HK1237253B - Thermosetting biophotonic compositions and uses thereof - Google Patents

Description

Thermosetting biophotonic compositions and uses thereof

Background Art

Recently, phototherapy has been recognized for a wide range of applications in medicine and cosmetics, including for surgery, treatment, and diagnosis. For example, phototherapy has been developed for the treatment of cancers and tumors with reduced invasiveness, as an antimicrobial treatment to disinfect target areas, to promote wound healing, and for facial skin rejuvenation.

Photodynamic therapy (PDT) is a type of light therapy that involves administering a photosensitizer to a target tissue and then, after a defined period of time (during which the photosensitizer is absorbed by the target tissue), exposing the target tissue to a light source. However, this therapy is often associated with undesirable side effects, including systemic or local toxicity to the patient or damage to non-target tissues. Furthermore, the efficacy of this existing therapy is often low, for example due to poor selectivity of the photosensitizer administered to the target tissue.

It is an object of the present invention to provide improved compositions and methods for phototherapy.

Summary of the Invention

The present invention provides thermosetting biophotonic compositions and methods that can be used for phototherapy.

From one aspect, the thermosetting biophotonic composition of the present invention comprises a block copolymer and at least one chromophore dissolved or solubilized within the copolymer.

In another aspect, the present invention provides a thermosetting biophotonic composition comprising a block copolymer and at least one chromophore dissolved in the copolymer, wherein the block copolymer is present in the composition at a concentration such that the composition can be thermally cured or thermally gelled within one minute upon contact with human or animal tissue. In certain embodiments, the composition can be thermally cured or thermally gelled within about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 70 seconds, about 80 seconds, about 90 seconds, about 100 seconds, about 110 seconds, or about 120 seconds. In certain embodiments, the composition can be thermally cured or thermally gelled upon contact with target tissue. In certain embodiments, the composition is in liquid form at room temperature (e.g., 22°C), and is heat cured when heated to or above 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C or 40°C. In certain embodiments, the block copolymer is present in the composition at a concentration of about 21%, 22%, 23%, 24% or 25% weight/volume. In certain embodiments, the block copolymer is poloxamer (poloxomer), such as Pluronic (Pluronic) F127, which is present in the composition at a concentration greater than 20% by weight, such as 21% by weight, 22% by weight, 23% by weight, 24% by weight or 25% by weight of the weight/volume. Advantageously, at these concentrations, the composition is in liquid form at room temperature, and gels when or after contact with the skin at body temperature.

From another aspect, the present invention provides a thermosetting biophotonic composition comprising a block copolymer and at least one chromophore dissolved in the copolymer, wherein the block copolymer is a poloxamer, such as Pluronic F127, and wherein the block copolymer is present in the composition at a concentration greater than 20 wt%.

In certain embodiments of any of the foregoing or following aspects, the block copolymer comprises at least one sequence of (PEG)-(PPG). In another embodiment, the block copolymer has the formula (PEG)-(PPG)-(PEG). In another embodiment, the block copolymer is a poloxamer such as Pluronic F127.

In certain embodiments of any of the foregoing or following aspects, the block copolymer comprises at least one (PEG)-(PLA) sequence. In some embodiments, the block copolymer comprises at least one (PEG)-(PLGA) sequence. In some embodiments, the block copolymer comprises at least one (PEG)-(PCL) sequence. In another embodiment, the block copolymer is a triblock copolymer or poloxamer of formula A-B-A or B-A-B, wherein A is PEG and B is PLA or PLGA or PCL.

In certain embodiments of any of the foregoing or following aspects, at least one chromophore is water-soluble. In certain embodiments of any of the foregoing or following aspects, at least one chromophore is negatively charged. At least one chromophore may be a fluorophore. In certain embodiments, the chromophore can absorb and/or emit light. In some embodiments, the light absorbed and/or emitted by the chromophore is within the visible light range. In some embodiments, the light absorbed and/or emitted by the chromophore has a peak wavelength within the range of about 400 nm to about 750 nm. In certain embodiments, the chromophore can emit light from about 500 nm to about 700 nm. In some embodiments, the chromophore or fluorophore is a xanthene dye. The xanthene dye can be selected from Eosin Y, Eosin B, Erythrosine B, Fluorescein, Rose Bengal, and Phloxin B. The chromophore can be a combination of one or more fluorophores, such as a variety of xanthene dyes.

In certain embodiments of any of the foregoing or following aspects, the block copolymer is present in the composition at a concentration greater than about 20% weight/volume of the composition. This provides a composition that can be heat-cured at body temperature, for example, when applied to a human or animal body. In some embodiments, the block copolymer is present in the composition at a concentration of about 21%, 22%, 23%, 24% or 25% weight/volume of the composition. In some embodiments, the block copolymer is present in the composition at a concentration that allows the composition to be heat-cured or thermally gelled within 1 minute of contact with human or animal tissue. In certain embodiments, the block copolymer is a poloxamer, such as Pluronic F127, which is present in the composition at a concentration greater than 20% by weight, such as 21%, 22%, 23%, 24% or 25% by weight of the weight/volume of the composition. Advantageously, at these concentrations, the composition is in liquid form at room temperature and gels when or after contact with the skin at body temperature.

In certain embodiments of any of the foregoing or following aspects, the composition further comprises an oxidizing agent. The oxidizing agent can include a peroxide, such as hydrogen peroxide, urea peroxide, and benzoyl peroxide, or any other oxidizing agent that can modulate the light absorption and/or emission properties of at least one chromophore, or can generate oxygen radicals in the presence of a chromophore.

In certain embodiments of any of the foregoing or following aspects, the thermoset biophotonic composition is at least substantially translucent. The thermoset biophotonic composition may be transparent. In some embodiments, the thermoset biophotonic composition has a translucency of at least about 40%, about 50%, about 60%, about 70%, or about 80% in the visible light range, for example, from 400 to 700 nm. In some embodiments, the thermoset composition has a light transmittance of about 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, or 90-100% when measured in the visible light range, for example, from 400 to 700 nm. The light transmittance through the thermoset biophotonic composition may be measured in the absence of at least one chromophore. In certain embodiments of any of the foregoing or following aspects, the thermoset biophotonic composition has a thickness of about 0.1 mm to about 50 mm, about 0.5 mm to about 20 mm, about 1 mm to about 10 mm, or about 1 mm to about 5 mm when applied to skin or a wound.

In certain embodiments of any of the foregoing or following aspects, the thermosetting biophotonic composition further comprises a stabilizer. The stabilizer may be selected from gelatin, HEC, and CMC or any other thickening agent.

The thermosetting biophotonic compositions of the present invention may be used for cosmetic or medical treatment of tissue. In some embodiments, cosmetic treatments include skin rejuvenation, skin conditioning, and/or promotion of collagen synthesis. In some embodiments, medical treatments include wound healing or tissue repair, treatment of skin conditions such as acne, eczema, psoriasis, or dermatitis, including prevention or reduction of scarring, and/or prevention or treatment of bacterial, viral, or fungal infections. In some embodiments, thermosetting biophotonic compositions are used to modulate inflammation, modulate collagen synthesis, or promote angiogenesis.

The present invention also provides a method for biophotonic therapy comprising applying the thermosetting biophotonic composition of the present invention to a target tissue and irradiating the composition with light.

Thus, in one aspect, the present invention provides a method for biophotonic treatment of a skin condition, wherein the method comprises applying a thermosetting biophotonic composition to or over target skin tissue, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer; and irradiating the biophotonic composition with light having a wavelength that overlaps with the absorption spectrum of the at least one chromophore dissolved in the thermosetting biophotonic composition; wherein the method promotes healing of the skin condition. The skin condition may be selected from acne, eczema, psoriasis, or dermatitis.

From another aspect, the present invention provides a method for biophotonic treatment of acne, comprising: applying a thermosetting biophotonic composition on or over target skin tissue, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer, and irradiating the biophotonic composition with light having a wavelength that overlaps with an absorption spectrum of the at least one chromophore; wherein the method treats acne.

From another aspect, the present invention provides a method of biophotonic therapy for wound healing, comprising: applying a thermosetting biophotonic composition to or over target skin tissue, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer, and irradiating the biophotonic composition with light having a wavelength that overlaps with an absorption spectrum of the at least one chromophore; wherein the method promotes wound healing.

In another aspect, the present invention provides a method for promoting skin renewal, comprising: applying a thermosetting biophotonic composition to or over target skin tissue, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer; and irradiating the biophotonic composition with light of a wavelength that overlaps with an absorption spectrum of the at least one chromophore; wherein the biophotonic composition emits fluorescent light at a wavelength and intensity that promotes skin renewal.

From another aspect, the present invention provides a method for preventing or treating scarring, comprising: applying a thermosetting biophotonic composition on or over target skin tissue, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer, and irradiating the biophotonic composition with light having a wavelength that overlaps with an absorption spectrum of the at least one chromophore; wherein the method reduces scarring.

Preferably, the composition is thermally cured or thermally gelled within about 1 minute after being applied to the target skin tissue. In some embodiments, the composition is thermally cured or thermally gelled within 50 seconds, 40 seconds, 30 seconds, 20 seconds, 10 seconds or 5 seconds after being applied to the target skin tissue.

In certain embodiments of any of the foregoing or following aspects, the thermosetting composition remains in place after irradiation, for example, for re-irradiation. In certain embodiments of any of the foregoing or following aspects, the thermosetting composition is removed after the treatment period. The composition can be removed by reducing the temperature of the composition and wiping or soaking the composition in a liquid state, thereby causing the composition to undergo a phase change to a liquid phase.

In some embodiments, the chromophore is at least partially photobleached during or after irradiation.In certain embodiments, the biophotonic composition is irradiated until the chromophore is at least partially photobleached.

In some embodiments, applying the thermosetting composition comprises spraying the composition onto the target skin tissue.In certain embodiments, the thermosetting composition is in a liquid phase and is sprayed and gelled upon or after contact with the skin.

In certain embodiments of any of the foregoing or following aspects, the light has a peak wavelength of about 400 nm to about 750 nm. The light may have a peak wavelength of about 400 nm to about 500 nm.

In certain embodiments of any of the foregoing or following aspects, the light comes from a direct light source such as a lamp. The lamp can be an LED lamp. In certain embodiments, the light comes from an ambient light source.

In certain embodiments of any of the foregoing or following aspects, the thermosetting composition is irradiated with a direct light source for about 1 minute to greater than 75 minutes, about 1 minute to about 75 minutes, about 1 minute to about 60 minutes, about 1 minute to about 55 minutes, about 1 minute to about 50 minutes, about 1 minute to about 45 minutes, about 1 minute to about 40 minutes, about 1 minute to about 35 minutes, about 1 minute to about 30 minutes, about 1 minute to about 25 minutes, about 1 minute to about 20 minutes, about 1 minute to about 15 minutes, about 1 minute to about 10 minutes, or about 1 minute to about 5 minutes.

From another aspect, the present invention provides uses of the above-mentioned thermosetting composition: for tissue repair; for wound healing; for preventing or treating scars; for skin renewal; for treating skin diseases such as acne, eczema, psoriasis or dermatitis; for regulating inflammation; for regulating angiogenesis; for regulating collagen synthesis; or for treating bacterial, viral or fungal infections.

In another aspect, the present invention provides a container comprising a chamber for holding a thermosetting biophotonic composition and an outlet in communication with the chamber for discharging the biophotonic composition from the container, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer. The thermosetting composition can be sprayed onto target tissue using the container and thermally cured upon contact with the tissue.

In some other embodiments, the thermosetting biophotonic composition defined herein has a gelation index in the range of about -3 to -0.1, about -2 to about -0.1, or about -0.2, -0.3, -0.4, -0.5, 0.6, -0.7, -0.8, or -0.9. As used herein, the expression "gelation index" refers to the gelation rate or solidification rate of the thermosetting biophotonic composition. The gelation rate is obtained by determining the time required for the thermosetting biophotonic composition to gel at 23°C and 25°C, or at 25°C and 27°C, or at 27°C and 29°C, or at 29°C and 31°C, or at 31°C and 33°C, or at 33°C and 35°C, or at 35°C and 37°C, and obtaining the gelation rate at the two temperature points.

