WO2019190531A1 - Rapidly dissolving collagen-based cosmetic films and methods thereof - Google Patents

Abstract

The invention provides novel cosmetic films having crosslinked derivatized collagen and one or more cosmetic ingredients, and methods for their preparation and use for applying collagen one or more cosmetic agents to skin.

Description

RAPIDLY DISSOLVING COLLAGEN-BASED COSMETIC FILMS AND

METHODS THEREOF

Technical Field of the Invention

[0001] The invention generally relates to collagen compositions and preparations. More particularly, the invention relates to novel collagen-based films, and compositions and preparations thereof, and methods of their fabrication and use in cosmetic and/or therapeutic applications.

Background of the Invention

[0002] Facial masks and cosmetic films comprising a collagen component have been described in the art, for example, in CN 1219505, CN 100460516, CN 101721342, US

4,294,241, US 4,131,650, US 4,389,487, and JPS62145006A. Various collagen-containing facial masks are commercially available ( e.g ., from Mijin Cosmetics, Global Beauty Care, and

LuccaBlu). For many of these masks, users wear the mask for a period of time, for example, 15- 20 min, and then remove and throw away the mask. Because the mask is removed from the skin, some of the collagen and other beneficial ingredients not yet absorbed into the skin are removed, leading to waste of materials and less than optimal benefits.

[0003] Thus, novel compositions and methods for effectively applying collagen and delivery of cosmetic and/or therapeutic ingredients to skin are needed that address the shortcomings of prior art collagen-containing masks.

Summary of the Invention

[0004] The invention provides collagen-based films for topical application of collagen and, optionally, one or more cosmetic or other active ingredients to skin. Cosmetic ingredients can be included in the films of the invention to improve skin appearance, enhance skin rejuvenation, increase moisturization, prevent wrinkle formation or minimize the appearance of wrinkles, increase skin whiting, and/or provide anti-aging care.

[0005] In one aspect, the invention generally relates to a cosmetic film. The cosmetic film includes crosslinked derivatized collagen and, preferably, one or more cosmetic ingredients. , Preferably, the cosmetic film is suitable for placement on human skin and dissolves or disassociates within 30 minutes of placement on skin.

[0006] In yet another aspect, the invention generally relates to a cosmetic product including the cosmetic film disclosed herein.

[0007] In yet another aspect, the invention generally relates to a method of making the cosmetic film, comprising exposing a solution comprising derivatized collagen and one or more cosmetic ingredients to a crosslinking condition to crosslink the derivatized collagen.

[0008] In yet another aspect, the invention generally relates to a cosmetic film made by a method disclosed herein.

[0009] In yet another aspect, the invention generally relates to a method for delivering one or more cosmetic agents to the skin of a subject. The method includes applying the cosmetic film of the invention to the skin of a subject.

[0010] In yet another aspect, the invention generally relates to a method for delivering collagen and/or one or more cosmetic ingredients to skin. The method includes applying a cosmetic film of the invention to the skin of the subject.

Brief Description of the Drawings

[0011] FIG. 1A and FIG. IB show exemplary embodiments of the cosmetic film of the invention.

Definitions

[0012] Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Thus, as used herein and in the claims, the singular forms include the plural reference and vice versa unless the context clearly indicates otherwise.

[0013] Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term“about.” The term“about” when used in connection with a value can mean 5% of the value being referred to. For example, about 100 means from 95 to 105.

[0014] As used herein, the term“average molecular weight” refers to weight average molecular weight as determined by size exclusion chromatography, e.g., multi-angle laser light scattering-size exclusion chromatography (MALLS-SEC).

[0015] As used herein, the term a“cosmetic ingredient” refers to a compound or composition intended to improve the appearance of skin.

[0016] As used herein, the term“dry weight” refers to the weight of the non-water components of a collagen film of the disclosure. Collagen content can be measured using a hydroxyproline assay. Content of other components can be measured by other means known in the art, for example by high performance liquid chromatography (HPLC) or gas

chromatography-mass spectrometry (GC/MS).

[0017] Unless required otherwise by context, the term“derivatized collagen” as used herein refers to derivatized soluble collagen.

[0018] Unless required otherwise by context, the term“hyaluronic acid or HA” as used herein includes hyaluronic acid, salts thereof, (e.g., sodium hyaluronate, potassium hyaluronate, magnesium hyaluronate, calcium hyaluronate), and combinations of any of the foregoing.

Detailed Description of the Invention

[0019] The invention provides collagen-based cosmetic films that can be used to deliver one or more ingredients included in the film to skin. The cosmetic films of the invention provide a number of advantages. For example, the cosmetic films can be used to deliver a cosmetic ingredient where both an effective dose and accurate placement are desired. The dose of an ingredient can be adjusted, for example, by adjusting the concentration of the ingredient in the film and/or by adjusting the size of the film, and the solid nature of the film allows its placement at any site on the body where skin rejuvenation is desired. In addition, the cosmetic films, after being applied to the skin, can be rubbed into the skin, thereby allowing for all or substantially all of the ingredients in the cosmetic films to remain on the skin following application. The amount of time that a cosmetic film of the invention needs to be worn before the film dissolves or disassociates and can be rubbed into the skin can be adjusted by controlling the amount of crosslinking of the derivatized collagen in the film.

[0020] In one aspect, the invention generally relates to a cosmetic film. The cosmetic film includes crosslinked derivatized collagen and, preferably, one or more cosmetic ingredients. Preferably, the cosmetic film is suitable for placement on human skin and dissolves or disassociates within 30 minutes of placement on skin.

[0021] In certain embodiments, the cosmetic film includes crosslinked derivatized collagen and one or more cosmetic ingredients, wherein, the cosmetic film dissolves or disassociates within about 30 min ( e.g ., within about 25 min, 20 min, 15 min, 10 min, or 5 min) of placement on human skin.

[0022] The cosmetic film of the invention may have any suitable thickness, for example, having a thickness in the range from about 0.4 mm to about 2 mm (e.g., from about 0.4 mm to about 1.6 mm, from about 0.4 mm to about 1.2 mm, from about 0.4 mm to about 1.0 mm, from about 0.4 mm to about 0.8 mm, from about 0.6 mm to about 2 mm, from about 0.8 mm to about 2 mm, from about 1.0 mm to about 2 mm, from about 1.2 mm to about 2 mm). In preferred embodiments, the film has a uniform thickness.

[0023] The cosmetic film may have any suitable amount of derivatized collagen. In certain embodiments, derivatized collagen may account for at least about 20%, about 30%, about 50%, about 60%, about 75% of the dry weight of the film. In certain embodiments, derivatized collagen may account for up to about 60%, about 70%, about 80%, about 90%, about 99% of the dry weight of the film.

[0024] The cosmetic films of the invention preferably include water, although films that do not include water are also within the scope of the invention. The concentration of derivatized collagen in the films can be varied, but is preferably in the range of 0.2% to 10% w/v of the cosmetic film. In some embodiments, the concentration of derivatized collagen in the film is 0.2% to 5% w/v, 0.2% to 2% w/v, 0.2% to 1% w/v, 1% to 10% w/v, 1% to 5% w/v, 5% to 10% w/v, or any range bounded by any two of the foregoing values. [0025] Various types of derivatized collagen described in the art can be used in the cosmetic films of the invention. U.S. Patent Nos. 4,851,513 (DeVore et al.), 6,197,934 (DeVore et al.), 6,780,840 (DeVore et al), 9,289,396 (DeVore et al), the contents of each of which are incorporated herein by reference in their entireties, describe derivatized collagen and processes for its preparation. Derivatized collagen can be made by reacting collagen with agents that react with deprotonated amines to alter the net charge on the collagen molecules to adjust its pKa, thereby producing a collagen that remains in solution at physiological pH.

