Compositions and methods for invasive and non-invasive procedural skin care

Abstract

This invention relates to compositions and methods for invasive and non-invasive procedural skincare. In particular, it relates to topical compositions for promoting skin repair, anhydrous topical compositions, methods for preparing a skin bed prior to dermatological treatment, and methods for promoting skin repair after dermatological treatment. More generally, this invention relates to skin care treatments, and more specifically to compositions and methods for promoting skin health, skin regeneration, skin repair, skin bed preparation, and enhanced wound healing.

Description

[0001] This application is a divisional application of the application filed on February 2, 2017, with application number 201780022293.5 and invention title "Compositions and methods for invasive and non-invasive procedural skin care". Incorporation of relevant applications

[0002] Any and all priority claims identified in the application data sheet, or any correction thereof, are incorporated herein by reference in accordance with 37 CFR 1.57. This application claims the benefits of U.S. Provisional Application No. 62 / 291,376, filed February 4, 2016, and U.S. Provisional Application No. 62 / 303,332, filed March 3, 2016. Each of the foregoing applications is incorporated herein by reference in its entirety, and each application is expressly incorporated in this specification. Invention Field

[0003] This invention provides compositions and methods for skin care treatments, including compositions and methods for skin repair, promoting skin health, skin regeneration, and enhancing wound healing. Background Technology

[0004] It is estimated that the invasive dermatological industry, including cosmetic surgery, facial aesthetics, and medical lasers, will expand to $3 billion by 2017. Due to this significant industry growth, the demand for skincare treatments that effectively promote skin regeneration and mitigate the negative side effects of invasive dermatological procedures is also rapidly increasing. These negative side effects are often the result of slow and ineffective skin regeneration or wound healing, leading to prolonged inflammation, skin allergies, scarring, bruising, dry skin, infections, and other unfortunate skin conditions.

[0005] Traditional skincare treatments for alleviating these negative side effects primarily rely on a variety of common over-the-counter medications. These common post-operative medications and over-the-counter products include skincare products such as facial and body lotions, creams, petroleum jelly or petroleum jelly-based products, butters, skin moisturizers, and various other treatment products. However, these common medications may often be primarily effective in treating one specific side effect, such as dry skin, while having little effect on treating other negative side effects. Furthermore, many widely available skincare products on the market focus on treating symptoms of slow wound healing but do not enhance or promote the skin's own regeneration. Summary of the Invention

[0006] This invention provides compositions and methods for promoting skin health, enhancing skin regeneration, promoting wound healing, and treating other skin conditions. These compositions preferably comprise combinations of two different peptides: a dipeptide, tripeptide, or tetrapeptide, and a pentapeptide, hexapeptide, or heptapeptide. The peptide combinations effectively promote skin health, skin regeneration, wound healing, and treat or improve various other skin conditions. This invention also provides methods for preparing and using the compositions.

[0007] Therefore, in a first aspect, a topical composition for promoting skin repair is provided, the topical composition comprising: one or more dipeptides, tripeptides or tetrapeptides; and one or more pentapeptides, hexapeptides or heptapeptides.

[0008] In a second aspect, a method for preparing a skin bed prior to treatment of a dermatological condition is provided, the method comprising: applying a topical composition of the first aspect to the skin bed, thereby preparing the skin bed for subsequent treatment of the dermatological condition, wherein applying the topical composition to the skin bed prior to the treatment of the dermatological condition promotes healing of damaged skin after the treatment of the dermatological condition.

[0009] In one embodiment of the second aspect, the topical composition of the first aspect is applied to the skin bed at least once a day.

[0010] In one embodiment of the second aspect, the topical composition of the first aspect is applied to the skin bed at least twice daily.

[0011] In one embodiment of the second aspect, the topical composition of the first aspect is applied for at least two weeks prior to the treatment of the skin condition.

[0012] In one embodiment of the second aspect, the topical composition of the first aspect is applied for at least four weeks prior to the treatment of the skin condition.

[0013] In a third aspect, a method for promoting skin repair after treatment for a dermatological disease is provided, the method comprising: applying the topical composition of the first aspect to skin damaged by the treatment for a dermatological disease, thereby promoting the healing of the damaged skin.

[0014] In one embodiment of the third aspect, the topical formulation of the first aspect is applied to the skin bed at least once a day.

[0015] In one embodiment of the third aspect, the topical formulation of the first aspect is applied to the skin bed at least twice daily.

[0016] In one embodiment of the third aspect, the topical formulation of the first aspect is applied for at least two weeks after the treatment of the skin disease.

[0017] In one embodiment of the third aspect, the topical formulation of the first aspect is applied for at least four weeks after the treatment of the skin disease.

[0018] In one embodiment of the third aspect, the method further comprises applying the topical composition of the first aspect to a skin bed prior to treatment of the dermatitis. For example, the topical composition of the first aspect is applied to the skin bed at least once daily, or at least twice daily, or initially applied for at least two weeks prior to treatment of the dermatitis, or initially applied for at least four weeks prior to treatment of the dermatitis.

[0019] In one embodiment of the method of the second or third aspect, or in any of their respective embodiments, the skin disease treatment is laser treatment.

[0020] In one embodiment of the method of the second or third aspect, or in any of their respective embodiments, the skin disease treatment is a chemical peel.

[0021] In one embodiment of the method of the second or third aspect, or in any of their respective embodiments, the skin disease treatment is for actinic keratosis.

[0022] In one embodiment of the method of the second or third aspect, or in any of their respective embodiments, the dermatological treatment is a treatment for reducing signs of aging.

[0023] In a fourth aspect, a topical composition for promoting skin repair is provided, the topical composition comprising palmitoyl tripeptide-1 and palmitoyl hexapeptide-12.

[0024] In one embodiment of the fourth aspect, the topical composition further comprises heptyl undecenoate.

[0025] In a fifth aspect, an anhydrous local composition is provided comprising phosphatidylserine, oleuropein, and octanoyl polymethylsiloxane, wherein the viscosity of the local composition is from 10,000 cPs to 25,000 cPs, and wherein the anhydrous local composition has the ability to maintain stability in three temperature test cycles from -10°C to 25°C.

[0026] In a sixth aspect, a topical composition for promoting skin repair is provided, the topical composition comprising: 82-92 wt.% of cyclopentasiloxane and polydimethylsiloxane crosspolymer; 1-4 wt.% of heptyl undecenoate; 0.01-0.06 wt.% of palmitoyl hexapeptide-12; 0.01-0.06 wt.% of palmitoyl tripeptide-1; 0.25-1 wt.% of octanoyl polymethylsiloxane; 0.05-0.1 wt.% of phosphatidylserine / lecithin; and 0.05-0.1 wt.% of oleuropein.

[0027] In one embodiment of the sixth aspect, the topical composition comprises: 2-5 wt.% of a first carrier containing palmitoyl hexapeptide-12, the first carrier further comprising pentaerythritol tetraisostearate, caprylic / capric triglyceride, propylene carbonate, and silachlorite, wherein the concentration of palmitoyl hexapeptide-12 in the carrier is 100 ppm; and 2-5 wt.% of a second carrier containing palmitoyl tripeptide-1, the second carrier further comprising pentaerythritol tetraisostearate, caprylic / capric triglyceride, propylene carbonate, and silachlorite, wherein the concentration of palmitoyl tripeptide-1 in the carrier is 100 ppm.

[0028] In one embodiment of the sixth aspect, the topical composition further comprises: 1-4 wt.% panthenol triacetate / naringenin; 1-4 wt.% arnica montana extract; and 0.5-2 wt.% Dunaliella salina extract.

[0029] In a seventh aspect, a method for preparing a skin bed prior to treatment of a dermatological condition is provided, the method comprising: applying a topical composition of any one of the fourth to sixth aspects to the skin bed, thereby preparing the skin bed for treatment of a dermatological condition affecting damaged skin, thereby promoting healing of the damaged skin following the treatment of the dermatological condition.

[0030] In one embodiment of the seventh aspect, the topical composition is applied to the skin bed at least once daily for at least two weeks prior to treatment of the dermatitis.

[0031] In one embodiment of the seventh aspect, the topical composition is applied to the skin bed at least twice daily for at least four weeks prior to treatment of the dermatitis.

[0032] In an eighth aspect, a method for promoting skin repair after treatment for a dermatological disease is provided, the method comprising: applying a topical composition of any one of the fourth to seventh aspects to skin damaged by the treatment for a dermatological disease, thereby promoting the healing of the damaged skin.

[0033] In one embodiment of the eighth aspect, the topical composition is applied to the skin bed at least once daily for at least two weeks prior to treatment of the dermatitis.

[0034] In one embodiment of the eighth aspect, the topical composition is applied to the skin bed at least twice daily for at least four weeks prior to treatment of the dermatitis.

[0035] In a ninth aspect, a method for preparing a skin bed prior to dermatological treatment is provided, the method comprising: applying a topical composition of any one of the fourth to seventh aspects to the skin bed, thereby preparing the skin bed for dermatological treatment of damaged skin; and subsequently applying the topical composition to the skin damaged by the dermatological treatment, thereby promoting the healing of the damaged skin.

[0036] In one embodiment of the ninth aspect, the topical composition is applied to the skin bed at least once daily for at least two weeks prior to treatment of the dermatitis.

[0037] In one embodiment of the ninth aspect, the topical composition is applied to the skin bed at least twice daily for at least four weeks prior to treatment of the dermatitis.

[0038] In any of the seventh to ninth aspects, the skin disease treatment is laser treatment.

[0039] In one embodiment of any of the seventh to ninth aspects, the skin disease treatment is a chemical peel.

[0040] In any embodiment of the seventh to ninth aspects, the skin disease treatment is a treatment for actinic keratosis.

[0041] In any embodiment of the seventh to ninth aspects, the skin disease treatment is a treatment for reducing signs of aging.

[0042] Any feature of any embodiment of any aspect applies to all other aspects and embodiments identified herein. Furthermore, any feature of any embodiment of any aspect may be combined, in whole or in part, independently with other embodiments described herein; for example, one, two, three, or more embodiments may be combined wholly or partially. Additionally, any feature of any embodiment of any aspect may be optional for other aspects or embodiments. Attached Figure Description

[0043] This patent or application document contains at least one color drawing. A copy of this patent or application disclosure with color drawings will be provided by the competent authority upon request and payment of the necessary fees.

[0044] Figure 1 This is a schematic diagram illustrating the mechanism by which exemplary combinations of peptides work to stimulate and restore elastin and collagen levels in the skin.

[0045] Figure 2 The effects of the tripeptide on promoting skin regeneration are schematically depicted through increased collagen and elastin synthesis, blocking the release of iron oxide from ferritin, attracting healing cells such as capillary cells and macrophages, and re-establishing new blood flow to the site of injury.

[0046] Figure 3 The effects of hexapeptide on promoting skin regeneration and wound healing are schematically depicted by inducing elastin and collagen production, fibroblast proliferation, extracellular matrix regeneration, and fibroblast-keratinocyte migration.

[0047] Figures 4A-4C The study depicted the progress of wound healing in a human subject with a 3 mm erbium CO2 laser spot on his or her forearm in response to treatment with the dipeptide therapy versus a control therapy.

[0048] Figure 5A Data on crusting / scab formation from a laser wound study comparing dipeptide treatment with Aquaphor® treatment and control treatments are provided. Figures 5B-5D These are photos of the appearance of three different wounds on day 4, each wound treated with a dipeptide ( Figure 5B ), control treatment ( Figure 5C ) or Aquaphor® Figure 5D ) to process.

[0049] Figures 6A-6B Provided information about the appearance of the wound ( Figure 6A ) and crust / scab ( Figure 6B Based on data from the study, the study evaluated the wound healing benefits of pretreatment with dipeptide two weeks prior to laser skin surgery and post-operative treatment with dipeptide or Aquaphor®. Figure 6C This is an image of the wound on day 9 after wound peptide treatment. Figure 6D This is an image of the wound on day 9 after Aquaphor® treatment.

[0050] Figures 7A-7C These are patient photos before, 4 days after, and 9 days after laser treatment, showing the use of dipeptide pretreatment and posttreatment in conjunction with laser treatment using an Encore UltraPulse® ActiveFX™ CO2 laser manufactured by Lumenis, Inc., Santa Clara, CA.

[0051] Figure 8The results of a study comparing elastin mRNA in fibroblast monolayers exposed to a combination of dipeptides or a control were described.

[0052] Figure 9 This is a graph showing the change in gelatin-degrading activity (peak area / µg protein) against MMP-2 over time.

[0053] Figure 10 A graph showing the change in gelatin degradation activity (peak area / µg protein) over time is shown.

[0054] Figure 11 The VEGF concentration (pg VEGF / ng DNA) is shown as a function of GHK concentration (ng / mL).

[0055] Figures 12A-12B Provides skin treated with dual peptides ( Figure 12B ) and untreated control ( Figure 12A Photographs (100×) of skin biopsy samples. Samples were stained with H / E, and the photographs demonstrate that treatment with dipeptide pretreatment resulted in a more organized distribution and arrangement of collagen.

[0056] Figures 13A-13B Provides skin treated with dual peptides ( Figure 13B ) and untreated control ( Figure 13A Photographs (100×) of skin biopsy samples. The samples were stained to target elastin (brown), and the photographs demonstrate a significant increase in elastin levels associated with dipeptide treatment.

[0057] Figures 14A-14B Provides skin treated with dual peptides ( Figure 14B ) and untreated control ( Figure 14A Photographs (100×) of skin biopsy samples. Samples were stained by IHC to target procollagen, and the photographs demonstrate a significant increase in procollagen levels associated with dipeptide treatment.

[0058] Figures 15A-15B Skin biopsy samples were provided from the skin treated with the dual peptide three weeks after application. Figure 15B ) and baseline samples ( Figure 15A Photographs (100×) were observed. Increased collagen formation in the upper dermis was observed within three weeks of local application, accompanied by a reduction in sun-induced elastin degeneration via new collagen, thus improving the appearance of the epidermis.

[0059] Figures 16A-16C Provided three weeks after application ( Figure 16B ), eight weeks after application ( Figure 16C Skin biopsy samples and baseline samples of facial skin treated with dipeptides. Figure 16APhotograph (100×). In patients with elastin in elastic tissues damaged by photorejuvenation, local application of the dipeptide resulted in less aggregation of elastin material and significant distribution into deeper dermal layers over eight weeks.

[0060] Figures 17A-17D Skin biopsy samples of skin treated with the dual peptide three weeks after application were provided at 40× ( Figure 17B Baseline samples at 40× ( Figure 17A Skin biopsy samples from skin treated with the dipeptide three weeks after application were taken at 100× ( Figure 17D ) and baseline samples at 100× ( Figure 17C (Photographs). A decrease in MMP1 staining was observed in the preauricular region over a three-week period.

[0061] Figures 18A-18B Skin biopsy samples were provided from the skin treated with the dual peptide three weeks after application. Figure 18B ) and baseline samples ( Figure 18A Photograph (100×). Increased core proteoglycan staining was observed in the preauricular region over a three-week period.

[0062] Figures 19A-19I Data plots generated during a randomized, single-blind trial of the Alastin Procedure Enhancement System are provided, comparing standard care following IPL and / or PDL with Q-switched alexandrite laser and fractional CO2 laser facial resurfacing.

[0063] Detailed Implementation of the Preferred Scheme The following description and examples illustrate preferred embodiments of the invention. Those skilled in the art will recognize that many variations and modifications of the invention are included within its scope. Therefore, the description of preferred embodiments should not be considered as limiting the scope of the invention.

[0064] definition As used herein, the terms “pharmaceutically acceptable salt” and “its pharmaceutically acceptable salt” are broad terms and will be given their common and conventional meaning (and not limited to specific or custom meanings) to those skilled in the art, and refer to, but not limited to, salts prepared from pharmaceutically acceptable non-toxic acids or bases. Suitable pharmaceutically acceptable salts include metal salts (e.g., aluminum, zinc salts), alkali metal salts such as lithium, sodium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts; organic salts such as salts of lysine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucosamine), procaine salts, and tris(hydroxymethyl)aminomethane; salts of free acids and bases; inorganic salts such as sulfates, hydrochlorides, and hydrobromic acids; and other salts currently widely used for pharmaceutical purposes and listed in sources well known to those skilled in the art (e.g., The Merck Index). Any suitable ingredient can be chosen to prepare the salt of the therapeutic agent discussed herein, provided that it is non-toxic and does not substantially interfere with the desired activity. In addition to salts, pharmaceutically acceptable precursors and derivatives of the compound can also be used. Pharmaceutically acceptable amides, lower alkyl esters, and protected derivatives are also suitable for the compositions and methods of the preferred embodiments. Although the compounds of the preferred embodiments can be administered in the form of pharmaceutically acceptable salts, it is generally preferred to administer the compounds in a neutral form.

[0065] It should be understood that in any compound described herein having one or more chiral centers, unless the absolute stereochemistry is explicitly specified, each center may independently be an R-configuration or an S-configuration, or a mixture thereof. Therefore, the compounds provided herein may be enantiomerically pure, enantiomerically enriched, racemic mixtures, diastereomerically pure, diastereomerically enriched, or stereoisomeric mixtures. Furthermore, it should be understood that in any compound described herein having one or more double bonds that generate geometric isomers that can be defined as E or Z, each double bond may independently be E or Z, or a mixture thereof.

[0066] Similarly, it should be understood that all tautomer forms are intended to be included in any of the compounds described. For example, all tautomers of phosphate groups are intended to be included. Furthermore, all tautomers of heterocyclic bases known in the art are intended to be included, including tautomers of natural and non-natural purine and pyrimidine bases.

[0067] It should be understood that when the compounds disclosed herein have unfilled valences, the valences will be filled with hydrogen or its isotopes (e.g., hydrogen-1 (protium) and hydrogen-2 (deuterium)).

[0068] It should be understood that the compounds described herein may be labeled with isotopes. Substitution with isotopes such as deuterium may provide certain therapeutic advantages due to greater metabolic stability, such as prolonged half-life in vivo or reduced dose requirements. Each chemical element represented by a compound structure may include any isotope of that element. For example, in a compound structure, a hydrogen atom may be explicitly disclosed or understood to be present in the compound. At any position in the compound where a hydrogen atom may be present, the hydrogen atom may be any isotope of hydrogen, including but not limited to hydrogen-1 (protium) and hydrogen-2 (deuterium). Therefore, unless the context clearly specifies otherwise, the compounds mentioned herein include all potential isotopic forms.

[0069] It should be understood that the methods and combinations described herein include crystalline forms (also known as polymorphs, which comprise different crystalline arrangements of the same elemental composition of the compound), amorphous phases, salts, solvates, and hydrates. In some embodiments, the compounds described herein are present in a solvated form with a pharmaceutically acceptable solvent such as water, ethanol, etc. In other embodiments, the compounds described herein are present in a non-solvated form. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and can be formed during crystallization with a pharmaceutically acceptable solvent such as water, ethanol, etc. Hydrates are formed when the solvent is water, or alcohols are formed when the solvent is an alcohol. Additionally, the compounds provided herein can be present in both non-solvated and solvated forms. Generally, for the purposes of the compounds and methods provided herein, the solvated form is considered equivalent to the non-solvated form.