BRIEF DESCRIPTION OF THE DRAWINGS

Other aspects and advantages of the present invention will be better understood with reference to the following description, in which:

FIG1 shows the light emission spectrum of a thermosetting biophotonic composition during 0-5 minutes of irradiation according to one embodiment of the present invention.

2 shows TGF-β-stimulated collagen production in DHF cells after irradiation with the biophotonic thermal gel of FIG. 1 , according to one embodiment of the present invention.

DETAILED DESCRIPTION

(1) Overview

The present invention provides thermosetting biophotonic films and uses thereof. Biophotonic therapies using these compositions combine the benefits of thermosetting compositions with the photobiostimulation effects of fluorescence generated upon illumination of the compositions. Furthermore, in certain embodiments, phototherapy using the thermosetting biophotonic compositions of the present invention will, for example, renew skin by promoting collagen synthesis; promote wound healing or tissue repair; prevent or treat scars; treat skin diseases such as acne, eczema, psoriasis, or dermatitis; or treat periodontitis.

(2) Definition

Before proceeding to describe the present invention in further detail, it should be understood that the present invention is not limited to the specific compositions or process steps, as these may vary. It must be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

The term "about" used in the present invention when a numerical value or range is given means that the numerical value or range is within 20%, preferably within 10% and more preferably within 5% of the given numerical value or range.

It is convenient to note that as used herein, "and/or" will be considered to specifically disclose each of the two specified features or components, both of which may be present simultaneously or individually. For example, "A and/or B" will be considered to specifically disclose each of (i) A, (ii) B, and (iii) A and B, just as if each were individually listed herein.

"Biophotonic" refers to the generation, manipulation, detection, and application of photons in a biologically relevant context. In other words, a biophotonic composition exerts its physiological effects primarily due to the generation and manipulation of photons.

"Thermosetting" refers to a liquid composition that undergoes a phase change to a solid or semi-solid state (e.g., a gel) either spontaneously (e.g., upon contact with a target tissue) or after exposure to some form of energy (e.g., heat). Thermal energy can be provided by contact with a warm body or by a light source. In some embodiments, the phase change to a solid or semi-solid state can occur upon contact with tissue at a temperature higher than the ambient temperature. The terms "thermosetting" and "thermogelling" are used interchangeably in the present invention. The terms "thermoset" and "thermogel" are also used interchangeably in the present invention.

"Topical administration" or "topical use" refers to application to body surfaces such as the skin, mucous membranes, vagina, mouth, internal surgical wound sites, and the like.

In the present invention, the terms "chromophore" and "photoactivator" are used interchangeably. A chromophore is a compound that absorbs light when irradiated with light. Chromophores are easily photoexcited and transfer their energy to other molecules or emit light (e.g., fluorescence).

"Photobleaching" or "photobleaching" refers to the photochemical destruction of a chromophore. A chromophore can be fully or partially photobleached.

The term "actinic light" refers to light energy emitted by a particular light source (e.g., a lamp, LED, or laser) and capable of being absorbed by a substance (e.g., a chromophore or photoactivator). In a preferred embodiment, actinic light is visible light. As used herein, the terms "actinic light" and "light" are used interchangeably.

"Skin renewal" refers to the process of reducing, eliminating, delaying or reversing one or more signs of skin aging or generally improving the condition of the skin. For example, skin renewal can include increasing skin brightness, reducing pore size, reducing fine lines or wrinkles, improving skin thinning and clarity, improving firmness, improving skin laxity (such as that caused by bone loss), improving dry skin (which may be itchy), reducing or reversing freckles, age spots, spider veins, reducing or preventing the appearance of rough and leathery skin, fine wrinkles that disappear when stretched, reducing sagging skin or improving blotchy skin. According to the present invention, one or more of the conditions described above can be improved or one or more signs of aging can be reduced, eliminated, delayed or even reversed by certain embodiments of the compositions, methods and uses of the present invention.

"Wound" refers to any tissue injury, including, for example, acute, subacute, slow-healing or difficult-to-heal wounds, and chronic wounds. Examples of wounds may include open wounds and closed wounds. Wounds include, for example, amputations, burns, incisions, excisions, injuries, lacerations, abrasions, puncture or penetrating wounds, surgical wounds, amputations, contusions, hematomas, pressure injuries, ulcers (such as, for example, pressure ulcers, diabetic ulcers, venous or arterial ulcers), and wounds caused by periodontitis (periodontal inflammation).

Features and advantages of the present subject matter will become more apparent upon reference to the following detailed description of selected embodiments, accompanied by accompanying drawings. As will be appreciated, the subject matter disclosed and claimed is susceptible of modifications in various respects, all without departing from the scope of the claims. The drawings and description are therefore to be regarded as illustrative in nature and not restrictive, with the full scope of the subject matter being indicated in the claims.

(3) Thermosetting biophotonic compositions

The present invention broadly provides thermosetting biophotonic compositions and methods of using such compositions. Biophotonic compositions are broadly activatable by light of a specific wavelength (e.g., photons). Thermosetting biophotonic compositions according to various embodiments of the present invention comprise block copolymers in which at least one chromophore is dissolved. The chromophore in the thermosetting biophotonic composition can be activated by light, which accelerates the dispersion of light energy, resulting in the light itself having a therapeutic effect, and/or photochemical activation of other agents included in the composition (e.g., accelerating the decomposition process of peroxides (oxidants or oxidizing agents) when such compounds are present in or in contact with the composition, resulting in the formation of oxygen radicals, such as singlet oxygen).

When a chromophore absorbs a photon of a certain wavelength, it becomes excited. This is an unstable state, and the molecule will attempt to return to its ground state, releasing excess energy. For some chromophores, releasing excess energy in the form of light is preferred when returning to the ground state. This process is known as fluorescence. Because energy is lost during this conversion process, the peak wavelength of the emitted fluorescence shifts toward longer wavelengths compared to the absorption wavelength. This phenomenon is known as the Stokes shift. In the right environment (e.g., in a biophotonic composition), much of this energy is transferred to other components of the biophotonic composition or directly to the treatment site.

Without wishing to be bound by theory, it is believed that the fluorescence emitted by photoactivated chromophores leads to beneficial biomodulation due to their femtosecond, picosecond, or nanosecond emission characteristics recognized by biological cells and tissues. Furthermore, the emitted fluorescence has a longer wavelength and therefore penetrates deeper into tissue than the activating light. Irradiating tissue with such a wide range of wavelengths (including, in certain embodiments, the activating light transmitted through the composition) has distinct and complementary effects on cells and tissues. In other words, chromophores are used in the biophotonic compositions of the present invention to produce therapeutic effects on tissues. In one aspect, and without wishing to be bound by any particular theory, the emitted fluorescence can have a mechanical effect by disrupting precipitates during electron flow that destabilize bacterial membranes, leading to a loss of structural integrity of the bacterial membrane. This represents a fundamentally different application of these photoactivators, distinct from the use of chromophores as simple stains or as catalysts for photopolymerization.

The thermosetting biophotonic compositions of the present invention may have topical applications. In some embodiments, the thermosetting biophotonic compositions are viscous when they are thermally cured. The viscous nature of these thermosetting biophotonic compositions facilitates their removal from the treatment site, and thus allows for quicker and less messy treatment. Once thermally cured, the adhesive nature of these thermosetting biophotonic compositions may also provide for less messy treatment. Furthermore, the thermosetting biophotonic compositions may limit contact between the chromophores and the tissue.

The thermosetting compositions of the present invention are designed to thermoset upon contact with target skin tissue. In some embodiments, the thermosetting biophotonic compositions thermoset at 32 degrees Celsius or higher. In some embodiments, the compositions thermoset within 1 minute at 32 degrees Celsius. In other embodiments, the compositions thermoset within 50 seconds, 40 seconds, 30 seconds, 20 seconds, 10 seconds, 5 seconds, or less than 5 seconds at 32 degrees Celsius. In other embodiments, the compositions thermoset spontaneously upon contact with target tissue.

These compositions can be described in terms of the components that comprise them. Additionally or alternatively, the compositions of the present invention possess functional and structural properties, and these properties can also be used to define and describe the compositions. The various components of the biophotonic compositions of the present invention, including the chromophore, block copolymer, and other optional components, are described in detail below.

(a) Chromophore

Suitable chromophores can be fluorescent compounds (or dyes) (also known as "fluorescent dyes" or "fluorophores"). Other dye groups or dyes (biological and histological dyes, food colorants, carotenoids, and other dyes) can also be used. Suitable photoactivators can be those that are generally regarded as safe (GRAS). Advantageously, photoactivators that are not well tolerated by the skin or other tissues can be included in the thermosetting biophotonic compositions of the present invention, such as in certain embodiments, when the photoactivator is dissolved in the block copolymer.

In certain embodiments, the thermosetting biophotonic compositions of the present invention comprise a first chromophore that partially or completely photobleachs upon application of light. In some embodiments, the first chromophore absorbs at a wavelength within the visible spectrum, such as at a wavelength of approximately 380-800 nm, 380-700 nm, 400-800 nm, or 380-600 nm. In other embodiments, the first chromophore absorbs at a wavelength of approximately 200-800 nm, 200-700 nm, 200-600 nm, or 200-500 nm. In one embodiment, the first chromophore absorbs at a wavelength of approximately 200-600 nm. In some embodiments, the first chromophore absorbs light at a wavelength of about 200-300 nm, 250-350 nm, 300-400 nm, 350-450 nm, 400-500 nm, 450-650 nm, 600-700 nm, 650-750 nm, or 700-800 nm.

It will be understood by those skilled in the art that the optical properties of a particular chromophore may vary depending on the medium surrounding the chromophore. Thus, as used herein, the absorption and/or emission wavelengths (or spectra) of a particular chromophore correspond to the wavelengths (or spectra) measured in the thermosetting biophotonic compositions of the present invention.

The biophotonic compositions disclosed herein may include at least one additional chromophore. Combining chromophores can increase light absorption by combining dye molecules and enhance the selectivity of absorption and photo-biomodulation. This creates several possibilities for creating novel photosensitivity and/or selective chromophore mixtures. Therefore, in certain embodiments, the biophotonic compositions disclosed herein include more than one chromophore. When such a multi-chromophore composition is illuminated with light, energy transfer can occur between the chromophores. This process, known as resonance energy transfer (RET), is a widely used photophysical process by which an excited "donor" chromophore (also referred to herein as the first chromophore) transfers its excitation energy to an "acceptor" chromophore (also referred to herein as the second chromophore). The efficiency and directionality of RET depend on the spectral characteristics of the donor and acceptor chromophores. Specifically, the energy flow between the chromophores depends on the spectral overlap, which reflects the relative position and shape of the absorption and emission spectra. More specifically, for energy transfer to occur, the emission spectrum of the donor chromophore must overlap with the absorption spectrum of the acceptor chromophore (Figure 1).

Energy transfer is manifested by a reduction or quenching of donor emission and a decrease in excited-state lifetime (accompanied by an increase in acceptor emission intensity). To enhance energy transfer efficiency, the donor chromophore should possess good photon absorption and emission capabilities. Furthermore, the greater the overlap between the donor chromophore's emission spectrum and the acceptor chromophore's absorption spectrum, the better the donor chromophore is at transferring energy to the acceptor chromophore.

In certain embodiments, the thermosetting biophotonic composition of the present invention further comprises a second chromophore. In some embodiments, the first chromophore has an emission spectrum that overlaps with the absorption spectrum of the second chromophore by at least about 80%, 50%, 40%, 30%, 20%, or 10%. In one embodiment, the first chromophore has an emission spectrum that overlaps with the absorption spectrum of the second chromophore by at least about 20%. In some embodiments, the first chromophore has an emission spectrum that overlaps with the absorption spectrum of the second chromophore by at least 1-10%, 5-15%, 10-20%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 50-60%, 55-65%, or 60-70%.

As used herein, % spectral overlap refers to the percent overlap of the emission wavelength range of the donor chromophore and the absorption wavelength range of the acceptor chromophore, measured at one quarter full width maximum (FWQM) of the spectrum. In some embodiments, the second chromophore absorbs at a wavelength within the visible spectrum. In some embodiments, the second chromophore has a relatively longer absorption wavelength than the absorption wavelength of the first chromophore, within the range of about 50-250 nm, 25-150 nm, or 10-100 nm.