[0026] Acylation agents can be used to change the net charge of collagen from positive to negative and to make derivatized collagen that can be used in the films of the invention.

Exemplary acylation agents that can be used to derivatize collagen include anhydrides, acid chlorides, sulfonyl chlorides, and sulfonic acids. Exemplary anhydrides include maleic anhydride, succinic anhydride, glutaric anhydride, citraconic anhydride, methyl succinic anhydride, itaconic anhydride, methyl glutaric anhydride, dimethyl glutaric anhydride, and phthalic anhydride. Exemplary acid chlorides include oxalyl chloride, and malonyl chloride. Exemplary sulfonyl chlorides include chlorosulfonylacetyl chloride, chlorosulfonylbenzoic acid, 4-chloro-3-(chlorosulfonyl)-5-nitrobenzoic acid, and 3-(chlorosulfonyl)-P-anisic acid. An exemplary sulfonic acid is 3-sulfobenzoic acid.

[0027] The derivatized collagen used in the cosmetic films of the invention can be a single type of derivatized collagen ( e.g ., glutarylated collagen or succinylated collagen) or a mixture of multiple types of derivatized collagens (e.g., a mixture of glutarylated collagen and succinylated collagen). In a preferred embodiment, the derivatized collagen used in the collagen films of the invention comprises or consists of glutarylated collagen.

[0028] The derivatized collagen can be made from collagen from any suitable source. For example, the collagen can be bovine collagen, porcine collagen, or human collagen. The collagen can be animal derived (e.g., from cow or pig hides) or can be recombinant (e.g., recombinant human collagen). The derivatized collagen can consist of collagen from a single species or can comprise a mixture of collagens from different species. For example, the derivatized collagen can consist of derivatized bovine collagen, derivatized porcine collagen, or derivatized human collagen. As another example, the derivatized collagen can be a mixture of bovine collagen and porcine collagen. [0029] In certain embodiments, the derivatized collagen in the cosmetic film of the invention includes derivatized bovine collagen, derivatized porcine collagen, derivatized human collagen, derivatized recombinant collagen, or a combination thereof. In certain embodiments, the derivatized collagen includes acylated collagen. In certain embodiments, the derivatized collagen includes glutarylated collagen and/or succinylated collagen.

[0030] Derivatized collagen can be the major non-water component of the collagen film. The weight of derivatized collagen in a film can be greater than any single other non-water component. Alternatively, or in addition, the amount by weight of derivatized collagen in a film can greater than all other non-water components of the film combined. For example, the derivatized collagen can comprise at least 50% of the dry weight of the film. In some

embodiments, the derivatized collagen comprises at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90%, or at least 95%, at least 97%, or at least 99% of the dry weight of the film. In some embodiments, the derivatized collagen comprises up to 80%, up to 85%, up to 90%, up to 95%, up to 97%, or up to 99% of the dry weight of the film.

[0031] Alternatively, the amount of the derivatized collagen in the cosmetic films of the invention can be less than the weight of at least one other non-water component.

[0032] The cosmetic film may include one or more cosmetic ingredients, for example, moisturizing ingredients, wrinkle preventative ingredients, anti-aging ingredients, skin whitening ingredients, or a combination thereof. The cosmetic films of the invention can include one or more ingredients from one or more of a combination of the foregoing categories of cosmetic agents. For example, a cosmetic film can comprise at least one moisturizing ingredient, at least one wrinkle preventative ingredient, and at least one anti-aging ingredient. In some

embodiments, a cosmetic film of the invention includes a single cosmetic ingredient. In other embodiments, the cosmetic film includes more than one cosmetic ingredient. For example, the cosmetic film can include 2 to 10, 2 to 5, 2 to 4, or 2 to 3 cosmetic ingredients. In some embodiments, the cosmetic film includes at least 2, at least 3, at least 4, at least 5, or more than 5 cosmetic ingredients.

[0033] Exemplary moisturizing ingredients that can be included in the cosmetic films include hyaluronic acid (HA), hydrolyzed collagen, glycerin, aloe vera juice, lipids ( e.g ., lecithin), oils e.g ., argan nut oil, chia seed oil, flaxseed oil, rosehip oil), butters (e.g., tamanu, almond), and combinations thereof. Exemplary wrinkle preventative ingredients include acetyl hexapeptide-3 (Argireline), HA, glycerin, retinol, ilomastat (ilomostat), ceramide, and combinations thereof. Exemplary anti-aging ingredients comprises epidermal growth factor, acetyl hexapeptide-3 (Argireline), ceramide, HA, retinol, soluble collagen, and combinations thereof. Exemplary skin whitening ingredients comprises dimethicone, nicotinamide, tetrahydrocurminoids, glutathione, and combinations thereof. Identification of a cosmetic ingredient herein as a moisturizing ingredient, wrinkle preventative ingredient, anti-aging ingredient, or skin-whitening ingredient does not preclude its use for another purpose. Other cosmetic ingredients known in the art can also be included in the cosmetic films of the invention, for example, those listed in Table 1 of ET.S. Patent No. 8,673,345 (Green Tea Extract and isolates; Licorice Extract;

Phytosphingosine/Biosine; Ethylbisiminomethylguaiacol; Manganese Chloride; White Birch extract; Hinokitiol; Coffee extract; Hoelen Mushroom Extract; Ascorbic Acid/Ascorbyl;

Glucoside; Siegesbeckia; Rosemary Extract; Silymarin; Boswellia Extract; Ubiquinone; Retinol; Resveratrol; Potassium Cholesterol; Sulfate; Enzymes (proteases, lipases, etc.); UVA/UVB absorbers; Apigenin; Vitamin E; Grape Seed Extract; Lutein; Licochalcone; Luteolin; Ursolic Acid; Centella Asiatica Extract; Ximenynic Acid; Ferulic Acid; Amentoflavone;

Dihydroxyacetone; FD&C Blue # l(Acid Blue) and others; Conjugated linoleate(CLA); Various Fragrance; Components such as essential oils and aromatic oils; Salicylic Acid and derivatives;

1, 3-Beta Glucan(l'6'); Triclosan), the contents of which are incorporated herein in their entireties by reference thereto.

[0034] In certain embodiments, the cosmetic film includes one or more moisturizing ingredients selected from hyaluronic acid (HA), hydrolyzed collagen, glycerin, aloe vera juice, lipids, oils, butters, or a combination thereof.

[0035] In certain embodiments, the cosmetic film includes one or more wrinkle preventative ingredients are selected from acetyl hexapeptide-3 (Argireline), HA, glycerin, retinol, ceramide, or a combination thereof.

[0036] In certain embodiments, the cosmetic film includes one or more anti-aging ingredients are selected from epidermal growth factor, acetyl hexapeptide-3 (Argireline), ceramide, HA, retinol, soluble collagen, or a combination thereof. [0037] In certain embodiments, the cosmetic film includes one or more skin whitening ingredients are selected from dimethicone, nicotinamide, tetrahydrocurminoids, glutathione, or a combination thereof.

[0038] The one or more cosmetic ingredients, e.g., moisturizing ingredients, wrinkle preventative ingredients, anti-aging ingredients, skin whitening ingredients, or a combination thereof, may be present in the cosmetic film in any suitable form. In certain embodiments, the one or more cosmetic ingredients are present in the form of nanoparticles or liposomes, or a combination thereof.

[0039] In certain embodiments, the cosmetic film includes HA in an amount accounting for from about 0.1% to about 15% (e.g., from about 0.5% to about 15%, from about 1% to about 15%, from about 3% to about 15%, from about 5% to about 15%, from about 0.1% to about 10%, from about 0.1% to about 5%, from about 0.1% to about 2%, from about 0.1% to about 1%) of the weight of the film.