[0070] Therapeutic uses The compositions described herein are suitable for skin treatments, and more specifically, for skin care treatments, promoting skin regeneration, and enhancing wound healing. These compositions are suitable for use in any surgical procedure involving skin punctures, incisions, or other skin injuries (e.g., chemical injuries, cryotherapy injuries, mechanical injuries, phototherapy, electrical injuries, thermal injuries, injuries due to skin deformation in stretch marks, etc.), where enhanced wound healing (e.g., shortened healing time, minimization of the appearance of resulting scars, etc.) is desirable. These compositions are suitable for use in enhancing wound healing (e.g., acne scars, burn scars, other skin scars, chronically non-healing wounds). These compositions are suitable for use in conjunction with various dermatological or medical treatments or procedures affecting the dermis (e.g., phototherapy, removal of exocrine glands, radiation therapy for cancer). The compositions described herein are applicable to skin treatment procedures, including but not limited to cosmetic laser resurfacing, laser hair removal, brow lift surgery, chemical peels (e.g., glycolic acid-alpha hydroxy acid peel, trichloroacetic acid peel, phenol peel, etc.), abdominoplasty, wrist arthroplasty, blepharoplasty, breast augmentation, breast fixation, wrinkle removal or lower facelift, rhinoplasty, thigh lift surgery, melanocytic nevus removal, dermabrasion and microdermabrasion, retinoic acid treatment (e.g., isotretinoin, all-trans retinoic acid treatment, etc.), hyaluronic acid injections, botulinum toxin injections, filler treatments (e.g., wrinkle filling treatment, etc.), sclerotherapy, buttock augmentation surgery, and microneedling. Therapies, tattoos and any and all other hair removal treatments, hair growth and regrowth treatments (e.g., in combination with oral or topical minoxidil), infection treatments (in combination with topical, oral or injectable antibiotics), skin rejuvenation or resurfacing, acne removal or reduction, treatment of ruptured capillaries, rosacea treatment, wrinkle reduction, pore reduction, cellulite ablation and other dermal lipid deposits, wart and fungal removal, fading or removal of scars including proliferative scars and keloids, treatment of abnormal pigmentation (e.g., port-wine stains), tattoo removal, treatment of skin inconsistencies (e.g., texture, color, tone, elasticity, hydration, etc.), and general lotions such as hand creams, facial lotions, body lotions, etc.

[0071] Various surgical treatments and procedures involving damage to skin tissue can be performed. Outpatient venectomy is an outpatient procedure performed by a dermatologist who removes superficial veins (such as capillaries, spider veins, or varicose veins) through small, narrow incisions in the skin. This treatment can cause temporary bruising, swelling, and inflammation due to small segments of veins remaining in the skin. Blepharoplasty is a surgical procedure that can restore a youthful appearance to the eye area. The upper and lower eyelids are lifted, and loose or excess skin and fat tissue is removed from the eye area. Mild swelling and bruising may occur. Cryolipolysis (also known as "fat freezing" or the product name CoolSculpting) is a procedure that involves non-invasively freezing body fat to break down fat cells, resulting in fat reduction without damaging other tissues. Results may take several months to appear. Redness and localized bruising may occur. In cryolipolysis, liquid nitrogen is used to remove skin growths, fade age spots, and treat early-stage basal cell carcinoma and squamous cell carcinoma. The doctor will apply the freezing nitrogen using cotton swabs or a spray device. The goal is to rapidly freeze the skin and then allow it to thaw slowly to cause maximum damage to the target skin cells. In some cases, additional application may be necessary. When treating skin cancer, a doctor can insert a small needle containing a thermometer into the treatment area to ensure it has been adequately cooled. Potential complications include redness, swelling, bleeding, blisters, and healing problems. Dermabrasion is a procedure that uses a wire brush or a diamond abrasive wheel with rough edges to remove and smooth the upper layer of skin. The treated area heals, allowing new skin to grow in its place. It is often used for facial surgery. It can cause swelling and infection. Dermal fillers—Juvederm, Restylane, Belotero, and Voluma—contain some form of hyaluronic acid, a substance naturally found throughout the body and in the skin, with the highest concentrations in the fluids of the eyes and joints. Radiesse is a calcium hydroxyapatite filler, while Sculptra contains L-polylactic acid. Fillers can be used temporarily to plump lips, lift sunken scars, and smooth wrinkled skin. First, the doctor marks the areas to be injected. Local or injectable anesthetic may be used to increase comfort. The filler is then injected through a syringe using a very fine needle. The effect typically lasts about six months. Afterward, additional injections are needed to maintain the effect. Potential side effects include swelling, bruising, bleeding, blisters, cysts, and inflammatory reactions. Hair transplantation involves piercing the scalp, where a plug containing hair follicles is removed from a section of scalp where hair is denser and transplanted to the treatment area. Hair transplantation has a high success rate, provided there is sufficient donor hair. This procedure can cause irritation or damage to the skin where hair is removed or transplanted. Laser / phototherapy is a non-invasive procedure that uses light energy to repair and regenerate damaged skin.During this procedure, patients may feel a squeezing sensation similar to a snapping rubber band. Local anesthetic or cryogel may be applied to prevent discomfort. The treated area may appear pink or red after treatment, lasting four to eight hours. A slight tingling sensation, similar to mild sunburn in the treated area, may be observed, which usually subsides within four to six hours. Mild swelling of the treated area, which typically disappears within a few days, may also occur. Potential complications include swelling and scabbing. In liposuction, a surgeon uses a cannula to remove pockets of excess fat from multiple areas of the body. This cannula is inserted through small incisions made in the skin. In some procedures, the fat is loosened with water or liquefied using a laser to facilitate its removal. This procedure is minimally invasive and is usually performed under local tumescent anesthesia. After fat removal, the incision is usually left open to allow drainage. This procedure can result in tissue damage, skin necrosis, bruising, and swelling. In microdermabrasion, a slightly abrasive applicator tip is applied to the surface of the skin on the face or neck to remove the outermost layer of skin. This results in a smoother skin texture. This procedure is painless and non-invasive, but can cause skin irritation or infection. Microinjection involves transferring or recirculating fat from one area of ​​the body to another. Potential complications include bruising and swelling. Tattooing (including what is also called permanent makeup) is the process of inserting colored pigment under the surface of the skin. This procedure can cause inflammation or infection. A neck lift is a surgical option to improve the appearance of the neck. The results are usually permanent. There are two main types of neck lifts, including cervicoplasty (where excess skin is surgically removed from the neck) and platysma myoplasty (a procedure that reduces the band-like appearance of the neck by removing, tightening, or repositioning the neck muscles). This procedure can cause bruising, infection, and swelling. Neuromodulators are wrinkle-relieving injections of botulinum toxin—commercially known as Botox Cosmetic, Dysport, or Xeomin—used to treat wrinkles, frown lines, and crow's feet. A small amount of neuromodulator is injected directly into the deep muscles, causing them to relax and gradually smooth the appearance of the overlying skin. The effects typically last about three months. Potential complications include bruising and soreness. Non-ablative skin rejuvenation uses lasers to improve the appearance of wrinkles, age spots, and minor scars by generating heat within the skin without damaging the skin's surface. The heat generated by the laser promotes collagen production, making the skin firmer and looking younger and healthier. Non-ablative lasers are typically fractional, meaning they deliver heat to the skin through thousands of tiny, dark columns called microthermal treatment zones, intervening in normal, untreated skin. This fractional approach allows the skin to heal faster than when treating an entire area. This method shortens the recovery period and reduces the number of potential complications. In most cases, multiple segments are required. This procedure can cause redness, swelling, and infection.Non-invasive body contouring treatments include low-intensity laser therapy (which delivers cold laser energy to body tissues that are absorbed by fat cells, causing them to break down and be absorbed into the body) and ultrasound (which uses high-intensity focused sound waves to destroy fat cells, causing them to gradually dissolve over time). Radiofrequency delivers controlled energy to areas of excess fat, generating heat deep within the fat cells and subsequently destroying them. Radiofrequency therapy is frequently used in dermatological body contouring. It can cause redness and bruising.

[0072] In each of the aforementioned treatments, some degree of skin damage may occur, which can be mitigated by using peptide compositions as described herein. The peptide compositions disclosed herein may be applied as a pretreatment of the treatment (e.g., once daily for 1-31 days or longer before the start of treatment, such as once daily for one, two, three, or four weeks) and / or as a posttreatment after the treatment (e.g., once daily for 1-31 days or longer after the completion of treatment, such as once daily for one, two, three, or four weeks).

[0073] Actinic keratosis The compositions described herein are suitable for use in conjunction with the treatment of actinic keratosis or certain other forms of cancerous skin lesions. Actinic keratosis, also known as solar keratosis, is a scaly, hardened growth or lesion caused by damage from ultraviolet radiation from the sun. They typically appear in sun-exposed areas such as the face, bald scalp, lips, and backs of the hands, and are usually raised, rough in texture, and wart-like. Most turn red, but some may turn brown, pink, and / or flesh-colored. Without treatment, up to ten percent of actinic keratosis may develop into squamous cell carcinoma. In rare cases, actinic keratosis may also transform into basal cell carcinoma. Almost all actinic keratosis can be eliminated if treated early before they become skin cancer (benign or malignant growths or tumors). Several treatment options are available, depending on the characteristics of the growth and the patient's age and health condition.

[0074] Cryotherapy is the most common treatment when a limited number of actinic keratosis lesions are present. Treatment can be performed in a doctor's office and does not require incision or anesthesia. Liquid nitrogen is applied using a spray device or cotton swab applicator to freeze the growth. The lesion then shrinks and / or blisters, becomes crusted, and falls off. Temporary redness and swelling may occur after treatment, and in some patients, loss of pigmentation may occur, leaving white patches.

[0075] When actinic keratosis is extensive and widespread, commercially available topical creams, gels, and solutions for treating actinic keratosis can be used in combination with the compositions described herein. One of the most commonly used topical medications for actinic keratosis is 5-fluorouracil (5-FU) cream or solution. Apply 5-FU in cream or lotion form to the affected area once or twice daily for two to four weeks. It can be used on all affected areas. 5-FU is available in various formulations at concentrations ranging from 0.5% to 5%. Temporary side effects include redness, swelling, and crusting. Aldara or Zyclara (imiquimod) creams are also used to treat actinic keratosis; they work by stimulating the immune system to produce interferon, a chemical that destroys cancer cells and precancerous cells. Topical formulations (e.g., at concentrations of 5%, 3.75%, or 2.5%) are applied to the affected area two or three times a week for weeks or months. This cream is generally well tolerated, but some individuals may experience redness and ulceration. Actinic keratosis. A gel combining hyaluronic acid with the nonsteroidal anti-inflammatory drug diclofenac may also be effective for individuals whose skin is overly sensitive to other topical treatments. The gel is applied twice daily for two to three months. Picato® (Gastrodin methylbutenoate) is available in 0.015% and 0.05% concentrations, treating actinic keratosis with just two or three days of application, including a 0.015% concentration for three consecutive days (for face and scalp) and a more concentrated 0.05% gel for two consecutive days (for trunk and extremities). Skin redness, peeling / scabbing, crusting, and swelling are the most common side effects. These adverse reactions (redness, crusting, swelling) can be reduced by using peptide compositions as described herein. The peptide compositions disclosed herein can be applied as a pretreatment (e.g., once daily for 1-31 days or longer, such as once daily for one, two, three, or four weeks, before the initiation of application of 5-FU, imiquimod cream, hyaluronic acid / diclofenac or megateryl methylbutenoate), and / or as a topical formulation identical to 5-FU, imiquimod cream, hyaluronic acid / diclofenac or megateryl methylbutenoate, and / or as a posttreatment following the application of 5-FU, imiquimod cream, hyaluronic acid / diclofenac or megateryl methylbutenoate (e.g., once daily for 1-31 days or longer, such as once daily for one, two, three, or four weeks, after application of 5-FU, imiquimod cream, hyaluronic acid / diclofenac or megateryl methylbutenoate has been discontinued).

[0076] Photodynamic therapy is also used to treat actinic keratosis and Bowen's disease (a form of squamous cell carcinoma that presents as persistent reddish-brown scaly patches). Photodynamic therapy is particularly useful for extensive lesions on the face and scalp. A photosensitizer (topically 5-aminolevulinic acid or methyl aminolevulinate) is applied to the lesion. These treated areas are then activated by intense blue or red light, which selectively destroys the actinic keratosis. Redness, pain, and swelling may occur. After the procedure, patients must strictly avoid sun exposure for at least 48 hours, as UV exposure will increase drug activation and may cause severe sunburn. The adverse effects of this treatment (redness, pain, swelling) can be mitigated by using peptide compositions as described herein. The peptide compositions disclosed herein can be applied as a pretreatment for photodynamic therapy (e.g., once daily for 1-31 days or longer before the start of photodynamic therapy, such as once daily for one, two, three, or four weeks) and / or as a posttreatment after photodynamic therapy (e.g., once daily for 1-31 days or longer after the completion of photodynamic therapy, such as once daily for one, two, three, or four weeks).

[0077] Cryotherapy, 5-FU, imiquimod cream, hyaluronic acid / diclofenac, megateryl methylbutyrate, and photodynamic therapy can be combined to treat actinic keratosis, along with the use of the peptide compositions disclosed herein to promote skin repair. Treatment regimens may include cryotherapy in combination with photodynamic therapy or topical agents such as imiquimod (5-FU) or diclofenac. Topical medications and photodynamic therapy may also be used continuously every three months, six months, or one year, as determined by a physician during a routine skin examination. This approach can improve cure rates and reduce side effects. One to two weeks of 5-FU followed by cryotherapy can shorten the healing time of 5-FU and reduce the likelihood of vitiligo after cryotherapy.

[0078] Scraping and electrodrying can be used to remove partial actinic keratosis, accompanied by electrocautery or trichloroacetic acid for hemostasis, optionally under local anesthesia. The peptide formulations disclosed herein can be advantageously used as pretreatment or posttreatment. The peptide compositions disclosed herein can be applied as a pretreatment for scraping and electrodrying (e.g., once daily before initiating scraping and electrodrying for 1-31 days or longer, e.g., once daily for one, two, three, or four weeks) and / or as a posttreatment after scraping and electrodrying (e.g., once daily after completion of scraping and electrodrying for 1-31 days or longer, e.g., once daily for one, two, three, or four weeks).

[0079] Chemical peels, commonly used to reverse signs of photoaging, are also used to remove actinic keratosis on the face, especially when other techniques have failed. Trichloroacetic acid and / or similar chemicals are applied directly to the skin, causing the top layer of skin to peel off. New skin typically regenerates within a few weeks. This technique may require local anesthesia and may cause temporary discoloration and irritation. The peptide formulations disclosed herein can be advantageously used as pretreatments or posttreatments. Peptide compositions disclosed herein can be used as a pretreatment for chemical peels (e.g., once daily for 1–31 days or longer before initiating a chemical peel, such as once daily for one, two, three, or four weeks) and / or as a posttreatment after a chemical peel (e.g., once daily for 1–31 days or longer after the completion of a chemical peel, such as once daily for one, two, three, or four weeks).

[0080] Laser surgery can also be used to treat actinic keratosis. As described elsewhere in this document, carbon dioxide lasers or erbium YAG lasers are used to remove the outer layer of skin and, depending on the depth, deeper layers. Lasers are effective in removing actinic cheilitis of the lips as well as actinic keratosis of the face and scalp. They provide good control over the depth of tissue removed. Lasers are also used as a secondary therapy when topical medications or other techniques are unsuccessful. However, local anesthesia may be required. The risk of scarring and loss of pigmentation is slightly higher than with other techniques. Therefore, the peptide formulations disclosed herein can be advantageously used as pretreatment or posttreatment. The peptide compositions disclosed herein can be applied as a pretreatment for laser treatment (e.g., once daily for 1–31 days or longer before the start of laser treatment, such as once daily for one, two, three, or four weeks) and / or as a posttreatment after laser treatment (e.g., once daily for 1–31 days or longer after the completion of laser treatment, such as once daily for one, two, three, or four weeks).

[0081] Chemical peeling The peptide compositions described herein can be used in conjunction with chemical peeling treatments. Chemical peeling is a technique used to improve the appearance of skin on the face, neck, or hands. A chemical solution is applied to the skin, causing it to exfoliate and eventually peel off. The new skin is typically smoother and has fewer wrinkles than the old skin. The new skin is also temporarily more sensitive to the sun. There are three basic types of chemical peels. Surface peels can use alpha-hydroxy acids or another mild acid to penetrate only the outer layer of the skin, gently exfoliating it. This treatment is used to improve the appearance of mild skin discoloration and rough skin, as well as to rejuvenate the face, neck, chest, or hands. In moderate peels, glycolic acid or trichloroacetic acid is applied to penetrate the outer and middle layers of the skin to remove damaged skin cells. This treatment is used to improve age spots, fine lines and wrinkles, freckles, and moderate skin discoloration. It can also be used to smooth rough skin and treat some precancerous skin growths, such as actinic keratosis described elsewhere in this article. In deep peels, trichloroacetic acid or phenol is applied to penetrate deep into the middle layer of the skin to remove damaged skin cells. This treatment eliminates moderate wrinkles, age spots, freckles, and superficial scars. Patients will see a significant improvement in the appearance of their skin; however, this treatment can cause varying degrees of damage to the skin layers. The peptide compositions disclosed herein can be used as a pretreatment for chemical peels (e.g., once daily for 1–31 days or longer before initiating a chemical peel, such as once daily for one, two, three, or four weeks) and / or as a posttreatment after a chemical peel (e.g., once daily for 1–31 days or longer after the completion of a chemical peel, such as once daily for one, two, three, or four weeks).

[0082] One type of chemical peel is pulsed peel, which involves applying glycolic acid for peeling, followed by the application of an agent targeting cancer cells (e.g., 5-fluorouracil). This treatment can be repeated four to six times over a two-week period. Pretreatment with the formulation described in the embodiment can advantageously be used in conjunction with pulsed peel, or can advantageously be used to promote healing when applied post-operatively.

[0083] Laser skin resurfacing In particular, the compositions described herein are suitable for use before and / or after ablative and nonablative laser resurfacing treatments, which may be fractional or nonablative. In ablative laser treatments, the laser procedure removes the outer layer of skin from a specific target area. These procedures require a longer wound healing process than nonablative laser treatments, which does not result in the removal or vaporization of skin. Nonablative laser treatments act on the entire projection surface area of ​​the treated skin, while fractional laser treatments act on uniformly separated portions of the target area to provide untouched skin areas for accelerated healing. Therefore, fractional laser treatments result in fewer side effects, with a lower reported amount of scarring. See, for example, Preissig, J., Hamilton, K., Markus, R., Current laser resurfacing technologies: A review that delves beneath the surface, Seminars in Plastic Surgery 2012, Vol. 26(3), pp. 109-116. Ablative laser treatments may include, but are not limited to, CO2, Er:YAG (erbium-doped yttrium aluminum garnet), combined erbium / CO2, and fractional laser photothermal resurfacing. CO2 lasers emit light at a wavelength of 10,600 nm with pulses ranging from 0.2 μs to 80 μs for high-intensity lasers and up to 10 ms for lower-intensity lasers. Er:YAG lasers emit light at a wavelength of 2,940 nm with pulses ranging from 0.25 to 5 ms. Combinations of CO2 and Er:YAG lasers rely on the combined treatment of both types of lasers. Shorter pulses promote higher energy delivery and facilitate deeper ablation of the skin. Non-ablative laser treatments may include diode, erbium glass, thulium fiber, and Nd:YAG (neodymium-doped yttrium aluminum garnet) lasers. These treatments use lasers that emit light at wavelengths ranging from 1319 to 1927 nm with pulse ranges from 450 μs to 210 ms. The peptide compositions disclosed herein can be used as a pretreatment for laser therapy (e.g., once daily for 1-31 days or longer before the start of laser therapy, such as once daily for one, two, three, or four weeks) and / or as a posttreatment after laser therapy (e.g., once daily for 1-31 days or longer after the completion of laser therapy, such as once daily for one, two, three, or four weeks).

[0084] Skin repair The active ingredient of the composition comprises two or more peptides. The first peptide in the composition is one or more dipeptides, tripeptides, and / or tetrapeptides, and the second peptide in the composition is one or more pentapeptides, hexapeptides, and / or heptapeptides. The composition can be used in cosmetics, medicated cosmetics, and general skincare compositions, or provided as a pharmaceutical composition. Methods for promoting skin health, skin regeneration, and enhanced wound healing using compositions comprising dipeptides, tripeptides, or tetrapeptides and pentapeptides, hexapeptides, or heptapeptides are also described.

