrated by FIG. 8, increased GLUT4 translocation was seen in samples stimulated with both 0.5 μg / ml and 5 μg / ml of peptide.

[0330] conclusion The peptide showed a trend for a stimulatory effect on skeletal muscle GLUT4 translocation and its ability to facilitate glucose transport in skeletal muscle.

[0331] Example 6 Muscle protein synthesis (phospho-mTOR assay) Study description The mammalian target of rapamycin (mTOR) is a Ser / Thr protein kinase that functions as an ATP and amino acid sensor to balance nutrient availability and cell proliferation. mTOR activity is regulated by insulin, amino acids, physical exercise, oxidative stress, and growth factors, and consistently, its phosphorylation promotes protein synthesis pathways. However, dysregulation of mTOR causes accelerated aging, among many other pathologies. In muscle, mTOR, when activated (phosphorylated), leads to skeletal muscle hypertrophy. This fact has been widely reported in the literature, both in rodents and humans. Amino acids such as leucine enhance the phosphorylation of mTOR, and its activation upregulates protein translation through the phosphorylation of eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and ribosomal protein S6 kinase (S6K), resulting in cell growth and proliferation.

[0332] The phospho-mTOR sandwich ELISA detects endogenous levels of phosphorylated mTOR at Ser2448.

[0333] method Day 1: HSkMC cells are seeded in 500 uL HSkMC growth medium per well and allowed to adhere to the wells for 16-24 hours. Day 2: Remove growth medium and add 500 uL HSkMC differentiation medium per well and allow them to differentiate for 5-7 days. Day 7: Cells are treated for 2 hours. Cell lysates are collected in lysis buffer. Day 8: BCA (protein determination) is performed and the assay is run. In the first incubation step, mTOR antibody coated on ELISA microwells is used to capture mTOR (phospho and non-phospho) from cell lysates. Then, after extensive washing, a specific phospho-mTOR detection antibody is added to detect only the already captured phospho-mTOR protein. Finally, an HRP-linked antibody is used to recognize the detection antibody and the addition of TMB (HRP substrate) develops color proportional to the amount of mTOR phosphorylated at Ser2448.

[0334] result The results are illustrated in Figure 9. Samples stimulated with 0.5 μg / ml and 5 μg / ml showed an increase in mTOR.

[0335] conclusion These results support the effectiveness of the peptide in facilitating muscle protein synthesis and muscle growth activity.

[0336] (Examples 7 to 15) formulation In embodiments, the composition is formulated as a topical composition, which in one embodiment can be an emulsion or cream, or a rub, such as a muscle rub.

[0337] In an embodiment, the cream comprises an excipient or diluent, a suspending agent, a preservative, and a variable amount of at least one peptide of the invention.

[0338] In an embodiment, the cream comprises alcohol, a carbomer, a sorbate, water, and at least one peptide of the invention.

[0339] Preferably, the alcohol is butylene glycol (1,3-butanediol). The alcohol may be present in an amount between 1 and 10%, preferably between 2 and 6%, preferably 4% of the composition. The sorbate may be polysorbate 20. The sorbate may be present in an amount between 0.01 and 1%, preferably between 0.05 and 0.5%, preferably 0.10% of the composition. Water may be present in an amount between 10% and 90%, preferably 30% to 75%. The carbomer may be present in an amount between 0.05% and 1%, preferably between 0.1% and 0.5%, preferably 0.15%. The carbomer may be Ultrez 10.

[0340] The peptides of the invention may be present in an amount of 0.5% to 10%, preferably 1% to 5%, preferably 3%.

[0341] The compositions may further include one or more of a sugar alcohol such as glycerin, a paraben, a silicone such as cyclohexasiloxane, a fatty alcohol or phosphate or a mixture of fatty alcohols and phosphates (such as cetearyl alcohol, dicetyl phosphate, and Cereth 10 phosphate, or combinations thereof), a polyoxyethylene stearyl ether (such as Steareth 2 and 10), and a fragrance.

[0342] Example 7 An exemplary rub or emulsion is as follows in Example 7:

[0343] [Table 1]

[0344] The resulting emulsion is suitable as a rub. In addition, the rub is suitable for fragile aging skin. The emulsion is suitable for improving fine lines, wrinkles, dryness, reducing redness and irritation.

[0345] It will be understood that the percentages are examples only and any suitable percentages may be used depending on the application.

[0346] The emulsion is prepared in the following manner: Phase A: Ultrez 10 (carbomer) is dispersed in water and allowed to swell for 20 minutes, then Phase B is added and heated to 75° C. Phase C is heated separately to 75° C. The two phases are mixed under stirring, homogenized, Phase D is added, neutralized with Phase E, cooled to reach 30° C., then Phase F and Phase G are added and the pH is adjusted to 6 with NaOH. It will be understood that this is only an example and any suitable method known in the art may be used.

[0347] In further embodiments, the composition may include one or more of the following active agents: water, carbomer, a sorbate such as potassium sorbate, a sugar alcohol such as glycerin, an alcohol such as 2-(2-ethoxyethoxy)ethanol, a polyoxyethylene stearyl ether such as Steareth 2, a fatty alcohol or phosphate, or a mixture of fatty alcohols and phosphates such as cetearyl alcohol, dicetyl phosphate, and Cereth 10 phosphate, or a combination thereof, a siloxane such as cyclomethicone, a skin lightening agent such as caprylic / capric triglyceride, sorbitan stearate, a paraben, sodium hydroxide, glycerin (and) butylene glycol (and) a mixture of Arcostaphylus Uva Ursi leaf extract (and) Mitracarpus Scaber extract (ETIOLINE®), or a muscle health agent, and a peptide of the present invention.

[0348] Example 8 The following compositions in Example 8 are examples of emulsions or creams or rubs.

[0349] [Table 2]

[0350] The active agent may be ETIOLINE® [glycerin (and) ethylene glycol (and) bearberry leaf extract and Mitracarpus scaber extract]. ETIOLINE® is a skin lightening ingredient sold by SEDERMA (WO 98 / 05299, November 19, 1996).

[0351] It will be appreciated that ETIOLINE may be replaced with any active agent, such as an agent that aids in muscle health, development, or recovery.

[0352] This formulation can be made following the procedure generally outlined in Example 7. The composition can be an emulsion or cream or rub that includes one or more of water, a carbomer such as Ultrez 10, a sugar alcohol such as glycerin, an alcohol such as Fenova (phenoxyethanol and mixed parabens), a fatty acid ester such as ethylhexyl palmitate, a fatty alcohol such as cetearyl alcohol, a lactate ester such as myristyl lactate, a sorbate such as polysorbate 20 and / or potassium sorbate, a polymeric emulsifier such as acrylates (C10-30 alkyl acrylates) and a cross-linked polymer, a siloxane such as cyclomethicone, sodium hydroxide, a muscle health agent, or an agent for the treatment of stretch marks such as siegesbeckia Orientalis extract (Darutoside), and at least one active agent such as a peptide.

[0353] Example 9 The following compositions in Example 9 are examples of emulsions or creams. In embodiments, the emulsion or cream is an anti-stretch mark cream.

[0354] [Table 3]

[0355] The active agent can be Darutoside (Horsetail extract). Darutoside is a molecule sold by SEDERMA for the treatment of stretch marks. It will be understood that Darutoside can be replaced with any active agent, such as any agent that aids in muscle health, development, or recovery.