The first chromophore may be present in an amount of about 0.001-40% by weight of the biophotonic composition.When present, the second chromophore may be present in an amount of about 0.001-40% by weight of the biophotonic composition. In some embodiments, the first chromophore is present in an amount of about 0.001-3%, 0.001-0.01%, 0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40% by weight of the biophotonic composition. In some embodiments, the second chromophore is present in an amount of about 0.001-3%, 0.001-0.01%, 0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40% by weight of the biophotonic composition. In some embodiments, the total weight per weight of the chromophore or combination of chromophores may be in an amount of about 0.005-1%, 0.05-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40.001% per weight of the biophotonic composition.

The concentration of the chromophore used can be selected based on the desired intensity and duration of the biophotonic activity of the biophotonic composition, as well as the desired medical or cosmetic effect. For example, some dyes, such as xanthene dyes, reach a "saturation concentration," after which further increases in concentration do not provide significantly higher levels of emitted fluorescence. Increasing the chromophore concentration above the saturation concentration can reduce the amount of activating light that passes through the matrix. Therefore, if a particular application requires more fluorescence than activating light, a high concentration of the chromophore can be used. However, if a balance is desired between emitted fluorescence and activating light, a concentration close to or below the saturation concentration can be selected.

Suitable chromophores that can be used in the thermosetting biophotonic compositions of the present invention include, but are not limited to, the following:

Chlorophyll dye

Exemplary chlorophyll dyes include, but are not limited to, chlorophyll a; chlorophyll b; chlorophyllin; bacteriochlorophyll a; bacteriochlorophyll b; bacteriochlorophyll c; bacteriochlorophyll d; protochlorophyll; protochlorophyll a; amphiphilic chlorophyll derivative 1; and amphiphilic chlorophyll derivative 2.

Xanthene derivatives

Exemplary xanthene dyes include, but are not limited to, Eosin B; Eosin B (4',5'-dibromo-2',7'-dinitro-fluorescein dianion); Eosin Y; Eosin Y (2',4',5',7'-tetrabromo-fluorescein dianion); Eosin (2',4',5',7'-tetrabromo-fluorescein dianion); Eosin (2',4',5',7'-tetrabromo-fluorescein dianion); Eosin (2',4',5',7'-tetrabromo-fluorescein dianion) methyl ester; Eosin (2',4',5',7'-tetrabromo-fluorescein monovalent anion); ion) p-isopropylbenzyl ester; Eosin derivatives (2',7'-dibromo-fluorescein dianion); Eosin derivatives (4',5'-dibromo-fluorescein dianion); Eosin derivatives (2',7'-dichloro-fluorescein dianion); Eosin derivatives (4',5'-dichloro-fluorescein dianion); Eosin derivatives (2',7'-diiodo-fluorescein dianion); Eosin derivatives (4',5'-diiodo-fluorescein dianion); Eosin derivatives (tribromofluorescein dianion); Eosin derivative (2',4',5',7'-tetrachloro-fluorescein dianion); Eosin; Eosin dihexadecylpyridinium chloride pair; Erythrosine B (2',4',5',7'-tetraiodo-fluorescein dianion); Erythrosine; Erythrosine dianion; Erythrosine B; Fluorescein; Fluorescein dianion; Fluorescein B (2',4',5',7'-tetrabromo-3,4,5,6-tetrachloro-fluorescein dianion); Fluorescent Fluorescent pink B (tetrachloro-tetrabromo-fluorescein); Fluorescent pink B; Rose Bengal (3,4,5,6-tetrachloro-2',4',5',7'-tetraiodofluorescein divalent anion); Pyronine G, Pyronine J, Pyronine Y; Rhodamine dyes, such as rhodamines including 4,5-dibromo-rhodamine methyl ester; 4,5-dibromo-rhodamine n-butyl ester; Rhodamine 101 methyl ester; Rhodamine 123; Rhodamine 6G; Rhodamine 6G hexyl ester; Tetrabromo-rhodamine 123; and Tetramethyl-rhodamine ethyl ester.

Methylene blue dye

Exemplary methylene blue derivatives include, but are not limited to, 1-methylmethylene blue; 1,9-dimethylmethylene blue; methylene blue; methylene violet; bromomethylene violet; 4-iodomethylene violet; 1,9-dimethyl-3-dimethyl-amino-7-diethyl-amino-phenothiazine; and 1,9-dimethyl-3-diethylamino-7-dibutyl-amino-phenothiazine.

Azo dyes

Exemplary azo (or disazo-) dyes include, but are not limited to, methyl violet, neutral red, para red (Pigment Red 1), amaranth (Azorubine S), acid red (Azorubine, Food Red 3, Acid Red 14), allura red AC (FD&C 40), tartrazine (FD&C Yellow 5), Orange G (Acid Orange 10), Ponceau 4R (Food Red 7), methyl red (Acid Red 2), and murexanthin-purpurogenum.

In some aspects of the present invention, the one or more chromophores of the thermosetting biophotonic composition disclosed herein can be independently selected from any one of the following chromophores: acid black 1, acid blue 22, acid blue 93, acid fuchsin, acid green, acid green 1, acid green 5, acid magenta, acid orange 10, acid red 26, acid red 29, acid red 44, acid red 51, acid red 66, acid red 87, acid red 91, acid red 92, acid red 94, acid red 101, acid red 103, acid roseine, acid fuchsin ... red rubin), Acid Violet 19, Acid Yellow 1, Acid Yellow 9, Acid Yellow 23, Acid Yellow 24, Acid Yellow 36, Acid Yellow 73, Acid Yellow S, Acridine Orange, Acridine Yellow, Alcian Blue, Alcian Yellow, Eosin, Alizarin, Alizarin Blue 2RC, Alizarin Carmine, Alizarin Cyanine BBS, Alizarin Cyanine R, Alizarin Red S, Alizarin Red Violet, Tryptophan, Amido Black 10B, Amidoschwarz, Aniline Blue WS, Anthracene Blue SWR, Auramine O, Azocannine B, Azocarmine G, Azodiazo (Azoicdiazo) 5, Azodiazo 48, Sky Blue A, Sky Blue B, Sky Blue C, Basic Blue 8, Basic Blue 9, Basic Blue 12, Basic Blue 15, Basic Blue 17, Basic Blue 20, Basic Blue 26, Basic Brown 1, Basic Fuchsin, Basic Green 4, Basic Orange 14, Basic Red 2, Basic Red 5, Basic Red 9, Basic Violet 2, Basic Violet 3, Basic Violet 4, Basic Violet 10, Basic Violet 14, Basic Yellow 1, Basic Yellow 2, Biebrich Scarlet, Bismarck Brown Y, Brilliant Crystal Scarlet 6R, Calcium Red, Carmine, Carminic Acid, Lapis Lazuli Blue B, China Blue, Wormwood Red, Coelestine Blue, Chrome Violet CG, Chrome Variation 2R, Chromoxane Cyanine R, Congo Corinth, Congo Red, Cotton Dye Blue, Cotton Red, Croceine Scarlet, Crocin, Crystal Ponceau 6R, Crystal Violet, Dahlia Violet, Diamond Green B, Direct Indigo 14, Direct Blue 58, Direct Red, Direct Red 10, Direct Red 28, Direct Red 80, Direct Yellow 7, Eosin B, Blue Eosin, Eosin, Eosin Y, Yellow Eosin, Eosinol, Erie Pomegranate Red B, Chrome Cyanine R, Erythrosine B, Ethyl Eosin, Ethyl Green, Ethyl Violet, Evans Blue, Fast Blue B, Fast Green FCF, Fast Red B, Fast Yellow, Fluorescent Yellow, Food Green 3, Pyrophthalein, Galamin Blue, Betacyanine, Gentian Violet, Oxidized Hematoxylin, Hematoxylin, Hematoxylin Violet, Daylight Fast Magenta BBL, Methyl Blue, Hematoxylin, Hematoxylin, Hematoxylin Violet, Hoffman Violet, Royal Red, Indocyanine Green, Alcian Blue, Alcian Blue 1, Alcian Yellow 1, INT, Magenta, Carminic Acid, Daylight Ruby Red, Lac, Lacic Acid, Routh's Violet, Light Green, Lissamine Green SF, Luxol Fast Blue, Magenta 0, Magenta I, Magenta II, Magenta III, Malachite Green, Manchester Brown, Matthew Yellow, Mercurochrome, Mercurochrome, Acid Metamin Yellow, Methylene Sky Blue A, Methylene Sky Blue B, Methylene Sky Blue C, Methylene Blue, Methyl Blue, Methyl Green, Methyl Violet, Methyl Violet 2B, Methyl Violet 10B, Mordant Blue 3, Mordant Blue 10, Mordant Blue 14, Mordant Blue 23, Mordant Blue 32, Mordant Blue 45, Mordant Red 3, Mordant Red 11, Mordant Violet 25, Mordant Violet 39, Naphthol Blue Black, Naphthol Green B, Naphthol Yellow S, Natural Black 1, Natural Green 3 (Chlorophyll), Natural Red, Natural Red 3, Natural Red 4, Natural Red 8, Natural Red 16, Natural Red 25, Natural Red 28, Natural Yellow 6, NBT, Natural Red, New Fuchsin, Niagara Blue 3B, Night Blue, Nitro BT, Nitro Blue Tetrazolium , Nuclear Fast Red, Orange G, Lichen Red, Paracarmine, Fluorescent Pink B, Picric Acid, Ponceau 2R, Ponceau 6R, Ponceau B, Xylidine Ponceau, Ponceau S, Primrose, Purpurin, Pyronine B, Phycocyanin, Phycocyanin, Phycoerythrin, Phycoerythrocyanin (PEC), Phthalocyanine, Pyronine G, Pyronine Y, Quinine, Rhodamine B, Rose Aniline, Rose Bengal, Safranine, Safranin O, Scarlet R, Scarlet Red, Scarlet R, Lac, Sirius Red F3B, Sarachrome Cyanine R, Soluble Blue, Elf Soluble Eosin, Sulfur S, Swiss Blue, Tartrazine, Thioflavin S, Thioflavin T, Thioflavin, Toluidine Blue, Toluidine Red, Trophium Orange G, Acridine Yellow, Trypan Blue, Fluorescein Sodium, Victoria Blue 4R, Victoria Blue B, Victoria Green B, Vitamin B12, Water-Soluble Blue I, Water-Soluble Eosin, Xylidine Ponceau or Yellow Eosin.

In certain embodiments, the thermosetting biophotonic compositions of the present invention include any of the chromophores listed above, or combinations thereof, to provide a synergistic biophotonic effect at the site of application.

Without being limited to any particular theory, a synergistic effect of a chromophore combination refers to a biophotonic effect that is greater than the sum of the individual effects. Advantageously, this can translate into increased reactivity of the thermosetting biophotonic composition, resulting in faster or improved treatment times. Furthermore, treatment conditions, such as the duration of light exposure, the power of the light source used, and the wavelength of the light used, need not be modified to achieve the same or better treatment results. In other words, using a synergistic chromophore combination can achieve the same or better treatment without necessarily requiring longer exposure to the light source, increasing the power of the light source, or using a light source of a different wavelength.

In some embodiments, the composition includes Eosin Y as the first chromophore and one or more of Rose Bengal, Fluorescein, Erythrosine, Fluorescent Pink B, and Chlorophyll as the second chromophore. It is believed that these combinations have a synergistic effect because, when they are activated in part because their absorption and emission spectra overlap or are close to each other, they can transfer energy from one chromophore to another chromophore. The energy of this transfer is then emitted as fluorescence or causes the generation of reactive oxygen species. This absorbed and re-emitted light is believed to be transmitted throughout the composition and also transmitted into the treatment site.

In other embodiments, the composition includes the following synergistic combinations: Eosin Y with fluorescein; fluorescein with Rose Bengal; Erythrosine with Eosin Y, Rose Bengal, or fluorescein; and Phosphorin B with one or more of Eosin Y, Rose Bengal, fluorescein, and Erythrosine. Other synergistic chromophore combinations may also be used.

Through the synergistic effect of the chromophore combination in the thermosetting biophotonic composition, chromophores that are not normally activated by activating light (such as blue light from an LED) can be activated by energy transfer from chromophores activated by the activating light. In this way, light-activated chromophores with different properties can be utilized and customized according to the desired cosmetic or medical treatment.