[0085] Skin regeneration and wound healing typically involve a complex and poorly understood process that relies on the synergistic action of numerous different tissue and cell types. Successful wound healing can be considered a form of tissue remodeling, occurring when tissue remodeling processes reduce the inflammatory response of the innate immune system and minimize scar formation as fibrous tissue replaces normal skin after injury. This process is most efficient in young children but decreases with age. Less efficient wound healing often results in unsightly, irritating, and even painful scars, such as keloids or hypertrophic scars. Given that patients undergoing invasive skin surgery are rarely children, there is a particular need for skincare treatments that promote wound healing after invasive skin procedures. The combination of two peptides demonstrates excellent efficacy in promoting wound healing and skin regeneration.

[0086] In addition, the compositions help treat or prevent skin conditions such as dry, dull, inelastic, lackluster skin, exaggerated lines and wrinkles, stretch marks, spider veins, or red spots. In some embodiments, the compositions described herein can be used to improve marionette lines, smile lines, deep nasolabial folds, crow's feet, fine lines / wrinkles, frown lines, horizontal forehead wrinkles, sagging thin / fragile skin, and the appearance of red and dull skin. The compositions can also be used to prevent and treat photodamaged skin, the appearance of fine lines and wrinkles, hyperpigmentation, age spots, and aging skin. The disclosed compositions may also help increase the suppleness of the stratum corneum, increase the content of collagen and / or glycosaminoglycans in the skin, increase skin hydration, reduce transepidermal water loss, and generally improve skin quality.

[0087] The composition can also be used in conjunction with mucous membranes, particularly the lips and vaginal mucosa. When applied to the vaginal mucosa, a commercially available vaginal applicator can be used. Suitable applicators may take the form of a pre-filled syringe, a tube attached to a pre-filled compressible reservoir, a pre-packaged stick containing a pre-selected amount of the composition, or a universal vaginal applicator including perforations along its length for dispensing the composition through the perforations.

[0088] Drug delivery use Some of the described compositions can be used as carriers for drug delivery. In this regard, anhydrous compositions such as those disclosed herein are particularly suitable for delivering topical drugs suitable for delivery, such as antibacterial agents, antiprotozoal agents, antifungal agents, antiviral agents, spermicides, prostaglandins, and steroids. Drugs suitable for delivery include bromocriptine, sildenafil, oxytocin, calcitonin, luteinizing hormone-releasing hormone and analogues, insulin, human growth hormone, oxybutynin, and steroids used for hormone replacement therapy or contraception. Antifungal agents include clotrimazole, econazole, miconazole, terbinafine, fluconazole, ketoconazole, and amphotericin B. Antibiotics include amoxicillin, doxycycline, cephalexin, ciprofloxacin, clindamycin, metronidazole, azithromycin, sulfamethoxazole / trimethoprim, amoxicillin / clavulanic acid, and levofloxacin. Antibiotics include penicillins, tetracyclines, cephalosporins, quinolones, lincomycins, macrolides, sulfonamides, glycopeptides, aminoglycosides, and carbapenems. Hormones include 5-alpha reductase inhibitors, corticosteroids, adrenocorticotropic hormone (ACTH), glucocorticoids, mineralocorticoids, corticosteroid inhibitors, antiandrogens, antidiuretic hormones, antigonadotropins, antithyroid drugs, aromatase inhibitors, calcitonin, estrogen receptor antagonists, gonadotropin-releasing hormone antagonists, growth hormone receptor blockers, growth hormone, insulin-like growth factor, parathyroid hormone and analogues, progesterone receptor modulators, prolactin inhibitors, selective estrogen receptor modulators, sex hormones, androgens and anabolic steroids, contraceptives, estrogens, gonadotropin-releasing hormone, gonadotropins, progestins, sex hormone combinations, somatostatin and somatostatin analogues, synthetic ovulation stimulants, and thyroid drugs. Antiviral agents include adamantane antiviral drugs, antiviral enhancers, antiviral drug combinations, antiviral interferons, chemokine receptor antagonists, integrase chain transfer inhibitors, other antiviral drugs, neuraminidase inhibitors, NNRTIs, NS5A inhibitors, nucleoside reverse transcriptase inhibitors (NRTIs), protease inhibitors, and purine nucleosides.

[0089] Medications for treating skin conditions that can be used with the selected composition as a delivery device include acne medications (isotretinoin), atopic dermatitis medications (topical steroids), herpes zoster medications (antiviral drugs such as valacyclovir), urticaria medications (antihistamines such as loratadine or fexofenadine, omalizumab), sunburn medications (lidocaine), contact dermatitis medications (antihistamines, topical steroids), diaper rash medications (zinc oxide), rosacea medications (metronidazole, doxycycline, azelaic acid, isotretinoin, beta-blockers, estrogens), athlete's foot medications (antifungal drugs), and basal cell carcinoma medications (imiquimod, fluorouracil, vismodegib).

[0090] Formulation type The peptide compositions of the embodiments described herein can be used in a variety of formulations. Topical formulations are provided comprising dipeptides, tripeptides, or tetrapeptides, as well as pentapeptides, hexapeptides, or heptapeptides, in combination with at least one excipient. The excipients may include non-aqueous or aqueous carriers, and one or more agents selected from humectants, pH adjusters, deodorants, fragrances, chelating agents, preservatives, emulsifiers, thickeners, solubilizers, penetration enhancers, anti-irritants, colorants, surfactants, beneficial agents, pharmaceutical reagents, and other components known in the art for use in combination with topical formulations for treating the skin. Preferably, the formulation is an anhydrous formulation to prevent skin irritation, such as water-based irritant contact dermatitis or stinging sensation when applied to damaged skin. In another embodiment, the composition is formulated such that the use of preservatives is unnecessary (e.g., a preservative-free formulation) to avoid skin irritation associated with certain preservatives.

[0091] For ease of application, the composition can be provided as an ointment, oil, lotion, paste, powder, gel, or cream. The composition may also include additional components such as protectants, emollients, astringents, humectants, sunscreens, tanning agents, UV absorbers, antibacterial agents, antifungal agents, antiviral agents, antiprotozoal agents, antiacne agents, anesthetics, steroidal anti-inflammatory agents, non-steroidal anti-inflammatory agents, antipruritic agents, additional antioxidants, chemotherapeutic agents, antihistamines, vitamins or vitamin complexes, hormones, anti-dandruff agents, anti-wrinkle agents, anti-skin atrophy agents, skin whitening agents, cleansers, additional peptides, additional modified peptides, and combinations thereof. In a further embodiment, the composition avoids animal- or cell-based materials to prevent skin irritation. The composition can be applied to the dermis or mucous membranes.

[0092] Methods for promoting skin health, skin regeneration, and enhanced wound healing using topical peptide formulations are provided. The compositions can also be applied to treat skin conditions such as inflammation, redness, pain, skin sensitivity, dry skin, bruising, and similar conditions. Application of peptide compositions containing a first dipeptide, tripeptide, or tetrapeptide and a second pentapeptide, hexapeptide, or heptapeptide can also be used to prevent scarring (e.g., in facial contouring surgery or other cosmetic procedures involving skin incisions), accelerate epithelial fusion, and limit scab formation and crusting during wound healing. Increased collagen production and / or increased elastin production can also be induced by applying compositions containing a first dipeptide, tripeptide, or tetrapeptide and a second pentapeptide, hexapeptide, or heptapeptide. Suitable methods for objectively measuring improvements in skin redness and inflammation may include tricolor excitation colorimetry, narrow-band reflectance spectroscopy, diffuse reflectance spectroscopy, skin reflectance spectroscopy, and / or UV photography.

[0093] Some embodiments include the application of the peptide compositions provided herein in the form of a topical formulation; however, other routes of administration (e.g., mucosal, subcutaneous, oral, etc.) are also considered. Considered routes of administration include, but are not limited to, topical, mucosal, and subcutaneous. Suitable liquid forms include suspensions, emulsions, solutions, etc. Unit dosage forms may also be provided, such as individually packaged formulations containing a predetermined amount of the formulation, prepared for application to the face or other body sites at predetermined times before and after surgery. Unit dosage forms prepared for application two or three times daily before and after surgery are particularly preferred; however, in some embodiments, it may be necessary to prepare the unit dosage form for application once daily, four times daily, or more.

[0094] In some embodiments, the topical and other formulations typically contain about 0.001 wt.% or less to about 50 wt.% or more of an active ingredient, such as a peptide, preferably about 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1 wt.% to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 or 45 wt.%.

[0095] Compositions and formulations for topical application may include transdermal patches, ointments, lotions, creams, gels, drops, sprays, liquids, aerosols, and powders. Conventional drug carriers, aqueous, powdery, or oily bases, thickeners, etc., can be used. In some applications, ointments, lotions, creams, gels, or similar formulations that can be applied to the skin using fingers may be provided. Such formulations are typically provided in squeeze tubes or bottles or cans, or in roll-on containers (where a ball is fixed to the top of the formulation container, allowing the ball to roll). By rolling the ball over the skin surface, the liquid in the container is transferred to the skin in a controlled manner. Another delivery mechanism includes containers with perforated caps that have a mechanism for advancing the extrudable formulation through the cap. In another form, a gel formulation with sufficient structural integrity to maintain its shape is provided, which is pushed upwards through a tube and applied to the skin (e.g., in stick form). The advantage of this stick form is that only the formulation comes into contact with the skin during application, rather than with the fingers or part of the container. Applicators (e.g., sticks, sponges, syringes, or other suitable methods) may also be used to place liquids or gels.

[0096] Components of the formulation peptides Formulations comprising combinations of two or more peptides are provided for the purpose of promoting skin health, skin regeneration, and enhanced wound healing, for example, in patients undergoing dermatological procedures such as laser treatment, chemical peels, dermabrasion, microneedling, and other such procedures; in patients undergoing any other treatment or resulting in skin damage, inflammation, or irritation (e.g., sunburn, eczema, psoriasis, herpes zoster, shingles, allergic reactions, contact dermatitis, etc.); or in any skin condition in which stimulation of collagen and / or elastin is beneficial. In topical formulations comprising a combination of two peptides, the first peptide (e.g., a tripeptide) is present in the composition either in pure form or as a peptide-containing carrier, for example, at 50 ppm or less up to 1000, 5000, 10000, 50000, 100000, 500000 ppm or more, such as 100 ppm of peptide. The topical formulation may contain 0.01 wt.% or less (e.g., 0.001 wt.%) to 10 wt.% or more, for example, 0.01 wt.% to 0.02 wt.%, 0.03 wt.%, 0.04 wt.%, 0.05 wt.%, 0.1 wt.%, 1 wt.% to 5 wt.% or 10 wt.% of a first peptide. A second peptide (e.g., a hexapeptide) is present in the topical formulation composition either in pure form or as a peptide-containing carrier, for example, 50 ppm or less to 1000, 5000, 10000, 50000, 100000, 500000 ppm or more, for example, 100 ppm of peptide or any other suitable amount. Topical formulations may contain 0.01 wt.% or less (e.g., 0.001 wt.%) to 10 wt.% or more, for example, 0.01 wt.% to 0.02 wt.%, 0.03 wt.%, 0.04 wt.%, 0.05 wt.%, 0.1 wt.%, 1 wt.% to 5 wt.% or 20 wt.%. The amount of peptide in the matrix can be adjusted up or down.

[0097] In exemplary embodiments, the weight ratio of the first peptide to the second peptide in the topical formulation is 1 part first peptide to 0.2 to 10 parts second peptide, or 1 to 10 parts second peptide, or 1 to 8 parts second peptide, or 1 to 5.5 parts second peptide. The following nomenclature is used herein to refer to various amino acids: alanine (also referred to herein as “Ala” or “A”), arginine (also referred to herein as “Arg” or “R”), asparagine (also referred to herein as “Asn” or “N”), aspartic acid (also referred to herein as “Asp” or “D”), cysteine ​​(also referred to herein as “Cys” or “C”), glutamic acid (also referred to herein as “Glu” or “E”), glutamine (also referred to herein as “Gln” or “Q”), glycine (also referred to herein as “Gly” or “G”), histidine (also referred to herein as “His” or “H”), isoleucine (also referred to herein as… The following amino acids are listed: leucine (also referred to as "Leu" or "L" in this article), lysine (also referred to as "Lys" or "K" in this article), methionine (also referred to as "Met" or "M" in this article), phenylalanine (also referred to as "Phe" or "F" in this article), proline (also referred to as "Pro" or "P" in this article), serine (also referred to as "Ser" or "S" in this article), threonine (also referred to as "Thr" or "T" in this article), tryptophan (also referred to as "Trp" or "W" in this article), tyrosine (also referred to as "Tyr" or "Y" in this article), and valine (also referred to as "Val" or "V" in this article).

[0098] In some embodiments, the first peptide is a dipeptide. Suitable dipeptides include, but are not limited to, dipeptides having the following amino acid sequences: KK, KP, CK, KC, KT, DF, NF, VW, YR, or TT. In other embodiments, the first peptide is a tripeptide. Suitable tripeptides include, but are not limited to, tripeptides having the following amino acid sequences: HGG, RKR, GHK, GKH, GGH, GHG, KFK, or KPK. In some embodiments, the first peptide is a tetrapeptide. Suitable tetrapeptides include, but are not limited to, tetrapeptides having the following amino acid sequences: GQPR, KTFK, AQTR, or RSRK. In some embodiments, the second peptide is a pentapeptide. Suitable pentapeptides include, but are not limited to, pentapeptides having the following amino acid sequences: KTTKS, YGGFX, or KLAAK. In some embodiments, the second peptide is a hexapeptide. Suitable hexapeptides include, but are not limited to, hexapeptides having the following amino acid sequences: VGVAPG or GKTTKS. In some embodiments, the second peptide is a heptapeptide. Suitable heptapeptides include, but are not limited to, heptapeptides or heptapeptide-6 (pre-sirtuin peptide) having the amino acid sequence RGYYLLE. The composition may include two or more peptides, for example, two dipeptides and one pentapeptide; one tripeptide and one hexapeptide; one dipeptide, one tripeptide, and one heptapeptide, etc., provided that the composition contains at least one dipeptide, tripeptide, or tetrapeptide and at least one pentapeptide, hexapeptide, or heptapeptide.

[0099] The peptide can be functionalized. For example, the peptide can be functionalized with fatty acids such as myristenoic acid, palmitoleic acid, cis-6-hexadecenoic acid (sapienic acid), oleic acid, trans oleic acid, isoleic acid, linoleic acid, trans linoleic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachidic acid, benzyl acid, ceric acid, etc. Examples include palmitoyl hexapeptide-12 (Pal-VGVAPG), palmitoyl tripeptide-1 (Pal-GHK), myristicoyl hexapeptide-12 (Myr-VGVAPG), and myristicoyl tripeptide-1 (Myr-GHK). In some embodiments, palmitoyl or myristicoyl functionalization may be desirable because it exhibits enhanced permeability compared to other fatty acids.

[0100] Some embodiments of the methods and compositions provided herein include glycine-histidine-lysine (GHK) as the first peptide. GHK is a peptide sequence that is generally rare in protein classes but common in extracellular matrix proteins. The small size of GHK allows its larger peptides to more easily access membrane receptors. Furthermore, its unique copper-binding structure enhances the transport of copper into and out of cells and promotes wound healing through several different but related pathways. Due to its strong copper-binding structure, GHK can be provided in the form of GHK-Cu (copper-bound GHK).

[0101] GHK-Cu acts as both an anti-inflammatory agent (see, for example, Pickart, L., The human tri-peptide GHK and tissue remodeling, J. Biomater. Sci. Polymer Edn. 2008, Vol. 19, pp. 969-988, 972-973; Pickart et al., The Human Tripeptide GHK-CU in Prevention of Oxidative Stress and Degenerative Conditions of Aging: Implications for Cognitive Health, Oxid. Med. Cell Longev. 2012, Vol. 2012, pp. 1-8, 3) and an antioxidant. GHK-Cu promotes wound healing by inhibiting the “acute phase response,” which generates both inflammation and induces scar formation. This biological response prevents bacterial invasion, promotes the arrival of immune cells, stops bleeding, and provides coverage to the injured area. GHK-Cu also inhibits the acute phase response by suppressing the production of molecules called cytokines. Cytokines are immune cell signaling molecules that attract immune cells and trigger the production of other molecules that promote inflammation and fibrosis (leading to scar tissue formation). Specifically, GHK-Cu inhibits the production of cytokines, including tumor necrosis factor-α (TNFα), interleukin-1 (IL-1), interleukin-6 (IL-6), and transforming growth factor-β-1 (TGF-β1) (several key drivers of inflammation and apoptotic cell death in the wound area). Since TGF-β1 is an important component of the sustained acute response, GHK-Cu's inhibition of TGF-β1 also helps shorten its duration after the onset of the acute response. GHK-Cu acts as an antioxidant by blocking the release of iron oxide from ferritin, preventing further inflammation or microbial infection (as invading microorganisms require iron to survive).

[0102] GHK-Cu also stimulates angiogenesis, increases collagen production, and promotes extracellular matrix regeneration. GHK-Cu acts as an inducer for cells crucial for the regeneration of damaged tissues, such as capillary cells that rebuild blood vessels. It also upregulates the production of various enzymes that remove damaged proteins and rebuild the extracellular matrix (ECM)—a critical external scaffold essential for intercellular communication and support. Specifically, GHK-Cu induces the production of messenger RNA (mRNA) necessary for ECM regeneration, namely collagen, proteoglycans, glycosaminoglycans, chondroitin sulfate, and dermatan sulfate. The increased collagen production induced by GHK-Cu also plays a key role in promoting skin regeneration. GHK-Cu further stimulates blood flow to damaged tissues through three processes: angiogenesis, anticoagulation, and vasodilation. First, GHK-Cu induces angiogenesis or new blood vessel formation by increasing the production of growth factor proteins essential for angiogenesis, such as basic fibroblast growth factor (BFGF) and vascular endothelial growth factor (VEGF). Secondly, GHK-Cu increases blood flow to the wound area by increasing the number of red blood cells (through the production of erythropoietin) and by downregulating the blood clotting molecule thromboxane through its anticoagulant effect. Thirdly, GHK-Cu promotes vasodilation by binding to the vasoconstrictor protein angiotensin II, thereby preventing angiotensin from constricting blood vessels and reducing blood flow.

[0103] GHK-Cu promotes stem cell proliferation (see, for example, Ito et al., Is the Hair Follicle Necessary for Normal Wound Healing, J. Invest. Dermatol. 2008, Vol. 128, pp.1059-1061, 1059). Wound healing studies have shown that, in mouse experiments, the addition of GHK-Cu significantly increased the generation of hair follicles around wounds. Dermal hair follicles are an important source of stem cells, which are crucial for skin healing. Studies on dermal hair follicles have shown that areas containing hair tend to heal more quickly, and cells from multiple parts of the hair follicle can also contribute to the replacement of both dermal and epithelial cells.

[0104] Therefore, by reducing inflammation, acting as an antioxidant, stimulating the growth of new blood vessels, regenerating the extracellular matrix, enhancing collagen production, and promoting stem cell proliferation, GHK can greatly enhance skin regeneration and promote wound healing.

[0105] Some embodiments of the methods and compositions provided herein include valine-glycine-valine-alanine-proline-glycine (VGVAPG) as a second peptide. VGVAPG is a hexapeptide derived from elastin (see, for example, Blanchevoye et al., Interaction between the Elastin Peptide VGVAPG and Human Elastin Binding Protein, J. Biol. Chem. 2012, Vol. 288, pp. 1317-1328, 1317-1318). Elastin is a protein found in connective tissues, such as skin, and is essential for tissues to regain their original shape and size after undergoing temporary expansion or contraction. Due to its importance in providing elasticity and resilience, elastin plays a crucial role in the resistance of skin cells to injury and their recovery from injury. The skin's ability to regain its original shape after being stretched or pulled depends on cross-linked elastin (the tropoelastin in the human body) which acts as a "flexible fiber". The disruption of the elastic fiber system during wound healing is closely related to the formation of scar tissue (see, for example, Rnjak-Kovacina et al., Severe Burn Injuries and the Role of Elastin in the Design of Dermal Substitutes, Tissue Eng. Part B. Rev. 2011, pp. 81-91, 85-86). Due to these and other properties, elastin is a key component in the effective wound healing process.