[0356] The emulsion or rub is prepared in the following manner: Phase A: Ultrez 10 (Carbomer) is dispersed in water and allowed to swell for 20 minutes, then Phase B is added and heated to 75° C. Phase C is heated separately to 75° C. The two phases are mixed under stirring, homogenized, Phase D is added, neutralized with Phase E, cooled to reach 30° C., then Phase F and Phase G are added and the pH is adjusted to −6 with NaOH.

[0357] In an embodiment of the invention, the composition may be an emulsion, cream, or gel, preferably a gel comprising one or more of water, a carbomer such as Ultrez 10, a sugar alcohol such as glycerin, an alcohol such as Fenova (phenoxyethanol and mixed parabens), a sorbate such as polysorbate 20 and / or potassium sorbate, a polymeric emulsifier such as acrylates (C10-30 alkyl acrylates), a siloxane such as cyclomethicone, sodium hydroxide, peptides, and at least one agent such as an agent for muscle health, recovery, or development, or an agent comprising Imperata Cylindrica (root) extract, water, glycerin, PEG-8, and a carbomer (MOIST 24), etc. In an embodiment, the gel is a moisturizing gel.

[0358] Example 10 The following composition in Example 10 is an example of a gel. In an embodiment, the gel is a moisturizing gel.

[0359] [Table 4]

[0360] Example 11 This formulation can be made following the procedures generally outlined in Example 11.

[0361] The active ingredient may be MOIST-24® (Imperata cylindrica (root) extract (and) water (and) glycerin (and) PEG-8 (and) carbomer). It will be understood that any agent may be used to replace MOIST-24, for example, any agent that aids in muscle health, development, or recovery.

[0362] In an embodiment of the invention, the composition may be an emulsion, cream, or rub comprising one or more of water, a carbomer such as Ultrez 10, a sorbate such as polysorbate 20, polysorbate 60, and / or potassium sorbate, Fenova (phenoxyethanol and mixed parabens), an alcohol such as butylene glycol (1,3-butanediol), lanolin alcohol, and / or cetearyl alcohol, sorbitan stearate, polydimethylsiloxane such as dimethicone, isotridecyl isononanoate, caprylic / capric triglyceride, cetyl esters, sodium hydroxide, water, an agent for muscle health, recovery, and / or development, or an anti-aging agent such as butylene glycol, water, laureth 3, hydroxyethylcellulose, and an agent comprising acetyl-dipeptide-1-cetyl ester (CALMOSENSINE®).

[0363] The following compositions in Example 11 are examples of creams or rubs.

[0364] [Table 5]

[0365] This formulation can be made following the procedure generally outlined in Example 9. The active ingredient can be CALMOSENSINE® [butylene glycol (and) water (and) Laureth-3 (and) hydroxyethylcellulose (and) acetyl-dipeptide-1-cetyl ester]. Calmosensine® is an analgesic peptide provided by SEDERMA (WO 98 / 07744, Feb. 26, 1998). It will be understood that any agent can be used to replace CALMOSENSINE, for example any agent that aids in muscle health, development, or recovery.

[0366] In an embodiment of the invention, the composition may be an emulsion, cream, or rub comprising one or more of water, a carbomer such as Ultrez 10, a sugar alcohol such as glycerin, a sorbate such as potassium sorbate, steareth such as Steareth 10, a fatty alcohol or phosphate, or a mixture of fatty alcohols and phosphates such as cetearyl alcohol, dicetyl phosphate, and Cereth 10 phosphate, or combinations thereof, ethylhexyl disuccinate, mixed parabens, sorbitan stearate, sodium hydroxide, water, peptides, and active agents such as agents for muscle health, recovery, and / or development, or agents for mature skin such as agents comprising Trifolium Pratense (clover) petal extract (and) glycerin (and) butylene glycol (and) lecithin (TEROCARE®).

[0367] Example 12 The following compositions in Example 12 are examples of creams or rubs.

[0368] [Table 6]

[0369] This formulation can be made according to the procedure generally outlined in Example 9. The active agent can be STEROCARE™ [Red clover (clover) petal extract (and) glycerin (and) butylene glycol (and) lecithin]. Sterocare® is offered by SEDERMA as an active ingredient for mature skin (FR 2769502, 14 April 2000, WO 99 / 18927, 22 April 1999). It will be understood that any agent can be used to replace STEROCARE®, for example any agent that contributes to muscle health, development or recovery.

[0370] Example 13 The following compositions in Example 13 are examples of rubs or tonics.

[0371] [Table 7]

[0372] Example 14 The following compositions in Example 14 are examples of creams or rubs.

[0373] [Table 8]

[0374] The agent may be deferoxamine and / or berberine. These are used as skin thickening agents. Deferoxamine and berberine can be replaced with another active agent, such as an agent for muscle health, recovery, and development.

[0375] Example 15 The following compositions in Example 15 are examples of gels or rubs.

[0376] [Table 9]

[0377] This gel can be prepared in the following manner: Homogenize Part B and pour into Part A. Heat Part (A+B) to 75°C. Heat Part C and Part D to 75°C. Pour Part C into Part (A+B) with helical mixing, then pour Part D into Part (A+B+C). Add Part F and Part E. Pour Part G at approximately 35°C.

[0378] The active agent can be rutin and / or Bowman-Birk inhibitor (BBI). These agents are used for tissue regeneration. Rutin and Bowman-Birk inhibitor (BBI) can be replaced with another active agent, such as an agent for muscle health, recovery, and development.

[0379] Example 16 Puromycin ELISA Study description Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. It is a structural analogue of aminoacyl-tRNA and therefore can be incorporated into growing peptide chains via the formation of peptide bonds. However, whereas aminoacyl-tRNA contains a hydrolyzable ester bond, puromycin does not have one at the equivalent position. Thus, binding of puromycin to a growing peptide chain prevents the formation of a new peptide bond by the next aminoacyl-tRNA. As a consequence, puromycin binding results in the termination of peptide elongation and the release of the puromycin-bound cleaved peptide from the ribosome. At very high concentrations, puromycin effectively blocks the elongation phase of translation and thus inhibits protein synthesis. However, at very low concentrations, it does not inhibit the overall translation rate, and the rate at which puromycin-labeled peptides are formed reflects the overall protein synthesis rate. This latter property makes puromycin a potential tool for measuring changes in protein synthesis rates.

[0380] method 1. Dispense 100 μl of coating buffer per well into the required number of wells in a 96-well ELISA plate. Dilute and mix protein extracts to 5 μg / μl (in the same buffer that the samples are being taken in). Immediately dispense 1 μL per well into the plate. 2. Incubate the plates at 4° C. for 16-24 hours. 3. Shake contents vigorously. Wash once with 200 μl PBS. 4. Add 200 μl of blocking solution (5% BSA in PBS). Incubate at room temperature for 4-6 hours. 5. Puromycin antibody is diluted in blocking solution and incubated overnight at 4°C. 8. Wash twice with 200 μl of PBS. 9. Add 100 μl of secondary antibody diluted 1:1000 in blocking solution. Incubate at room temperature for 1 hour. 10. Wash 4 times with 200 μl PBS. Shake contents vigorously. 11. Add 100 μl of TMB substrate and incubate for 15-20 minutes. 12. Stop solution is added and plates are read at 450 nm.

[0381] result FIG. 10 illustrates the increase in protein synthesis.

[0382] (Example 17) toxicity method 100 μl of HSkMC cells were plated in collagen-precoated 96-well plates at 1 × 10 cells per well. 4 The cells were seeded at a concentration of 100 μl. The cells were allowed to attach overnight at 37° C., 5% CO2, and the next day the growth medium was replaced with 100 μl of cell differentiation medium and returned to the incubator at 37° C., 5% CO2. The cell differentiation medium was changed every 2 days for 6 days until the HSkMC cells were fully differentiated into myotubes. On day 7, the cell differentiation medium was replaced with 100 μl of basal medium and starved overnight at 37° C., 5% CO2.