For example, when Rose Bengal is activated in the presence of molecular oxygen, it can produce high levels of singlet oxygen. However, Rose Bengal has a low quantum yield in terms of fluorescence emission. Rose Bengal has a peak absorption near 540 nm, and therefore can be activated with green light. Eosin Y has a high quantum yield and can be activated with blue light. Combining Rose Bengal and Eosin Y produces a composition that, when activated with blue light, emits therapeutic fluorescence and produces singlet oxygen. In this case, the blue light photoactivates Eosin Y, which transfers some of its energy to Rose Bengal, emitting some of the energy as fluorescence.

In some embodiments, one or more chromophores are selected such that upon photoactivation, the fluorescent light emitted is one or more of the green, yellow, orange, red, and infrared portions of the electromagnetic spectrum, for example, with a peak wavelength in the range of about 490 nm to about 800 nm. In some embodiments, the fluorescent light emitted has a power density of 0.005 to about 10 mW/cm ² , or about 0.5 to about 5 mW/cm ² .

(b) Block copolymers

The thermosettable biophotonic compositions of the present invention comprise a block copolymer. The block copolymer is present in an amount greater than 20% by weight/volume of the total composition. In some embodiments, the block copolymer is present in an amount of at least 21%, 22%, 23%, 24%, or 25%. In some embodiments, the block copolymer is present in an amount of 21%, 22%, 23%, 24%, or 25%.

The term "block copolymer" used in the present invention refers to a copolymer consisting of blocks (or segments) of two or more different homopolymers. The term homopolymer refers to a polymer consisting of a single monomer. Many variants of block copolymers are possible, including simple diblock polymers with AB structure and triblock polymers with ABA, BAB or ABC structure, and more complex block copolymers are known. In addition, unless otherwise specified in the present invention, the number of repetitions and types of monomers or repeating units constituting the block copolymer are not particularly limited. For example, when monomer repeating units are represented as "a" and "b", in the present invention, it refers to that the copolymer not only includes a random copolymer with an average composition of (a) _m (b) _n , but also includes a diblock copolymer of a composition (a) _m (b) _n , and a triblock copolymer of a composition (a) _l (b) _m (a) _n . In the above formula, l, m and n represent the number of repeating units and are positive numbers.

In certain embodiments of any of the foregoing or following aspects, the block copolymer is biocompatible. A polymer is "biocompatible" because the polymer and its degradation products are substantially non-toxic to cells or organisms, including being non-carcinogenic and non-immunogenic, and are cleared or otherwise degraded in a biological system, such as an organism (patient), without substantial toxic effects.

In certain embodiments, the block copolymer is derived from a triblock copolymer designated as a poloxamer. A poloxamer is an A-B-A block copolymer in which the A segment is a hydrophilic polyethylene glycol (PEG) homopolymer and the B segment is a hydrophobic polypropylene glycol (PPG) homopolymer. PEG is also known as polyethylene oxide (PEO) or polyoxyethylene (POE), depending on its molecular weight. Additionally, PPG is also known as polypropylene oxide (PPO), depending on its molecular weight. Poloxamers are commercially available from BASF. Poloxamers produce inverse-phase thermogelatin compositions, characterized by an increase in viscosity with increasing temperature until a point at which the viscosity decreases again. Depending on the relative sizes of the blocks, the copolymer can be a solid, liquid, or paste. In certain embodiments of the present invention, the poloxamer is F127 (also known as Poloxamer 407). In some embodiments of the thermosetting biophotonic compositions of the present invention, F127 is included in an amount exceeding 20% by weight/volume of the composition. In other embodiments, pluronic is present in an amount of at least 21%, 22%, 23%, 24%, or 25%.Other poloxamers may also be used.

Since the PEG block contributes to the hydrophilicity of the polymer, increasing the length of the PEG block or the total amount of PEG in the polymer will tend to make the polymer more hydrophilic. Depending on the amounts and ratios of the other components of the polymer, the desired overall hydrophilicity, and the nature and chemical functionality of any drugs or therapeutic agents that may be included in the polymer formulation, one skilled in the art can readily adjust the length (or MW) of the PEG block used and/or the total amount of PEG incorporated into the polymer in order to obtain a polymer with the desired physical and chemical properties.

The total amount of PEG in the polymer can be about 80% by weight or less, 75% by weight or less, 70% by weight or less, 65% by weight or less, about 60% by weight or less, about 55% by weight or less, or about 50% by weight or less. In specific embodiments, the total amount of PEG is about 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69% or about 70% by weight. Unless otherwise indicated, the weight percentage of a particular component of a polymer means that the total weight of the polymer is made up of the monomers of the specified percentage of that component. For example, 65% by weight of PEG means that 65% of the weight of the polymer is made up of PEG monomers, and the monomers are connected into blocks of different lengths that are distributed along the length of the polymer, including being randomly distributed.

The presence of surfactants can enhance the fluorescence of chromophores. However, complementary surfactant and chromophore combinations can be tested and selected based on non-electro-repulsive combinations. For example, negatively charged chromophores can be used with ionic or nonionic surfactants, and vice versa.

The block copolymers may also be mixed with thickeners or stabilizers such as gelatin and/or modified celluloses (such as hydroxyethylcellulose (HEC) and carboxymethylcellulose (CMD)), and/or polysaccharides such as xanthan gum, guar gum and/or starch and/or any other thickeners. In certain embodiments, the stabilizer or thickener may comprise gelatin. For example, the surfactant phase may comprise about 0-5% by weight, about 5-25% by weight, about 0-15% by weight, or about 10-20% by weight of gelatin.

Thickeners and/or stabilizers may be selected based on their effect on the optical clarity of the biophotonic film or on the stabilization effect of the block copolymer.The thermosetting biophotonic composition should be capable of transmitting sufficient light to activate at least one chromophore.

(c) Oxidants/antimicrobial agents

According to certain embodiments, the thermosetting biophotonic compositions of the present invention may optionally include one or more additional components, such as oxygen-rich compounds that serve as a source of oxygen free radicals ("oxidants"). Peroxide compounds are oxidants containing peroxide radicals (R-O-O-R), which are chain-like structures containing two oxygen atoms, each linked to another oxygen atom and a radical or element. When the thermosetting biophotonic compositions of the present invention are irradiated with light, the chromophores are excited to a higher energy state. When the chromophore's electrons return to a lower energy state, they emit photons of lower energy, resulting in the emission of light at a longer wavelength (Stokes shift). In the appropriate environment, some of this energy is transferred to the oxidant (if present), such as peroxide or oxygen, and can result in the formation of oxygen free radicals, such as singlet oxygen. It is believed that the singlet oxygen and other reactive oxygen species generated by the activation of the biophotonic composition operate in an excitatory manner. That is, by stimulating and modulating stress response pathways in target tissue cells, beneficial health effects can be achieved with reduced exposure to typically toxic stimuli (e.g., reactive oxygen species). The endogenous response to exogenously generated free radicals (reactive oxygen species) is modulated, increasing defenses against exogenous free radicals and inducing accelerated healing and regeneration processes. Furthermore, this can also produce an antimicrobial effect. Bacteria are extremely sensitive to exposure to free radicals, making the thermosetting biophotonic composition of the present invention a potentially bactericidal composition.

Antimicrobial agents kill microorganisms or inhibit their growth or accumulation and are optionally included in the thermosetting biophotonic compositions of the present invention. Suitable antimicrobial agents for use in the methods of the present invention include, but are not limited to, hydrogen peroxide, urea peroxide, benzoyl peroxide, phenolic and chlorinated phenolic compounds, resorcinol and its derivatives, bisphenol compounds, benzoic acid esters (parabens), halogenated carbanilides, polymeric antimicrobials, thiazolines, trichloromethylthioimide, natural antimicrobials (also known as "natural essential oils"), metal salts, and broad-spectrum antibiotics.

Hydrogen peroxide ( _H2O2 ) is a powerful oxidant and decomposes into water and oxygen without forming any persistent toxic residual compounds. Suitable concentrations of hydrogen peroxide _that can be used in the biophotonic composition are in the range of about 0.1% to about 3%, about 0.1 to 1.5%, about 0.1% to about 1%, about 1%, or less than about 1%.

Carbamide peroxide (also known as urea peroxide, percarbamide) is soluble in water and contains approximately 35% hydrogen peroxide. Suitable concentrations of carbamide peroxide that can be used in biophotonic compositions range from less than about 0.25%, or less than about 0.3%, 0.001 to 0.25%, or about 0.3% to about 5%. Carbamide peroxide decomposes into urea and hydrogen peroxide in a slow-release manner, a process that can be accelerated by heat or photochemical reactions.

Benzoyl peroxide consists of two benzoyl groups attached to a peroxide group (the carboxylic acid H of benzoic acid has been removed). Benzoyl peroxide has been found to be useful in the treatment of acne at concentrations of 2.5% to 10%. The released peroxide groups are very effective in killing bacteria. Benzoyl peroxide also promotes skin turnover and pore clearance, which helps further reduce bacteria counts and alleviate acne. Benzoyl peroxide decomposes upon contact with the skin into benzoic acid and oxygen, both of which are non-toxic. Suitable concentrations of benzoyl peroxide for use in thermosetting biophotonic compositions are from about 2.5% to about 5%.

Specific phenolic and chlorophenolic antimicrobial agents that may be used in the present invention include, but are not limited to, phenol; 2-methylphenol; 3-methylphenol; 4-methylphenol; 4-ethylphenol; 2,4-dimethylphenol; 2,5-dimethylphenol; 3,4-dimethylphenol; 2,6-dimethylphenol; 4-n-propylphenol; 4-n-butylphenol; 4-n-pentylphenol; 4-tert-pentylphenol; 4-n-hexylphenol; 4-n-heptylphenol; mono- and poly-alkyl and aromatic halogenated phenols; p-chlorophenol; methyl p-chlorophenol; ethyl p-chlorophenol; n-propyl p-chlorophenol; n-butyl p-chlorophenol; n-pentyl-p-chlorophenol; sec-pentyl-p-chlorophenol; n- Hexyl p-chlorophenol; cyclohexyl p-chlorophenol; n-heptyl p-chlorophenol; n-octyl p-chlorophenol; o-chlorophenol; methyl o-chlorophenol; ethyl o-chlorophenol; n-propyl o-chlorophenol; n-butyl o-chlorophenol; n-pentyl o-chlorophenol; tert-pentyl o-chlorophenol; n-hexyl o-chlorophenol; n-heptyl o-chlorophenol; o-benzyl p-chlorophenol; o-benzyl-m,m-dimethyl p-chlorophenol; o-phenylethyl p-chlorophenol; o-phenylethyl-m-methyl p-chlorophenol; 3-methyl p-chlorophenol; 3,5-dimethyl p-chlorophenol; 6-ethyl-3-methyl p-chlorophenol, 6- n-Propyl-3-methyl-p-chlorophenol; 6-iso-propyl-3-methyl-p-chlorophenol; 2-ethyl-3,5-dimethyl-p-chlorophenol; 6-sec-butyl-3-methyl-p-chlorophenol; 2-iso-propyl-3,5-dimethyl-p-chlorophenol; 6-diethylmethyl-3-methyl-p-chlorophenol; 6-iso-propyl-2-ethyl-3-methyl-p-chlorophenol; 2-sec-pentyl-3,5-dimethyl-p-chlorophenol; 2-diethylmethyl-3,5-dimethyl-p-chlorophenol; 6-sec-octyl-3-methyl-p-chlorophenol; p-chloro-m-cresol p-bromophenol; methyl p-bromophenol; ethyl p-bromophenol; n-propyl p-bromophenol; n-butyl p-bromophenol Phenol; n-pentyl-p-bromophenol; sec-pentyl-p-bromophenol; n-hexyl-p-bromophenol; cyclohexyl-p-bromophenol; o-bromophenol; tert-pentyl-o-bromophenol; n-hexyl-o-bromophenol; n-propyl-m,m-dimethyl-o-bromophenol; 2-phenylphenol; 4-chloro-2-methylphenol; 4-chloro-3-methylphenol; 4-chloro-3,5-dimethylphenol; 2,4-dichloro-3,5-dimethylphenol; 3,4,5,6-tetrabromo-2-methylphenol; 5-methyl-2-pentylphenol; 4-isopropyl-3-methylphenol; p-chloro-m-xylenol (PCMX); chlorothymol; phenoxyethanol; phenoxyisopropyl alcohol; and 5-chloro-2-hydroxydiphenylmethane.