[0106] VGVAPG plays a crucial role in enhancing elastin's ability to prevent skin damage and promote skin regeneration (see, for example, Floquet et al., Structural Characterization of VGVAPG, an Elastin-Derived Peptide, Biopolymers (Peptide Science) 2004, Vol. 76, 266-280, 267). First, its ability to attract monocytes and fibroblasts has been demonstrated (see, for example, Senior et al., Val-Gly-Val-Ala-Pro-Gly, a Repeating Peptide in Elastin, Is Chemotactic for Fibroblasts and Monocytes, J. Cell Biol. 1984, Vol. 99, pp. 870-874, 870). Monocytes are essential for infection resistance, while fibroblasts are necessary for collagen production (the most abundant protein in the skin) and extracellular matrix regeneration. Second, VGVAPG provides binding sites for elastin-binding proteins (an intrinsic component of mature elastic fibers). Third, VGVAPG provides binding sites for elastin and extracellular matrix degrading enzymes such as matrix metalloproteinases (MMPs), which promote the replacement and regeneration of elastic fibers and extracellular matrix proteins.

[0107] Figure 1 The diagram illustrates the mechanism by which the peptide composition of the described embodiment acts to stimulate and restore elastin and collagen levels in the skin. An exemplary composition contains a tripeptide and a hexapeptide. This schematic diagram demonstrates the beneficial effects of the tripeptide and hexapeptide, which synergistically promote skin regeneration and wound healing by attracting healing cells, increasing elastin and collagen production, enhancing fibroblast proliferation, exhibiting antioxidant behavior (preventing the release of iron oxide), and inducing extracellular matrix regeneration. As a result, the combination of the two peptides exhibits synergistic and superior performance far exceeding that expected from either peptide alone.

[0108] Figure 2This diagram illustrates the effects of the tripeptide on promoting skin regeneration by enhancing collagen and elastin synthesis, blocking ferritin's release of iron oxide, attracting healing cells such as capillary cells and macrophages, and restoring new blood flow to the site of injury. The tripeptide acts as an antioxidant, stimulating collagen, elastin, and hyaluronic acid. It is formulated to penetrate the stratum corneum. In the extracellular matrix (ECM), it is an antioxidant that attracts capillary cells and macrophages, promoting wound healing. Intracellularly, it reduces inflammatory cytokines and increases collagen, elastin, dermal stem cell proliferation, and hyaluronic acid.

[0109] Figure 3 A schematic diagram is provided illustrating the effects of the hexapeptide on promoting skin regeneration and wound healing by inducing elastin and collagen production, fibroblast proliferation, extracellular matrix regeneration, and fibroblast-keratinocyte migration. The hexapeptide is formulated to penetrate the stratum corneum and mimics the elastin-binding sequence to stimulate elastin. It specifically binds to EBP receptors on fibroblasts and keratinocytes. This binding triggers intracellular signal transduction.

[0110] In topical formulations, the tripeptide is typically present in amounts of about 50 ppm or less to about 100, 200, 300, 400 or 500 ppm or higher (e.g., 50 ppm to 150 ppm).

[0111] In topical formulations, the hexapeptide is typically present in amounts of about 50 ppm or less to about 100, 200, 300, 400 or 500 ppm or higher (e.g., 50 ppm to 150 ppm).

[0112] The peptide may be advantageously provided in a matrix to be suitable for combination with other components of a topical formulation. The matrix may include one or more components such as thickeners / binders (e.g., pentaerythritol tetraisostearate), emollients / dispersants (e.g., caprylic / capric triglycerides), solvents (e.g., propylene carbonate), and / or rheology modifiers / antisettling agents (e.g., distearate lithium dimethylammonium montmorillonite).

[0113] Olive bitter glycoside In some embodiments, polyphenols such as oleuropein may be added to the composition. Oleuropein is a polyphenol isolated from olive leaves (see, for example, Omar SH. Oleuropein in olive and its pharmacochemical effects. Sci Pharm 2010; 78(2): 133-54; Al-Rimawi F, Yateem H, Afaneh I. Formulation and evaluation of a moisturizing day cream containing olive leaves extract. International Journal of Development Research 2014; 4(10): 1996-2000; Kontogianni VG, Charisiadis P, Margianni E, Lamari FN, Gerothanassis IP, Tzakos AG. Olive leaf extracts are a natural source of advanced glycation end product inhibitors. Journal of medicinal food 2013; 16(9): 817-22). Oleuropein exhibits its main anti-inflammatory effects by inhibiting lipoxygenase activity and leukotriene production. More specifically, researchers have demonstrated that oleuropein enhances in vitro proteasome activity more effectively than other known chemical activators, possibly through conformational changes in the proteasome. In this respect, it reduces reactive oxygen species (ROS), decreases the amount of oxidized proteins by enhancing proteasome-mediated degradation and autophagy pathways, and preserves proteasome function during replicative aging. Inhibiting AGE formation by blocking sugar-protein adhesion, clearing reactive intermediates, or disrupting established AGE-induced cross-links constitutes an attractive therapeutic / preventive target. Oleuropein has been shown to inhibit AGE formation and degrade AGE products through its proteasome-enhancing function. When oleuropein is used in topical formulations, it is preferably present in amounts of about 0.005% by weight or less to about 10.0% by weight or more, typically in amounts of about 0.01% by weight to about 5.0% by weight (e.g., about 0.05% by weight to about 0.1% by weight). Oleuropein can be used in compositions that promote healing. Oleuropein is not typically used in anti-aging compositions because its effects tend to be incompatible with plumping, but it can be advantageously used in formulations for pretreatment of the skin prior to procedures described herein (e.g., laser resurfacing, chemical peeling, etc.).

[0114] Phosphatidylserine In some implementations, phospholipids such as phosphatidylserine (a highly enriched membrane phospholipid component) may be added. Phosphatidylserine is known to have several physiological functions, such as activating signaling enzymes and antioxidant activity (see, for example, Draelos, Z., Pugliese, P. Glycation and Skin Aging: A Review. Cosmetics & Toiletries Magazine 2011; June 2011:1-6; Lee, S., Yang, J., Park Y. et al., Protective effect and mechanism of phosphatidylserine in UVB-induced human dermal fibroblasts. European Journal of Lipid Science and Technology 2013; 115(7):783-90; He, M., Kubo, H., Morimoto, K. et al., Receptor for advanced glycation end products binds to phosphatidylserine and assists in the clearance of apoptotic cells. EMBO reports 2011; 12(4):358-64). Phosphatidylserine has been found to reduce MMP-1 in a dose-dependent manner, increase procollagen formation, and act as a substrate for AGE targets, thereby reducing the damage caused by glycosylation. The clearance of apoptotic cells is essential for tissue development, homeostasis, and inflammation resolution. Phosphatidylserine provides an "eat me" signal on the cell surface, and phagocytes recognize this signal using specific receptors such as the receptor for advanced glycation end products (RAGE). It then binds to PS and helps clear apoptotic cells and AGE end products. When phosphatidylserine is used in topical formulations, it is preferably present in amounts of about 0.005% by weight or less to about 10.0% by weight or more, typically in amounts of about 0.01% by weight to about 5.0% by weight (e.g., about 0.05% by weight to about 0.1% by weight).

[0115] Phosphatidylserine can be advantageously used in formulations for pretreatment of the skin prior to procedures described herein (e.g., laser resurfacing, chemical peeling, etc.).

[0116] carrier system Liquids and gels containing the peptides and other components described herein can be prepared using techniques known in the field of cosmetic manufacturing. See, for example, Handbook of Cosmetic Science and Technology, 4th Edition, edited by André O. Barel, Marc Paye, Howard I. Maibach, CRC Press, 2014, the contents of which are incorporated herein by reference in their entirety. A variety of formulations are possible. As an example, a transparent cosmetic gel stick composition may comprise 60% to about 90% of an aliphatic polyol (e.g., a C2-6 alcohol containing 2 to 6 hydroxyl groups); 1-10% of soap; and 1-10% of a water-soluble emollient, such as a polyoxyethylene ether of a C8-22 fatty alcohol, as a main component combined with the peptide composition of the preferred embodiment. Aqueous squeezeable gels are based on water-oil emulsion technology. To minimize the amount of water introduced into the squeezeable gel formulation, the concentration of the active solution is adjusted. Ideally, a high concentration of active peptide solution (45-50%) can be used. Carrier systems for AP solids are typically based on volatile cyclosiloxanes because they evaporate rapidly and leave no residue on the skin. Alternatives to volatile cyclosiloxanes include isohexadecane or C13-15 isoalkanes. Solidification systems are employed to develop solid bars that do not melt under typical storage or consumption conditions but provide an elegant skin feel and are easily transferable. Combinations of cyclopentasiloxane and stearyl alcohol with varying degrees of additional waxes such as hydrogenated castor wax, hydrogenated vegetable oils, and polyethylene can be used.

[0117] For liquid formulations (e.g., gels or lotions), siloxanes (e.g., cyclosiloxanes) or linear siloxanes (e.g., siloxane elastomers) can be used as carriers. One suitable type of carrier is a polydimethylsiloxane crosspolymer gel, such as a polydimethylsiloxane crosspolymer in cyclopentane. Other suitable polydimethylsiloxane crosspolymers include cyclopentane, polydimethylsiloxane / vinyl polydimethylsiloxane crosspolymers; polydimethylsiloxane, polydimethylsiloxane / vinyl polydimethylsiloxane crosspolymers; and isodecanane polydimethylsiloxane / vinyl polydimethylsiloxane crosspolymers.

[0118] Typically, the carrier is present in amounts of about 80 wt.% to about 95 wt.% or 82 wt.% to 92 wt.%, for example, in topical formulations for application to the skin or mucous membranes.

[0119] Penetration enhancer Heptyl undecanoate can be used to enhance peptide penetration and provide a silky feel to formulations. Other fatty acid esters can also be used, such as formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, myristic acid, isovaleric acid, palmitoleic acid, cis-6-hexadecenoic acid, oleic acid, trans-oleic acid, isoleic acid, linoleic acid, trans-linoleic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachidic acid, benzyl acid, ceric acid, and medium-chain fatty acids (e.g., C64-C ... 6-12 Fatty acids, etc. When used in topical formulations, the typical amount is 1% to 4% by weight. Although heptadecenoate can be advantageously used in most formulations, it can be considered an optional component of lubricant formulations used in conjunction with microneedles, and this use can be omitted.

[0120] Anti-irritant Panthenol triacetate / naringenin is a natural plant extract that reduces skin redness and moisture loss. When used in topical formulations, the typical amount of the anti-irritant is 1% to 4% by weight.

[0121] anti-inflammatory agents Arnica montana extract comprises components such as essential oils, fatty acids, thymol, pseudoguaiac lactones (sesquiterpene lactones), and flavanone glycosides. It exhibits anti-inflammatory effects. When used in topical formulations, the typical amount of the anti-inflammatory agent is 1% to 4% by weight.

[0122] antioxidants Dunaliella salina extract includes components such as beta-carotene. It can exhibit antioxidant activity. When used in topical formulations, the typical amount of anti-inflammatory agent is 0.1% to 2% by weight.

[0123] Solubility enhancer Certain components of the formulation tend to be difficult to dissolve in conventional formulations. For example, phosphatidylserine and oleuropein are known to exhibit solubility problems. Siloxane polymers (e.g., octanoyl polymethylsiloxane) have been found to be particularly effective in dissolving these two components in anhydrous formulations. For topical compositions containing about 0.05 wt% to about 0.1 wt% phosphatidylserine and / or about 0.05 wt% to about 0.1 wt% oleuropein, an amount of about 0.5 wt% to 1 wt% octanoyl polymethylsiloxane can dissolve these components in anhydrous formulations.

[0124] Lithium montmorillonite clay Lithium montmorillonite clay, such as modified lithium montmorillonite clay, can be used in combination with peptides to impart permeability and adsorption properties to the composition and can help stabilize emulsions. The chemical formula of lithium montmorillonite is Na. 0.3 (Mg,Li)3Si4O 10 (OH)2. Other clays, such as bentonite and magnesium aluminum silicate, can also be used.

[0125] Lithium montmorillonite or other clays can be modified to produce organically modified clay compounds. Salts of fatty acids (e.g., hydrogenated fatty acids) (e.g., quaternary ammonium salts) can react with lithium montmorillonite or other clays. As provided herein, fatty acids are referred to and described using conventional nomenclature employed by those skilled in the art. Saturated fatty acids do not contain carbon-carbon double bonds. Unsaturated fatty acids contain at least one carbon-carbon double bond. Monounsaturated fatty acids contain only one carbon-carbon double bond. Polyunsaturated fatty acids contain two or more carbon-carbon double bonds. Double bonds in fatty acids are typically cis; however, trans double bonds are also possible. The position of a double bond can be indicated by Δn, where n represents the lower-numbered carbon atom in each pair of double-bonded carbon atoms. A specification of total #carbons:#double bond Δ can be used. 双键位置 Simplified notation. For example, 20:4Δ 5,8,11,14 This refers to fatty acids with 20 carbon atoms and four double bonds, where the double bonds are located between carbon atoms 5 and 6, 8 and 9, 11 and 12, and 14 and 15, with carbon atom 1 being the carbon of a carboxylic acid group. Stearic acid (octadecanoic acid) is a saturated fatty acid. Oleic acid (cis-Δ9-octadecenoic acid) is a monounsaturated fatty acid, and linolenic acid (fully cis-Δ9,12,15-octadecanetrienoic acid) is a polyunsaturated fatty acid. Suitable fatty acids can contain 5 to 30 carbon atoms, for example, 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, or 30 carbon atoms. Fatty acids can be fully saturated or can include as many double bonds as feasible in terms of chain length. Fatty acids suitable for functionalizing lithium montmorillonite or other clays include palmitic acid and stearic acid. Dialkyl quaternary ammonium salt cationic modifiers include dipalmitoyldimethylammonium chloride and distearyldimethylammonium chloride. Amide quaternary ammonium salt cationic modifiers include palmitamidopropyltrimethylhexadecyl alcohol chloride and palmitamidopropyltrimethylammonium chloride.

[0126] Other excipients and reagents In some embodiments, the peptide may be mixed with a suitable carrier, diluent, or excipient, and may contain auxiliary substances such as wetting agents or emulsifiers, pH buffers, gelling or viscosity-enhancing additives, preservatives, flavorings, colorings, etc., depending on the route of administration and the desired formulation. See, for example, “Remington: The Science and Practice of Pharmacy”, Lippincott Williams & Wilkins; 20th edition (June 1, 2003) and “Remington's Pharmaceutical Sciences,” Mack Pub. Co.; 18th and 19th editions (December 1985 and June 1990, respectively). These formulations may include complexing agents, metal ions, polymers (such as polyacetic acid, polyglycolic acid, hydrogels, dextran, etc.), liposomes, microemulsions, micelles, monolayer or multilayer vesicles, erythrocyte shadows, or spheroids. Suitable lipids for liposome formulations include, but are not limited to, monoglycerides, diglycerides, thioglycosides, lysophosphatidylcholine, phospholipids, saponins, bile acids, etc. The presence of these additional components can affect physical state, solubility, stability, release rate, clearance rate, and penetration of the active ingredient.

[0127] Compositions for topical application comprise a peptide composition as described herein and a dermatologically acceptable medium. This medium may be aqueous or non-aqueous. Dermatologically acceptable mediums for topical compositions may be in the form of lotions, gels, ointments, liquids, creams, or emulsions. If the medium is an emulsion, the emulsion may have a continuous aqueous phase and a discontinuous non-aqueous or oil phase (oil-in-water emulsion), or a continuous non-aqueous or oil phase and a discontinuous aqueous phase (water-in-oil emulsion). When applied topically in liquid or gel form, a liquid carrier may be added to the active ingredient, such as water, petroleum, animal or plant-derived oils such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils. Physiological saline solutions, glucose or other sugar solutions, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol are also suitable liquid carriers. Pharmaceutical compositions may also be in the form of oil-in-water emulsions. The oil phase may be vegetable oils such as olive oil or peanut oil, mineral oils such as liquid paraffin, or mixtures thereof. Suitable emulsifiers include naturally occurring gums such as gum arabic and tragacanth, naturally occurring phospholipids such as soybean lecithin, esters or metaesters derived from fatty acids and hexitan anhydrides such as sorbitan monooleate, and condensation products of these metaesters with ethylene oxide such as polyoxyethylene sorbitan monooleate. Emulsions may also contain colorants and flavorings.

[0128] In some implementations, siloxane elastomers (e.g., polydimethylsiloxane cross-linked polymers) are used to increase peptide delivery and penetration into the skin. An alternative to increasing the molecular weight (e.g., with silicone) or adding fillers (e.g., with siloxane compounds) is to partially cross-link the siloxane polymer and disperse the material in a suitable siloxane carrier fluid. The resulting polydimethylsiloxane cross-linked polymers (also known as siloxane elastomers in the personal care industry) differ from basic polydimethylsiloxane (PDMS) due to the cross-linking between linear polymers. These materials can be used in peptide formulations and also offer benefits such as scar treatment, wound periarthritis protection, and enzyme delivery. In skin care applications, the cosmetic properties of siloxane elastomers (including those with functional groups) and their ability to absorb various oils (e.g., with polydimethylsiloxane / vinyl polydimethylsiloxane cross-linked polymers such as Dow Corning® 9506 elastomer powder) are two desirable properties of elastomers. Siloxane elastomers have a skin feel different from any siloxane fluid and are described as “smooth,” “velvety soft,” and “powder-like.” It can be altered by controlling the amount of liquid phase in the formulation, and therefore by controlling the degree of swelling. Due to the film-forming properties of polydimethylsiloxane crosspolymers, they can be used as delivery systems for active ingredients (such as peptides as described herein) or other formulation components (such as oil-soluble vitamins and sunscreens). Sunscreens such as octyl methoxycinnamate can be delivered more efficiently from formulations containing siloxane elastomers, resulting in higher sun protection factors (SPF). Siloxane elastomer blends can be used to enhance the SPF in oil-in-water formulations containing organic sunscreens. For example, in tests of SPF, adding 4% siloxane elastomer blends to a sunscreen formulation containing organic sunscreens increased the SPF from 5.7 to 18. This property of siloxane elastomers maximizes the effectiveness of sunscreens in the formulation while reducing the amount required to achieve the desired SPF. As a result, formulation costs can be reduced along with potential irritation caused by the activity of sunscreens. Therefore, higher SPFs can be achieved with the same amount of UV absorbers, thereby improving performance without increasing formulation costs. Siloxane elastomers can be prepared from linear siloxane polymers via a variety of crosslinking reactions (e.g., hydrosilylation reactions of vinyl groups with silane compounds). General methods involve linear siloxane polymers, wherein a crosslinking agent reacts with reactive sites along the polymer chain. Polydimethylsiloxane crosslinked polymers can be prepared as gels (e.g., mixtures of high molecular weight siloxane elastomers in cyclopentasiloxane, such as Dow Corning® 9040 siloxane elastomer blends) made from suspensions of elastomer particles swollen in a carrier fluid, or as spray-dried powders (polydimethylsiloxane / vinyl polydimethylsiloxane crosslinked polymers such as Dow Corning® 9506 elastomer powder).The gel form with the desired properties is cyclodimethylsiloxane, but low-viscosity polydimethylsiloxanes and organic fluids can also be used. Examples of polydimethylsiloxane crosslinked polymers in suspension or gel form are high molecular weight siloxane elastomers (12%) in decamethylcyclopentane siloxane (e.g., Dow Corning® ST-elastomer 10) and mixtures of high molecular weight siloxane elastomers in cyclopentane siloxane (e.g., Dow Corning® 9040 siloxane elastomer blends), with elastomer content typically 10-20% by weight.