[0383] On the day of the MTT assay, the starvation medium was removed and replaced with 100 μl of basal medium containing the peptide of SEQ ID NO:1. The cells were incubated during this treatment for 20 minutes at 37° C., 5% CO2, at which point the treatment medium was removed and replaced with 110 μl of MTT solution and incubated for 2 hours at 37° C., 5% CO2. After this incubation step, the MTT solution was removed and replaced with 110 μl of DMSO. The plate was covered with tin foil and placed in a shaker for 5 minutes at room temperature. The plate was read using a spectrophotometer (SpectraMax M3, Molecular devices, Sunnyvale, CA 94089 USA) set at 570 nm, and the cytotoxic effect of the peptide of SEQ ID NO:1 was evaluated based on the optical density of the samples. The results are illustrated by FIG. 11.

[0384] (Example 18) This set of experiments was designed to understand the mechanism of action of the peptide of SEQ ID NO:1.

[0385] Phosphoacetyl CoA method The Phosphoacetyl-CoA Carboxylase (Ser79) Sandwich ELISA Kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects the endogenous level of acetyl-CoA carboxylase (ACC) when phosphorylated at Ser79. Phospho-ACC (Ser79) rabbit antibody was coated onto the microwells. After incubation with cell lysate, phospho-ACC protein was captured by the coated antibody. After extensive washing, ACC mouse detection mAb was added to detect the captured ACC protein. Anti-mouse IgG HRP-linked antibody was then used to recognize the bound detection antibody. HRP substrate TMB was added to develop color. The magnitude of absorbance for color development was proportional to the quantity of ACC phosphorylated at Ser79. These results are illustrated by FIG. 12.

[0386] Acetyl-CoA carboxylase (ACC) catalyzes the carboxylation of acetyl-CoA to malonyl-CoA. It is a key enzyme in fatty acid biosynthesis and oxidation. In rodents, the 265 kDa ACC1 (ACCα) form is predominantly expressed in lipogenic tissues, whereas the 280 kDa ACC2 (ACCβ) is the major isoform in oxidative tissues. However, in humans, ACC2 is the predominant isoform in both lipogenic and oxidative tissues. Phosphorylation by AMPK at Ser79 or PKA at Ser1200 inhibits the enzymatic activity of ACC. ACC is a potential target for anti-obesity drugs.

[0387] Phospho-Akt1 method The phospho-Akt1 (Ser473) sandwich ELISA kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects the endogenous levels of phospho-Akt1 (Ser473) protein. Phospho-Akt (Ser473) rabbit mAb was coated onto the microwells. After incubation with cell lysate, phospho-Akt (Ser473) protein was captured by the coated antibody. After extensive washing, Akt1 mouse antibody was added to detect the captured phospho-Akt1 (Ser473) protein. Anti-mouse IgG HRP-linked antibody was then used to recognize the bound detection antibody. HRP substrate TMB was added to develop color. The magnitude of absorbance for this color development is proportional to the amount of phospho-Akt1 (Ser473) protein.

[0388] The results are illustrated in FIG.

[0389] Akt, also called PKB or Rac, plays a crucial role in the control of survival and apoptosis. This protein kinase is activated by insulin and various growth and survival factors and functions in a wortmannin-sensitive pathway involving PI3 kinase. Akt is activated by phospholipid binding and activation loop phosphorylation by PDK1 at Thr308, as well as phosphorylation within the carboxy terminus at Ser473. The previously unknown PDK2, which contributes to phosphorylation of Akt at Ser473, has been identified as the mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1. Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad, forkhead transcription factors, c-Raf, and caspase 9. PTEN phosphatase is a major negative regulator of the PI3 kinase / Akt signaling pathway. LY294002 is a specific PI3 kinase inhibitor. Another important Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β. Akt may also play a role in insulin stimulation of glucose transport. In addition to its role in survival and glycogen synthesis, Akt is also involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 and negatively regulating the cyclin-dependent kinase inhibitors p27 Kip1 and p21 Waf1 / Cip1. Akt also plays a pivotal role in cell proliferation by directly phosphorylating mTOR in the rapamycin-sensitive complex containing raptor. More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR, in the mTOR-raptor complex.

[0390] Phospho-AMPKα method The phospho-AMPKα (Thr172) sandwich ELISA kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects the endogenous level of AMPKα when phosphorylated at Thr172. AMPKα rabbit antibody was coated onto the microwells. After incubation with cell lysate, AMPKα (phospho and non-phospho) was captured by the coated antibody. After extensive washing, phospho-AMPKα (Thr172) mouse detection antibody was added to detect the phosphorylation of the captured AMPKα protein at Thr172. Then, anti-mouse IgG HRP-linked antibody was used to recognize the bound detection antibody. HRP substrate TMB was added to develop color. The magnitude of absorbance for this color development was proportional to the quantity of AMPKα phosphorylated at Thr172. The results are illustrated in FIG. 14.

[0391] AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in regulating energy homeostasis. AMPK is a heterotrimeric complex composed of a catalytic α subunit and regulatory β and γ subunits, each of which is encoded by two or three separate genes (α1, 2; β1, 2; γ1, 2, 3). The kinase is activated by an increase in the AMP / ATP ratio resulting from cellular and environmental stresses such as heat shock, hypoxia, and ischemia. The tumor suppressor LKB1, which associates with the accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 within the activation loop, and this phosphorylation is required for AMPK activation. AMPKα is also phosphorylated at Thr258 and Ser485 (for α1; Ser491 for α2). The upstream kinases and biological significance of these phosphorylation events remain to be elucidated. The β1 subunit is post-translationally modified by myristoylation and multiple phosphorylation sites, including Ser24 / 25, Ser96, Ser101, Ser108, ​​and Ser182. Phosphorylation at Ser108 of the β1 subunit is required for the activation of the AMPK enzyme, whereas phosphorylation at Ser24 / 25 and Ser182 affects AMPK localization. Several mutations in the AMPKγ subunit have been identified, most of which are located in the putative AMP / ATP binding site (CBS or Bateman domain). Mutations at these sites result in reduced AMPK activity, causing glycogen accumulation in the heart or skeletal muscle. Accumulating evidence indicates that AMPK not only regulates fatty acid and glycogen metabolism, but also modulates protein synthesis and cell proliferation via the EF2 and TSC2 / mTOR pathways, as well as blood flow via eNOS / nNOS.

[0392] conclusion The peptide with the sequence of SEQ ID NO:1 acts through the AMPK pathway. In skeletal muscle, activation (phosphorylation) of AMPK leads to phosphorylated ACC, which promotes fatty acid oxidation. This is a pathway that is quite different from the insulin pathway, as shown in the gene expression profile in skeletal muscle cells described below.

[0393] (Example 19) Gene Expression Studies method The top 50 genes showing over- and under-expression relative to the negative control were searched for insulin and peptide of SEQ ID NO:1.

[0394] HSkMC cells were cultured in a 2 ml volume in a collagen-precoated 6 cm plate at 2 × 10 cells / cm. 5 The cells were seeded at a concentration of 100x100 / mL. The cells were allowed to attach overnight at 37°C, 5% CO2, and the next day the growth medium was replaced with 2ml of cell differentiation medium and returned to the incubator at 37°C, 5% CO2. The cell differentiation medium was changed every 2 days for 6 days until the HSkMC cells were fully differentiated into myotubes. On day 7, the cell differentiation medium was replaced with 2ml of basal medium and starved overnight at 37°C, 5% CO2.