[0040] In certain embodiments, the HA is characterized by a weight average molecular weight in the range of about 1 kDa to about 4 MDa (e.g., about 10 kDa to about 4 MDa , about 100 kDa to about 4 MDa , about 250 kDa to about 4 MDa, about 1 MDa to about 4 MDa, about 1 kDa to about 1 MDa, about 1 kDa to about 500 kDa, about 1 kDa to about 250 kDa, about 1 kDa to about 100 kDa).

[0041] The cosmetic films can be made with a mixture of two or more populations of HA having different average molecular weights selected, for example, from the molecular weight ranges or values identified in the preceding paragraph. For example, the cosmetic film can comprise a mixture of two HA populations in a 1 :5 to 5: 1 weight ratio. In some embodiments, the ratio of the two HA populations is 1 :3 to 3: 1. Including high average molecular weight HA (e.g., 500 kDa) in a film with low average molecular weight HA (e.g., 5 kDa) can allow for the inclusion of a greater amount of the low average molecular weight HA than would be possible in the absence of the high average molecular weight HA. In some embodiments, at least 25%, at least 50% or at least 75% of the HA in the film has a weight average molecular weight of at least 100 kDa. In some embodiments, at least 25%, at least 50% or at least 75% of the HA in the film has a weight average molecular weight of at least 500 kDa. In some embodiments, at least 25%, at least 50% or at least 75% of the HA in the film has a weight average molecular weight of at least 1 MDa. In some embodiments, the weight average molecular weight of the HA in the film is at least lOOkDa, at least 250 kDa, at least 500 kDa, or at least 1 MDa. In some embodiments, the weight average molecular weight of the HA in the film is no more than 4 MDa.

[0042] In certain embodiments, the one or more cosmetic ingredients include hydrolyzed collagen. Any amount of hydrolyzed collagen may be used, for example, accounts for about 0.1% to about 2% (e.g., about 0.1% to about 2%, about 0.2% to about 2%, about 0.5% to about 2%, about 1% to about 2%, about 0.1% to about 1.5%, about 0.1% to about 1%, about 0.1% to about 0.5%) of the weight of the film.

[0043] In certain embodiments of the cosmetic film, the one or more cosmetic ingredients include one or more lipids, one or more oils, one or more butters, or a combination thereof.

[0044] In certain embodiments, the one or more lipids, one or more oils, one or more butters are selected from argan nut oil, chia seed oil, flaxseed oil, rosehip oil, tamanu butter, lecithin, or a combination thereof. In certain embodiments, the one or more lipids, one or more oils, one or more butters, or a combination thereof, account for up to about 3% w/v, about 5% w/v, about 10% w/v, or about 15% w/v of the weight of the film.

[0045] In certain embodiments, the cosmetic film of the invention also includes a

photoinitiator, especially when the films are produced by a process that entails crosslinking of a solution of collagen (with or without additives such as cosmetic ingredients) via irradiation (e.g., UV irradiation). Photoinitiators can be used to promote crosslinking of the derivatized collagen in the collagen films. Suitable photoinitiators include sodium persulfate and riboflavin.

[0046] In certain embodiments, the photoinitiator is riboflavin. Riboflavin can be included in the cosmetic films in amounts ranging from, for example, 0.0001% to 0.02% (w/v). In some embodiments, the amount of riboflavin ranges from 0.001% to 0.01% (w/v). In some

embodiments, the amount of riboflavin ranges from 0.001% to 0.008% (w/v).

[0047] In certain embodiments, the photoinitiator is sodium persulfate. Sodium persulfate can be included in the films in amounts ranging from, for example, 0.1% to 15%. In some embodiments, the amount of sodium persulfate ranges from 0.1% to 2 % w/v. In some embodiments, the amount of sodium persulfate ranges from 0.25% to 1%.

[0048] In certain embodiments, the photoinitiator is a combination riboflavin and sodium persulfate. [0049] The cosmetic film of the invention may take any suitable shape or geometrical dimensions. In certain embodiments, the shape of the cosmetic film is selected from (i) face shaped, optionally having eye, mouth, and nose cutouts, (ii) rectangular with rounded edges, (iii) rectangular without rounded edges, (iv) circular, (v) oval, (vi) elliptical, or (vi) kidney shaped.

[0050] Preferred substrates are those that allow the film to be peeled off of a supporting substrate without tearing. The substrate can be the substrate on which the cosmetic film was formed or can be a substrate onto which the film has been transferred after being formed.

Substrates on which the cosmetic film is formed are preferably non-reactive during the crosslinking step to avoid crosslinking the derivatized collagen to the substrate. Substrates on which the cosmetic film is formed are preferably also hydrophilic so as to avoid dewetting of the substrate surface. Exemplary substrates that can be used in the cosmetic products of the invention include aluminum foil, substrates having a non-silicone coating, substrates having a polyester surface ( e.g ., Fisher Super Polyfoil™), polyester films (e.g., polyethylene terephthalate films), and polypropylene films.

[0051] In certain embodiments, the cosmetic film is transferrable from a coated or non- coated aluminum foil, a polyester film, a non-silicone coating, or a combination thereof) onto facial skin without tearing.

[0052] The cosmetic film may be rubbed into skin after application to the skin for about 10 min, 15 min, or about 20 min or longer or optionally wetted with water after about 10 min, about 15 min, about 20 min or longer.

[0053] The cosmetic film is preferably being capable of adhering to facial skin of an upright human for at least about 5 min., about 10 min, about 15 min or about 20 min.

[0054] In yet another aspect, the invention generally relates to a cosmetic product including the cosmetic film disclosed herein.

[0055] In certain embodiments, the cosmetic product includes a substrate that the cosmetic film of the invention is supported by and is capable of being peeled off of the substrate without tearing. In certain embodiments, the substrate comprises aluminum foil, a polyester film, a non silicone coating or a combination thereof. [0056] The cosmetic products of the invention can further comprise packaging. For example, the cosmetic product can comprise a pouch or wrapper to hold the cosmetic film, a rigid container to hold one or more films, a box, or a combination thereof.

[0057] In yet another aspect, the invention generally relates to a method of making the cosmetic film, comprising exposing a solution comprising derivatized collagen and one or more cosmetic ingredients to a crosslinking condition to crosslink the derivatized collagen.

[0058] Cosmetic films of the invention can be prepared by drying a solution of derivatized collagen (e.g., by using a laminar flow hood or similar air stream to“dry” the solution or by placing the solution in an oven, optionally with controlled air circulation) and/or by exposing the solution or dried solution to crosslinking conditions to crosslink the derivatized collagen. In a preferred embodiment, the derivatized collagen solution is not dried before crosslinking.

[0059] The solution of derivatized collagen can optionally be heated to up to 45°C prior to forming the cosmetic film (e.g., to a temperature ranging from 20°C to 25 °C, from 25 °C to 30 °C, from 30 °C to 35 °C, from 35 °C to 40 °C or from 40 °C to 45 °C).

[0060] One or more cosmetic ingredients can be included in the derivatized collagen solution prior to drying and/or crosslinking. Alternatively, or in addition, one or more cosmetic ingredients can be added to the cosmetic film after drying and/or crosslinking (e.g., by adding one or more cosmetic ingredients to the surface of the cosmetic film). The crosslinking can be performed using ultraviolet (UV) radiation, preferably using a wavelength of about 254 nm.

[0061] Crosslinking can be performed under normal atmospheric conditions. Alternatively, crosslinking can be performed in an oxygen free environment, such as in a nitrogen atmosphere. The length of the UV radiation can range, for example, from 10 sec to 30 min (e.g., 20 sec to 5 min, 5 min to 10 min, 10 min to 20 min, or 20 min to 30 min). Shorter crosslinking times can be used when using a photoinitiator.