[0129] The pharmaceutical excipients for topical formulations of peptide compositions may be selected from solvents, emollients and / or emulsifiers, oil bases, preservatives, antioxidants, tension modifiers, penetration enhancers and solubilizers, chelating agents, buffers, surfactants, one or more polymers, and combinations thereof.

[0130] Suitable solvents for aqueous or hydrophilic topical formulations include water; ethanol; isopropanol; mixtures of water and ethanol and / or isopropanol; glycerin; ethylene glycol, propylene glycol, or butylene glycol; DMSO; and mixtures thereof. Suitable solvents for hydrophobic topical formulations include mineral oil, vegetable oil, and silicone oil. If desired, the peptide composition as described herein can be dissolved or dispersed in a hydrophobic oil phase, and the oil phase can subsequently be emulsified in an aqueous phase containing water, alone or in combination with lower alcohols, glycerin, and / or ethylene glycol. Anhydrous compositions are generally preferred because the presence of water can cause stinging when applied to skin that has undergone laser treatment, chemical peels, dermabrasion, etc. Anhydrous formulations also serve to prevent the development of water-based irritant contact dermatitis in damaged or sensitive skin, which can produce rashes and skin irritation that can delay wound healing and skin quality improvement. Tsai, TF, Maibach, HI How irritant is water? An overview. Contact Dermatitis 41(6) (1999): 311-314 (describes contact dermatitis caused by water as an irritant). However, in some embodiments, providing a water-based composition or allowing the presence of a limited amount of water may be acceptable. For example, water may be present, but in amounts below a threshold that could cause a stinging sensation when applied to damaged skin. Osmotic shock or osmotic stress is a sudden change in the concentration of solutes around cells, resulting in a rapid change in the movement of water across its cell membrane. Under conditions of high concentrations of any solute in salt, substrate, or supernatant, water is drawn from the cell by osmosis. This also inhibits the transport of substrates and cofactors into the cell, resulting in cellular “shock.” Alternatively, in low concentrations of solutes, large amounts of water enter the cell, causing it to swell and rupture or undergo apoptosis. If it is desirable to minimize osmotic shock, certain formulations as described herein may be advantageously employed.

[0131] Pharmaceutically acceptable thickeners can be used to maintain the viscosity of the composition at a selected level. Suitable viscosity enhancers or thickeners that can be used to prepare viscous gels or creams with an aqueous matrix include sodium polyacrylate, xanthan gum, polyvinylpyrrolidone, acrylic polymers, carrageenan, hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose, ethyl cellulose, propyl cellulose, hydroxypropyl methylcellulose, polyethoxylated polyacrylamide, polyethoxylated acrylates, and polyethoxylated alkane thiols. Methylcellulose is preferred because it is readily available, economical, and easy to use. Other suitable thickeners include, for example, xanthan gum, carboxymethyl cellulose, hydroxypropyl cellulose, carbomer, etc. The preferred concentration of the thickener will depend on the thickener selected. It is preferred to use an amount that will achieve the selected viscosity. Viscous compositions are typically prepared from solution by adding these thickeners or by using a matrix with an acceptable viscosity level.

[0132] Suitable emollients include hydrocarbon oils and waxes such as mineral oil, petrolatum, paraffin, mineral wax, ceresin, microcrystalline wax, polyethylene, squalene, squalane, silicone oil, triglycerides, acetylated glycerides such as acetylated monoglyceride; ethoxylated glycerides such as ethoxylated monostearate; and alkyl esters of fatty acids or dicarboxylic acids.

[0133] Suitable silicone oils for use as emollients include dimethylpolysiloxane, methyl (phenyl)polysiloxane, and water-soluble and alcohol-soluble siloxane glycol copolymers. Suitable triglycerides for use as emollients include vegetable and animal fats and oils, including castor oil, safflower oil, cottonseed oil, corn oil, olive oil, cod liver oil, almond oil, avocado oil, palm oil, sesame oil, and soybean oil.

[0134] Suitable esters of carboxylic acids or diacids for use as emollients include methyl, isopropyl, and butyl esters of fatty acids. Specific examples of alkyl esters include hexyl laurate, isohexyl laurate, isohexyl palmitate, isopropyl palmitate, decyl oleate, isodecyl oleate, hexadecyl stearate, decyl stearate, isopropyl isostearate, dilauryl lactate, myristyl lactate, and hexadecyl lactate; and alkenyl esters of fatty acids such as oleyl myristate, oleyl stearate, and oleyl oleate. Specific examples of alkyl esters of diacids include diisopropyl adipate, diisohexyl adipate, bis(hexyldecyl) adipate, and diisopropyl sebacate.

[0135] Other suitable types of emollients or emulsifiers that can be used in topical formulations include fatty acids, fatty alcohols, fatty alcohol ethers, ethoxylated fatty alcohols, fatty acid esters of ethoxylated fatty alcohols, and waxes.

[0136] Specific examples of fatty acids used as emollients include nonanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, hydroxystearic acid, oleic acid, linoleic acid, ricinoleic acid, arachidic acid, benzyl acid, and erucic acid. Specific examples of fatty alcohols used as emollients include lauryl alcohol, myristic alcohol, cetyl alcohol, hexadecyl alcohol, stearyl alcohol, isostearyl alcohol, hydroxystearyl alcohol, oleyl alcohol, ricinoleyl alcohol, benzyl alcohol, and erucic acid alcohol, as well as 2-octyldodecyl alcohol.

[0137] Specific examples of waxes suitable for use as emollients include lanolin and its derivatives, including lanolin oil, lanolin wax, lanolin alcohol, lanolin fatty acids, lanolin isopropyl ester, ethoxylated lanolin, ethoxylated lanolin alcohol, ethoxylated cholesterol, propoxylated lanolin alcohol, acetylated lanolin, acetylated lanolin alcohol, lanolin alcohol linoleate, lanolin alcohol ricinoleate, lanolin alcohol ethyl ricinoleate, lanolin alcohol ethyl ricinoleate, ethoxylated alcohol ester acetates, hydrogenated lanolin, hydrogenated lanolin, ethoxylated hydrogenated lanolin, ethoxylated sorbitan lanolin, and liquid and semi-solid lanolin. Waxes can also be used, including hydrocarbon waxes, ester waxes, and amide waxes. Useful waxes include wax esters such as beeswax, cetearyl, myristyl myristate, and stearyl stearate; beeswax derivatives such as polyoxyethylene sorbitan beeswax; and plant waxes, including carnauba wax and candelilla wax.

[0138] Polyols and polyether derivatives can be used as solvents and / or surfactants in topical formulations. Suitable polyols and polyethers include propylene glycol, dipropylene glycol, polypropylene glycol 2000 and 4000, poly(ethylene oxide-co-propylene oxide) glycol, glycerol, sorbitol, ethoxylated sorbitol, hydroxypropyl sorbitol, polyethylene glycol 200-6000, methoxy polyethylene glycol 350, 550, 750, 2000 and 5000, poly[ethylene oxide] homopolymers (100,000-5,000,000), polyalkylene glycols and their derivatives, hexanediol, 2-methyl-2,4-pentanediol, 1,3-butanediol, 1,2,6-hexanetriol, 2-ethyl-1,3-hexanediol, ortho-diols having 15 to 18 carbon atoms, and polyoxypropylene derivatives of trimethylolpropane.

[0139] Polyol esters can be used as emulsifiers or emollients. Suitable polyol esters include ethylene glycol mono- and di-fatty acid esters, diethylene glycol mono- and di-fatty acid esters, polyethylene glycol (200-6000) mono- and di-fatty acid esters, propylene glycol mono- and di-fatty acid esters, polypropylene glycol 2000 monooleate, polypropylene glycol 2000 monostearate, ethoxylated propylene glycol monostearate, glyceryl mono- and di-fatty acid esters, polyglycerol polyfatty acid esters, ethoxylated glyceryl monostearate, 1,3-butanediol monostearate, 1,3-butanediol distearate, polyoxyethylene polyol fatty acid esters, sorbitan fatty acid esters, and polyoxyethylene sorbitan fatty acid esters.

[0140] Suitable emulsifiers for topical formulations include anionic surfactants, cationic surfactants, nonionic surfactants, and amphoteric surfactants. Preferred ionic emulsifiers include phospholipids, such as lecithin and its derivatives.

[0141] Lecithin and other phospholipids can be used to prepare liposomes containing peptide compositions as described herein. When phospholipids (such as lecithin) are placed in water and thus form a bilayer or a series of bilayers (each bilayer separated by water molecules), lipid vesicles form once sufficient energy is provided. Liposomes can be produced by sonicating phospholipids in water. Low shear rates produce multilayered liposomes. Sustained high-shear sonication tends to form smaller monolayer liposomes. Hydrophobic chemicals can dissolve in the phospholipid bilayer membrane. The lipid bilayer of liposomes delivers peptide compositions as described herein.

[0142] The topical formulation may contain micelles, or aggregates of surfactant molecules dispersed in an aqueous solution. Micelles can be prepared by dispersing an oil solvent in an aqueous solution containing a surfactant, wherein the surfactant concentration exceeds a critical micelle concentration. The resulting formulation contains micelles, i.e., spherical oil droplets surrounded by a film of polar surfactant molecules, dispersed in an aqueous solvent.

[0143] Sterols include, for example, cholesterol and cholesterol fatty acid esters; amides such as fatty acid amides, ethoxylated fatty acid amides and fatty acid alkanolamides can also be used as emollients and / or penetration enhancers.

[0144] Pharmaceutically acceptable preservatives can be used to extend the shelf life of the composition. Other suitable preservatives and / or antioxidants for topical formulations can be used, including benzalkonium chloride, benzyl alcohol, phenol, urea, parabens, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tocopherol, thimerosal, chlorobutanol, and mixtures thereof. If a preservative, such as an antioxidant, is used, the concentration is typically from about 0.02% to about 2% based on the total weight of the composition, but may require larger or smaller amounts depending on the agent chosen. As described herein, reducing agents can be advantageously used to maintain a good shelf life of the formulation. Anhydrous formulations of the embodiments are generally observed to exhibit satisfactory stability, making it possible to omit preservatives from the formulation.

[0145] Suitable chelating agents for topical formulations include ethylenediaminetetraacetic acid, its alkali metal salts, its alkaline earth metal salts, its ammonium salts, and its tetraalkylammonium salts.

[0146] The pH of the carrier is preferably from about 4.0 to 10.0, more preferably from about 6.8 to about 7.8. The pH can be controlled using buffer solutions or other pH adjusters. Suitable pH adjusters include phosphoric acid and / or phosphates, citric acid and / or citrates, hydroxides (i.e., calcium hydroxide, sodium hydroxide, potassium hydroxide), and amines such as triethanolamine. Suitable buffer solutions include buffers containing potassium dihydrogen phosphate and dipotassium hydrogen phosphate solutions to maintain a pH between 5.8 and 8; and buffers containing sodium dihydrogen phosphate and disodium hydrogen phosphate solutions to maintain a pH between 6 and 7.5. Other buffers include citric acid / sodium citrate and disodium hydrogen phosphate / citric acid. The peptide composition of the embodiments described is preferably isotonic with the recipient's blood or other bodily fluids. Sodium tartrate, propylene glycol, or other inorganic or organic solutes can be used to obtain the isotonicity of the composition. Sodium chloride is particularly preferred. Buffers such as acetic acid and acetates, citric acid and citrates, boric acid and borates, and phosphoric acid and phosphates can be used. It may be necessary to include reducing agents, such as vitamin C, vitamin E, or other reducing agents known in the pharmaceutical industry, in the formulation.

[0147] Surfactants can also be used as excipients, for example, anionic detergents such as sodium dodecyl sulfate, sodium dioctyl sulfosuccinate and sodium dioctyl sulfonate, cationic detergents such as benzalkonium chloride or benzyl chloride, or nonionic detergents such as polyoxyethylene hydrogenated castor oil, glyceryl monostearate, polysorbate, sucrose fatty acid esters, methylcellulose or carboxymethylcellulose.

[0148] When the peptide formulation of the embodiments described above is administered via subcutaneous injection, it is preferably in the form of a pyrogen-free, parenterally acceptable aqueous solution or oily suspension, emulsion, or solution. The suspension can be formulated using suitable dispersants or wetting agents and suspending agents according to methods well known in the art. The preparation of acceptable aqueous or non-aqueous solutions with suitable properties (e.g., pH, isotonicity, stability, etc.) is within the scope of the art. For example, isotonic carriers such as 1,3-butanediol, water, isotonic sodium chloride solution, Ringer's solution, glucose solution, glucose and sodium chloride solution, lactated Ringer's solution, or other media known in the art can be used, or fixed oils, such as synthetic monoglycerides or diglycerides, fatty acids, etc., can be conventionally used as solvents or suspension media. The peptide formulation may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those skilled in the art.

[0149] In some implementations, including other pharmacologically active agents may be advantageous. Anti-infective agents include, but are not limited to, anthelmintics (mebendazole), antibiotics including aminoglycosides (gentamicin, neomycin, tobramycin), antifungal antibiotics (amphotericidal b, fluconazole, griseofulvin, itraconazole, ketoconazole, nystatin, miconazole, tonaphalate), cephalosporins (cefaclor, cefazolin, cefotaxime, ceftazidime, ceftriaxone, cefuroxime, cefalexin), β-lactam antibiotics (cefotiam, meropenem), chloramphenicol, macrolides (azithromycin, clarithromycin, erythromycin), and penicillins (penicillin G sodium, amoxicillin, ampicillin, dicloxacillin, nafcillin, piperacillin). The following are listed as antiviral agents: lincomycin, ticarcillin, tetracyclines (doxycycline, minocycline, tetracycline), bacitracin, clindamycin, polymyxin E methanesulfonate sodium, polymyxin B sulfate, vancomycin; antiviral agents including acyclovir, amantadine, norinosine, efavirenz, foscarnet, ganciclovir, indinavir, lamivudine, nelfinavir, ritonavir, saquinavir, stavudine, valacyclovir, valganciclovir, zidovudine; quinolones (ciprofloxacin, levofloxacin); sulfonamides (sulfadiazine, sulfamethoxazole); sulfones (dapsone); furazolidone; metronidazole; pentamivir; crystalline sulfonamides; gatifloxacin; and sulfamethoxazole / trimethoprim. Anesthetic agents may include, but are not limited to, ethanol, bupivacaine, chloroprocaine, levobupivacaine, lidocaine, mepivacaine, procaine, ropivacaine, tetracaine, desflurane, isoflurane, ketamine, propofol, sevoflurane, codeine, fentanyl, hydromorphone, macaine, meperidine, methadone, morphine, oxycodone, remifentanil, sufentanil, butorphanol, nalbuphine, tramadol, benzocaine, debupivacaine, chloroethane, lidocaine, and phenapyridine. Anti-inflammatory agents include, but are not limited to, nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, celecoxib, magnesium trisalicylate, diclofenac potassium, diclofenac sodium, diflunisal, etodoxacin, fenprofen, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, mefenamic acid, nabumetone, naproxen, naproxen sodium, oxapazine, piroxicam, rofecoxib, and disalicylate. Acetates, sulindac, and tometetin; and corticosteroids such as cortisone, hydrocortisone, methylprednisolone, prednisone, prednisolone, betamethasone, beclomethasone dipropionate, budesonide, dexamethasone sodium phosphate, flunisolone, fluticasone propionate, triamcinolone, betamethasone acetate, betamethasone dipropionate, betamethasone valerate, desonide, deshydroxymethasone, fluocinolone, triamcinolone, clobetasol propionate, and dexamethasone.

[0150] In some embodiments, the addition of emollients, emulsion stabilizers, humectants, excipients, and other compounds can enhance the sensory properties of the topical formulation, including but not limited to: skin feel (smooth, light, creamy, etc.), absorbability (the time required for the product to lose its moist feel and no longer be perceived on the skin), consistency, firmness, spreadability (e.g., viscosity, flow initiation, shear rate), tackiness, shape integrity, gloss, hydrophilicity, or hydrophobicity. Preferably, the composition will have high spreadability and low viscosity. Compositions with these properties have been shown to have enhanced “smooth” or “light” skin feel evaluations (see, for example, Bekker, M. Webber, G., Louw, N. Relating rheological measurements to primary and secondary skin feeling when mineral-based and Fischer-Tropsch wax-based cosmetic emulsions and jellies are applied to the skin, International Journal of Cosmetic Science 2013, 35(4), pp. 354-61).

[0151] The composition can be advantageously used as a carrier of platelet-rich plasma (PRP), which is plasma rich in platelets. As a concentrated source of autologous platelets, PRP contains several different growth factors and other cytokines that can stimulate soft tissue healing. Platelet-rich plasma therapy utilizes growth factors present in platelet alpha granules in an autologous manner, for example, for the treatment of androgenetic alopecia, wound healing, facial rejuvenation, etc. Various protocols are used to prepare PRP, and there is no standard protocol, but the main principle essentially involves concentrating platelets to a concentration 3-5 times the physiological value and subsequently injecting the concentrated plasma into the tissue requiring healing or effect. Such preparations may be particularly advantageous in vaginal regeneration applications.

[0152] Stability test The stability test of topical formulations can be performed as follows.

[0153] High-temperature testing is now commonly used as a predictor of long-term stability. High-temperature testing can be conducted at 37°C (98°F) and 45°C (113°F). If a product is stored at 45°C for three months (and exhibits acceptable stability), then it should be stable at room temperature for two years. Of course, the product must be stored at 25°C (77°F) for one year. A good controlled temperature is 4°C (39°F), where most products will exhibit good stability. The product should also withstand -10°C (14°F) for three months.

[0154] The product should pass three temperature cycles from -10°C (14°F) to 25°C (77°F). Place the product at -10°C for 24 hours, then at room temperature (25°C) for 24 hours. This completes one cycle. If the product passes three cycles, you can have good confidence in its stability. A more rigorous test is a five-cycle test from -10°C to 45°C. This puts the emulsion under tremendous stress; if it passes this test, it indicates a highly stable product.

[0155] In oil-in-water emulsions, the dispersed phase tends to separate and rise to the top of the emulsion, forming an oil droplet layer. This phenomenon is called emulsion creaming. Emulsion creaming is one of the earliest signs of impending emulsion instability. A test method to predict emulsion creaming is centrifugation. Heat the emulsion to 50°C (122°F) and centrifuge it at 3000 rpm for 30 minutes. Then check the resulting product for signs of emulsion creaming.

[0156] Both the formulation and packaging are sensitive to UV radiation. The product was placed in a glass container, and the actual packaging was placed in a light box with broad-spectrum output. Another glass jar completely covered with aluminum foil served as a control. Discoloration of the product was observed.

[0157] For all of the above tests, color, odor / fragrance, viscosity, pH value, and (if applicable) particle size uniformity and / or particle agglomeration can be observed under a microscope.

[0158] Kits for non-invasive use and for use with invasive procedures Some embodiments of the methods and compositions provided herein include kits containing the peptides provided herein. In some embodiments, the kits may be provided to administering physicians, other healthcare professionals, patients, or caregivers. In some embodiments, the kit includes a container containing the peptide composition in a suitable topical formulation and instructions for administering the peptide composition to a subject. The kit may also optionally contain one or more additional therapeutic agents or other pharmaceutical agents. For example, a kit containing a topical form of the peptide composition may be provided together with other skin care agents (such as cleansers, occlusive moisturizers, penetrating moisturizers, sunscreens, etc.). The kit may contain the peptide composition in bulk form or may contain individual doses of the peptide composition for continuous or sequential administration. The kit may optionally contain one or more diagnostic tools, application tools, and / or instructions for use. The kit may contain a suitable delivery device, such as a syringe, pump dispenser, single-dose package, etc., along with instructions for administering the peptide composition and any other therapeutic or beneficial agents. The kit may optionally contain instructions for storing, reconstitute (if applicable), and administering any or all of the included therapeutic or beneficial agents. The kit may include multiple containers that reflect the number of administrations to be given to the subject, or different products to be administered to the subject.