[0395] On the day of the experiment, the starvation medium was removed and replaced with 2 ml of basal medium containing 0.5 μg / ml of peptide of SEQ ID NO: 1. In parallel, untreated cells were tested. Cells were incubated in this treatment for 20 minutes at 37° C., 5% CO2, at which point the treatment medium was removed and the cells were scraped in 1 ml of PBS. The cell suspension was pelleted in a microcentrifuge at 1500 rpm for 5 minutes and the supernatant was removed. The cells were immediately flash frozen in liquid nitrogen and transferred to a −80° C. freezer for storage until shipment to the CRO.

[0396] To determine gene expression, Agilent Single Color experiments were performed by Elda Biotech using an Agilent G2565CA microarray scanner system. RNA was extracted from cells and then hybridized to Agilent Human Gene Expression Microarray. The following kits were used: Qiagen Rneasy, Agilent RNA 6000 Nano, Lowlnput QuickAmp labeling, RNA Spike In (single color), Hi RPM GE Hybridisation Kit Large, Gene Expression Wash Pack. The experiment used SurePrint G3 Human Gene Expression v3 8x60K microarrays. Each sample was in triplicate. Samples passed RNA quality and quantity tests. Subsequent data analysis was performed using the Bioconductor package "limma".

[0397] The intensity raw data was background corrected by the normexp method and offset of 50, and quartile normalized, resulting in a log2 expression scale. Batch-effect correction was applied to adjust for separation by the number of replicates revealed by principal component analysis using ComBat. Empirical Bayes analysis was then applied. Differential expression was calculated based on comparison with untreated samples and evaluated using moderated t-statistics. To correct for multiple testing, the resulting p-values ​​were corrected using the Benjamini and Hochberg method. Differential expression was not detected under an adjusted p-value threshold of 0.05. Instead, a raw p-value threshold of 0.001 was used. Genes with absolute fold change values ​​above 1.3 are listed below.

[0398] List of significantly up- and down-regulated genes and their fold changes (microarray gene expression)

[0399] Genes upregulated upon insulin treatment: ABCA9、ACIN1、ADAP2、AKIRIN2、ALG3、AMN、ANKS3、ANP32A-IT1、ARGLU1、ARHGEF35、ARPC5L、ASAP1-IT1、ATP5H、ATP8、AURKAPS1、BMS1P6、BOD1L1、BRD3、 BROX、C12orf65、C14orf169、C20orf96、C21orf59、C4orf33、C5orf24、C5orf58、C6orf47、CACNA1A、CASC15、CATSPER2P1、CBX5、CCDC102B、CCNL1、CCNT2 -AS1、CD86、CDK11B、CENPC、CFAP36、CHD3、CIAO1、CLASRP、CMTM8、COPG2IT1、CRHR1-IT1、CWC22、DKFZP586B0319、DKFZp686M1136、DPP9-AS1、DRAP1、DRD 4、DYX1C1、EIF4G3、EIF5、EPSTI1、EXOC1、FAM127C、FAM155A-IT1、FAM173A、FAM212A、FAM71F1、FASTKD1、FIP1L1、FLJ11292、FLJ13773、FTSJ3、GABPB1-A S1、GADD45B、GATA2、GBP2、GOLGA2P6、GOLGA6L4、GPATCH11、GPR135、GRIN2D、GSK3A、HEBP2、HMGB1、HMGB3P1、HNRNPA1L2、HOXB-AS1、HOXC9、HOXD8、ID1、 IER2、IFI44、IFT172、IGF1R、IGF2BP2、KANK3、KCTD3、KIAAI654、K1Z、KLF17、KRTAP19-2、KRTAP4-11、LDHAL6A、LINC00083、LINC00504、LRRC37A2、MALAT 1、MAP3K10、MAP3K3、MBNL1、MIDN、MIR22HG、MIR612、MPV17L2、MRE11A、MTA2、MYO1C、MZF1、NAMPT、NBPF8、ND4L、NF1P2、NIPBL、NOL8、NOP58、NPIPB5、NR2C1、OLA1、PAK1IP1、PCDHGA2、PDCD6IP、PDGFRB、PFDN4、PGBD2、PGF、PGM5P2、PMAIP1、PNPLA8、POU2F1、PP12719、PPIG、PP6R1、PRKAG2、PRO2852、PRPF40A、PRR4, PSMA4, PTMA, RALBP1, RC3H2, RCC1, REP15, RHOB, RN7SL1, ROM1, RPLP1, RPS6KC1, SAMD11, SFSWAP, SH3KBP1, SLC25A34, SLFN5, SMARCC1, SMCR6, SMIM11, SNAI1, SNAR-A3, SNAR-B2, SNAR-D, SNAR-F, SNAR-G2, SNAR-H, SNHG9, SNORA10, SNORD97, SNO RD99, SOCS3, SOX8, SRP19, SRSF11, SSC5D, STAR, TAB2, TAF3, TCEAL7, TCF7L1, TFPI2, THOC2, TINAGL1, TNFAIP8L1, TOPORS, T RANK1, TRAPPC10, TXLNG, UBE2Q2P1, UBN2, VASP, VHL, VPS37D, YBX3, ZBTB2, ZCCHC17, ZMYND11, ZNF503-AS2, ZSWIM4, ZSWIM6,