[0062] The amount of time required for crosslinking to obtain a film having desirable properties can be adjusted depending on preheating conditions, collagen concentration, presence of photoinitiator, and the presence, amounts and types of different cosmetic ingredients. For example, inclusion of an oil based ingredient, low molecular weight HA or hydrolyzed collagen may require a longer crosslinking time (see, for example, Tables 7 and 12 (to compare films made without and with oils), Table 14 (comparing films made with different molecular weight HA), and Table 17 (comparing films made with different amounts of hydrolyzed collagen)). On the other hand, using a photoinitiator and preheating the derivatized collagen can shorten the required crosslinking time (see, for example, Table 6 (comparing films made with different amounts of sodium persulfate) and Tables 4 and 5 (comparing films made with different collagen preheating conditions and crosslinking times)).

[0063] Exemplary photoinitiators include riboflavin and sodium persulfate. Riboflavin can be included in the derivatized collagen solution in amounts ranging from, for example, 0.0001% to 0.02% (w/v). In some embodiments, the amount of riboflavin ranges from 0.001% to 0.01% (w/v). In some embodiments, the amount of riboflavin ranges from 0.001% to 0.008% (w/v). Sodium persulfate can be included in the derivatized collagen solution in amounts ranging from, for example, 0.1% to 15%. In some embodiments, the amount of sodium persulfate ranges from 0.1% to 2 % w/v). In some embodiments, the amount of sodium persulfate ranges from 0.25% to 1%.

[0064] The methods of making the cosmetic films of the invention can further comprise forming the solution used to make the film by combining a solution of derivatized collagen with one or more cosmetic ingredients and, optionally, one or more photoinitiators.

[0065] Films of a uniform thickness can be made by using a film applicator ( e.g ., Gardco microm film applicator) to spread the solution of derivatized collagen on a suitable substrate prior to crosslinking or drying. When making the cosmetic films of the invention, the solution can be spread to a desired thickness, for example, a thickness in the range of 0.4 mm to 2 mm. In some embodiments, the solution is spread to a thickness of 0.5 mm to 1.5 mm. In other embodiments, the cosmetic film is spread to a thickness 0.6 mm to 1.2 mm.

[0066] Suitable substrates on which a cosmetic film of the invention can be formed include aluminum foil, some polyester films, and some substrates that have non-silicone coatings, such as the Cheever #520 substrate described in the Examples. The cosmetic film can optionally be transferred to a second substrate after being formed (e.g., transferred from an aluminum foil substrate to a polyester substrate).

[0067] In certain embodiments, the crosslinking conditions include exposing the solution to UV radiation (e.g., including a wavelength of about 254 nm). [0068] In certain embodiments, the solution of derivatized collagen with one or more cosmetic ingredients further includes one or more photoinitiators.

[0069] In certain embodiments, the photoinitiator is selected from riboflavin, sodium persulfate, or a combination thereof. In certain embodiments, the amount of riboflavin is in the range from about 0.0001% to about 0.02% (w/v). In certain embodiments, the amount of sodium persulfate is in the range from about 0.1% to about 2% (w/v).

[0070] In certain embodiments, the solution is (i) exposed to the UV radiation in an oxygen free atmosphere, which is optionally a nitrogen atmosphere, or (ii) exposed to the UV radiation under ambient atmospheric conditions. In certain embodiments, the solution is exposed to the UV radiation for a period in the range of about 10 sec to about 30 min ( e.g ., about 10 sec to about 15 min, about 10 sec to about 10 min, about 10 sec to about 5 min, about 10 sec to about 1 min, about 30 sec to about 30 min, about 1 min to about 30 min, about 5 min to about 30 min, about 10 min to about 30 min).

[0071] In certain embodiments, the solution is applied to a substrate (e.g., aluminum foil, a polyester film, a non-silicone coating, or a combination thereof) prior to exposing the solution to crosslinking conditions. In certain embodiments, the solution is spread on the substrate to a uniform thickness using a film applicator.

[0072] In yet another aspect, the invention generally relates to a cosmetic film made by a method disclosed herein.

[0073] In yet another aspect, the invention generally relates to a method for delivering one or more cosmetic agents to the skin of a subject. The method includes applying the cosmetic film of the invention to the skin of a subject.

[0074] The cosmetic film of the invention may be applied to the face, neck, or another body part of the subject.

[0075] In certain embodiments, the method further includes rubbing the film into the skin of the subject. In certain embodiments, the method further includes wetting the film prior to rubbing the film into the skin of the subject. In certain embodiments, the method further includes the film is rubbed into the skin after being applied to the skin for at least 5 min (e.g., about 5 to 10 min, about 10 to 15 min). [0076] Films of the invention can be fabricated so that following application they form an adherent dry film that can be rubbed into skin. Preferably, the films of the invention form an adherent dry film after being applied to skin for a period of time, such as 20 or fewer min, 30 or fewer min, or 40 or fewer min. after application to the skin at room temperature and 30% or less humidity.

[0077] In some embodiments, the films are capable of maintaining contact with the dry facial skin of an upright individual for at least 20, 30 or 40 min. following application, for example at room temperature and 30% or less humidity, so that, for example, the user does not need to lay down until the cosmetic film dries sufficiently to be rubbed into the skin.

[0078] Films for cosmetic applications should ideally be able to be applied to skin, such as facial skin, without tearing. Accordingly, in some embodiments, the cosmetic films of the invention are capable of being transferred from a substrate ( e.g ., a non-coated aluminum foil substrate) onto skin (e.g., facial skin) without tearing.

[0079] The cosmetic films of the invention can be fabricated to have dimensions suited for cosmetic applications. For example, the films can be faced shaped so that the film can be used as a face mask. Face shaped films can further comprise eye, mouth, and nose cutouts to make the film more comfortable for the user. The films can be made in different sizes and shapes when intended to be used on different skin surfaces. For example, a film intended for application around the eyes can be kidney shaped and sized to match the area surrounding the eye where crow’s feet wrinkles typically form, and a film intended for application to a forehead or neck may be a rectangular shape and sized to cover the forehead or portion thereof or neck or portion thereof. Accordingly, the films of the invention can be a variety of shapes, including face shaped, rectangular with rounded edges, rectangular without rounded edges, circular, oval, or elliptical.

[0080] In yet another aspect, the invention generally relates to a method for delivering collagen and/or one or more cosmetic ingredients to skin. The method includes applying a cosmetic film of the invention to the skin of the subject. The subject is preferably a human, e.g., an adult over the age of 20, over the age of 30, over the age of 40, over the age of 50, or over the age of 60. In some embodiments, the subject is female.

[0081] The cosmetic films can be applied to any skin surface to which delivery of collagen and/or the one or more cosmetic ingredients is desired. For example, the cosmetic film can be applied to the face or one or more parts thereof (such as the area around the eyes, forehead, or nasolabial folds), the neck, or any site on the body. After the film has been applied to the surface for a period of time, for example 10 to 60 min, 10 to 30 min, 10 to 20 min, or 20 to 30 min, the film can be rubbed into the skin of the subject. In some embodiments, the film can be left on the skin for at least 10 min, at least 20 min, at least 30 min. Preferably, the films are rubbed into the skin after the film has formed an adherent dry film. By rubbing the film into the skin of the subject rather than removing the film, all or substantially all of the collagen and cosmetic ingredients in the film remain on the skin of the subject. Prior to rubbing the film into the skin, the film can be wetted, for example with water or a liquid cosmetic product such as a lotion, to aid in spreading the ingredients in the film over the skin surface.

Examples

[0082] The following examples are meant to be illustrative of the practice of the invention, and not limiting in any way.

[0083] The following abbreviations are used in the Examples: second (sec); minute (min); hours (hr); Polyethylene terephthalate (PET); biaxially oriented polypropylene (BOPP);

hyaluronic acid (HA); molecular weight (MW); and room temperature (Room Temp or r.t.) about 20 °C to 25 °C.