[0159] In addition to tripeptides and hexapeptides such as palmitoyl hexapeptide-12 and palmitoyl tripeptide-1 as described herein, the topical peptide formulation may contain other ingredients. For example, other ingredients may include cyclopentasiloxane, polydimethylsiloxane crosspolymer, pentaerythritol tetraisostearate, heptyl undecenoate, wild soybean (soybean) oil, panthenol triacetate, naringenin, arnica montana extract, Dunaliella salina extract, lithium distearate dimethylammonium montmorillonite, tocopherol, squalane, caprylic / capric triglyceride, silachlorite, and propylene carbonate. In some embodiments, the formulation is configured to support the skin before, during, and after cosmetic surgery and also to act on the skin's own natural regeneration processes and contribute to improving the skin's appearance. The topical peptide formulation may be applied immediately after surgery for faster recovery or generally for making the skin look healthier. The peptide formulation may increase the natural levels of elastin in the skin, improve the quality of existing elastin, stimulate increased collagen production, and exhibit high antioxidant activity to reduce inflammation, redness, and irritation. The topical peptide formulation is suitable for all skin types and post-operative skin. The topical formulation may be provided to the patient in bulk form to allow the patient to self-apply an appropriate amount of peptide. For example, the patient may apply an amount sufficient to provide a uniform coating on the affected area, or as directed by a physician. In some embodiments, it may be necessary to incorporate additional therapeutic agents or active agents into the topical formulation. Alternatively, adjunctive therapies or agents may be applied alone. For example, cleansers, sunscreens, sunblocks, penetrating moisturizers, and / or occlusive moisturizers may be provided for application before or after the topical composition of the embodiments described.

[0160] The localized peptide composition can be used in conjunction with a mild cleanser. A mild, self-foaming cleanser removes impurities from the skin without drying or irritating it. This mild cleanser may contain a mixture of moisturizers and vitamins designed to cleanse, soothe, and soften the skin without disrupting its delicate moisture balance. In use, the cleanser softens, soothes, and moisturizes the skin, thoroughly removing environmental pollutants and cosmetics. It is sulfate-free and suitable for post-treatment use on sensitive skin. In one embodiment, the mild cleanser comprises water, sodium C14-16 olefin sulfonate, cocamidopropyl betaine, acrylate copolymer, PEG / PPG-8 / 3 diisostearate, oat kernel extract, panthenol, glycerin, beta-glucan, bisabolol, lavender oil, sweet orange peel oil, titanium dioxide, tin oxide, synthetic fluorophlogopite, disodium EDTA, phenoxyethanol, ethylhexylglycerin, citric acid, and sodium hydroxide.

[0161] The localized peptide composition can be used in conjunction with an occlusive moisturizer. This occlusive moisturizer is an ointment formulated to moisturize the skin and works immediately after cosmetic surgery in conjunction with the body's spontaneous natural regeneration process. It protects and enhances the post-operative results of the skin. The occlusive moisturizer softens, soothes, and moisturizes the skin, supports post-operative skin renewal, helps restore the skin's moisture balance, moisturizes dry and damaged skin, and is suitable for post-operative skin and extremely dry skin. In one embodiment, the occlusive moisturizer comprises petrolatum, microcrystalline wax, physalis extract, caprylic / capric triglycerides, shea butter extract, bisabolol, and tocopherol.

[0162] The localized peptide composition can be used in conjunction with a penetrating moisturizer. This moisturizer is a ceramide-rich hydrating agent formulated to help rebuild the skin's natural barrier function, which may be impaired after cosmetic surgery. The moisturizer contains nourishing ingredients, antioxidants, and soothing phytonutrients to help promote and maintain the skin's barrier function. It helps restore moisture balance, softens, soothes, and moisturizes the skin, inhibits free radicals, and replenishes and brightens dull, dry skin. This moisturizer is suitable for all skin types and post-operative skin. In one embodiment, the penetrating moisturizer comprises water, caprylic / capric triglyceride, cetyl ethylhexanoate, cetearyl alcohol, squalane, niacinamide, dimethicone, cetearyl glucoside, glyceryl stearate, PEG-100 stearate, propylene glycol, ceramide 3, phytosterols, shea butter extract, shea butter, olive oil, sodium hyaluronate, beta-glucan, hydrolyzed pea protein, Dunaliella salina extract, xylitol glucoside, dehydrated xylitol, xylitol, glycerin, lecithin, caprylyl glycol, capryloyl oxime acid, xanthan gum, disodium EDTA, ethylhexylglycerin, and phenoxyethanol.

[0163] The topical peptide composition can be used in combination with a sunscreen. The sunscreen may have a suitable SPF, such as SPF 10+, SPF 15+, SPF 20+, SPF 30+, SPF 40+, SPF 50+, or higher. The sunscreen may be broad-spectrum and waterproof, such as a sunscreen with a duration of 80 minutes or longer. The sunscreen may include antioxidants, moisturizers, and skin-soothing phytonutrients. The sunscreen can be used daily or immediately after facial treatments. The sunscreen provides broad-spectrum UVA / UVB sun protection, moisturizes the face with UV protection, is non-comedogenic, suitable for post-operative use, fragrance-free, and preservative-free. The active ingredients of the sunscreen may include zinc oxide (e.g., 10% by weight) and octyl methoxycinnamate (e.g., 7.5% by weight). In one embodiment, the light-blocking agent comprises water, ethylhexyl palmitate, cyclodimethylsiloxane, polydimethylsiloxane, lauryl methyl polysiloxane copolyol, butylene glycol, tocopheryl acetate, sodium chloride, kukui (cocoa nut) seed oil, tea (green tea) extract, cucumber (cucumber) fruit extract, aloe vera (aloe) leaf extract, tetrahexyldecyl ascorbate, allantoin, sodium hyaluronate, disodium EDTA, methylisothiazolinone, ethylhexylglycerin, and fragrance.

[0164] In one embodiment, a kit is provided for use in conjunction with invasive skin procedures as described herein. This kit (referred to as the “invasive kit”) comprises a topical peptide composition, an occlusive moisturizer, a mild cleanser, a penetrating moisturizer, and a broad-spectrum SPF 30+ sunscreen.

[0165] In another embodiment, a kit is provided for use in conjunction with improving skin health but not with invasive skin procedures. This kit (referred to as the "non-invasive kit") comprises a topical peptide composition, a gentle cleanser, a penetrating moisturizer, and a broad-spectrum SPF 30+ sunscreen.

[0166] Various examples of creams, ointments, lotions, solutions, gels, sprays, and patches can be incorporated as active ingredients into peptide compositions as described herein, combined with penetration enhancers and other active agents that synergistically act on the skin for the purpose of promoting wound healing or wound closure or the treatment of chronic skin wounds.

[0167] oral supplements While topical application of the peptides disclosed herein can be advantageous, systemic administration may be necessary in some embodiments. In such embodiments, the peptides are formulated into compositions suitable for oral administration, but other routes of administration are also considered.

[0168] The peptide compositions described herein can be administered to a subject independently or mixed in the composition with other active agents, such as in combination therapy, or with a carrier, diluent, excipient, or a combination thereof. The formulation depends on the chosen route of administration. The formulations and administration techniques of the compounds described herein are known to those skilled in the art (see, for example, “Remington: The Science and Practice of Pharmacy”, Lippincott Williams & Wilkins; 20th edition (June 1, 2003) and “Remington's Pharmaceutical Sciences,” MackPub. Co.; 18th and 19th editions (December 1985 and June 1990, respectively).

[0169] The peptide compositions disclosed herein can be prepared into an applicable form by methods known per se, such as conventional mixing, dissolving, granulating, coating, grinding, emulsifying, encapsulating, embedding, tableting, or extraction methods.

[0170] Various techniques for administering compounds exist in the art, including but not limited to oral, rectal, topical, aerosol, injection, and parenteral delivery, with injection and parenteral delivery including intramuscular, subcutaneous, intravenous, intramedullary, intrathecal, direct intracardiac, intraperitoneal, intranasal, and intraocular injection. This document considers any combination of the foregoing or other methods known to those skilled in the art (see, for example, any combination of “Remington: The Science and Practice of Pharmacy”, Lippincott Williams & Wilkins; 20th edition (1 June 2003) and “Remington's Pharmaceutical Sciences,” Mack Pub. Co.; 18th and 19th editions (December 1985 and June 1990, respectively).

[0171] In practice, according to conventional pharmaceutical compounding techniques, the peptides can be tightly mixed with a drug carrier as the active ingredient. The proportions of the peptides are maintained as in topical formulations; however, it is preferable to minimize excipients to ensure adequate peptide delivery in a compact form. In its simplest form, the peptides can be directly added to, for example, gelatin capsules or soft capsules for patient administration. In other embodiments, a carrier can be used. The carrier can take various forms depending on the desired form of the formulation. Thus, the peptide compositions provided herein can exist as discrete units suitable for oral administration, such as capsules, pouches, or tablets, each containing a predetermined amount of the active ingredient. Furthermore, the peptide compositions can exist as oils, powders, granules, solutions, suspensions in aqueous liquids, non-aqueous liquids, oil-in-water emulsions, or water-in-oil emulsions, similar to topical formulations described elsewhere herein, but using components suitable for human administration. In addition to the common dosage forms listed above, the peptide compositions provided herein can also be administered via controlled-release and / or delivery devices. The peptide compositions can be prepared by any pharmaceutical method. Typically, these methods involve the step of binding the active ingredient with a carrier constituting one or more essential components. Typically, peptide compositions are prepared by uniformly and tightly mixing the peptide components with a liquid carrier or a finely chopped solid carrier, or both. The product can then be easily shaped into the desired appearance.

[0172] Peptide formulations can also be administered locally rather than systemically, for example, by injecting the peptide composition directly into the target area, such as in long-acting or sustained-release formulations. Furthermore, targeted drug delivery systems for the peptides can be used, for example, in liposomes coated with tissue-specific antibodies.

[0173] The peptide composition may contain an amount of peptide effective for the desired therapeutic effect. In some embodiments, the peptide composition is a unit dosage form, and each unit dosage form contains about 0.1 mg or less to about 5000 mg or more of peptide. In further embodiments, the peptide composition contains about 1 to about 500 mg of peptide per unit dosage form or about 500 to 5000 mg of peptide per unit dosage form. These dosage forms may be solid, semi-solid, liquid, emulsion, or suitable for delivery by aerosol, etc.

[0174] The carrier used can be, for example, a solid, a liquid, or a gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, and stearic acid. Examples of liquid carriers are syrup, peanut oil, olive oil, lower alcohols, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0175] The peptide compositions provided herein can be prepared as solutions or suspensions of peptides in water or non-aqueous liquids. Suitable surfactants, such as hydroxypropyl cellulose, may be included. The dispersion can also be prepared in glycerol, liquid polyethylene glycol, and mixtures thereof in oil. Furthermore, preservatives may be included to, for example, prevent the harmful growth of microorganisms.

[0176] The peptide compositions provided herein for injectable applications include sterile aqueous solutions or dispersions. Furthermore, the peptide compositions may be in the form of sterile powders for the ad hoc preparation of such sterile injectable solutions or dispersions. The peptide compositions must be stable under manufacturing and storage conditions; therefore, contamination by microorganisms such as bacteria and fungi should preferably be prevented. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

[0177] In addition to the carrier components described above, the peptide formulations may suitably include one or more other carrier components such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. Furthermore, other adjuvants may be included to make the formulation isotonic with the blood or other bodily fluids of the intended recipient. The peptide compositions may also be prepared in powder or liquid concentrate form for dilution.

[0178] This document considers peptide compositions comprising a peptide as described herein combined with at least one additional active agent. The peptide and the at least one additional active agent may be present in a single formulation or in multiple formulations provided together, or may be unformulated. In some embodiments, the peptide may be administered in a single composition together with one or more additional agents. For example, the peptide may be administered in one composition, and at least one additional agent may be administered in a second composition. In a further embodiment, the peptide and the at least one additional active agent combination is packaged in a kit. For example, pharmaceutical manufacturers, drug distributors, physicians, multivitamin stores, or pharmacists may provide kits containing the peptide combined with another product or component for delivery to patients. These additional components may include anti-infective agents, anti-inflammatory agents, anesthetics, etc.

[0179] Some embodiments described herein relate to oral compositions of peptides, which may include a therapeutically effective amount of the peptide described herein and a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof. The peptide composition may include a peptide in an amount, for example, >1%, ≥2%, ≥3%, ≥4%, ≥5%, ≥6%, ≥7%, ≥8%, ≥9%, ≥10%, ≥20%, ≥30%, ≥40%, ≥50%, ≥60%, ≥70%, ≥80%, ≥90%, ≥95%, or ≥98%. As described elsewhere herein, the first and second peptides are present in the composition at a weight ratio of 1 part of the first peptide (or the first peptide) to about 1-2 parts of the second peptide (or the second peptide).

[0180] Example Example 1A: Peptide Formulation Several topical formulations containing peptides, comprising a first peptide and a second peptide combined with excipients, were prepared. The suitability of these formulations for use as topical preparations was evaluated, including skin sensation and stability. The formulations were prepared as shown in the table below.

[0181] Formula 1A

[0182] Formula 2A

[0183] Formula 3A

[0184] Formula 4A

[0185] Formula 5A

[0186] Formula 6A

[0187] Formula 7A

[0188] Formula 8A

[0189] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0190] Formula 9A

[0191] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0192] Formula 10A

[0193] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0194] Formula 11A

[0195] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0196] Formula 12A

[0197] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0198] It exists in the formulation at 0.01 wt.% and in the carrier at 100 ppm.

[0199] Formula 13A

[0200] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0201] It exists in the formulation at 0.01 wt.% and in the carrier at 100 ppm.

[0202] Formula 14A

[0203] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0204] It exists in the formulation at 0.01 wt.% and in the carrier at 100 ppm.

[0205] Formula 15A

[0206] *Present in the formulation at 0.055 wt.%; present in the carrier at 100 ppm.

[0207] It exists in the formulation at 0.03 wt.% and in the carrier at 100 ppm.

[0208] Formula 16A

[0209] *Present in the formulation at 0.03 wt.%; present in the carrier at 100 ppm.

[0210] It exists in the formulation at 0.03 wt.% and in the carrier at 100 ppm.

[0211] Example 1B: Peptide Formulation A series of formulations were prepared to evaluate the effects of various components on viscosity and stability. As mentioned above, phosphatidylserine and oleuropein are poorly soluble.

[0212] The starting formulation—Formulation 1B—has a viscosity of approximately 5200 cPs. Formulation 2B has a viscosity of approximately 6100 cPs. Formulation 3B has a viscosity of approximately 7000 cPs. Formulations 2B and 3B each use different matrices, with cyclopentasiloxane providing the higher viscosity. The results of this comparative experiment indicate that cyclopentasiloxane can provide a more desirable viscosity level in formulations than isododecane and polydimethylsiloxane crosslinked polymer-3.

[0213] Formulation 4B has a viscosity of approximately 8000 cPs. Formulation 5B (which includes cyclopentasiloxane as a matrix) exhibits a viscosity of approximately 9500 cPs. Formulation 6B (which uses cyclopentasiloxane, polydimethylsiloxane crosspolymer (DC9045), and heptyl undecenoate) exhibits a viscosity of approximately 28000 cPs. The results of this comparative experiment demonstrate that cyclopentasiloxane can provide a more desirable viscosity level in formulations than isododecane and polydimethylsiloxane crosspolymer-3. This experiment further demonstrates that the combination of cyclopentasiloxane, polydimethylsiloxane crosspolymer, and heptyl undecenoate exhibits an acceptable viscosity level.

[0214] Formula 7B is similar to Formula 6B, but also includes 1% palmitoyl tripeptide, exhibiting a viscosity of approximately 16,000 cPs—lower than Formula 6B, but still acceptablely high. In Formula 8B, the amount of palmitoyl tripeptide is increased to 3%, and phosphatidylserine (lipid PSP 70) and 0.025% oleuropein 80% are added. Lipid PSP 70 and oleuropein were found to exhibit poor solubility in heptyl undecenoate. In Formula 9B, the concentration of heptyl undecenoate is reduced to 2.5%, and octanoyl polymethylsiloxane is added to dissolve lipid PSP 70, increasing it to 0.05%, and oleuropein 80% is increased to 0.05%. The viscosity of Formula 9B is approximately 5,000 cPs. The data indicate that octanoyl polymethylsiloxane effectively dissolves phosphatidylserine and oleuropein; however, greater stability is achieved at the expense of viscosity.

[0215] Experiments were conducted to evaluate the feasibility of adding additional heptadecenoate to the formulation. In Formulation 10B, the concentration of heptadecenoate was increased from 2.5% to 8.75%. Formulation 10B exhibited separation and instability, with a viscosity of approximately 1800 cPs. In Formulation 11B, the concentration of heptadecenoate was increased from 8.75% to 16.75%. Formulation 11B exhibited separation and instability, with a viscosity of approximately 400 cPs. The results of these experiments confirm that, despite the presence of octanoyl polymethylsiloxane, increasing the level of heptadecenoate leads to formulation instability due to solubility issues with phosphatidylserine and oleuropein.

[0216] Other strategies were investigated to increase viscosity while maintaining stability. In Formulation 12B, the concentration of the matrix containing palmitoyl hexapeptide-12 was reduced from 5.5% to 3%, with a corresponding increase in the cyclopentasiloxane and polydimethylsiloxane crosspolymers, resulting in a formulation with a viscosity of approximately 24,000 cPs. In Formulation 13B, the concentration of palmitoyl hexapeptide-12 was reduced from 3% to 2%, and heptyl undecenoate was removed to increase viscosity and stability, yielding a formulation with a viscosity of approximately 47,000 cPs. In Formulation 14B, the concentration of palmitoyl hexapeptide-12 was increased back to 3%, and heptyl undecenoate was removed, resulting in a formulation with a viscosity of approximately 34,000 cPs. These results indicate that even in the presence of heptyl undecenoate, reducing the matrix containing palmitoyl hexapeptide-12 is beneficial for the cyclopentasiloxane and polydimethylsiloxane crosspolymers to increase viscosity to the desired level, thereby obtaining a stable formulation.

[0217] For topical formulations, viscosity levels of 400 cPs to 50,000 cPs are generally preferred, such as 1,000 cPs to 30,000 cPs, or 10,000 cPs to 25,000 cPs or 20,000 cPs to 25,000 cPs.

[0218] Formula 1B

[0219] Formula 2B

[0220] Formula 3B

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231] Example 1C: Prepare an exemplary formulation.

[0232] Exemplary Recipe 1C

[0233] Exemplary Recipe 2C

[0234] Exemplary Formula 3C

[0235] Exemplary Recipe 4C

[0236] Exemplary Recipe 5C

[0237] Exemplary formula 6C

[0238] Exemplary Recipe 7C

[0239] *Present in the carrier at 100 ppm.

[0240] It exists in the carrier at 100 ppm.

[0241] Exemplary formula 8C

[0242] *Present in the carrier at 100 ppm.

[0243] It exists in the carrier at 100 ppm.