Resorcinol and its derivatives can also be used as antimicrobial agents. Specific resorcinol derivatives include, but are not limited to, methylresorcinol; ethylresorcinol; n-propylresorcinol; n-butylresorcinol; n-pentylresorcinol; n-hexylresorcinol; n-heptylresorcinol; n-octylresorcinol; n-nonylresorcinol; phenylresorcinol; benzylresorcinol; phenylethylresorcinol; phenylpropylresorcinol; p-chlorobenzylresorcinol; 5-chloro-2,4-dihydroxydiphenylmethane; 4'-chloro-2,4-dihydroxydiphenylmethane; 5-bromo-2,4-dihydroxydiphenylmethane; and 4'-bromo-2,4-dihydroxydiphenylmethane.

Specific bisphenol antimicrobial agents that can be used in the present invention include, but are not limited to: 2,2'-methylenebis-(4-chlorophenol); 2,4,4'-trichloro-2'-hydroxy-diphenyl ether, a product sold under the trade name PANTRADE by Ciba Geigy, Florham Park, N.J.; 2,2'-methylenebis-(3,4,6-trichlorophenol); 2,2'-methylenebis-(4-chloro-6-bromophenol); bis-(2-hydroxy-3,5-dichloro-p-hexyl) sulfide; and bis-(2-hydroxy-5-chlorobenzyl) sulfide.

Specific benzoates (parabens) that can be used in the present invention include, but are not limited to: methylparaben; propylparaben; butylparaben; ethylparaben; isopropylparaben; isobutylparaben; benzylparaben; sodium methylparaben; and sodium propylparaben.

Specific halogenated carbanilides that may be used in the present invention include, but are not limited to: 3,4,4′-trichlorocarbanilide, such as 3-(4-chlorophenyl)-1-(3,4-dichlorophenyl)urea, sold under the trade name 3-(4-chlorophenyl)-1-(3,4-dichlorophenyl)urea by Ciba-Geigy, Florham Park, N.J.; 3-trifluoromethyl-4,4′-dichlorocarbanilide; and 3,3′,4-trichlorocarbanilide.

Specific polymeric antimicrobial agents that may be used in the present invention include, but are not limited to, polyhexamethylene biguanide hydrochloride; and poly(imidoimidecarbonylimidoimidecarbonylimidohexamethylene hydrochloride), sold under the trade name IB.

Specific thiazolines that may be used in the present invention include, but are not limited to, products sold under the trade name DIDP; and 2-n-octyl-4-isothiazolin-3-one, sold under the trade name IT-3000DIDP.

Specific trichloromethylthioimides that may be used in the present invention include, but are not limited to, N-(trichloromethylthio)phthalimide, sold under the trade name PHENOL®; and N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide, sold under the trade name PHENOL®.

Specific natural antimicrobial agents that can be used in the present invention include, but are not limited to, oils of aniseed, lemon, orange, rosemary, wintergreen, thyme, lavender, clove, hops, tea tree, maple grass, wheat, barley, rue, cedar leaf, fir, cinnamon, fleagrass, geranium, sandalwood, violet, cranberry, eucalyptus, verbena, mint, benzoin, basil, fennel, fir, impatiens, menthol, oregano (ocmea origanuin), coptisine, carradensis, Berberidaceae (Daceae), Ratanhiae longa, and turmeric. Also included in the category of natural antimicrobial agents are the key chemical components found in plant oils that provide antimicrobial benefits. These chemicals include, but are not limited to: anethole; catechol; camphene; thymol; eugenol; eucalyptol; ferulic acid; farnesol; hinokitiol; tropolone; limonene; menthol; methyl salicylate; carvacrol; terpineol; verbenone; berberine; ratanhiae root extract; caryophyllene oxide; citronellal; turmeric; nerolidol; and geraniol.

Specific metal salts that can be used in the present invention include, but are not limited to, salts of metals from Groups 3a-5a, 3b-7b, and 8 of the Periodic Table of the Elements. Specific examples of metal salts include, but are not limited to, salts of the following metals: aluminum; zirconium; zinc; silver; gold; copper; lanthanum; tin; mercury; bismuth; selenium; strontium; scandium; yttrium; cerium; praseodymium; neodymium; promethium; samarium; europium; gadolinium; terbium; dysprosium; holmium; erbium; thallium; ytterbium; lutetium; and mixtures thereof. An example of a metal ion antimicrobial is an antimicrobial manufactured by HealthShield Technology, Wakefield, Mass., and sold under the trade name ANTIBODYNAMIC®.

Specific broad-spectrum antimicrobial agents that may be used in the present invention include, but are not limited to, those antimicrobial agents cited in the other antimicrobial agent classes of the present invention.

Other antimicrobial agents that may be used in the methods of the present invention include, but are not limited to, pyrithiones, particularly pyrithione-including zinc complexes, such as the products sold under the trade name pyrithione; dimethyldimethylol hydantoin, the products sold under the trade name pyrithione; methylchloroisothiazolinone/methylisothiazolinone, the products sold under the trade name Kathon; sodium sulfite; sodium bisulfite; imidazolidinyl urea, the products sold under the trade name Germall; diazolidinyl urea, the products sold under the trade name Germall; benzyl alcohol v 2-bromo-2-nitropropane-1,3-diol, the products sold under the trade name pyrithione; formalin or formaldehyde; iodopropenyl butylcarbamate, the products sold under the trade name Polyphase; chloroacetamide; methanamine; methyldibromonitrile glutaronitrile (1,2-dibromo-2 ,4-dicyanobutane), sold under the trade name; glutaraldehyde; 5-bromo-5-nitro-1,3-dioxane, sold under the trade name; phenylethyl alcohol; o-phenylphenol/sodium o-phenylphenol hydroxymethylglycine sodium salt, sold under the trade name Suttocide; a polymethoxybicyclic oxazolidine, sold under the trade name Nuosept; dimethicone; thimerosal; dichlorobenzyl alcohol; captan; chlorphenenesin; dichlorophen; chlorobutanol; glyceryl laurate; halogenated diphenyl ethers; 2,4,4'-trichloro-2'-hydroxy-diphenyl ether, sold under the trade name and available from Ciba-Geigy, Florham Park, N.J.; and 2,2'-dihydroxy-5,5'-dibromo-diphenyl ether.

(4) Optical properties of thermosetting biophotonic compositions

In some embodiments, the biophotonic compositions of the present invention are substantially transparent or translucent. For example, the percent transmittance of the biophotonic compositions can be measured using a Perkin-Elmer Lambda 9500 Series UV-Visible Spectrophotometer over a wavelength range of 250 nm to 800 nm. In some embodiments, transmittance is measured over the visible range and the data averaged. In some other embodiments, the transmittance of the thermoset biophotonic composition is measured without the chromophore. Because transmittance depends on thickness, the thickness of each sample is measured using a caliper gauge before loading the sample into the spectrophotometer. Transmittance data can be normalized according to the following formula:

Wherein, t ₁ = actual thickness of the sample, t ₂ = thickness normalized for transmittance measurement. In the art, transmittance measurement is usually normalized to 1 cm.

In some embodiments, the biophotonic composition has a transmittance of greater than about 20%, 30%, 40%, 50%, 60%, 70%, or 75% in the visible range. In some embodiments, the transmittance in the visible range is greater than 40%, 41%, 42%, 43%, 44%, or 45%. In some embodiments, the composition has a transmittance of about 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, or 95-100%.

(5) Usage

The thermosetting biophotonic compositions of the present invention may have cosmetic and/or medical benefits. They can be used to promote skin renewal and conditioning, promote the treatment of skin conditions such as acne, eczema, or psoriasis, promote tissue repair, promote wound healing, including periodontal pockets, prevent or treat scars, and prevent or treat bacterial, fungal, or viral infections. They can be used to treat acute inflammation. Acute inflammation can manifest itself as pain, heat, redness, swelling, and loss of function. This includes those seen in allergic reactions, such as insect bites; for example, mosquito, bee, or wasp stings, poison ivy stings, or after ablative surgery.

Thus, in certain embodiments, the present invention provides a method of treating acute inflammation.

In certain embodiments, the present invention provides a method for providing skin renewal or for improving skin condition, treating skin diseases, preventing or treating scarring, and/or accelerating wound healing and/or tissue repair, comprising applying a thermosetting biophotonic composition of the present invention to an area of skin or tissue in need of treatment, and irradiating the biophotonic composition with light having a wavelength that overlaps with the absorption spectrum of a chromophore present in the biophotonic composition.

In the method of the present invention, any light source of actinic light can be used. Any type of halogen lamp, LED lamp or plasma arc lamp or laser is suitable. The main feature of a suitable light source of actinic light is that it emits light of one (or more) wavelengths suitable for activating one or more photoactivators present in the composition. In one embodiment, an argon laser is used. In another embodiment, a potassium titanyl phosphate (KTP) laser (e.g., a GreenLight ^™ laser) is used. In another embodiment, the light source of actinic light is an LED lamp, such as a light curing device. In yet another embodiment, the light source of actinic light is a light source with a wavelength of about 200 to 800 nm. In another embodiment, the light source of actinic light is a visible light source with a wavelength of about 400 to 600 nm. In another embodiment, the light source of actinic light is a visible light source with a wavelength of about 400 to 800 nm. In another embodiment, the light source of actinic light is a visible light source with a wavelength of about 400 to 700 nm. In yet another embodiment, the light source of actinic light is blue light. In yet another embodiment, the light source of actinic light is red light. In another embodiment, the light source of actinic light is green light. In addition, the light source of actinic light should have a suitable power density. Suitable power densities for non-collimated light sources (LED lamps, halogen lamps, or plasma lamps) are about 0.1 mW/ ^cm2 to about 200 mW/ ^cm2 . Suitable power densities for laser light sources are about 0.5 mW/ ^cm2 to 0.8 mW/ ^cm2 .

In some embodiments of the present methods, the light has an energy at the surface of the subject's skin of about 0.1 mW/ ^cm2 to about 500 mW/ ^cm2 , or 0.1-300 mW/ ^cm2 , or 0.1-200 mW/ ^cm2 , wherein the energy applied depends at least on the condition to be treated, the wavelength of the light, the distance between the skin and the light source, and the thickness of the biophotonic composition applied to the target skin or wound. In some embodiments, the light at the subject's skin is about 1-40 mW/ ^cm2 , or 20-60 mW/ ^cm2 , or 40-80 mW/ ^cm2 , or 60-100 mW/ ^cm2 , or 80-120 mW/ ^cm2 , or 100-140 mW/ ^cm2 , or 30-180 mW/ ^cm2 , or 120-160 mW/ ^cm2 , or 140-180 mW/ ^cm2 , or 160-200 mW/ ^cm2 , or 110-240 mW/ ^cm2 , or 110-150 mW/ ^cm2 , or 190-240 mW/ ^cm2 .

Activation of the chromophores in the biophotonic composition can occur almost immediately upon irradiation (femtosecond or picosecond). Extended exposure periods can be beneficial for exploiting the synergistic effects of the absorption, reflection, and re-emitted light of the biophotonic compositions of the present invention and their interaction with the tissue being treated. In one embodiment, the tissue or skin or biophotonic composition is exposed to actinic light for a period of 0.01 to 90 minutes. In another embodiment, the tissue or skin or biophotonic composition is exposed to actinic light for a period of 1 to 5 minutes. In some other embodiments, the biophotonic composition is irradiated for a period of 1 to 3 minutes. In certain embodiments, the light is applied for a period of 1-30 seconds, 15-45 seconds, 30-60 seconds, 0.75-1.5 minutes, 1-2 minutes, 1.5-2.5 minutes, 2-3 minutes, 2.5-3.5 minutes, 3-4 minutes, 3.5-4.5 minutes, 4-5 minutes, 5-10 minutes, 10-15 minutes, 15-20 minutes, or 20-30 minutes. The treatment time may not exceed about 90 minutes, about 80 minutes, about 70 minutes, about 60 minutes, about 50 minutes, about 40 minutes, or about 30 minutes. It should be understood that the treatment time can be adjusted by adjusting the rate of energy density (fluence) delivered to the treatment site to maintain the dosage. For example, the energy density delivered is about ⁴ to about 60 J/cm2, about 10 to about 60 J/ ^cm2 , about 4 to about 90 J/ ^cm2 , about 10 to about 90 J/ ^cm2 , about 10 to about 50 J/ ^cm2 , about 10 to about 40 J/ ^cm2 , about 10 to about 30 J/ ^cm2 , about 20 to about 40 J/ ^cm2 , about 15 J/ ^cm2 to about 25 J/ ^cm2 , or about 10 to about 20 J/ ^cm2 .