Example 1. Preparation of Chemically Derivatized Collagen

[0084] Two hundred milliliters of 3mg/mL purified, soluble collagen (Porcogen, Lot #531131080 or in-house prepared bovine collagen prepared from calf-hide) were filtered through 0.45 pm and 0.2 pm cartridge filters. The filtered collagen was place in a 500 mL beaker and adjusted pH 9.0 using 10N and lN NaOH. After stirring for 5 min. at room temperature, pulverized glutaric anhydride powder (Sigma, >95%) was slowly added to the stirring collagen solution at a concentration equal to 10% of the collagen (60 mg). The pH of the collagen solution was maintained at pH 9.0 by addition of drops of 10N NaOH. The glutaric anhydride reaction continued for 15-30 min at which point drops of 6N HC1 and 1N HC1 were added to reduce the pH top approximately 4.5 to precipitate the derivatized collagen. The derivatized collagen was then placed in 50 mL centrifuge tubes or centrifuge bottles and centrifuged at 10,000 rpm to precipitate the derivatized collagen. The recovered precipitate was then solubilized by adjusting the pH to 7.2 by adding drops of 10N NaOH and 1N NaOH. The pH was monitored as the NaOH was mixed with the derivatized collagen pellet. The neutralized, clear and transparent collagen gel was then placed in centrifuge tubes or bottle and centrifuged to remove air bubbles. The clear, de-aerated collagen gel was stored at refrigeration temperatures pending fabrication of dissolving, collagen-based, cosmeceutical films having active ingredients to improve skin appearance and enhance skin rejuvenation. If necessary, the derivatized collagen gels were diluted with neutral pH solutions to reduce concentration and viscosity.

Example 2. Fabrication of Collagen Films Not Exposed to UV Irradiation

[0085] Chemically derivatized collagen prepared as in Example 1 was removed from refrigeration storage and warmed to room temperature. Films without active ingredients were prepared in glass petri dishes or placed on glass plates to form a thin layer of liquid collagen. The glass petri dishes or glass plates were then placed in a laminar flow hood to dry the solution to a thin film. The films were removed from the glass plates, cut to desired sizes, thickness measured using a caliper, placed in plastic pouches, labeled, and stored at refrigerated temperatures.

Samples were evaluated for dissolution time by (1) placing the films on skin surfaces and examining dissolution over time or (2) by placing the films (stained with picrosirus red) on wet cheesecloth and placing the samples in a 35°C oven. Dissolution was observed as the film melted and the stain penetrated the wet cheesecloth base. Dissolution occurred within 30 min.

Example 3. Fabrication of Collagen Films Stabilized by Exposure to 254nm UV Irradiation in

Nitrogen

[0086] Chemically derivatized collagen solutions (6.7 mg/mL) prepared as in Example 1 were removed from refrigeration storage and cast into thin, wet sheets, by placing the collagen solution on a polyester support film (Cheever # 520) using a microm film applicator (knife bar) to provide a wet film thickness of 0.05 inches (approximately 1.3 mm). The wet collagen films were placed in a controlled atmosphere chamber that was flushed with nitrogen gas to eliminate oxygen. UV lights in the chamber were activated and films were exposed to 254 nm ultraviolet irradiation for times ranging from 10 min to 17 min. Resultant stabilized collagen films were removed from the chamber and evaluated for dissolution and skin moisturization by placing film samples on skin on the back of the hand. Films exposed to UV irradiation for less than 12 min provided a collagen film that moisturized and could be easily rubbed into the skin within 30 min. Films that were exposed to UV irradiation for more than 12 min. were more stable and less moisturizing and did not totally dissolve when finger pressure was applied to”rub” the film into the skin surface.

Example 4. Fabrication of Collagen Films Stabilized by Exposure to 254 nm UV Irradiation in Normal Atmosphere with a Photoinitiator

[0087] Three milliliters of 0.3 M sodium persulfate was added to chemically derivatized collagen prepared as in Example 1 to a total volume of 30 mL. Aliquots of this solution were placed in glass petri dishes to cover the bottom surface. The petri dish cover was removed and the thin collagen layer exposed to 254 nm ultraviolet irradiation for 30 sec. Firm and wet collagen films were removed and sections placed on the back of the hand to evaluate feel and dissolvability. Within 10 min the wet film contracted in area, leaving a small dry film that could be wetted with a drop of water and smoothed into the skin by finger motion. The treated site felt comfortably moistened.

Example 5. Fabrication of Collagen Films Stabilized by Exposure to 254 nm UV Irradiation in Nitrogen with a Photoinitiator, sodium persulfate

[0088] Chemically derivatized collagen solutions prepared as in Example 1 were removed from refrigeration storage. Sodium persulfate was mixed into a collagen solution prepared as in Example 1 at concentrations ranging from 2.5% to 10% (w/v), heated to 28°C or 30°C, and cast into thin, wet sheets, by placing the collagen solution on aluminum tins and spreading the solutions into a thin sheet. The sheets were exposed to 254 nm ultraviolet irradiation in nitrogen for 20 sec. Resulting films were examined for firmness. The films having 10% sodium persulfate were very firm, while those with 2.5% sodium persulfate were not cured. Films exposed to 5% sodium persulfate were firm and slightly wet.

Example 6. Fabrication of Collagen Films Stabilized by Exposure to 254 nm UV Irradiation in Nitrogen with a Photoinitiator, Riboflavin

[0089] Chemically derivatized collagen solutions (5.5 mg/mL) prepared as in Example 1 were removed from refrigeration storage. Riboflavin solution was mixed into the collagen solution at concentrations ranging from 0.001 to 0.008%, and cast into thin, wet sheets, by placing the collagen solution on aluminum foil and spreading the solutions into, small (l/2”xl- 1/2”) sheets using a pipette. The sheets were exposed to 254 nm ultraviolet irradiation in nitrogen for two 30 sec intervals. All resulting films appeared to cure well except for lower riboflavin concentration solution (0.001%). When placed on skin on the back of hands, samples with 0.002% and 0.004% riboflavin appeared to provide the best wetting and moisturizing effects.

Example 7. Fabrication of Collagen Films prepared from Hydrolyzed Collagen Preparations

Stabilized by Exposure to 254 nm UV Irradiation

[0090] Hydrolyzed solutions of chemically derivatized collagen solutions (5.5 mg/mL) prepared as in Example 1 were prepared by heating the solutions to 38°C for 30 min. Aliquots of 200 pL were placed on aluminum foil and exposed to 254 nm ultraviolet irradiation for 30 min. None of the wet collagen films were firm but when placed on the back of the hand were able to be rubbed in the skin by finger pressure.

Example 8. Fabrication of Collagen Films prepared from Chemically Derivatized Collagen

Having Hydrolyzed Collagen and Stabilized by Exposure to 254 nm UV

Irradiation

[0091] Hydrolyzed collagen solutions were prepared as described in Example 7. Three sets of hydrolyzed collagen in chemically derivatized collagen (non-hydrolyzed) prepared as in Example 1 were prepared: 3 mL hydrolyzed in 1 mL of chemically derivatized collagen, 2 mL hydrolyzed in 2 mL of chemically derivatized, and 1 mL of hydrolyzed collagen in 3 mL of chemically derivatized collagen. Films were prepared as described in Example 7 and examined for firmness and back of the hand feel and moisturization. None of the films completely cured in the center of the films. In a subsequent study, samples having 250 pL of hydrolyzed collagen mixed with 1570 pL of base derivatized collagen and 125 pL of hydrolyzed collagen mixed with 1875 pL of base derivatized collagen formed firm, wet films following exposure to 254 nm ultraviolet irradiation in nitrogen.