[0400] Genes downregulated upon insulin treatment: <h2 style=";text-align:left;direction:ltr">ABHD2, ACTA2, ACVR1, ADAMTS9, ADCY6, ADD3, ADPRHL2, AGTRAP, ALAD, AMZ2P1, ANXA2P1, AP2B1, ARMX6, ATP5A1, ATP5B, ATP5G1, ATP6V0E1, ATRAID, AXL 、BFAR、BOD1、BRF2、C1orf43、C5orf15、C9orf78、CA12、CAV1、CD47、CD82、CD 9、CDC123、CDC42BPA、CDC42EP4、CHST14、CHST3、CHSY3、CLMP、CLPTM1L、CLU、 CNPPD1、COLGALT1、COPG1、COPZ2、CRISPLD2、CRTC3、CSRP1、CTSLP8、CYB5D2 、CYB5R1、DAG1、DANCR、DEK、DENND4C、DFNA5、DHRS1、DLST、DNAJB9、DNASE1L1 、DNASE2、DPAGT1、EBF2、EI24、EIF2B4、EIF3I、EIF3K、EIF3L、EMC3、ENDOD1、 ENO2、ENPP4、EPRS、ESYT1、ESYT2、EXT1、EZH1、FAF1、FAM57A、FAM96A、FAS、FH OD1、FKBP14、FLII、FLNB、FN3KRP、G6PD、GABARAP、GALM、GARS、GBA、GBAP1、G DE1、GJA1、GOLM1、GPRC5A、GPX3、GREM1、GSTM2、HDDC2、HEPH、HEXB、HLA-A、HL A-DMA、HLA-DPA1、HLA-DPB1、HLA-DRB4、HOOK2、HTRA2、IDH1、IDS、IFIT3、IG F2R、IGFBP6、IL10RB、IL11RA、ILK、ITGA3、ITGAE、JKAMP、KIF3B、KLHDC2、LAM A2, LAPTM4B, LASP1, LUM, MAGED1, MAN2A1, MAN2B2, MANBA, MANSC1, MAP2K1, MFSD5, MKNK1, MLEC, MMGT1, MPHOSPH8, MRPL17, MRPL37, MRPS18A, MTFR1L, MT HFD1L、MUL1、MYD88、NBAS、NDRG1、NDUFA9、NIF3L1、NLRP1、NME6、NPC1、NSMC E1、NUDT2、OAT、OLFML3、OXA1L、P4HA2、P4HTM、PAMR1、PARP6、PCYOX1、PDE6D、PDLIM1, PDXP, PEA15, PEAR1, PEF1, PELO, PGAM1, PGM1, PGRMC1, PHB2, PIGH, PLEKHG4, PLP2, PLS3, PPAP2B, PPIAL4B, PP L, PRKAB1, PRMT5, PRNP, PRPF8, PRPS1, PRRC1, PSAP, PSMB8, PSMD10, PTGS1, QSOX2, RAB11FIP5, RAF1, RARG, RBM23, RER 1, RHBDD2, RP9, RPL29P2, RPL5, RPRD1B, RPUSD4, RTCB, SCAMP2, SCG2, SDC4, SDHB, SEC22C, SEC23A, SEC63, SENP3, SEP1 5, SERINC1, SERINC3, SF3B2, SFXN3, SGCE, SIAE, SLBP, SLC12A4, SLC1A1, SLC35B1, SLC35E2, SLC35E3, SLC39A1, SLC39 A7, SMPD1, SNORA70F, SNX11, SNX19, SPPL2A, SPRED2, SRPR, STAU1, STEAP1B, STIM1, STK38, STX18, SYPL1, SYT11, TAF1 1, TAPBPL, TBC1D22B, TCN2, TCTA, TES, TFG, TFRC, TGFB3, TM7SF2, TM9SF1, TMBIM1, TMCO1, TMCO3, TMED10, TMEM138, TM EM179B, TMEM185A, TMEM216, TMEM50A, TNS1, TNS3, TPI1P2, TRAM2, TRIM16L, TSPAN9, TUB, TUBA1A, UBAC2, UBE2E3, UNC 50, USO1, USP24, VCL, VOPP1, VPS25, VPS33B, WAS, WBP1, WNT5B, ZCCHC24, ZFP91, ZMAT3, ZMYM6NB, ZNF384, ZNF667-AS1,

[0401] Genes upregulated upon SEQ ID NO:1 treatment <h2 style=";text-align:left;direction:ltr">ACAA2, ACSL1, ACVR1, ADAR, ADD3, ADIPOR2, ADPRM, AGTR1, AKR1B10, AKR1B15, ALAD, ALG14, AMZ2, AMZ2P1, ANKHD1, APOL6, ASCC1, ASH2L, ATF5, ATG3, ATP 5A1、ATP5C1、ATP5J2、ATP6V0E1、ATP6V1B2、ATXN7L3B、B2M、B4GALT5、BBS4、 BIRC5、BLMH、BOD1、BRK1、BST1、BTF3、C11orf73、C16orf58、C17orf62、C19or f52、C1orf43、C4orf3、C5orf15、C5orf58、C7orf25、C8orf33、CALM1、CCNB1 IP1、CCNG2、CD47、CDCA8、CFDP1、CHAMP1、CHCHD3、CHURC1、CISD3、CKAP5、CLM P、CLTA、COA1、COG3、COG8、COPS8、COX14、CPD、CSTF1、CSTF2T、CTCF、CYB5R3 、CYCS、CYTB、DCTN6、DCTPP1、DDX21、DDX5、DDX60、DECR1、DERL1、DFNA5、DHX1 5, DHX32, DRG1, DSTYK, EFNB3, EMCN, EMG1, ENDOD1, ENPP2, ERICH1, ETFA, EZH1, F8A2, FAM96A, FH, FOS, FOSB, FUCA2, FUNDC2, GABARAPL2, GADD45B, GALNT 15、GAS1、GATS、GBF1、GBP1、GDE1、GGA3、GJA1、GLB1、GLUL、GOLM1、GPR89B、G RAMD3、GSTA4、GTF2E2、H2AFZ、HES1、HEXB、HIBADH、HIST1H4F、HIST2H2AA4、H MGN3、HMGN4、HNRNPM、HOOK2、HSPB8、IDH1、IER3、IFI44L、IFIT1、IFT88、IMM P2L、IMPA2、INTS12、KAT7、KCTD10、KDELR3、KIAA0196、KIF2C、KIFAP3、KLF10 、KLHL18、LAP3、LAPTM4B、LDHA、LDLR、LETMD1、LMAN2L、LMO4、LOXL4、LRP10、 LSM1、LTBP2、LYPD6B、LYSMD2、MAD2L1BP、MANBA、MAT2A、MCL1、MEDAG、MFAP1、<h2 style=";text-align:left;direction:ltr">MIF-AS1、MIOS、MKKS、MMGT1、MRFAP1、MROH5、MRPL20、MRPL33、MYD88、NDN、N DRG1、NDUFV2、NKAP、NOLC1、NT5C3B、NT5E、OAS1、OAT、OAZ2、ODF3L2、OLFML1 、OLFML2B、OSBP、OSTC、OTUD1、OXCT1、PAIP2、PARL、PARP9、PATL1、PCED1A、P DE1A、PEAR1、PELO、PFKL、PFKM、PGAM1、PGK1、PGM1、PIGH、PLEKHJ1、POGK、POL D2, POLR2B, POLR3F, PPA2, PPAP2B, PPAPDC2, PPL, PPP2CB, PPP2R2A, PQLC2L, PRC1, PRDM4, PSMA2, PSMA5, PSMD10, PSME4, PTGS1, PTP4A1, PTTG2, RAB31, R AB32、RAB9BP1、RABEP1、RAC1、RASL11B、RCN1、RHOA、RIOK1、RNASEL、RNF121 、RNF146、RNLS、RPA2、RPL15、RPL21P44、RPP38、RTCB、SEC22C、SEC23IP、SENP 2、SEP15、SERP2、SERTAD4、SETX、SFRP1、SFT2D1、SGCE、SIAE、SLBP、SLC25A1 2、SLC25A25、SLC30A9、SLC35B1、SLC44A1、SLC9A3R1、SMAD7、SMIM19、SNAPC 5、SNHG3、SNORD116-19、SNORD96A、SNX19、SRRD、SRSF2、STARD7、STK16、STO M、STOML2、SUCLA2、SUCLG1、SYNGR4、SYT11、TASP1、TCP1、TFRC、TGFBR2、TIMM 17A、TIMMDC1、TINF2、TMCO1、TMEM173、TMEM203、TMEM230、TMEM50A、TMEM50 B、TMEM60、TMX2、TNC、TNS3、TOMM22、TOR1A、TOR3A、TP53I3、TPBG、TPI1、TRIB 1、TRIL、TUBB3、TUG1、TUSC3、UBE2C、UBE2E3、UBE2L3、UGCG、UNC119、UQCRFS 1、USP18、USP24、USP34、VAMP7、VDAC3、VIM-AS1、VOPP1、WARS、WDR36、WDR48、WDR61, WDYHV1, WLS, WRB, XPNPEP1, XRCC6BP1, YIPF6, ZFP91, ZMAT2, ZNF157, ZNF281, ZNF384, ZNF426, ZNF564, ZNF696, ZNF75D, ZSCAN32,