In some embodiments, the thermosetting biophotonic composition can be re-irradiated at intervals. In another embodiment, the source of actinic light is continuously moved over the treatment site for an appropriate exposure time. In another embodiment, the thermosetting biophotonic composition is irradiated until the biophotonic composition is at least partially photobleached or completely photobleached.

In certain embodiments, the chromophore can be photoexcited by ambient light, including that from the sun and overhead lighting. In certain embodiments, the chromophore can be photoactivated by light in the visible range of the electromagnetic spectrum. Such light can be emitted by any light source, such as sunlight, a light bulb, an LED device, an electronic display such as a television, a computer, a phone, a mobile device, or a flashlight on a mobile device. In the methods of the present invention, any light source can be used. For example, a combination of ambient light and direct or artificial direct light can be used. Ambient light can include overhead lighting, such as LED bulbs, fluorescent lights, and the like, as well as indirect sunlight.

In the methods of the present invention, the biophotonic composition can be cleared from the skin after application of light. In other embodiments, the biophotonic composition remains on the tissue for an extended period of time and is reactivated with direct or ambient light at an appropriate time to treat the condition.

In certain embodiments of the present methods, the biophotonic composition may be applied to tissue, such as the face, once a week, twice a week, three times a week, four times a week, five times a week, or six times a week, daily, or at any other frequency. The total treatment time may be one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve weeks, or any other length of time deemed appropriate. In certain embodiments, the total tissue area to be treated may be divided into separate areas (cheeks, forehead), and each area treated separately. For example, the composition may be topically applied to a first portion of skin, irradiated with light, and then removed. The biophotonic composition may then be applied to a second portion of skin, irradiated, and removed. Finally, the biophotonic composition may be applied to a third portion of skin, irradiated, and removed.

In certain embodiments, the biophotonic composition may be used after wound closure to optimize scar repair. In such cases, the biophotonic composition may be administered at regular intervals, such as once a week, or at intervals deemed appropriate by the physician.

In certain embodiments, the biophotonic composition may be used after acne treatment to maintain the condition of the treated skin. In such cases, the biophotonic composition may be applied at regular intervals, such as once a week, or at intervals deemed appropriate by the physician.

In certain embodiments, the biophotonic composition may be used after an ablative skin rejuvenation treatment to maintain the condition of the treated skin. In such cases, the biophotonic composition may be applied at regular intervals, such as once a week, or at intervals deemed appropriate by the physician.

In certain embodiments, the thermosetting biophotonic composition can be used to treat eczema or psoriasis. In such cases, the biophotonic composition can be applied at regular intervals, such as once a week, or at intervals deemed appropriate by the physician. Spraying the composition onto the affected area may be less painful for the patient.

In the compositions and methods of the present invention, additional components may optionally be included in or used in combination with the thermosetting biophotonic compositions. Such additional components include, but are not limited to, healing factors, antimicrobial agents, oxygen enriching agents, wrinkle fillers such as botulinum toxin, hyaluronic acid, and polylactic acid, antifungal agents, antibacterial agents, antiviral agents, and/or agents that promote collagen synthesis. These additional components may be topically applied to the skin before, simultaneously with, and/or after topical application of the biophotonic compositions of the present invention. Suitable healing factors include compounds that promote or enhance the healing or renewal process of tissue at the site of application. During photoactivation of the thermosetting biophotonic compositions of the present invention, they can increase the absorption of molecules of such additional components by the skin or mucous membranes at the treatment site. In certain embodiments, increased blood flow at the treatment site is observed over a period of time. Increased lymphatic drainage and potential changes in osmotic equilibrium caused by the dynamic interaction of the free radical cascade are improved or even enhanced by the addition of healing factors. Healing factors can also modulate the biophoton yield of the biophotonic composition, such as the duration and characteristics of photobleaching, or modulate the bleaching of certain components of the composition. Suitable healing factors include, but are not limited to, glucosamine, allantoin, saffron, agents that promote collagen synthesis, antifungal agents, antibacterial agents, antiviral agents, and wound healing factors, such as growth factors.

(i) Skin Renewal

The thermosetting biophotonic composition of the present invention can be used to promote skin renewal or improve skin condition and appearance. The dermis is the second layer of the skin and contains connective tissue, the structural element of the skin. There are various types of connective tissue with different functions. Elastic fibers give the skin elasticity, while collagen provides strength.

The junction between the dermis and epidermis is a crucial structure. The dermal-epidermal junction interlocks to form finger-like epidermal ridges. The cells of the epidermis receive their nutrients from blood vessels within the dermis. These ridges increase the surface area of the epidermis exposed to these vessels and the nutrients they need.

Skin aging is accompanied by significant physiological changes. The production of new skin cells slows, and the epidermal ridges at the dermal-epidermal junction flatten. While the number of elastic fibers increases, their structure and coherence decrease. Furthermore, the amount of collagen and the thickness of the dermis decrease with aging.

Collagen is the main component of the skin's extracellular matrix, providing its structural framework. During the aging process, collagen synthesis decreases and collagen fibers become insoluble, leading to dermal thinning and loss of the skin's biomechanical properties.

Physiological changes in the skin lead to the appearance of noticeable symptoms of aging, commonly referred to as chronoaging, intrinsic aging, and photoaging. The skin becomes drier, rougher, and more flaky, with a duller appearance, most notably the appearance of fine lines and wrinkles. Other symptoms of skin aging include, but are not limited to: thinning and translucency of the skin, loss of underlying fat (resulting in sunken cheeks and undereyes, as well as a noticeable loss of firmness in the hands and neck), bone loss (due to bone loss, the bone shrinks away from the skin, causing the skin to relax), dry skin (which may be itchy), an inability to adequately perspire to cool the skin, unwanted facial hair, freckles, age spots, spider veins, rough and leathery skin, fine wrinkles that disappear when stretched, and loose or blotchy skin.

At the dermal-epidermal junction lies the basement membrane, which separates the keratinocytes of the epidermis from the extracellular matrix beneath it. This membrane consists of two layers: the basal lamina, which contacts the keratinocytes, and the underlying reticular lamina, which contacts the extracellular matrix. The basal lamina is rich in type IV collagen and laminin, molecules that play a role in providing a structural network and bioadhesive properties for cell attachment.

Laminin is a glycoprotein found only within the basement membrane. It is composed of three polypeptide chains (α, β, and γ) arranged in an asymmetric cross-shaped structure and held together by disulfide bonds. These three chains exist as different isoforms, resulting in twelve different isomers of laminin, including laminin-1 and laminin-5.

The dermis is anchored to the keratinocytes of the basement membrane by type VII collagen fibers at hemidesmosomes. Hemidesmosomes are specialized junctions on keratinocytes composed of α-integrin and other proteins. Laminins, particularly laminin-5, form the actual anchor between the hemidesmosomal transmembrane proteins and type VII collagen in basal keratinocytes.

It has been shown that in aging skin, laminin-5 synthesis and type VII collagen expression decrease. This leads to a loss of contact between the dermis and epidermis, causing the skin to lose elasticity and become loose.

More recently, another type of wrinkle, often called expression lines, has gained recognition. Expression lines arise from a loss of elasticity, particularly in the dermis, because when facial muscles produce facial expressions, the skin is no longer able to return to its original state.

The thermosetting biophotonic compositions and methods of the present invention promote skin renewal. In certain embodiments, the thermosetting biophotonic compositions and methods of the present invention promote skin conditions such as skin brightness, pore size reduction, spot reduction, even skin tone, reduced dryness, and skin firming. In certain embodiments, the thermosetting biophotonic compositions and methods of the present invention promote collagen synthesis. In certain embodiments, the thermosetting biophotonic compositions and methods of the present invention can reduce, eliminate, slow, or even reverse one or more signs of skin aging, including, but not limited to, the appearance of fine lines or wrinkles, thinning and translucency of the skin, loss of underlying fat (leading to sunken cheeks and sunken eyes, as well as a noticeable loss of firmness in the hands and neck), bone loss (due to bone loss, bone shrinks away from the skin, causing skin to loosen), dry skin (which may be itchy), inability to adequately perspire to cool the skin, unwanted facial hair, freckles, age spots, spider veins, rough and leathery skin, fine wrinkles that disappear when stretched, and sagging or blotchy skin. In certain embodiments, the biophotonic compositions and methods of the present invention can induce pore size reduction, enhance sculpturing of skin subsegments, and/or enhance skin translucency.

In certain embodiments, the thermosetting biophotonic composition can be used in combination with a collagen promoter. Agents that promote collagen synthesis (ie, procollagen synthesis agents) include amino acids, peptides, proteins, lipids, small chemical molecules, natural products, and extracts from natural products.

For example, it has been found that taking vitamin C, iron, and collagen can effectively increase the amount of collagen in the skin or bones. See, for example, U.S. Patent Application Publication 20090069217. Examples of vitamin C include derivatives of ascorbic acid, such as L-ascorbic acid or sodium L-ascorbate, ascorbic acid preparations obtained by applying ascorbic acid with an emulsifier, and mixtures containing two or more vitamin Cs in any proportion. In addition, natural products containing vitamin C, such as acerola and lemon, can also be used. Examples of iron preparations include: inorganic iron, such as ferrous sulfate, sodium ferrous citrate, or ferric pyrophosphate; organic iron, such as heme iron, ferritin iron, or lactoferrin iron; and mixtures containing two or more of these iron preparations in any proportion. In addition, iron-containing natural products, such as spinach or liver, can also be used. Examples of collagen include extracts obtained by treating bones, skin, etc. of mammals (such as cows or pigs) with acid or alkali; peptides obtained by hydrolysis with proteases such as pepsin, trypsin, or chymotrypsin; and mixtures containing two or more of these collagens in any ratio. Collagen extracted from plant sources can also be used.

(ii) Skin diseases

The thermosetting biophotonic compositions and methods of the present invention can be used to treat skin diseases including, but not limited to, erythema, telangiectasia, actinic telangiectasia, basal cell carcinoma, contact dermatitis, cutaneous fibrosarcoma, genital warts, hidradenitis suppurativa, melanoma, Merkel cell carcinoma, nummular dermatitis, infectious diseases, psoriasis, psoriatic arthritis, rosacea, scabies, scalp psoriasis, sebaceous gland carcinoma, squamous cell carcinoma, seborrheic dermatitis, seborrheic keratosis, herpes zoster, varicella, erythema, erythema, dermatitis ... Skin cancer includes dermatitis, rash, eczema, rashes, impetigo, herpes simplex, nasopharyngitis, perioral dermatitis, pseudofolliculitis, drug eruption, erythema multiplex, erythema nodosum, granuloma annulare, actinic keratosis, purpura, alopecia, aphthous stomatitis, dry skin, chapped skin, xerosis, ichthyosis vulgaris, fungal infection, herpes simplex, cataract, keloid, keratosis, miliaria, mumps, pityriasis rosea, pruritus, urticaria and vascular tumor and malformation. Dermatitis includes contact dermatitis, atopic dermatitis, seborrheic dermatitis, nummular dermatitis, exfoliative dermatitis and static dermatitis (statisdermatitis). Skin cancer includes melanoma, basal cell carcinoma and squamous cell carcinoma.