Example 9. Fabrication of Collagen Films prepared from Chemically Derivatized Collagen having Hyaluronic Acid and Stabilized by Exposure to 254 nm UV Irradiation

[0092] Chemically derivatized collagen solutions (5.5 mg/mL) prepared as in Example 1 were removed from refrigerator storage and supplemented with different concentrations and molecular weights of hyaluronic acid (LifeCore Biomedical). Molecular weights included 5,000, 20,000, 100,000, and 500,000 Dalton hyaluronic acid. Concentrations of each ranged from lmL per 6ml of derivatized collagen to 3 mL per 6 mL of derivatized collagen. Samples of mixed molecular weights and concentrations in base derivatized collagen were placed on aluminum foil and exposed to 254 nm ultraviolet irradiation for 30 min in nitrogen. All samples cured well producing a moisturizing, cool to the feel film when placed on skin on the back of the hand. Samples having a mixture of high and low molecular weight hyaluronic acid appeared most desirable for skin moisturization and comfort.

Example 10. Fabrication of Collagen Films prepared from Chemically Derivatized Collagen

Having Active Ingredients to Provide Skin Moisturization, Wrinkle Prevention or Removal, Skin Whitening, and Provide Anti-aging Activity

[0093] A: Collagen films were prepared by exposure to 254 nm ultraviolet irradiation in a nitrogen atmosphere, as described above. Prior to forming films, active ingredients were added to the collagen gel before casting thin layers on a solid backing, aluminum foil. Active ingredients included 250 pL of argan nut oil and chia seed oil with 500 pL of lecithin solution per 5 mL collagen solution. Samples were spread onto a solid backing and exposed to 254 nM ultraviolet irradiation for 2 min. Resultant films were formed.

[0094] B: Collagen films were prepared by exposure to 254 nm ultraviolet irradiation in a nitrogen atmosphere, as described above. Prior to forming films, active ingredients were added to the collagen gel before casting thin layers on a solid backing. Ingredients included rosehip oil (2.4-7% with in collagen solution) and argan nut oil (2.4-7% w/v in collagen solution). Samples were placed on aluminum backing and spread into a wet sheet using the knife bar set at 0.025 inches (approximately 0.6 mm). Sheets were place in the nitrogen chamber and exposed to 254 nm ultraviolet irradiation for 10 min. All samples formed films with a slight white color.

Example 11. Fabrication and Testing of a Variety of Collagen Films

[0095] Collagen films were prepared under a variety of conditions and with variety of ingredients. In each of the films, a solution of derivatized collagen prepared as described in Example 1 was used. Crosslinking was performed under nitrogen atmosphere with a wavelength of 254 nm, without a photoinitiator, and without allowing the collagen solution to dry prior to crosslinking, unless otherwise noted. In each experiment, the collagen solution was warmed to room temperature prior to crosslinking, unless otherwise noted. Results of an experiment were judged as positive (+) or negative (-) on the basis of the observed properties of the film.

Varying collagen concentration and crossl inkin time

[0096] Collagen films were prepared by dispensing 1,000 pL of an undiluted or diluted derivatized collagen solution to cover a 0.5 inch by 0.5 inch (about 13 mm by 13 mm) square of Reynolds Wrap® aluminum foil, followed by crosslinking for the length of time shown in Table 1

Table 1. Varying crosslinking time and collagen concentration

[0097] Collagen films were also prepared by dispensing 500 pL of a derivatized collagen solution to cover a 0.75 inch (approximately 19 mm) diameter circle of Reynolds Wrap® aluminum foil, followed by crosslinking for the length of time shown in Table 2.

Table 2. Effect of crosslinking time

[0098] These experiments show that collagen concentration and crosslinking time can be used to modulate the properties of the collagen films.

Varying film thickness

[0099] A collagen film was prepared on a Reynolds Wrap® aluminum foil substrate having a variable thickness by using a knife-bar set at 0.075 inches (approximately 1.9 mm) on the left side of the film and set to 0.0000 inches (0 mm) at the right side of the film. Crosslinking was performed for 20 min. Material on the right 1/3 of foil was too thin, material on the left 1/4 of foil did not coat the foil (de-wetted), while material in the middle of the foil was a good consistency and formed a nice film. From this experiment, the ideal thickness appeared to be at about 0.045 inch (about 1.1 mm). The film at this thickness was observed to have nice elasticity and was neither too wet nor too dry.

[00100] In a second set of experiments, different volumes of collagen solution were dispensed onto a 9.365 cm diameter circle of Reynolds Wrap® aluminum foil and crosslinked for 25 min.

Table 3. Effect of film thickness on crosslinkability

[00101] From these experiments, it appeared that results from earlier experiments were scalable from 0.75 inch (approximately 19 mm) to 9.365 cm diameters and that crosslinkability was not thickness dependent, even for heated samples. [00102] In a third set of experiments, films having a thickness of 0.025 inches (approximately 0.6 mm) and 0.05 inches (approximately 1.3 mm) were made by dispensing a volume of derivatized collagen solution onto silicone free Super Polyfoil™ (Fisher Scientific), spreading the solution to the desired thickness using a microm film applicator (Gardco), followed by crosslinking for 30 min. Both films cured beautifully, were gel-like, were able to be peeled off the foil easily, and unrolled on the back of the hand easily.

[00103] In a fourth set of experiments, films having a thickness of 0.025 inches

(approximately 0.6 mm) were made by dispensing a volume of derivatized collagen solution onto silicone free Super Polyfoil™ (Fisher Scientific), spreading the solution to the desired thickness using a film applicator, followed by crosslinking for 3, 5, or 10 min. The film that was cured for 5 min cured well, transferred to skin from the foil easily, dried on the skin in about 30 min, and was able to be peeled off of the skin easily.

Varying collagen preheating and crossl inkin time

[00104] Collagen films were prepared by dispensing 500 pL of a preheated collagen solution to cover a 0.75 inch (approximately 19 mm) diameter circle of Reynolds Wrap® aluminum foil, followed by crosslinking for the length of time shown in Table 4.

Table 4. Varying preheating and crosslinking time

[00105] A crosslinking time of 15 to 30 min produced the best films when the collagen solution was preheated to 35 °C for 30 min. Following these experiments, additional experiments were performed varying the preheating conditions and crosslinking times from 15 to 30 min.

Table 5. Varying preheating and crosslinking time

Sodium persulfate and riboflavin as photoinitiators

[00106] Collagen films were made using varying amounts of sodium persulfate as a photoinitiator. All films were made by dispensing 3.125 mL of collagen solution having an amount of sodium persulfate identified onto a 9.365 cm diameter circle of Reynolds Wrap® aluminum foil prior to crosslinking. All films were crosslinked wet, except for the films of experiments 12 and 13, which were crosslinked after drying.

Table 6. Sodium persulfate as photoinitiator

[00107] A second set of experiments was performed using varying amounts of riboflavin as photoinitiator. All derivatized collagen solutions were preheated to room temperature prior to dispensing 8 drops on a 0.5 inch by 0.5 inch (about 13 mm by 13 mm) square of silicone free Super Polyfoil™ (Fisher Scientific) and crosslinking for 1 min. Table 7. Riboflavin as photoinitiator

[00108] These experiments confirm that photoinitiators can be used to reduce the amount of time required to crosslink derivatized collagen films.

Different substrates

[00109] Experiments were performed to evaluate the suitability of a variety of substrates. In a first set of experiments, films were made by dispensing three drops of a derivatized collagen solution on each substrate followed by crosslinking for 20 min.

Table 8. Comparison of alternative substrates

[00110] In a second set of experiments, films were made by dispensing an amount of a derivatized collagen solution on each substrate followed by crosslinking for 30 min. The film of experiment number 7 was dried before crosslinking.

Table 9. Comparison of alternative substrates

[00111] In a third set of experiments, 0.025 inch thick films (about 0.6 mm) were made by dispensing an amount of a derivatized collagen solution onto the substrate and using a film applicator to spread the solution to a uniform thickness, followed by crosslinking.