[0402] Genes downregulated upon SEQ ID NO:1 treatment ABLIM1、ACTR2、ADAM20、AHSA2、ALDH1L2、AMN、APC、APOLD1、ARGLU1、ARPC5、ASAP1-IT1、ATXN2、ATXN3、AURKAPS1、BCL2L11、BCLAF1、BRD1、BTG3、C14orf169、C1QTNF5、C20orf141、C20orf96、C5orf28、C6orf203、CACNA2D1、CCDC102B、CCDC125、CCDC6、CCNG1、CCNL1、CD46、CEACAM19、CENPC、CHML、CIRBP、CN2、COL1A1、COL4A1、CRYBG3、CWC22、DDX3X、DEND4B、DHRS3、DIAPH2、DLG1、DNAJB4、DNAJC3、DOCK6、DPEP3、DSE、DST、DYNLT3、DZIP3、EDIL3、EGLN1、EIF4G3、ELF2、ELP2、EPC1、FAM111A、FAM177A1、FBXO32、FERMT2、FGF7、FNIP1、FOSL2、FOXN2、FRYL、GABPB1-AS1、GABRE、GLS、GOLGA6L4、GOLGA8R、GOLIM4、GPATCH11、GPR135、GTF2H5、HCG11、HCG18、HIF1A、HNRNPA1、HOOK3、HOXC6、HSP90AA1、HSP90AA2P、HSPA1B、HSPA4、HSPH1、HTT、IGF2R、ITGB3、JMJD4、KCNMB2-AS1、KCTD3、KDM2A、KIAA1143、KIZ、KRAS、KRT8P12、LCE1D、LINC01506、LOX、LPGAT1、LPP、LRP12、LRRC3C、MACF1、MAMDC2、MAP1B、MAP2K3、MAP3K10、MATR3、MBNL1、MBTPS2、MDM2、MDM4、MED15、METTL15、MGC24103、MUSTN1、MYH3、MYH8、MYLPF、MYO6、MZF1、NAB1、NADK、NADK2、NBEAL1、NCKAP5、ND6、NEK7、NKTR、NR2C1、NRN1、OFD1、PAPOLA、PAWR、PAXBP1、PBRM1、PCDHGA2、PDE1C、PDZD8、PEAK1、PFDN2、PGBD2、PHF1、PJA2、PLOD3、POLR2J2、PPME1、PPP1R14A、PPP1R16B、PPP1R2, PPP1R3B, PRO2852, PRPF40A, PRUNE2, PTPN11, PUM2, PXK, RAB12, RAB2A, RBM10, RCN2, REEP3, REST, RHOBTB3, RHOQ, RIOK3, RNF19B, RNF215, RNPC3, ROBO1, RO CK1, RPL26L1, SAMD14, SDPR, SEPT7P2, SERP1, SFSWAP, SGIP1, SH3KBP1, SKI, SLC35A5, SLC46A3, SLC7A5, SMARCC1, SMARCC2, SMN1, SNAR-H, SNORA16A, SNORD89, SOX6, SPHAR, SREK1, SRSF11, STT3B, STX2, TAB2, TFPI, THBS1, THOC2, TINAGL1, TMA16, TMEM106C, TMEM26-AS1, TNFAIP8, TOB1, TTC28, TXLNG, UBE2D1, UBE2Q2P1, UBL3, UCHL 5, UPF2, UPF3A, USP15, USP33, UTRN, UVSSA, WHAMMP1, WNT3, YBX3, YWHAZ, ZBTB2, ZFC3H1, ZMYND11, ZNF22, ZNF37A, ZNF429, ZNF525, ZNF532, ZNF626, ZRANB2, ZSWIM8,

[0403] (Example 20) Proteomics method Protein levels were obtained using LC / MS / MS. Statistical analysis on protein counts yielded under- and over-abundant proteins. Only absolute protein level fold change values ​​greater than 0.379 were retained. For all these proteins, microarray analysis provides the associated transcript fold change.

[0404] Mass spectrometry was performed by MSBioworks. Cell pellets were lysed in 100 μL of modified RIPA buffer by sonication. Quantification was performed using Qubit fluorescence. 20 μg of each sample was separated on a 4-12% Bis-Tris gradient gel using MOPS buffer. Gels were stained with Coomassie and each lane was cut into 20 equally sized segments.

[0405] Gel pieces were processed using a robot (ProGest, DigiLab) according to the following protocol: Washed with 25 mM ammonium bicarbonate followed by acetonitrile. Reduction with 10 mM dithiothreitol at 60 °C was followed by alkylation with 50 mM iodoacetamide at RT. Digestion with trypsin (Promega) was performed at 37°C for 4 hours. After quenching with formic acid, the supernatant was analyzed directly without further processing.

[0406] Digests were analyzed by nanoLC / MS / MS with a Waters NanoAcquity HPLC system interfaced to a ThermoFisher Q Exactive. Peptides were loaded onto a capture column and eluted on a 75 μm analytical column at 350 nL / min. Both columns were packed with Luna CI 8 resin (Phenomenex). A 30 min gradient was used (total 10 h per sample). The mass spectrometer was operated in data-dependent mode and MS and MS / MS were performed on an Orbitrap at a resolution of 70,000 FWHM and 17,500 FWHM, respectively. The 15 most abundant ions were selected for MS / MS. Data was searched using a local copy of Mascot with the following parameters: Enzyme: Trypsin; Database: Swissprot Human (forward and reverse with common contaminant sequences added); Fixed modification: Carbamidomethyl (C); Variable modifications: Oxidation (M), Acetyl (protein N-terminus), Pyro-Glu (N-terminus Q), Deamidation (NQ); Mass values: Monoisotopic Peptide mass tolerance: 10 ppm; Fragment mass tolerance: 0.02 Da; Maximum miscleavages: 2 Mascot DAT files were sent to Scafold software for validation, filtering, and generation of non-redundant lists per sample. Data were filtered at 1% protein and peptide level false discovery rate (FDR), requiring at least two unique peptides per protein. Up- and down-regulated proteins were then estimated.

[0407] mass spectrometry Digests were analyzed by nanoLC / MS / MS with a Waters NanoAcquity HPLC system interfaced to a ThermoFisher Q Exactive. Peptides were loaded onto a capture column and eluted at 350 nL / min on a 75 μm analytical column, both packed with Luna CI 8 resin (Phenomenex). A 30 min gradient was used (total 10 h per sample). The mass spectrometer was operated in data-dependent mode, and MS and MS / MS were performed on an Orbitrap at a resolution of 70,000 FWHM and 17,500 FWHM, respectively. The 15 most abundant ions were selected for MS / MS.

[0408] Example 21 QPCR for specific genes of interest and their fold changes Experimental procedure Quantitative PCR was performed using a TaqMan probe-based method to quantify target mRNA expression relative to the expression of the B2M housekeeping gene. A master mix containing primers / probes and Taqman® Gene Expression Master Mix (ABI Biosystems, CA, USA) was added to 1 μl of cDNA template. A final volume of 10 μl was pipetted in duplicate into wells of a 96-well Lightcycler plate (Sarstedt, Numbrecht, Germany) and real-time PCR was performed on a Roche Lightcycler 96 (Roche Diagnostics, Basal, Switzerland) real-time thermal cycler. The threshold cycle (Ct) of each well was calculated using the instrument software. Data analysis was based on the ΔΔCt method with raw data normalized by the B2M housekeeping gene incorporated on the plate. Results are expressed as fold change relative to control.