[0244] Exemplary formula 9C

[0245] Exemplary Recipe 10C

[0246] Exemplary Recipe 11C

[0247] Example 2: Laser Wound Study A laser wound study was conducted in which subjects received a 3 mm erbium CO2 laser spot on their forearm. Wound healing progress was observed 18 days post-injury. Half of the subjects treated their wounds at least once daily for 18 days with a topical formulation (referred to herein as the “dipeptide”) containing 1.0 wt.% GKH and 5.5 wt.% VGVAPGA in an anhydrous siloxane elastomer gel matrix. The other half of the subjects followed the same post-injury treatment regimen but used a control formulation instead of the dipeptide formulation. This control was either Aquaphore® (containing petrolatum, panthenol, glycerin, and bisabolol) from Beiersdorf Inc., Wilton, CT, or no treatment. Figure 4AThe chart depicts the wound appearance of subjects receiving dipeptide treatment and those receiving control treatment over 18 days post-injury. Subjects receiving dipeptide treatment showed significantly better wound appearance compared to the control group from days 2 to 14. Figure 4B The data showed that epithelial fusion was superior in subjects who received dipeptide treatment from day 2 to day 18, with significant differences between the two treatments on each day of these days. Figure 4C The data depicted showed that subjects who received dipeptide treatment from day 1 to day 18 had less crusting / scab formation, which was significantly different from subjects who received dipeptide treatment from day 1 to day 14.

[0248] Example 3: Laser Wound Study Studies were conducted following the same protocol as in Example 2, including trials of commercial wound care treatments (Aquaphor®) other than the dipeptide and control. Figure 5A Data on crusting / scab formation were provided, showing that the dipeptide treatment was superior to the control group from day 1 to day 18, and superior to Aquaphor® from day 2 to day 11. Figures 5B-5D These are photos of the appearance of three different wounds taken on day 4, each wound treated with a dipeptide (…). Figure 5B ), control treatment ( Figure 5C ) or Aquaphor® Figure 5D The treatment was performed. The photos show that the dipeptide treatment resulted in minimal crusting or scab formation on the wound.

[0249] Example 4: Laser Wound Study A laser wound treatment study was conducted in which a subject received a 1 mm erbium CO2 laser spot on his or her forearm, which penetrated the epidermis and reached the dermis. Subjects received local anesthesia prior to laser wound treatment. Each subject received a dipeptide multiple times daily for two weeks prior to laser wound treatment (days 1–14 pre-injury). Half of the subjects received dipeptide treatment at least once daily for 9 days after laser wound treatment, while the other half received Aquaphor® treatment multiple times daily for 9 days after laser wound treatment. Figures 6A-6B Provided with information about the appearance of the wound ( Figure 6A ) and crust / scab ( Figure 6B Data from [the study] demonstrates the benefits of pretreatment with dipeptides two weeks prior to laser skin surgery and post-operative treatment with dipeptides or Aquaphor® for wound healing. Figure 6A The study provides wound appearance data for dipeptide treatment versus Aquaphor® treatment on days 1 to 4, 7, and 9 post-injury, with a significant difference observed on day 9 post-injury. Figure 6BThe study compared the differences in crusting / scab formation between dipeptide treatment and Aquaphor® treatment on days 1 to 4, 7, and 9 post-injury, with significant differences observed on days 1 to 4. Figures 6C-6D Provided post-injury dipeptide treatment on day 9 ( Figure 6C ) and post-injury Aquaphor® treatment ( Figure 6D Images of the wound, taken by the researchers, demonstrate superior wound healing associated with pretreatment and posttreatment with the dipeptide.

[0250] Example 5: Research on Laser Aesthetic Treatment Human subjects underwent facial cosmetic laser treatment. This laser treatment utilized the Encore UltraPulse® ActiveFX™ CO2 laser. The treatment regimen included a 21-day pretreatment with a dual peptide prior to the facial cosmetic laser treatment. Post-treatment, patients received dual peptide treatment on days 1–14 (as described in this article, “invasive kit”) and after day 14 (as described in this article, “non-invasive kit”). The photograph shows the pre-treatment (…). Figure 7A ), 4 days after laser treatment ( Figure 7B ) and the 9th day after laser treatment ( Figure 7C The patient's image shows that by day 9 post-laser treatment, the patient's skin appeared to have fully recovered from the surgery.

[0251] Example 6: Study on collagen and elastin mRNA Fibroblast monolayers were exposed to the dipeptide for 48 hours. mRNA production was compared to that of a group of fibroblasts exposed to the control group. After 48 hours, the fibroblast monolayers were harvested and mRNA was extracted from the cell lysates. The elastin (ELN) mRNA level in the dipeptide-exposed fibroblasts was more than three times higher than that in the control fibroblasts. The dipeptide also stimulated fibroblasts to produce more than two times the amount of collagen (COL1A1) mRNA compared to the control. Figure 8 The results of a study involving the exposure of fibroblast monolayers to a dipeptide composition were described. One group of fibroblasts was exposed to the dipeptide for 48 hours, while a second group received a control treatment for 48 hours. Cells were then harvested, and mRNA was extracted from the cell lysates. The elastin (ELN) mRNA level in the dipeptide-treated cells was more than three times that of the control fibroblasts. Collagen (COL1A1) mRNA production was more than twice that of the control fibroblasts. The data indicate that the dipeptide significantly upregulated collagen and elastin mRNA.

[0252] Example 7: Pretreatment The concept of pretreatment is analogous to the concept of wound bed preparation implemented in wound healing. Pretreatment can be described as skin bed preparation. To maximize the healing power of therapeutic agents in chronic wounds, the bed must be cleaned and prepared to ensure that the concentrations of proteases and cytokines are balanced to prevent these therapeutic agents from dissolving or degrading from within the wound bed or from the corrosive wound fluid generated by these chronic wounds. See, for example, Wound repair and regeneration: official publication of the Wound Healing Society [and] the European Tissue Repair Society 2011; 19(3): 287-91.

[0253] Similarly, chronically photoaged skin requires adequate preparation to ensure that treatment procedures are optimized for its regenerative capacity. However, the context is different. Modulation of the extracellular matrix (ECM) is necessary before regeneration can occur. In the case of damaged skin, this typically involves excessive division and aggregation of anti-ECM proteins (primarily collagen, elastin, and some glycosaminoglycans), which collect in tissueless bundles resistant to protease digestion. The result is cell-cell dysfunction and cell-matrix interactions, as well as inefficient and persistent repair. See, for example, Fligiel S, Varani J, Datta S, Kang S, Fisher G, Voorhees JJ. Collagen Degradation in Aged / Photodamaged Skin InVivo and After Exposure to Matrix Metalloproteinase-1 InVitro. The Journal of investigative dermatology 2003; 120: 842-8. Skin resurfacing procedures often cause collagen denaturation, resulting in additional breakage and increasing the load on the ECM.

[0254] The theory behind pretreatment is based on the premise that processes that aid in the degradation of these ECM bundles help clear the ECM, thereby improving cell-matrix interactions. This allows the regenerative phase of repair to occur earlier in the process, thus promoting more effective healing.

[0255] Denatured collagen and elastin fragments exist in gelatin form, and the main proteases involved in clearing these fragments are metalloproteinases (MMPs) 2 and 9, namely gelatinases. See, for example, Simeon A, Monier F, Emonard H, et al. Expression and Activation of Matrix Metalloproteinases in Wounds: Modulation by the Tripeptide–Copper Complex Glycyl-L-Histidyl-L-Lysine-Cu2F. The Journal of investigative dermatology 1999; 112: 957-62. The balance between MMP function and activity is crucial for normal wound healing—chronic wounds exhibit excessive MMP activity, accompanied by the destruction of a large portion of the EMC, including matrikine fragments derived from collagen and elastin that stimulate regeneration. Therefore, pretreatment aims to temporarily increase the levels of MMPs (especially 2 and 9) to degrade EMC fragment bundles and subsequently allow newly denatured collagen fragments to be stimulated for regeneration in a relatively cleared EMC.

[0256] Clearly, the aim is to achieve a balance between inflammation and protease activity, and the tri-hex technology introduces higher levels of MMP 2 and 9, which are secreted at different times during the healing process. Studies have shown that GHK can modulate the expression and activation of MMPs in the wound. From day 3 to day 22 after implantation in the wound chamber, a continuous increase in MMP-2 was detected in the wound model, while the increase in MMP 9 expression was transient, consistent with neutrophil infiltration, monocyte and macrophage activity, and early phagocytosis. MMP2 then continues to clear the matrix over the next 3 weeks, clearing pathways for angiogenesis and revascularization. Figure 9 This includes a graph showing the change in gelatinase activity (peak area / µg protein) against MMP-2 over time. The dashed line represents the application of GHK to the wound cavity compared to the control (solid line). Starting from day 3, GHK activated MMP-2 (gelatinase, Y-axis) to a much greater extent than the control, peaking between days 18 and 22. Figure 10 Includes a graph showing the change in gelatin-degrading activity (peak area / µg protein) over time. The dashed line represents the application of GHK to the wound cavity compared to the control (solid line). Starting from day 12, GHK activated MMP-2 (gelatinase, Y-axis) to a much greater extent than the control, peaking between days 18 and 22. See Simeon et al., J Invest Dermatol 112: 957–964, 1999.

[0257] Peptides, such as the tripeptide disclosed herein, stimulate increased MMP-2 levels associated with extracellular matrix clearance prior to collagen and elastin regeneration. A direct relationship also exists between MMP-2 and lipogenesis—the generation of new, healthy fat. Lipogenesis is beneficial because it can provide a plumping effect that promotes anti-aging, plumpness, and a refreshed appearance. Adipose tissue (AT) expansion and regression are well-controlled processes in response to nutritional status. Expansion consists of hypertrophy (where the volume of existing adipocytes increases) and proliferation (where newly formed adipocytes differentiate from precursor cells (lipogenesis)). Tissue growth is also associated with angiogenesis and intense proteolytic remodeling of the extracellular matrix (ECM). All these processes are regulated in part by the matrix metalloproteinase (MMP) system. MMP deficiency has been observed to be associated with impaired AT development. Some studies have shown that gelatinase subsets (MMP-2 and MMP-9) are secreted by AT, and their activity is regulated during AT expansion / regression. Elevated plasma levels of MMP-2 have been observed in obese patients, and these levels decrease after bariatric surgery. Based on in vitro studies using rat, mouse (3T3-F442A and 3T3-L1 cell lines) and human preadipocytes, the role of gelatinase in adipocyte differentiation has been demonstrated.

[0258] The role of MMP-2 in in vitro preadipocyte differentiation and in in vivo de novo fat pad formation using precursor cells with genetic defects, gene silencing, or MMP-2 overexpression has been demonstrated. See Bauters et al., Biochimica et Biophysica Acta 1850 (2015) 1449–1456. Bauters et al. used an in vivo model of de novo adipogenesis in which 3T3-F442A preadipocytes transduced with MMP-2 shRNA constructs TRCN0000031228 or SHC002V negative controls were subcutaneously injected into the back of 8-week-old male athymic BALB / c nude mice. Bauters et al. examined the size and density of adipocytes and vascular density normalized to the number of adipocytes, and observed the differentiation of embryonic fibroblasts into mature adipocytes. A significant increase in lipid accumulation was observed during differentiation of 3T3-F442A cells with MMP-2 overexpression, and this accumulation was further enhanced by aP2 and Ppar- The significant increase in expression supports this. The de novo fat pads formed showed no difference in adipocyte size and density.

[0259] The differentiation of preadipocytes into new mature adipocytes can be divided into two main stages. The defining stage consists of progenitor cells directed into adipocyte lineages. In the terminal differentiation stage, preadipocytes acquire the specific characteristics of mature adipocytes. Differentiation of 3T3-F442A cells was reduced compared to control cells by selectively silencing MMP-2 levels. MMP-2 overexpression had the opposite effect and stimulated differentiation of 3T3-F442A preadipocytes. However, enhanced differentiation was observed even with minimal enhancement of MMP-2 levels. Therefore, MMP-2 appears to play a functional role in both the early and late stages of in vitro lipogenesis. Endothelial cells produce and secrete MMPs, and in vivo lipogenesis has been reported to be dependent on angiogenesis. Therefore, locally active MMP-2 (independent of its source) may be sufficient to illustrate its role in lipogenesis. Thus, increased levels of MMP-2 can be obtained by administering peptides (such as the tripeptide of the described embodiment) to patients, which in turn can stimulate lipogenesis. Stimulated lipogenesis can produce a plumping effect, resulting in a reduced appearance of signs of aging and fuller, fresher skin.

[0260] The second area related to ECM preparation involves the fundamental processes of angiogenesis, nutrient delivery, and exchange. This process is associated with VEGF stimulation, and vessel development and growth require several weeks to progress. Pre-vascularization is expected to optimize wound healing. It has been demonstrated that delivery methods used to induce angiogenesis must provide a prolonged (4 to 6 weeks) presence of the therapeutic agent at the site of desired vessel growth. See, for example, Simons M. Angiogenesis: where do we stand now? Circulation 2005 ; 111(12): 1556-66. Single-dose administration of protein or peptide cannot achieve this prolonged presence, but requires prolonged administration. GHK dose-dependently increases VEGF secretion and stimulates tube formation and angiogenesis in animal models. Figure 11 The VEGF concentration (pg VEGF / ng DNA) as a function of GHK concentration (ng / mL) is shown. See, for example, Jose S, Hughbanks ML, Binder BY, Ingavle GC, Leach JK. Enhanced trophic factor secretion by mesenchymal stem / stromal cells with Glycine-Histidine-Lysine (GHK)-modified alginate hydrogels. Acta biomaterialia2014; 10(5):1955-64. Therefore, the process of administering the dipeptide for 4 weeks prior to laser treatment ensures a robust start to angiogenic stimulation of the ECM.

[0261] To evaluate the effects of pretreatment with the dipeptide, subjects received the dipeptide in their forearms for 2–3 weeks, followed by biopsies from the treated forearms and another biopsy from the untreated (control) forearms. Results are shown in [link to results]. Figure 12A and Figure 12B . Figure 12A The baseline of an untreated biopsy sample stained with H / E is shown. Baseline sample ( Figure 12A The image shows irregularly aggregated collagen in an unorganized extracellular matrix; the basal cells at the dermal-epidermal junction are flattened, exhibiting reduced function. Biopsies were taken 2-3 weeks after treatment with the dipeptide. Figure 12B The extracellular matrix has been cleared of large aggregates of collagen bundles and replaced with a more organized distribution and arrangement of collagen; the basal stem cells at the dermal-epidermal junction are cubic in shape, representing greater functional activity, and the epidermis is thicker, with a healthier appearance.

[0262] Therefore, pretreatment is a useful procedure for optimizing wound healing and aesthetic outcomes in patients undergoing skin resurfacing surgery. This is based on the concept of wound bed preparation, ensuring early initiation of ECM remodeling. More specifically, early upregulation of MMP-9 and subsequent upregulation of MMP-2 promotes the degradation of accumulated ECM protein bundles seen in elastic, photoaged skin. Additionally, VEGF-based stimulation of angiogenesis over a period of time improves the ECM status associated with future wound healing. A duration of 2–3 weeks is generally the minimum satisfactory duration for pretreatment, but shorter pretreatment times can be used in certain cases. Therefore, pretreatment lasting at least two weeks leading to the procedure, such as two, three, four, or more weeks, is advantageously permitted. Daily pretreatment is ideal, e.g., at least once a day, e.g., two, three, four, or more applications per day. Twice-daily application, e.g., in the morning and evening, is conveniently possible. In some implementations, less than once a day is acceptable, e.g., every other day or every three days, or twice a week.

[0263] Similar to pretreatment, posttreatment can also be advantageously employed. Two weeks is generally the minimum satisfactory duration of posttreatment use, but shorter durations may be used in certain cases. Therefore, once the skin has sufficiently healed to allow for the application of a topical preparation, posttreatment use for at least two weeks post-surgery is advantageously chosen, for example, two, three, four, or longer periods, such as two to four weeks. Daily application of posttreatment is ideal, for example, at least once daily, such as two, three, four, or more times daily. Twice-daily application, such as in the morning and evening, is also convenient. In some embodiments, less than once-daily application is acceptable, for example, every other day or every three days, or twice a week.

[0264] In some cases, maintenance therapy may be required indefinitely to promote skin health, skin repair, and / or optimal skin bed condition. For maintenance, a frequency of application less than once daily is acceptable, such as once a week or every two or three days; however, twice-daily application, such as in the morning and evening, is conveniently possible.

[0265] When the peptide is formulated in combination with other therapeutic agents, the formulation can be administered according to the preferred administration regimen of the other therapeutic agents. Alternatively, the administration regimen can be the administration regimen of the composition as described herein, wherein the peptide is the only therapeutic agent present.

[0266] In various implementation schemes, one or more combinations of pretreatment, posttreatment application, and maintenance therapy may be employed.

[0267] Example 8: Elastin Production The effects of the dipeptide formulation on elastin production were investigated. Subjects received daily application of the dipeptide to their left forearm for 2–3 weeks, with biopsies obtained at the end of each treatment session. The treated portion (dipeptide, ...) was extracted from the left forearm. Figure 13B ) and from the untreated portion of the right forearm (control, Figure 13A Skin biopsy samples were obtained. Using immunohistochemistry (IHC), the presence of elastin was observed as brown staining in the obtained images at 20× and 100× magnification. As shown in the images, a significant increase in elastin levels was observed in skin treated with the dipeptide preparation compared to the control. Significantly lower levels of elastin were observed in the untreated areas.

[0268] Example 9: Increased procollagen levels The effects of dipeptide-treated skin on procollagen were investigated. Baseline biopsies of facial skin were obtained before starting dipeptide treatment. Subjects then received daily dipeptide application to their faces for two weeks, followed by a second biopsy. Figure 14A This is a 100× image of a skin biopsy showing precollagen hypostaining via IHC as baseline, while... Figure 14BThis is a skin biopsy photograph (100×) showing a significant increase in procollagen levels after 2 weeks of dipeptide treatment.

[0269] Example 10: Increased Collagen Figures 15A-15B Skin biopsy samples were provided from the skin treated with the dual peptide three weeks after application. Figure 15B ) and baseline samples ( Figure 15A Photographs (100×) were observed. Increased collagen formation in the upper dermis was observed within three weeks of local application, accompanied by a reduction in sun-induced elastin degeneration via new collagen, thus improving the appearance of the epidermis.

[0270] Example 11: Distribution of elastin Figures 16A-16C Provided three weeks after application ( Figure 16B ), eight weeks after application ( Figure 16C Skin biopsy samples and baseline samples of facial skin treated with dipeptides. Figure 16A Photograph (100×). In patients with elastin in elastic tissues damaged by photorejuvenation, local application of the dipeptide resulted in less aggregation of elastin material and significant distribution into deeper dermal layers over eight weeks.

[0271] Example 12: Reduced MMP1 Figures 17A-17D Skin biopsy samples of skin treated with the dual peptide three weeks after application were provided at 40× ( Figure 17B Baseline samples at 40× ( Figure 17A Skin biopsy samples from skin treated with the dipeptide three weeks after application were taken at 100× ( Figure 17D ) and baseline samples at 100× ( Figure 17C (Photographs). A decrease in MMP1 staining was observed in the preauricular region over a three-week period.

[0272] Example 13: Added core proteoglycan Figures 18A-18B Skin biopsy samples were provided from the skin treated with the dual peptide three weeks after application. Figure 18B ) and baseline samples ( Figure 18A A photograph (100×) shows increased staining of core proteoglycans in the preauricular region over a three-week period. Core proteoglycans are thought to influence fibrillation and also interact with fibronectin, platelet-reactive protein, complement component Clq, epidermal growth factor receptor (EGFR), and transforming growth factor-β (TGF-β). Core proteoglycans have been shown to enhance or inhibit TGF-β1 activity. The main function of core proteoglycans is involved in cell cycle regulation.

[0273] Example 14: Comparison of dipeptide formulations and standard care To evaluate the efficacy of Formulation 16A (the composition provided above) in accelerating wound healing and subject satisfaction, it was compared with the Vaniply regimen (Pharmaceutical Specialties, Inc., Rochester, MN) and with standard of care (SOC) following intense pulsed light (IPL) and / or pulsed dye laser (PDL) skin resurfacing with Q-switched alexandrite laser and fractional CO2 laser. Additionally, adverse events were analyzed depending on the treatment formulation—the mild, preservative-free formulation (Vaniply) versus other formulations containing active peptides and herbal medicines (Formulation 16A).