Table 10. Comparison of alternative substrates

[00112] From these experiments, it was concluded that optimal substrates are ETV-resistant (to avoid curing the collagen to the substrate) and hydrophilic (to limit amount of dewetting). Based on the positive results for films made using the Cheever #520 - 92g PET substrate, this substrate was selected for further evaluation. A series of 0.05 inch (approximately 1.3 mm) thick films were made using varying crosslinking times.

Table 11. Effect of crosslinking time on films made on Cheever #520 - 92g PET substrate

Different crosslinking wavelengths

[00113] Three derivatized collagen films were made on silicone free Super Polyfoil™ (Fisher Scientific) by crosslinking for 20 min using ETV radiation at 254 nm, 302 nm, or 365 nm. Only the film made using 254 nm ETV radiation cured well. Films with added ingredients

[00114] Derivatized collagen films were made with various different cosmetic ingredients. Concentrations of solid ingredients identified in this section were measured as weight percent (w/w) of the collagen solution used to make the films and concentrations of liquid ingredients were measured as volume percent (v/v) of the collagen solution used to make the films.

[00115] In a first set of experiments, collagen films on a Reynolds Wrap® aluminum foil substrate were made with 0.015%, 0.045%, or 0.09% hyaluronic acid having an average molecular weight of 1M Daltons. The films were crosslinked for 30 min. Each of the films cured nicely and felt squishy but dry when touched.

[00116] In a second set of experiments, collagen films having various ingredients were made on a silicone free Super Polyfoil™ (Fisher Scientific) substrate using riboflavin as a

photoinitiator.

Table 12. Films having ingredients and made using riboflavin as photoinitiator

[00117] In a third set of experiments, collagen films having various ingredients were made on a silicone free Super Polyfoil™ (Fisher Scientific) substrate without a photoinitiator.

Table 13. Films having ingredients and made without photoinitiator

[00118] In a fourth set of experiments, collagen films having various concentrations of hyaluronic acid of different molecular weights were made on silicone free Super Polyfoil™ (Fisher Scientific) using riboflavin as a photoinitiator.

Table 14. Films having different molecular weight HA made using riboflavin as photoinitiator

[00119] In a fifth set of experiments, derivatized collagen films having various concentrations of hyaluronic acid of different molecular weights were made on silicone free Super Polyfoil™ (Fisher Scientific) without a photoinitiator. All films were crosslinked for 30 min.

Table 15. Films having different molecular weight HA made without a photoinitiator

[00120] In a sixth set of experiments, derivatized collagen films having various blends of different molecular weight hyaluronic acid were made on silicone free Super Polyfoil™ (Fisher Scientific) without a photoinitiator. All films were crosslinked for 30 min.

Table 16. Films having blends of HA

[00121] In a seventh set of experiments, derivatized collagen films having different amounts of hydrolyzed collagen were made on silicone free Super Polyfoil™ (Fisher Scientific). All films were crosslinked for 30 min.

Table 17. Films made with hydrolyzed collagen

[00122] In an eighth set of experiments, derivatized collagen films having different amounts of ingredients were made on silicone free Super Polyfoil™ (Fisher Scientific). All films were crosslinked for 5 min. Films were then transferred to a second substrate.

Table 18. Films transferred to second substrate

[00123] In a ninth set of experiments, derivatized collagen films having different amounts of rosehip and argan nut oils were made on the Cheever #520 substrate. All films were crosslinked for 10 min.

Table 19. Films made with rosehip or argan nut oil

Example 11. Fabrication of Collagen Films Having Active Ingredients for Moisturization and

Wrinkle Prevention

[00124] Collagen films are fabricated as described in Examples 3-6. Prior to forming films, active ingredients are added to the collagen gel. In one preparation, Acetyl hexapeptide-3 (Argireline) is added to the liquefied collagen at 0.5% before forming the films. In another preparation ceramide is added to the liquefied collagen at 0.25% before forming the films. In a third preparation, glycerin and retinol are added to the liquefied collagen at 1% before forming the films. Following film fabrication samples are examined for skin moisturization and ability to be rubbed into skin after application by placing films on the back of hands. Films are able to be rubbed into skin following application for 30 min. Skin feels moisturized.

Example 12. Fabrication of Collagen Films Having Active Ingredients for Anti-aging

[00125] Collagen films are fabricated as described in Examples 3-6. Prior to forming films, active ingredients are added to the collagen gel. In one preparation, Epidermal Growth Factor is added to the liquefied collagen at 0.5% before forming the films. Following film fabrication samples are examined for skin moisturization and ability to be rubbed into skin after application by placing films on the back of hands. Films are able to be rubbed into skin following application for 30 min. Skin feels moisturized.

Example 13. Fabrication of Collagen Films Having Active Ingredients for Skin Whiting

[00126] Collagen films are fabricated as described in Examples 3-6. Prior to forming films, active ingredients are added to the collagen gel. In one preparation, Dimethicone is added to the liquefied collagen at 0.5% before forming the films, or nicotinamide is added to the liquefied collagen at 0.5% before forming the films, or Tetrahydrocurcuminoids are to the liquefied collagen at 0.5% before forming the films. Following film fabrication samples are examined for skin moisturization and ability to be rubbed into skin after application by placing films on the back of hands. Films are able to be rubbed into skin following application for 30 min. Skin feels moisturized.

Example 14. Fabrication of Collagen Films Having Nanoparticle Active Ingredients

[00127] Collagen films are fabricated as described in Examples 3-6. Prior to forming films, active ingredients in nanoparticle form are added to the collagen gel prior to forming films. Following film fabrication samples are examined for skin moisturization and ability to be rubbed into skin after application by placing films on the back of hands. Films are able to be rubbed into skin following application for 30 min. Skin feels moisturized.

[00128] While various specific embodiments have been illustrated and described, it will be appreciated that various changes can be made without departing from the spirit and scope of the invention.

[00129] Applicant’s disclosure is described herein in preferred embodiments with reference to the Figures, in which like numbers represent the same or similar elements. Reference throughout this specification to“one embodiment,”“an embodiment,” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases “in one embodiment,”“in an embodiment,” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

[00130] The described features, structures, or characteristics of Applicant’s disclosure may be combined in any suitable manner in one or more embodiments. In the description, herein, numerous specific details are recited to provide a thorough understanding of embodiments of the invention. One skilled in the relevant art will recognize, however, that Applicant’s composition and/or method may be practiced without one or more of the specific details, or with other methods, components, materials, and so forth. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the disclosure.

[00131] In this specification and the appended claims, the singular forms "a," "an," and "the" include plural reference, unless the context clearly dictates otherwise.

[00132] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described.

Methods recited herein may be carried out in any order that is logically possible, in addition to a particular order disclosed.

Incorporation by Reference [00133] References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made in this disclosure.

All such documents are hereby incorporated herein by reference in their entirety for all purposes. Any material, or portion thereof, that is said to be incorporated by reference herein, but which conflicts with existing definitions, statements, or other disclosure material explicitly set forth herein is only incorporated to the extent that no conflict arises between that incorporated material and the present disclosure material. In the event of a conflict, the conflict is to be resolved in favor of the present disclosure as the preferred disclosure.

Equivalents

[00134] The representative examples are intended to help illustrate the invention, and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including the examples and the references to the scientific and patent literature included herein. The examples contain important additional information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.

Claims

1. A cosmetic film comprising crosslinked derivatized collagen and one or more cosmetic ingredients, wherein optionally the cosmetic film is suitable for placement on human skin and dissolves or disassociates within 30 minutes of placement on skin.

2. The cosmetic film of claim 1, which has a thickness of about 0.4 mm to about 2 mm.

3. The cosmetic film of claim 1 or 2, wherein the derivatized collagen comprises at least about 75% of the dry weight of the film.