(iii) Acne and acne scars

The thermosetting biophotonic compositions and methods of the present invention can be used to treat acne. As used herein, "acne" refers to a skin disease caused by inflammation of the skin glands or hair follicles. The thermosetting biophotonic compositions and methods of the present invention can be used to treat acne in its early, pre-emergent stages, or in its later stages, when lesions from the acne are already visible. Embodiments of the biophotonic compositions and methods of the present invention can be used to treat mild, moderate, and severe acne. The early, pre-emergent stage of acne typically begins with excessive secretion of sebum, or skin oil, by sebaceous glands located within the pilosebaceous apparatus. Sebum reaches the skin surface through the follicular duct. Excess sebum in the duct and on the skin can block or stagnate the normal flow of sebum through the follicular duct, causing the sebum to thicken and solidify, forming a solid plug known as a comedone. In the normal sequence of acne development, excessive keratinization of the hair follicle opening is stimulated, completely blocking the duct. The typical result is a papule, pustule, or cyst, which often becomes contaminated with bacteria, leading to secondary infection. Acne is particularly characterized by the presence of a comedone, inflamed papule, or cyst. The appearance of acne can range from mild skin irritation to pitting, and can even develop into disfiguring scars. Therefore, the thermosetting photonic compositions and methods of the present invention can be used to treat one or more of the skin irritation, pitting, scarring, comedones, inflammatory papules, cysts, hyperkeratinization, and thickening and hardening of sebum associated with acne.

Some skin conditions present with different symptoms, including redness, redness, burning, scaling, bumps, papules, pustules, comedones, spots, nodules, cysts, blisters, telangiectasias, spider veins, sores, surface irritation or pain, itching, inflammation, red, purple, or blue spots or discoloration, moles, and/or tumors.

The thermosetting biophotonic compositions and methods of the present invention can be used to treat various types of acne. For example, some types of acne include acne vulgaris, cystic acne, atrophic acne, bromoacne, chloracne, conglobate acne, cosmetic acne, detergent acne, epidermal acne, summer acne, fulminant acne, halogen acne, nodular acne, iodine acne, keloid acne, mechanical acne, papular acne, hair wax acne, premenstrual acne, pustular acne, scurvy acne, tuberculous acne, urticarial acne, acne acne, toxic acne, propionic acid acne, artificial acne, Gram-negative acne, steroid acne, and nodulocystic acne.

In certain embodiments, the thermoset biophotonic compositions of the present invention are used in conjunction with systemic or topical antibiotic therapy. For example, antibiotics used to treat acne include tetracycline, erythromycin, minocycline, and doxycycline, which can also be used with the compositions and methods of the present invention. Use of the thermoset biophotonic compositions can shorten the duration of antibiotic therapy or reduce the dosage required.

(iv) Wound healing

The thermosetting biophotonic compositions and methods of the present invention can be used to treat wounds and promote wound healing. Wounds treatable with the biophotonic compositions and methods of the present invention include, for example, injuries to the skin and subcutaneous tissue induced in various ways and with varying characteristics (e.g., pressure ulcers from prolonged bed rest, wounds induced by trauma or surgery, burns, ulcers associated with diabetes or venous insufficiency, and wounds induced by conditions such as periodontitis). In certain embodiments, the present invention provides thermosetting biophotonic compositions and methods for treating and/or promoting the healing of wounds such as burns, incisions, excisions, injuries, lacerations, abrasions, puncture or penetrating wounds, surgical wounds, contusions, hematomas, pressure injuries, amputations, sores, and ulcers.

The thermosetting biophotonic compositions and methods of the present invention can be used to treat and/or promote the healing of chronic skin ulcers or wounds, which are wounds that fail to achieve durable structural, functional, and aesthetic closure through an orderly, timely series of events. Most chronic wounds can be categorized into three types based on their etiology: pressure ulcers, neuropathic (diabetic foot) ulcers, and vascular (venous or arterial) ulcers.

For example, the present invention provides thermosetting biophotonic compositions and methods for treating and/or promoting the healing of diabetic ulcers. Due to neurological and vascular complications, diabetic patients are prone to foot ulcers and other ulcers. Peripheral neuropathy can lead to altered or complete loss of sensation in the feet and/or legs. Diabetic patients with advanced neuropathy completely lose the ability to distinguish between sharp and dull pain. In patients with neuropathy, any wound or trauma to the foot may go unnoticed for days or weeks. Patients with advanced neuropathy lose the ability to sense persistent pressure injuries, resulting in possible tissue ischemia and gangrene, which can lead to, for example, plantar ulcers. Microvascular disease is one of the significant complications of diabetes and can also lead to ulcers. In some embodiments, the present invention provides thermosetting biophotonic compositions and methods for treating chronic wounds, wherein the chronic wounds are characterized by diabetic foot ulcers and/or ulcers caused by neurological and/or vascular complications of diabetes.

In other examples, the present invention provides thermosetting biophotonic compositions and methods for treating and/or promoting the healing of pressure ulcers. Pressure ulcers, including bedsores, decubitus ulcers, and ischial tuberosity ulcers, can cause significant pain and discomfort to patients. Pressure ulcers are caused by prolonged pressure applied to the skin. Thus, pressure can be exerted on the patient's skin by the weight or mass of an individual. A pressure ulcer can form when blood flow to an area of skin is blocked or interrupted for more than two or three hours. The affected area of skin can become red, painful, and necrotic. If left untreated, the skin can break down and become infected. Thus, a pressure ulcer is a skin ulcer that develops on an area of skin under pressure from prolonged bed rest, wheelchair use, and/or the use of a cast. Pressure ulcers can develop when a person is bedridden, unconscious, unable to feel pain, or unable to move. Pressure ulcers commonly develop over bony prominences of the body, such as the buttocks (on the sacrum or iliac crest) or on the heel.

The wound healing process has three distinct phases. First, during the inflammatory phase, which typically lasts from the time the wound appears to the first two to five days, platelets aggregate to form granulation tissue, promote fibrin deposition, and stimulate the release of growth factors. White blood cells migrate to the wound site and begin to break down and remove debris from the wound. During this inflammatory phase, monocytes also transform into macrophages, which release growth factors that stimulate angiogenesis and the production of fibroblasts.

Next, during the proliferative phase, which typically occurs in two days to three weeks, granulation tissue forms and epithelialization and contraction begin. Fibroblasts are the key cell type in this phase; they fill the wound by proliferating and synthesizing collagen, providing a strong matrix for epithelial cells to grow into. As fibroblasts produce collagen, blood vessels are formed extending from nearby blood vessels, leading to the formation of granulation tissue. Granulation tissue typically grows from the base of the wound. Epithelialization involves the migration of epithelial cells from the wound surface, thereby sealing the wound. Epithelial cells are driven by the need to contact similar cell types and are guided by a network of fibrin strands that acts as a grid, along which they migrate. Contractile cells called myofibroblasts appear at the wound site and help the wound close. These cells display collagen synthesis and contractility and are commonly found in granulating wounds.

Then, during the remodeling phase, the final stage of wound healing, which lasts from three weeks to several years, the collagen in the scar undergoes repeated degradation and resynthesis. During this phase, the tensile strength of the newly formed skin increases.

However, when the rate of wound healing increases, scar formation usually increases accordingly. Scarring is the result of the healing process in most adult animals and human tissues. Scar tissue is different from the tissue it replaces and usually has poor functional quality. The types of scars include but are not limited to atrophic scars, hypertrophic scars and keloid scars, as well as scar contractures. Atrophic scars are flat, with their surface lower than the surrounding skin, forming a valley or hole. Hypertrophic scars are raised scars that remain within the boundaries of the original injury and usually contain excessive collagen arranged in an abnormal manner. Keloid scars are raised scars that spread beyond the original wound edge, invade near normal skin in a site-specific manner, and usually contain collagen spirals arranged in an abnormal manner.

In contrast, normal skin is composed of collagen fibers arranged in a basket-weave pattern, which contributes to the strength and elasticity of the dermis. Therefore, to facilitate the wound healing process, a method is needed that not only stimulates collagen production, but also stimulates collagen production in a way that reduces scarring.

The thermosetting biophotonic compositions and methods of the present invention promote wound healing by promoting substantially uniform epithelialization, promoting collagen synthesis, promoting controlled contraction, and/or reducing scar tissue formation. In certain embodiments, the thermosetting biophotonic compositions and methods of the present invention promote wound healing by promoting substantially uniform epithelialization. In some embodiments, the thermosetting biophotonic compositions and methods of the present invention promote collagen synthesis. In some other embodiments, the thermosetting biophotonic compositions and methods of the present invention promote controlled contraction. In certain embodiments, the thermosetting biophotonic compositions and methods of the present invention promote wound healing, for example, by reducing scar tissue formation.

In the methods of the present invention, the thermosetting biophotonic compositions of the present invention may also be used with negative pressure assisted closure devices and systems.

In certain embodiments, the thermosetting biophotonic composition is maintained in place for up to one, two, or three weeks and is irradiated with light (which may include ambient light) at various intervals. In such cases, the composition can be covered with an opaque composition or left exposed to light during the intervals between exposures to light.

(6) Test kit

The present invention also provides kits for preparing the biophotonic compositions of the present invention and/or providing any components required to form the biophotonic compositions of the present invention.

In some embodiments, a kit includes a container containing components or compositions for preparing the thermosetting biophotonic compositions of the present invention. In some embodiments, a kit includes a thermosetting biophotonic composition of the present invention. The different components comprising the biophotonic compositions of the present invention can be provided in separate containers. For example, a block copolymer can be provided in a container separate from the chromophore. Examples of such containers are dual-chamber syringes, dual-chamber containers with removable dividers, bags with pouches, and multi-compartment blister packs. Another example is one component provided in a syringe that can be injected directly into a container containing another component. In some embodiments, the composition is provided in a spray can or bottle.

In other embodiments, the kit includes systemic drugs for enhancing the therapeutic effect of the thermosetting biophotonic composition of the present invention. For example, the kit may include systemic or topical antibiotics, hormone therapy drugs (e.g., for acne treatment or wound healing), or negative pressure devices.

In other embodiments, the kit includes a device for applying the components of the thermosetting biophotonic composition of the present invention.

In certain aspects, the present invention provides a container comprising a chamber for holding a thermosetting biophotonic composition and an outlet in communication with the chamber for discharging the biophotonic composition from the container, wherein the thermosetting biophotonic composition comprises at least one chromophore dissolved in a block copolymer. The chamber can be partitioned to separate components that are unstable when mixed, such as an oxidizing agent and the chromophore. The container can further comprise a light source for activating the composition once discharged.

In certain embodiments of the kit, the kit may further comprise a light source, such as a portable lamp having a wavelength suitable for activating the chromophore of the thermosetting biophotonic composition. The portable lamp may be battery operated or rechargeable.

Written instructions on how to use the thermosetting biophotonic composition of the present invention may be included in the kit, or may be included on or associated with a container containing the composition or components comprising the thermosetting biophotonic composition of the present invention.

Identification of equivalent thermosetting biophotonic compositions, methods, and kits is well within the skill of one of ordinary skill in light of the teachings of the present invention and will require no more than routine experimentation.

For those skilled in the art, after reading the present invention, various changes and modifications can be made to the present invention. The disclosed features can be implemented with any combination and subcombination (including several dependent combinations and subcombinations) of one or more other features described herein. The various features described or illustrated above, including any components thereof, can be combined or integrated into other systems. In addition, some features can be omitted or not implemented. Those skilled in the art can change, replace and modify these examples without departing from the scope of the information disclosed by the present invention. All references cited in the present invention are incorporated into the present application in their entirety by reference and constitute a part of the present application.

The present invention will be more fully understood from the following examples, which are provided for illustration purposes only and are not to be construed as limiting the present invention in any way.

Examples

Example 1 - Preparation of an Exemplary Thermosetting Biophotonic Composition

According to one embodiment of the present invention, a thermosetting biophotonic composition comprising a poloxamer matrix in which a chromophore is incorporated is prepared. Specifically, F-127, a block copolymer of polyethylene glycol (PEG) and polypropylene glycol (PPG) (having a repeating unit comprising a PPG block linked to two PEG blocks), is used at a weight percentage that allows it to be thermally cured to an adhesive biophotonic composition at a temperature of about 20° C. to 39° C., for example, when applied to tissue or when applied to tissue and/or heated with a lamp.