[0409] QPCR Results (The method of treatment remained the same and the concentration of SEQ ID NO:1 remained at 0.5ug / ml)

[0410] [Table 10]

[0411] Example 22 Exploring the effects of a synthetic peptide (SEQ ID NO:1) on blood glucose control in mouse models of obesity and diabetes Experimental procedure Animal Description: Species: KK.Cg-Ay / J (KKAY mouse) ·Supplied by: Jackson Laboratories Age: 12 weeks ·Gender: Male Randomization: Based on baseline fasting glucose

[0412] Housing and Feeding: - Taming: 5 days or more Housing: Mice were housed on a 12-hour light / dark cycle. : No more than 4 mice per cage depending on size : Ventilated cage rack system Diet: Standard rodent chow and water (available ad libitum)

[0413] design: Route of administration: Subcutaneous injection Dosage: 10mL / kg Dose levels: Test drug: 100μM / kg, Metformin: 250mg / kg Number per group: 10 Total number of animals: 30

[0414] [Table 11]

[0415] Oral glucose tolerance test in KKAY mice Thirty minutes before the OGTT, mice were bled for baseline glucose and insulin (baseline). This was immediately followed by test drug administration. Thirty minutes later, at 0 minutes, all mice were administered a glucose solution (2 g / kg) via oral gavage. After the glucose challenge, blood glucose levels were measured according to the following schedule:

[0416] [Table 12]

[0417] The results are illustrated in Figures 18 to 20. Figure 18 illustrates the fasting blood glucose of KKAY mice after 5 days of treatment with the peptide of SEQ ID NO: 1. Figure 19 illustrates the fasting blood glucose of KKAY mice after 7 days of treatment with the peptide of SEQ ID NO: 1. Figure 20 illustrates the body weight of KKAY mice after 13 days of treatment with the peptide of SEQ ID NO: 1.

[0418] conclusion Treatment with peptide of SEQ ID NO:1 significantly reduces blood glucose after 7 days of treatment.Treatment with peptide of SEQ ID NO:1 does not cause weight gain, as many antidiabetic treatments do.SEQ ID NO:1 reduces the weight of treated group compared to non-treated group.

[0419] (Example 23) Antioxidant assay (DPPH radical scavenging activity assay) DPPH tests were performed using the method described in Lin SY, Wang J, Zhao P, Pang Y, Ye HQ, Yuan Y, Liu JB and Jones G, Optimized antioxidant peptides fractions preparation and secondary structure analysis by MIR. Int J Biol Macromol 59: 151-157 (2013).

[0420] DPPH radical was dissolved in ethanol to a final concentration of 0.1 mM. Peptides and ascorbic acid (positive control) were dissolved in demineralized water to obtain the tested concentrations. In a 96-well plate, samples were prepared as follows: 100 μL of 0.1 mM DPPH solution, 100 μL of peptide solution, and 100 μL of ethanol were mixed per well. Ascorbic acid was prepared as follows: 100 μL of 0.1 mM DPPH solution, 100 μL of ascorbic acid, and 100 μL of ethanol were mixed per well. A blank was prepared as follows: 100 μL of 0.1 mM DPPH solution, 200 μL of ethanol were mixed per well. Triplicate wells were used for each test condition.

[0421] The plate was incubated in the dark for 30 minutes at 25° C. The absorbance at 517 nm was then measured using a plate reader.

[0422] The DPPH radical scavenging activity (%) is as follows: DPPH radical scavenging activity (%) = [1-(absorbance in sample / absorbance in blank)] x 100 It was calculated as follows.

[0423] Data were presented as percentage of the mean value of DPPH radical scavenging activity ± standard deviation.

[0424] (Example 24) Inflammatory response assay TNFα is secreted by macrophages in response to stimulation by endotoxins such as lipopolysaccharide (LPS). TNFα is thought to be involved in systemic inflammation, and dysregulation of TNFα production is thought to be involved in many diseases. The Biolegend assay is a sandwich ELISA kit designed for accurate quantification of human TNFα from cell culture supernatants, serum, or plasma.

[0425] THP-1 monocytes were seeded at 10,000 cells per well in RPMI containing 10% fetal calf serum (FCS), 1% Pen / strep, 1% L-glutamine, 100 nM PMA in 96-well plates and allowed to differentiate for 72 hours before experimentation. After differentiation, cells were incubated with 100 ng / ml, 10 ng / ml or 1 ng / ml of each of the synthetic peptides for 24 hours. After treatment, cells were stimulated with 10 ng / ml LPS for 5 hours and the amount of TNFα in the supernatant was determined using a Biolegend assay ELISA kit.

[0426] Results were calculated as a percentage of untreated controls. An increase in optical density reading indicates an increase in the amount of TNFα released into the cell culture supernatant. All experiments were performed in duplicate (6 wells per condition) on three plates. Significance was determined by Student's t test ( * p<0.05 compared with control; ** p<0.01 compared with control; *** p<0.001 compared to control) was used to calculate the mean mean ± SEM. Equivalent

[0427] The foregoing description details the presently preferred embodiments of the present invention. Numerous modifications and variations in its practice are anticipated by those skilled in the art upon review of the description. These modifications and variations are intended to be encompassed within the scope of the appended claims.

Claims

1. A composition comprising a peptide of SEQ ID NO:1 or a glucose transport enhancing variant of SEQ ID NO:1 having 1 to 3 amino acid mutations compared to SEQ ID NO:1, wherein the amino acid changes occur at one or more of positions 1, 2, 3, 9, 10 and 11 of SEQ ID NO:1, and the amino acid mutations in said variant include amino acid insertions, additions, deletions and substitutions.

2. The composition of claim 1, wherein the glucose transport-enhancing mutant has an amino acid change at position 1, an amino acid change at positions 1 and 2, an amino acid change at positions 1, 2 and 3, an amino acid change at position 11, an amino acid change at positions 10 and 11, or an amino acid change at positions 9, 10 and 11.

3. The composition of claim 2, wherein the amino acid change occurs at position 1 and is selected from an amino acid insertion, addition, deletion and substitution.

4. The composition of claim 2 , wherein the amino acid change is a deletion of an amino acid at positions 1, 2 and / or 3.

5. The composition of claim 4 , wherein the amino acid changes are deletions of amino acids at positions 1 and 2, or deletions of amino acids at positions 1, 2 and 3.

6. The composition of claim 2 , wherein the amino acid change is a deletion of an amino acid at position 9, 10 and / or 11.

7. The composition of claim 2 , wherein the amino acid change is a deletion or addition of an amino acid at position 11.

8. The composition of claim 6 , wherein the amino acid change is a deletion of amino acids at positions 10 and 11.

9. The composition of claim 6 , wherein the amino acid changes are deletions of amino acids at positions 9, 10 and 11.

10. A composition comprising a peptide of SEQ ID NO: 1 or a cell proliferation-promoting variant of SEQ ID NO: 1 having 1 to 3 amino acid mutations compared to SEQ ID NO: 1, wherein the amino acid changes occur at one or more of positions 1, 2, 3, 9, 10 and 11 of SEQ ID NO: 1, and the amino acid mutations in the variant include amino acid insertions, additions, deletions and substitutions.

11. The composition of claim 10 , wherein the cell proliferation-promoting mutant has an amino acid change at position 1, an amino acid change at positions 1 and 2, or an amino acid change at positions 10 and 11.

12. The composition of claim 11 , wherein the amino acid change occurs at position 1 and is an amino acid substitution.

13. The composition of claim 11 , wherein the amino acid change is a deletion of an amino acid at position 1 and / or 2.

14. The composition of claim 13 , wherein the amino acid change is a deletion of amino acids at positions 1 and 2.

15. The composition of claim 11 , wherein the amino acid change is a deletion of an amino acid at position 10 and / or 11.