4. The cosmetic film of any of claims 1-3, wherein the derivatized collagen comprises up to about 99% of the dry weight of the film.

5. The cosmetic film of any of claims 1-4, wherein the concentration of the derivatized

collagen in the film is in the range from about 0.2% to about 10% w/v.

6. The cosmetic film of any of claims 1-5, wherein the derivatized collagen comprises

derivatized bovine collagen, derivatized porcine collagen, derivatized human collagen, derivatized recombinant collagen, or a combination thereof.

7. The cosmetic film of any of claims 1-6, wherein the derivatized collagen comprises

acylated collagen.

8. The cosmetic film of claim 7, wherein the derivatized collagen comprises glutarylated collagen and/or succinylated collagen.

9. The cosmetic film of any of claims 1-6, wherein the one or more cosmetic ingredients comprises moisturizing ingredients, wrinkle preventative ingredients, anti-aging ingredients, skin whitening ingredients, or a combination thereof.

10. The cosmetic film of claim 9, wherein the one or more moisturizing ingredients are

selected from hyaluronic acid (HA), hydrolyzed collagen, glycerin, aloe vera juice, lipids, oils, butters, or a combination thereof.

11. The cosmetic film of claim 9, wherein the one or more wrinkle preventative ingredients are selected from acetyl hexapeptide-3 (Argireline), HA, glycerin, retinol, ceramide, or a combination thereof.

12. The cosmetic film of claim 9, wherein the one or more anti-aging ingredients are selected from epidermal growth factor, acetyl hexapeptide-3 (Argireline), ceramide, HA, retinol, soluble collagen, or a combination thereof.

13. The cosmetic film of claim 9, wherein the one or more skin whitening ingredients are selected from dimethicone, nicotinamide, tetrahydrocurminoids, glutathione, or a combination thereof.

14. The cosmetic film of any of claims 9-13, wherein the one or more cosmetic ingredients comprises moisturizing ingredients, wrinkle preventative ingredients, anti-aging ingredients, skin whitening ingredients, or a combination thereof are present the form of nanoparticles or liposomes, or a combination thereof.

15. The cosmetic film of any of claims 9-14, wherein the one or more cosmetic ingredients comprises HA in an amount accounting for at least 0.1% of the weight of the film.

16. The cosmetic film of any of claims 9-14, wherein the one or more cosmetic ingredients comprises HA in an amount accounting for up to about 15% of the weight of the film.

17. The cosmetic film of claim 15 or 16, wherein the HA is characterized by a weight

average molecular weight in the range of 1 kDa to 4 MDa.

18. The cosmetic film of claim 15 or 16, wherein the HA is characterized by a weight

average molecular weight in the range of 250 kDa to 4 MDa.

19. The cosmetic film of any of claims 1-18, wherein the one or more cosmetic ingredients comprises hydrolyzed collagen.

20. The cosmetic film of claim 19, wherein the hydrolyzed collagen accounts for about 0.1% to about 2% of the weight of the film.

21. The cosmetic film of any of claims 1-20, wherein the one or more cosmetic ingredients comprises one or more lipids, one or more oils, one or more butters, or a combination thereof.

22. The cosmetic film of claim 21, wherein the one or more lipids, one or more oils, one or more butters are selected from argan nut oil, chia seed oil, flaxseed oil, rosehip oil, tamanu butter, lecithin, or a combination thereof.

23. The cosmetic film of claim 22, wherein the one or more lipids, one or more oils, one or more butters, or a combination thereof account for up to about 15% of the weight of the film.

24. The cosmetic film of any of claims 1-23, further comprising a photoinitiator.

25. The cosmetic film of claim 24, wherein the photoinitiator is riboflavin, sodium persulfate, or a combination thereof.

26. The cosmetic film of claim 25, wherein the amount of riboflavin is in the range from

about 0.0001% to about 0.02% (w/v).

27. The cosmetic film of claim 25, wherein the amount of sodium persulfate is in the range from about 0.1% to about 2% (w/v).

28. The cosmetic film of any of claims 1-27, wherein the shape of the cosmetic film is

selected from (i) face shaped, optionally having eye, mouth, and nose cutouts, (ii) rectangular with rounded edges, (iii) rectangular without rounded edges, (iv) circular, (v) oval, (vi) elliptical, or (vi) kidney shaped.

29. The cosmetic film of any of claims 1-28, being transferrable from a non-coated aluminum foil substrate onto facial skin without tearing.

30. The cosmetic film of any of claims 1-29, being suitable for rubbing into skin after

application to the skin for 20 min or longer or optionally wetted with water after the 20 min or longer.

31. The cosmetic film of any of claims 1-30, being capable of adhering to facial skin of an upright human for at least 20 min.

32. The cosmetic film of any of claims 1-31, wherein the crosslinked derivatized collagen is characterized by a crosslinking density in the range from about _ to about _ .

33. The cosmetic film of any of claims 1-32, wherein the cosmetic film dissolves or

disassociates within 15 minutes of placement on skin of a subject.

34. A cosmetic product comprising the cosmetic film of any of claims 1-33.

35. The cosmetic product of claim 34, comprising a substrate wherein the film is supported by and is capable of being peeled off of the substrate without tearing.

36. The cosmetic product of claim 35, wherein the substrate comprises aluminum foil, a

polyester film, a non-silicone coating or a combination thereof.

37. A method of making the cosmetic film, comprising exposing a solution comprising derivatized collagen and one or more cosmetic ingredients to a crosslinking condition to crosslink the derivatized collagen.

38. The method of claim 37, wherein the crosslinking conditions comprise exposing the

solution to ultraviolet (UV) radiation.

39. The method of claim 38, wherein the UV radiation comprises a wavelength of about 254 nm.

40. The method of claim 38 or 39, wherein the solution further comprises a photoinitiator.

41. The method of claim 40, wherein the photoinitiator is riboflavin, sodium persulfate, or a combination thereof.

42. The method of claim 41, wherein the amount of riboflavin is in the range from about 0.0001% to about 0.02% (w/v).

43. The method of claim 41, wherein the amount of sodium persulfate is in the range from about 0.1% to about 2% (w/v).

44. The method of any of claims 37-43, wherein the solution is (i) exposed to the UV

radiation in an oxygen free atmosphere, which is optionally a nitrogen atmosphere, or (ii) exposed to the UV radiation under ambient atmospheric conditions.

45. The method of any of claims 37-43, wherein the solution is exposed to the UV radiation for a period in the range of about 10 seconds to about 30 minutes.

46. The method of any of claims 37-45, further comprising applying the solution to a

substrate prior to exposing the solution to crosslinking conditions.

47. The method of claim 46, wherein the substrate comprises aluminum foil, a polyester film, a non-silicone coating, or a combination thereof.

48. The method of any of claims 37-47, further comprising spreading the solution on the substrate to a uniform thickness using a film applicator.

49. A cosmetic film made by the method of any of claims 37-48.

50. A method for delivering one or more cosmetic agents to the skin of a subject, comprising applying the cosmetic film of any of claims 1-33 to the skin of the subject.

51. The method of claim 50, wherein the cosmetic film is applied to the face, neck, or body of the subject.

52. The method of claim 50 or 51, further comprising rubbing the film into the skin of the subject.

53. The method of any of claims 50-52, further comprising wetting the film prior to rubbing the film into the skin of the subject.

54. The method of claim 53, wherein the film is rubbed into the skin after being applied to the skin for at least 5 minutes.

55. A method for applying collagen and one or more cosmetic ingredients to skin, comprising applying a cosmetic film of any of claims 1-33 to skin of a subject.

56. The method of claim 55, further comprising rubbing the cosmetic film into the skin of the subject.

57. The method of claim 55 or 56, further comprising wetting the cosmetic film prior to rubbing the film into the skin of the subject.