The thermosetting (gelation) behavior of 20% w/v and 25% w/v Pluronic F-127 solutions at different temperatures was evaluated. Solutions were prepared by dissolving Pluronic F-127 in cold deionized water (approximately 4°C). For the 20% w/v Pluronic solution, 20g of Pluronic F-127 powder was dissolved in 100mL of cold water. For the 25% w/v Pluronic F-127 solution, 25g of Pluronic F-127 powder was dissolved in 100mL of cold water. The concentration of Pluronic F-127 is expressed as weight/volume of water. Gelation was evaluated by placing 2mL aliquots of cold Pluronic F-127 solution in test tubes and immersing the test tubes in a water bath preheated to a well-defined temperature. If no flow was observed in the composition when the tube was inverted, gelation was considered to have occurred. The results are summarized in Table 1.

Table 1 – Thermoset behavior of F-127 at various concentrations.

Next, the effect of carbamide peroxide on the gelation temperature of a 25% Pluronic composition including a chromophore was evaluated. The presence of carbamide peroxide, at least at a concentration of 25%, appears to affect gelation behavior by increasing the temperature at which gelation occurs, but well within the physiologically relevant temperature range. Therefore, these biophotonic compositions based on Pluronic F-127 may also include carbamide peroxide or other peroxides or peroxide precursors.

Table 2 - Exemplary Adhesive Biophotonic Compositions According to the Present Invention

It is noteworthy that once gelled, all of the Pluronic solutions in Table 2 formed transparent/translucent adhesive compositions (non-peelable). These adhesive biophotonic compositions can be made peelable by adding thickeners such as cellulosic compositions, e.g., alginate, methylcellulose, hydroxyethylcellulose, or carboxymethylcellulose. Thermosetting is reversible, as the thermogelled compositions can reliquefy if the temperature is lowered below their gelling temperature.

The solutions of Table 2 are placed in a pump sprayer and can be sprayed onto the target tissue to form a gel upon contact with the target tissue (at approximately 37°C). They can be easily removed by washing or wiping. Alternatively, a cold absorbent composition can be placed over the gel to return the gel to liquid form and then soaked in the composition. The composition can be a sponge or cloth. The composition only needs to be at a temperature below the gelling temperature of the gel.

When activated by blue light, gels 1-3 of Table 2 absorb and emit light.

It will be clear to those skilled in the art that any other poloxamer can be used in place of the Pluronic F-127 of this example, such as P-123, L-122, L-61, L-121, and P-65, in a weight percentage that will allow them to gel at or near body temperature, at or near skin temperature or lower.

Example 2 - Biophotonic Evaluation of the Thermoset Biophotonic Composition of Example 1

The emission spectrum of Gel 3 of Example 1 (containing 0.01 wt% Eosin Y + 0.01 wt% Fluorescein) is shown in Figure 1. When irradiated with light having a peak wavelength of approximately 400-470 nm and a power density of approximately 30-150 mW/cm ² for 5 minutes, the emitted fluorescence was measured using an SP-100 spectroradiometer (SP-100, ORB Optronix). As can be seen in Figure 1, the chromophore was not completely photobleached after 10 minutes of irradiation.

Example 3 - Modulation of IL6 and IL8 of HaCaT human keratinocytes by the thermosetting biophotonic composition of Example 2.

The thermoset biophotonic composition of Example 2 was evaluated for its ability to modulate inflammation, specifically the cytokines IL6 and IL8. HaCaT human keratinocytes serve as an acceptable in vitro model for evaluating the modulation of these inflammatory cytokines. Excessive, uncontrolled inflammation is observed in many skin conditions and wounds and can be detrimental to the host, such as by impairing wound healing processes. Therefore, downregulation of IL6 and IL8 secretion may be beneficial for wound healing and the alleviation of other conditions, such as eczema and psoriasis.

Non-toxic concentrations of IFNγ were used to modulate IL6 and IL8 secretion by HaCaT cells. Dexamethasone (final concentration 5 uM) was used as a positive control (a potent inhibitor of proinflammatory cytokine production). The potential toxic effects of light on HaCaT cells were evaluated using an in vitro toxicology assay kit (based on XTT), which was a spectrophotometric assessment of viable cell counts.

The cell culture was irradiated with light emitted and transmitted by the thermosetting biophotonic composition of Example 2. The thermosetting biophotonic composition was placed 5 cm above the cell culture and irradiated with blue light having a peak wavelength of 400-470 nm (average 460 nm) and a power density of approximately 30-150 mW/ ^cm2 for 90 seconds.

According to the manufacturer's instructions (DuoSet ELISA development kit from R&D Systems), culture supernatants were subjected to cytokine quantification by cytokine ELISA 24 hours after irradiation. The number of secreted cytokines was normalized to cell activity. Spectrophotometric evaluation of viable cell counts was used 24 hours after treatment to measure cell viability, and no toxic effects were observed in all test samples. All samples were screened in quadruplets. Three biological replicates were performed on the tested membranes.

It was found that light emitted by the thermosetting biophotonic composition of Example 2 produced downregulation of IL6 and IL8 on IFNγ-stimulated HaCaT cells. Table 3 summarizes the light treatment received by cultured cells during the irradiation time from the thermosetting biophotonic composition and the IL6 and IL8 expression after irradiation.

Table 3. Light treatment received by cultured HaCaT cells during illumination time from thermosetting biophotonic compositions and observed IL6 and IL8 regulation.

Note: *Statistically significant

Example 4 - Modulation of Inflammation by the Thermoset Biophotonic Composition of Example 2

To obtain a more detailed picture of the biological effects mediated by the thermosetting biophotonic composition of Example 2, a human cytokine antibody array (RayBio C Series, RayBiotech, Inc.) was performed. Cytokines are broadly defined as secreted cell-cell signaling proteins that play important roles in inflammation, innate immunity, apoptosis, angiogenesis, cell growth, and differentiation. Simultaneous detection of multiple cytokines provides a powerful tool for studying cellular activity. Cytokine regulation of cellular processes is a complex, dynamic process that often involves multiple proteins. Positive and negative feedback loops, multiple effects and redundant functions, spatial and temporal expression or synergistic interactions between multiple cytokines, and even regulation of membrane-bound receptors via the release of soluble forms are all common mechanisms of action that regulate cytokine signaling.

A human cytokine antibody array (RayBio C series from Raybiotech) was used to determine the effect of light transmitted through and from thermosetting biophotonic compositions on the cytokine secretion profile of HaCaT cells in culture. Briefly, HaCaT cells were irradiated as in Example 3. Supernatants were collected 24 hours after irradiation and incubated with array antibody membranes according to the manufacturer's instructions. The resulting signals were quantified using ImageJ software. For each experiment, an XTT assay was performed to normalize the amount of secreted cytokines to cell viability (cell viability exceeded 90% in all samples). All samples were run in quadruplets. The table below summarizes the expression of proteins modulated by treatment of cells with thermosetting biophotonic compositions.

Table 4. Modulation of protein expression in IFNg-stimulated HaCaT cells 24 hours after treatment with THERA Light and thermosetting biophotonic compositions compared to control untreated cells.

↓Less than 25% decrease ↑Less than 25% increase

↓↓25-50% decrease ↑↑25-50% increase

↓↓↓A decrease greater than 50% ↑↑↑An increase greater than 50%

in conclusion

The thermoset biophotonic composition of Example 2 allowed blue light transmission (up to 6 J/cm ² of energy flux delivered to cells) and produced fluorescence in the green and yellow spectrum (up to 0.3 J/cm ² of energy flux delivered to cells). Results showed that this light downregulated the proinflammatory cytokines IL-6 and IL-8 in HaCaT cells.

Protein array assays showed that the thermosetting biophotonic composition of Example 2 can also negatively regulate the production of pro-inflammatory cytokines (such as TNFα, IL6) and pro-inflammatory chemokines (such as MCP-3, TARC). Interestingly, the thermosetting biophotonic composition can also downregulate the secretion of growth factors (such as EGF, IGF-1, ANG, VEGF).

Thus, certain embodiments of the thermoset biophotonic compositions of the present invention may be used in the inflammatory phase of wound healing when rapid resolution of this phase is desired, or in other inflammatory diseases.

Example 5. Modulation of collagen production by thermosetting biophotonic compositions

The present invention herein uses human dermal fibroblast (DHF) cells as an in vitro model to study the effect of visible blue light in combination with the thermosetting biophotonic composition of the present invention to evaluate the effect on the secretion of collagen, a component of the extracellular matrix.

Collagen production can be used, for example, in wound healing, as well as for other indications such as skin disorders and renewal. In wound healing, within four to five days after injury, matrix-producing cells, i.e., fibroblasts, migrate into the granulation tissue. These fibroblasts degrade the temporary matrix via MMPs and respond to cytokines/growth factors by proliferating and synthesizing a new extracellular matrix (ECM) composed of collagens I, III, and V, proteoglycans, fibronectin, and other components. TGF-β simultaneously inhibits proteases and enhances protease inhibitors, favoring matrix accumulation.

Non-toxic concentrations of TGFβ-1 were added to the cells to simulate hyperproliferative conditions.The potential toxic effects of light on HaCaT cells were evaluated using an in vitro toxicology assay kit (based on XTT), which is a spectrophotometric assessment of viable cell number.

The cell culture was illuminated with light emitted and transmitted by the thermosetting biophotonic composition of Example 2. The gel was placed 5 cm above the cell culture and irradiated with blue light having a peak wavelength of 400-470 nm and a power density of approximately 30-150 mW/cm ² for 5 minutes. Vitamin C and TGFB1 were used as positive controls.

48 hours after treatment, collagen production was assessed using a microinfrared method. In short, collagen molecules rich in basic amino acids react strongly with acidic dyes. Sirius red is an elongated dye molecule that reacts with collagen (types I, II, and V), binds to it, and after several washes, the free dye is removed. The bound Sirius red is eluted with sodium hydroxide and quantified using a spectrophotometer. All samples were screened in quadruplets. Two biological replicates were performed for each matrix tested.

2 , collagen production can be stimulated by and from light irradiation with the thermosetting biophotonic composition of Example 2. A 4-fold increase in collagen production in the DHF cell culture supernatant was observed compared to the untreated control.

It should be understood that the present invention is not limited to the particular embodiments described and illustrated herein, but encompasses all modifications and alterations coming within the scope of the present invention as defined in the appended claims.

Claims (13)

1. A thermosetting biophotonic composition comprising a block copolymer of the composition at a concentration greater than 20% by weight/volume, and a xanthan dye dissolved in the block copolymer, said block copolymer being Pranic F-127, said xanthan dye being a combination of eosin Y and fluorescein, said xanthan dye being present in an amount of 0.001-3% by weight/volume of said thermosetting biophotonic composition, said thermosetting biophotonic composition being in liquid form at 22°C and thermosetting when heated to above 23°C. 2. The thermosetting biophotonic composition of claim 1, wherein the zeatin dye has a peak absorption or peak emission wavelength in the range of 400 nm to 750 nm. 3. The thermosetting biophotonic composition of claim 1, further comprising a stabilizer selected from gelatin, hydroxyethyl cellulose (HEC), and carboxymethyl cellulose (CMC). 4. The thermosetting biophotonic composition of claim 1, further comprising an oxidizing agent. 5. The thermosetting biophotonic composition of claim 4, wherein the oxidant is selected from hydrogen peroxide, urea peroxide, and benzoyl peroxide. 6. Use of the thermosetting biophotonic composition of claim 1 in the preparation of a medicament for treating target skin tissue, comprising: Applying the thermosetting biophotonic composition of claim 1 to the target skin tissue; and The thermosetting biophotonic composition is irradiated with light having a wavelength absorbed by the zeaton dye; The aforementioned uses include promoting the treatment of the skin tissue. 7. The use of claim 6, wherein the treatment comprises treating a skin condition selected from acne, eczema, psoriasis and dermatitis. 8. Use of the thermosetting biophotonic composition of any one of claims 1-5 in the preparation of a medicament for treating wounds, comprising: Applying the thermosetting biophotonic composition of claim 1 to the target skin tissue; and The composition is irradiated with light having a wavelength absorbed by the pyridine dye; The aforementioned uses include promoting wound healing. 9. The use of claim 6, wherein the thermosetting biophotonic composition is removed after irradiation. 10. The use of claim 6, wherein the thermosetting biophotonic composition remains in situ after irradiation. 11. The use of claim 6, wherein the thermosetting biophotonic composition is irradiated until the zeatin dye is at least partially photobleached. 12. The use of claim 6, wherein the skin treatment includes promoting skin renewal. 13. The use of claim 8, wherein the use includes treating or preventing scar formation.