[0274] In this single-blind, randomized study, 15 female participants aged 45–70 years with Fitzpatrick skin types I–III and moderate to moderate photodamage underwent IPL and / or PDL facial resurfacing with Q-switched alexandrite laser and 10600 nm fractional CO2 laser. Participants were randomized 2:1 to either formulation 16A or standard care (Vaniply ointment regimen). Pretreatment was performed 3 weeks prior to the resurfacing and continued for 8 weeks post-procedure. Participants were assessed on days 1, 3, 4, 7, 28, and 84 post-procedure. Endpoints included investigator- and participant-reported signs of healing and symptoms, as well as participant-reported satisfaction questionnaires (p<0.05).

[0275] Assessing healing within the first two weeks post-surgery was considered the most important endpoint. The overall analysis reported by the investigators showed that skin healing and patient experience were superior in the Formulation 16A group during the first 7 days (and all time points up to that point) after laser resurfacing treatment, compared to expected healing outcomes. The Formulation 16A group demonstrated better skin healing than standard care at all time intervals, and by day 7, this group showed statistically significant superior healing compared to SOC (P=0.015). Furthermore, for all post-operative time points, investigator-assessed healing ratings and reported patient experience were significantly higher in the Formulation 16A group than in the SOC group, again showing statistical significance on day 7 (P=0.02). The most significant differences in healing turnaround time and the day when the differences between the groups began to emerge were day 3, when signs such as erythema (P=0.02) and exudation (P=0.01) and symptoms such as skin tenderness (P=0.02), burning, and stinging (P=0.03) showed statistically significant differences between the two groups. As the study progressed, subjective assessments improved, and by the end of the study (day 84), significantly better results were achieved in formulation 16A, correlated with the following three statements: On day 28 (P=0.08) and day 84 (P=0.02), it "made me more confident about how my skin looks." On day 28 (P=0.08) and day 84 (P=0.03), the response was, "I will continue with this treatment regimen." On day 28 (P=0.08) and day 84 (P=0.03), “I would recommend this treatment to others.”

[0276] Formulation 16A appeared to improve the healing experience following IPL and / or PDL facial resurfacing with Q-switched alexandrite laser and fractional CO2 laser. The significantly reduced healing time with formulation 16A allowed subjects to resume normal daily activities more quickly. Furthermore, subjects were more satisfied with formulation 16A than with standard care, and no adverse events were reported.

[0277] Example 21: Comparison of dipeptide formulations and standard care A study was conducted to evaluate the efficacy of the Alastin Procedure Enhancement System (“Alastin”, or 16A) in accelerating wound healing and patient satisfaction, comparing it, particularly during the first week of treatment, with standard skincare following IPL and / or PDL with Q-switched alexandrite laser and fractional CO2 laser resurfacing (Pharmaceutical Specialties, Inc., Rochester, MN). The aim was to objectively assess whether the Alastin group healed faster over 7 days, experienced any adverse events, experienced greater symptom relief during the same period, and whether clients were able to return to the workforce or society earlier than the SOC group. In summary, this study was an assessment of downtime and patient experience following invasive resurfacing procedures. Endpoints included investigator- and participant-rated signs and symptoms of healing, as well as participant-rated questionnaires on comfort and satisfaction with the procedure.

[0278] When analyzing the experimental results, given the relatively small number of cases, a one-sided Student-t test was used, with statistical significance defined as ≤0.05.

[0279] Statistically significant difference The overall analysis reported by the researchers showed that skin healing and patient experience were superior in the Alastin group during the first 7 days (and all time points up to that point) after laser skin resurfacing treatment, compared to the expected healing outcome. The Alastin group demonstrated better skin healing than standard care at all time intervals, and by day 7, this group showed statistically significant superior healing compared to SOC (P=0.015). Furthermore, for all postoperative time points, blinded investigator-assessed healing ratings and reported patient experience were higher in the Alastin group than in the SOC group, again showing statistical significance on day 7 (P=0.02). Results from questionnaires including healing ratings and experience are presented separately. Figure 19A and Figure 19B middle.

[0280] The healing turnaround time and the day when the differences between the groups began to be most pronounced were day 3, when signs such as erythema (P=0.02) and exudation (P=0.01) and symptoms such as skin tenderness (P=0.02), burning, and stinging (P=0.03) showed statistically significant differences between the two groups. The analysis results of signs, symptoms, and participant satisfaction on day 3 postoperatively are presented in [the table below]. Figure 19C , Figure 19D and Figure 19E middle.

[0281] In terms of subjective ratings, the Alastin group consistently demonstrated superior results. By day 84, the Alastin group showed significantly higher long-term subject satisfaction than the SOC group (P=0.03). Regarding the overall cosmetic improvement in long-term skin quality, by the end of the study (day 84), the Alastin group showed statistically significant better results than the SOC group for the following three statements: "Makes me more confident about how my skin looks" - Day 84 (P=0.02) (Question 11) "I will continue with this treatment plan" - Day 84 (P=0.03) (Question 12) "I would recommend this treatment to others" - Day 84 (P=0.03) (Question 13) The answers to questions 11, 12, and 13 are presented as follows: Figure 19F , Figure 19G and Figure 19H middle.

[0282] No significant difference Researchers assessed the side effect—scabbing: For the SOC group, crusting was worse on days 1, 4, and 7; however, for the Alastin group, crusting was worse on day 3. Due to the healing properties of Aiastin's occlusive moisturizing properties, the crust adhered better to the face and sloughed off between days 3 and 4, thus showing a significant reversal on day 4, with a 35% improvement in appearance compared to SOC. The researchers' assessment of the side effect—scabbing—is presented in… Figure 19I middle.

[0283] The foregoing description presents the best mode for carrying out the invention, as well as the manner and process of making and using the invention, in complete, clear, concise, and precise terms, to enable any person skilled in the art to make and use the invention. However, the invention is readily adaptable to completely equivalent modifications and alternative structures discussed above. Therefore, the invention is not limited to the specific embodiments disclosed. Rather, the invention encompasses all modifications and alternative structures falling within the spirit and scope of the invention as generally indicated by the appended claims, which specifically point out and clearly claim protection for the subject matter of the invention. While the disclosure has been illustrated and described in detail in the accompanying drawings and the foregoing description, such illustrations and descriptions are intended to be illustrative or exemplary rather than restrictive.

[0284] All references cited herein are incorporated herein by reference in their entirety. If any publication or patent or patent application incorporated herein by reference contradicts the disclosure contained in this specification, this specification is intended to supersede and / or take precedence over any such contradictory material.

[0285] Unless otherwise defined, all terms (including technical and scientific terms) shall be given their common and customary meanings as understood by one of ordinary skill in the art, and are not limited to special or customary meanings unless expressly defined herein. It should be noted that the use of specific terms in describing certain features or aspects of this disclosure should not be construed as implying a redefinition of the term herein to limit it to include any particular feature or aspect of the disclosure associated with that term. Unless expressly stated otherwise, the terms and phrases and variations thereof used in this application, and especially in the appended claims, shall be interpreted as open-ended rather than restrictive. As an example of the foregoing, the term 'including' should be interpreted as 'including, but not limited to', 'including but not limited to', etc.; the term 'comprising' as used herein is synonymous with 'including', 'containing', or 'characterized as', and is inclusive or open-ended, and does not exclude additional unlisted elements or method steps; the term 'having' should be interpreted as 'at least'; the term 'including' should be interpreted as 'including but not limited to'; the term 'example' is used to provide exemplary instances of items under discussion, not an exhaustive or limiting list thereof; adjectives such as 'known', 'normal', 'standard', and similar terms should not be interpreted as limiting the described items to items available within a given time period or time, but should be interpreted as including known, normal, or standard techniques that may be available or known now or in the future; and the use of terms such as 'preferred', 'ideal', 'desired', or 'ideal', and similar terms should not be construed as implying that certain features are critical, necessary, or even important to the structure or function of the invention, but are merely intended to highlight alternative or additional features that may or may not be used in a particular embodiment of the invention. Similarly, unless otherwise explicitly stated, a group of items associated with the conjunction 'and' should not be interpreted as requiring each of these items to be present in the group, but rather as 'and / or'. Likewise, unless otherwise explicitly stated, a group of items associated with the conjunction 'or' should not be interpreted as requiring mutual exclusivity between the groups, but rather as 'and / or'.

[0286] When a series of numerical values ​​are provided, it should be understood that the upper and lower limits, as well as each intermediate value between the upper and lower limits of the range, are included in the implementation scheme.

[0287] Regarding the use of virtually any plural and / or singular terms herein, those skilled in the art may appropriately convert from plural to singular and / or from singular to plural depending on the context and / or application. For clarity, various singular / plural transformations may be explicitly stated herein. The indefinite article 'a' or 'an' does not exclude multiple. A single processor or other unit may perform the functions of several items recited in the claims. The mere fact that certain measures are recited in mutually different dependent claims does not imply that a combination of these measures cannot be used for benefit. Any reference numerals in the claims should not be construed as limiting the scope.

[0288] Those skilled in the art will further understand that if it is intended to introduce a specific number of claim enumerations, then such an intent will be explicitly enumerated in the claims, and without such enumerations, such an intent does not exist. For example, to aid understanding, the appended claims may contain the use of the introductory phrases 'at least one' and 'one or more' to introduce claim enumerations. However, even when the same claim includes the introductory phrases 'one or more' or 'at least one' and indefinite articles such as 'a' or 'an,' the use of these phrases should not be construed as implying that a claim enumeration introduced by the indefinite article 'a' or 'an' limits the specific claim containing such an introduced claim enumeration to embodiments containing only one such enumeration (e.g., 'a' and / or 'an' should generally be interpreted as meaning 'at least one' or 'one or more'); the same applies to the use of definite articles for introducing claim enumerations. Furthermore, even when a specific number of introduced claims are explicitly listed, those skilled in the art will recognize that such a list should generally be interpreted as meaning at least the number listed (e.g., a bare list of 'two lists' without other modifiers generally means at least two lists, or two or more lists). Additionally, in cases where the convention of 'at least one of A, B, and C' is used, such a structure is generally intended for those skilled in the art to understand the meaning of that convention (e.g., 'a system having at least one of A, B, and C' will include, but is not limited to, systems having a single A, a single B, a single C, A and B together, A and C together, B and C and / or A, B, and C together, etc.). In cases where the convention of 'at least one of A, B, or C' is used, such a structure is generally intended for those skilled in the art to understand the meaning of that convention (e.g., 'a system having at least one of A, B, or C' will include, but is not limited to, systems having a single A, a single B, a single C, A and B together, A and C together, B and C and / or A, B, and C together, etc.). Those skilled in the art will further understand that virtually any extractive word and / or phrase presenting two or more alternative terms, whether in the specification, claims, or drawings, should be understood to include the possibility of including one, any, or both of the terms. For example, the phrase 'A or B' will be understood to include the possibility of 'A' or 'B' or 'A and B'.

[0289] All figures used in this specification to indicate the amount of ingredients, reaction conditions, etc., should be understood to be modified by the term 'about' in all cases. Therefore, unless otherwise indicated, the numerical parameters described herein are approximate values ​​and may vary depending on the desired properties being sought. At least, and not at all, the application of the doctrine of equivalence should be limited to any claim in any application claiming priority to this application; each numerical parameter should be interpreted according to the number of significant figures and common rounding methods.

[0290] Furthermore, although the foregoing has been described in detail through illustration and examples for clarity and understanding, it will be apparent to those skilled in the art that certain changes and modifications can be made. Therefore, the description and examples should not be construed as limiting the scope of the invention to the specific embodiments and examples described herein, but rather encompass all modifications and alternatives that have the true scope and spirit of the invention.

[0291] This application also involves the following items: 1. A topical composition for promoting skin repair, comprising: One or more dipeptides, tripeptides, or tetrapeptides; and One or more pentapeptides, hexapeptides, or heptapeptides.

[0292] 2. The local composition as described in Item 1, wherein the one or more dipeptides, tripeptides or tetrapeptides comprise palmitoyl tripeptide-1, and wherein the one or more pentapeptides, hexapeptides or heptapeptides comprise palmitoyl hexapeptide-12.

[0293] 3. The topical composition as described in any one of items 1-2, further comprising heptyl undecenoate.

[0294] 4. An anhydrous topical composition comprising phosphatidylserine, oleuropein, and octanoyl polymethylsiloxane, wherein the viscosity of the topical composition is from 10,000 cPs to 25,000 cPs, and wherein the anhydrous topical composition has the ability to maintain stability in three temperature test cycles from -10°C to 25°C.

[0295] 5. A topical composition for promoting skin repair, comprising: 82-92 wt.% of cyclopentasiloxane and polydimethylsiloxane crosslinked polymers; 1-4 wt.% of heptyl undecenoate; 0.01-0.06 wt. % palmitoyl hexapeptide-12; 0.01-0.06 wt. % palmitoyl tripeptide-1; 0.25-1 wt.% of octanoyl polymethylsiloxane; 0.05-0.1 wt.% phosphatidylserine / lecithin; and 0.05-0.1 wt.% of oleuropein.

[0296] 6. The local composition as described in item 5, comprising: 2-5 wt.% of a first carrier containing palmitoyl hexapeptide-12, the first carrier further comprising pentaerythritol tetraisostearate, caprylic / capric triglyceride, propylene carbonate and silachlorite, wherein the concentration of palmitoyl hexapeptide-12 in the carrier is 100 ppm; 2-5 wt.% of a second carrier containing palmitoyl tripeptide-1, the second carrier further comprising pentaerythritol tetraisostearate, caprylic / capric triglyceride, propylene carbonate and silachlorite, wherein the concentration of palmitoyl hexapeptide-12 in the carrier is 100 ppm.

[0297] 7. The topical composition as described in item 5, further comprising: 1-4 wt.% panthenol triacetate / naringin; 1-4 wt.% Arnica montana extract; and 0.5-2 wt.% of Dunaliella salina extract.

[0298] 8. A method for preparing a skin bed prior to treatment of a skin disease, comprising: The topical composition of any one of items 1-7 is applied to a skin bed, thereby preparing the skin bed for the treatment of dermatological diseases involving damaged skin, thereby promoting the healing of damaged skin following the treatment of the dermatological disease.

[0299] 9. The method of Project 8, wherein the topical composition is applied to the skin bed at least once daily for at least two weeks prior to treatment of the dermatitis.

[0300] 10. A method for promoting skin repair after treatment of a skin disease, comprising: The topical composition of any one of items 1-7 is applied to skin damaged by dermatological treatment, thereby promoting the healing of the damaged skin.

[0301] 11. The method of Project 10, wherein the topical composition is applied to the skin bed at least once daily for at least two weeks after treatment of the dermatitis.

[0302] 12. A method for preparing a skin bed prior to treatment of a skin disease, comprising: The topical composition described in any one of items 1-7 is applied to a skin bed, thereby preparing the skin bed for the treatment of dermatological conditions involving damaged skin; subsequently... The topical composition is applied to skin damaged by dermatological treatment, thereby promoting the healing of the damaged skin.

[0303] 13. The method of item 12, wherein the topical composition is applied to the skin bed at least once daily for at least two weeks prior to treatment of the dermatitis and at least once daily for at least two weeks after treatment of the dermatitis.

[0304] 14. The method of any one of items 8-13, wherein the treatment of the skin disease is laser treatment.

[0305] 15. The method of any one of items 8-13, wherein the treatment of the skin disease is a chemical peel.

[0306] 16. The method of any one of items 8-13, wherein the skin disease treatment is for actinic keratosis.

[0307] 17. The method of any one of items 8-13, wherein the skin treatment is for reducing signs of aging.

Claims

1. Use of a topical composition comprising tripeptide-1 and hexapeptide-12 in the manufacture of a medicament for promoting skin repair or regeneration, wherein: The tripeptide-1 is present in the topical composition at a concentration of 1 ppm to 10 ppm; and The hexapeptide-12 is present in the topical composition at 1 ppm to 10 ppm.

2. The use according to claim 1, wherein the hexapeptide-12 and the tripeptide-1 are present in an amount sufficient to synergistically promote skin regeneration or wound healing.

3. The use according to claim 1, wherein the tripeptide-1 comprises palmitoyl tripeptide-1, myristoyl tripeptide-1, or a combination thereof.

4. The use according to claim 1, wherein the hexapeptide-12 comprises palmitoyl hexapeptide-12, myristoyl hexapeptide-12, or a combination thereof.

5. The use according to claim 1, wherein the hexapeptide-12 is palmitoyl hexapeptide-12 and the tripeptide-1 is palmitoyl tripeptide-1.

6. The use according to claim 1, wherein the local composition is anhydrous.

7. The use according to claim 1, wherein the local composition comprises a dipeptide, a tetrapeptide, a tripeptide having an amino acid sequence different from that of the tripeptide-1, or a hexapeptide having an amino acid sequence different from that of the hexapeptide-12.

8. The use according to claim 7, wherein the hexapeptide having an amino acid sequence different from that of the hexapeptide-12 is present in the topical composition at 1 ppm to 10 ppm.

9. The use according to claim 7, wherein the tetrapeptide is present in the topical composition at 1 ppm to 10 ppm.

10. The use according to claim 7, wherein the tetrapeptide comprises an amino acid sequence selected from the following: GQPR, KTFK, AQTR, or RSRK.

11. The use according to claim 1, wherein the topical composition comprises 0.01 wt.% to 5.0 wt.% of phosphatidylserine.

12. The use according to claim 1, wherein the topical composition comprises 0.05 wt.% to 0.1 wt.% oleuropein.

13. The use according to claim 1, wherein the local composition comprises 82 wt.% to 92 wt.% of cyclopentasiloxane, a polydimethylsiloxane crosslinked polymer.

14. The use according to claim 1, wherein the topical composition comprises 1 wt.% to 4 wt.% of heptyl undecenoate.

15. The use according to claim 1, wherein the topical composition comprises at least one selected from the group consisting of: caprylic / capric triglyceride; cetyl ethylhexanoate; cetearyl alcohol; squalane; nicotinamide; polydimethylsiloxane; cetearyl glucoside; glyceryl stearate; PEG-100 stearate; propylene glycol; ceramide 3; phytosterols; shea butter; olive oil; sodium hyaluronate; β-glucan; hydrolyzed pea protein; xylitol glucoside; dehydrated xylitol; xylitol; glycerin; lecithin; caprylyl glycol; capryloyl oxime acid; xanthan gum; disodium EDTA; ethylhexylglycerin; phenoxyethanol; panthenol triacetate; naringenin; arnica montana extract; Dunaliella salina extract; cyclopentasiloxane, polydimethylsiloxane crosspolymer; heptyl undecenoate; capryloyl polymethylsiloxane; or any combination thereof.

16. The use according to claim 1, wherein the topical composition comprises: 82-92 wt.% of cyclopentasiloxane and polydimethylsiloxane crosslinked polymers; 1-4 wt.% of heptyl undecenoate; 0.25-1 wt.% of octanoyl polymethylsiloxane; 0.01-0.1 wt.% phosphatidylserine / lecithin; and 0.01-0.1 wt.% of oleuropein.

17. The use according to claim 16, wherein the topical composition comprises: 85.9 wt.% of cyclopentasiloxane and polydimethylsiloxane crosslinked polymers; 2.5 wt.% of heptyl undecenoate; 0.25-1 wt.% of octanoyl polymethylsiloxane; 0.05-0.1 wt.% phosphatidylserine / lecithin; and 0.05-0.1 wt.% of oleuropein.

18. The use according to claim 1, wherein the topical composition synergistically promotes skin repair or wound healing.

19. The use according to claim 2, wherein the tripeptide-1 and the hexapeptide-12 synergistically promote skin repair or wound healing by attracting healing cells, increasing the production of elastin and collagen, enhancing fibroblast proliferation, antioxidant behavior, or inducing the regeneration of the extracellular matrix.

20. The use according to claim 2, wherein the tripeptide-1 and the hexapeptide-12 synergistically promote skin repair or wound healing by enhancing fibroblast proliferation.

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