16. The composition of claim 15 , wherein the amino acid change is a deletion of amino acids at positions 10 and 11.

17. The composition of any one of claims 1 to 16, wherein the peptide is a bioactive peptide.

18. 18. A composition according to any preceding claim, comprising a plurality of peptides.

19. 19. The composition of any one of claims 1 to 18, which is a pharmaceutical composition comprising a therapeutically effective amount of the peptide in combination with a suitable pharmaceutical carrier.

20. 20. The composition of claim 19, wherein the peptide consists of SEQ ID NO:

1.

21. 19. The composition of claim 17 or 18, formulated for oral or parenteral administration to a patient.

22. 22. The composition of any one of claims 1 to 21, wherein the peptide is a modified peptide.

23. The composition of claim 22, wherein the peptide is modified to extend its plasma half-life.

24. The composition of claim 22 or 23, wherein the peptide has been modified by side chain modifications, introduction of protecting groups, incorporation of up to five unnatural amino acids, cyclization, conjugation with a conjugation partner, covalent bonding with a binding partner, fusion with a fusion partner, PEGylation, amidation, lipidation, use of a cross-linking agent, or a method that imposes conformational constraints on the peptide.

25. 25. The composition of claim 24, wherein the peptide is modified with a modifying group selected from the group consisting of polyethylene glycol (PEG), monomethoxypolyethylene glycol, dextran, poly(N-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, polypropylene glycol, polyoxyethylated polyols (e.g., glycerol), and polyvinyl alcohol, colominic acid or other carbohydrate-based polymers, polymers of amino acids, and biotin derivatives.

26. 26. The composition of any one of claims 10 to 18 or 20 to 25, which is a cosmetic composition.

27. 26. The composition of any one of claims 10 to 18 or 20 to 25, wherein the composition is a formulation for a topical composition.

28. 28. The composition of claim 27, formulated for topical application to human skin in a formulation selected from the group including creams, multiple emulsions, anhydrous compositions, aqueous dispersions, oils, milks, balsams, foams, lotions, gels, cream gels, hydroalcoholic solutions, hydroglycolic solutions, cosmetics, personal care products, hydrogels, liniments, serums, soaps, dusting powders, pastes, semisolid formulations, liniments, serums, shampoos, conditioners, ointments, optional rinse formulations, talcs, mousses, powders, sprays, aerosols, solutions, suspensions, emulsions, syrups, elixirs, polysaccharide films, patches, gel patches, bandages, adhesive systems, water-in-oil emulsions, oil-in-water emulsions, and silicone emulsions.

29. 29. A composition according to any one of claims 26 to 28, wherein the peptide has cell proliferation promoting activity for use in delaying or inhibiting the onset of skin ageing.

30. 29. The composition of any one of claims 26 to 28, wherein the peptide has cell proliferation promoting activity for use in the treatment or care of one or more of the visible signs of wrinkles, stretch marks and dark circles under the eyes, dryness, fine lines, age spots, red spots, and sagging skin, as well as conditions caused by sun exposure including sunburn, stress, pollution, and diet.

31. 26. A composition according to any one of claims 19 to 25, for use in a method for treating or preventing diabetes or pre-diabetes, wherein the peptide has glucose transport promoting activity.

32. The composition of claim 31 , wherein the diabetes is type 2 diabetes.

33. The composition of claim 31 or 32, wherein the composition is to be administered together with one or more agents consisting of the group consisting of insulin, insulin analogues, insulin derivatives, glucagon-like peptide 1, glucagon-like peptide 2, derivatives or analogues of glucagon-like peptide 1 or glucagon-like peptide 2.

34. 34. A medical device comprising the composition of any one of claims 1 to 33.

35. A peptide of sequence SEQ ID NO:1 or a glucose transport enhancing variant of the sequence of sequence SEQ ID NO:1 having 1 to 3 amino acid mutations compared to SEQ ID NO:1, wherein the amino acid changes occur at one or more of positions 1, 2, 3, 9, 10 and 11 of SEQ ID NO:1, and the amino acid mutations in said variant include insertions, additions, deletions and substitutions of amino acids.

36. The peptide of claim 35, wherein the glucose transport-promoting mutant has an amino acid change at position 1, an amino acid change at positions 1 and 2, an amino acid change at positions 1, 2 and 3, an amino acid change at position 11, an amino acid change at positions 10 and 11, or an amino acid change at positions 9, 10 and 11.

37. 37. The peptide of claim 36, wherein the amino acid change occurs at position 1 and is selected from an amino acid insertion, addition, deletion and substitution.

38. 37. The peptide of claim 36, wherein the amino acid change is a deletion of an amino acid at positions 1, 2 and / or 3.

39. 39. The peptide of claim 38, wherein the amino acid change is a deletion of amino acids at positions 1 and 2, or a deletion of amino acids at positions 1 and 2.

40. 37. The peptide of claim 36, wherein the amino acid change is a deletion of an amino acid at positions 9, 10 and / or 11.

41. 37. The peptide of claim 36, wherein the amino acid change is a deletion or addition of an amino acid at position 11.

42. 41. The peptide of claim 40, wherein the amino acid change is a deletion of amino acids at positions 10 and 11.

43. 41. The peptide of claim 40, wherein the amino acid changes are deletions of amino acids at positions 9, 10 and 11.

44. A peptide which is a cell proliferation promoting variant of the sequence of SEQ ID NO: 1, having 1 to 3 amino acid mutations compared to SEQ ID NO: 1, wherein the amino acid changes occur at one or more of positions 1, 2, 3, 9, 10 and 11 of SEQ ID NO: 1, and the amino acid mutations in said variant include amino acid insertions, additions, deletions and substitutions.

45. The peptide of claim 44, wherein the cell proliferation-promoting mutant has an amino acid change at position 1, an amino acid change at positions 1 and 2, or an amino acid change at positions 10 and 11.

46. 46. ​​The peptide of claim 45, wherein the amino acid change occurs at position 1 and is an amino acid substitution.

47. 46. ​​The peptide of claim 45, wherein the amino acid change is a deletion of an amino acid at position 1 and / or 2.

48. 48. The peptide of claim 47, wherein the amino acid change is a deletion of amino acids at positions 1 and 2.

49. 46. ​​The peptide of claim 45, wherein the amino acid change is a deletion of an amino acid at position 10 and / or 11.

50. 50. The peptide of claim 49, wherein the amino acid change is a deletion of amino acids at positions 10 and 11.

51. 51. A peptide according to any one of claims 35 to 50, which is modified.

52. 52. The peptide of claim 51, which is modified with a protecting group.

53. 53. The peptide of claim 52, modified with an N-terminal protecting group or a C-terminal protecting group.

54. 52. The peptide of claim 51, modified by replacing at least one L-amino acid with a D-isomer.

55. 52. The peptide of claim 51, which is modified with a modifying group selected from polyethylene glycol (PEG), monomethoxypolyethylene glycol, dextran, poly(N-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymer, poly(propylene oxide / ethylene oxide) copolymer, polypropylene glycol, polyoxyethylated polyols (e.g., glycerol) and polyvinyl alcohol, colominic acid or other carbohydrate-based polymers, polymers of amino acids, and biotin derivatives.

56. 52. The peptide of claim 51, wherein at least one amino acid is modified by replacing with an amino acid analogue.

57. 52. The peptide of claim 51, which has been modified to increase the resistance of the modified peptide to proteolysis by mammalian gastrointestinal proteases.

58. 58. A peptide or modified peptide according to any one of claims 35 to 57 for use as a medicament.

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