Compositions and methods for cross-linking structural proteins

Abstract

The present application provides compounds, compositions, and methods for promoting photocrosslinking of structural proteins. In one example, the structural protein is collagen and is present in biological tissue. In one example, a pharmaceutical composition is provided that includes a compound of Formula (II) (wherein R6, R7, R8, and R9 are as described herein). In one example, the composition includes an organic vehicle, such as a semifluorinated alkane vehicle, a medium-chain triglyceride vehicle, or a combination thereof. Methods of using the compositions are also provided, including medical applications where tissue crosslinking is known to be beneficial. In one example, the present disclosure provides a method for corneal crosslinking using the compositions. TIFF2025542093000022.tif32128

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Application No. 63 / 434,487, filed December 22, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] Technical Field The present disclosure relates to pharmaceutical compositions comprising riboflavin or riboflavin derivatives and methods of using the compositions to crosslink structural proteins, such as collagen or keratin. [Background technology]

[0003] background Various biological tissues or structural proteins found in biological tissues can be modified by crosslinking. For example, mammalian corneal tissue can be modified by photocrosslinking collagen, a structural protein within the corneal tissue. Crosslinking of collagen within the cornea can stabilize pathogenic or degenerative eye conditions or correct visual disorders. Riboflavin-5-phosphate has been used as a photosensitizer to promote photocrosslinking of collagen within the corneal tissue. Summary of the Invention

[0004] overview The present disclosure relates to compositions and methods for crosslinking structural proteins, such as collagen. The structural proteins may, in some embodiments, be present in biological tissue. The tissue may, in some embodiments, be ex vivo or may be present in a subject, e.g., a mammal.

[0005] The present disclosure is based, at least in part, on the recognition that certain riboflavin derivatives can have the activity of promoting the photocrosslinking of structural proteins, such as collagen.Without being bound by theory, it is believed that the riboflavin derivatives described herein (1) can act as photosensitizers themselves, promoting the photocrosslinking of structural proteins, such as collagen, and / or (2) can be converted to riboflavin in biological tissues, and then act as photosensitizers promoting the photocrosslinking of structural proteins.The present disclosure is also based on the recognition that the compositions and derivatives described herein can be useful at least in the stable formulation of photosensitizers and in improving the penetration and delivery of compounds into biological tissues.

[0006] For example, one non-limiting example of tissue that can benefit from collagen cross-linking is corneal tissue. Generally, the cornea has a five-layer structure from the outside to the inside: (1) corneal epithelium, (2) Bowman's layer, (3) stroma, (4) Descemet's membrane, and (5) endothelium. In certain eye conditions, such as keratoconus, the corneal stroma thins or liquefies, which can ultimately lead to corneal protrusion and subsequent visual changes. Corneal cross-linking (CXL) is a procedure that cross-links collagen fibrils within the stroma, thereby increasing the mechanical strength of the stroma, modifying its shape, and / or providing relief from various eye symptoms, and can be used to treat or stop the progression of eye diseases, such as keratoconus.

[0007] The Dresden protocol, the first protocol developed for this procedure, is the standard CXL protocol in use and generally involves corneal de-epithelialization (removal of at least a portion of the corneal epithelium), application of 0.1% aqueous riboflavin-5-phosphate, and application of ultraviolet light. See, e.g., Wollensak G, Spoerl E, Seiler T. Riboflavin / ultraviolet-a-induced collagen crosslinking for the treatment of keratoconus. Am J Ophthalmol 2003;135:620-627. One reason for de-epithelializing the cornea for this procedure is that riboflavin and riboflavin-5-phosphate are hydrophilic molecules that do not penetrate well into the dense structure of the corneal epithelium, thereby reducing the amount of enhancer / photosensitizer that can reach the stroma for the crosslinking procedure. This procedure typically takes 30 minutes and requires multiple reapplications of riboflavin or riboflavin-5-phosphate due to limitations such as poor penetration into corneal tissue and easy decomposition. Furthermore, while this procedure provides successful improvement in corneal condition, it is not without side effects and complications. The epithelial removal procedure required to allow riboflavin to effectively penetrate the cornea due to crosslinking causes considerable discomfort to the patient, leading to delayed recovery times and the risk of infection.

[0008] Various formulations have been developed to improve stromal penetration of riboflavin without epithelial removal, but many of these formulations contain irritating or even damaging ingredients, such as ionic agents, that can irritate or damage ocular tissues. Furthermore, while these ingredients may improve riboflavin penetration, they do not alter riboflavin's highly hydrophilic nature. Therefore, even with the use of additional ingredients, riboflavin penetration efficiency is limited. Mechanical methods have also been proposed to avoid epithelial removal, but these often require specialized equipment and operating skills. Furthermore, riboflavin and riboflavin-5-phosphate are highly susceptible to photodegradation. There remains a need for improved compounds, compositions, and methods for crosslinking the cornea and other biological tissues.

[0009] The present disclosure provides compositions containing riboflavin or certain riboflavin derivatives that can be useful in cross-linking structural proteins, and methods for cross-linking structural proteins, for example, structural proteins in mammalian tissues. The compounds and compositions described herein provide improved stability and delivery of photosensitizers / accelerators. For example, in some embodiments, the compounds, compositions, and methods herein provide enhanced penetration of epithelial tissue without the inclusion of irritants, such as preservatives or ionic agents, thus avoiding the need for corneal epithelial removal in corneal cross-linking procedures.

[0010] In one aspect, Formula I: TIFF2025542093000002.tif41128[in the formula, R1, R2, and R3 are each independently hydrogen or -C(O)R 5 More selected; R4 is hydrogen, -C(O)R 5 , phosphoric acid, or salts thereof; R5 is a linear, branched or cyclic alkyl having 1 to 26 carbon atoms. with compounds of; a medium selected from medium-chain triglycerides (MCTs), semi-fluorinated alkanes, or a combination thereof; Described herein are compositions comprising:

[0011] In this and other embodiments, Formula I may optionally have the following features: R1, R2, R3, and R4 are each independently hydrogen or -C(O)R 5 R5 can be a linear, branched, or cyclic alkyl having 1 to 26 carbon atoms; at least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 may be -C(O)R 5 or at least three of R1, R2, R3, and R4 may be -C(O)R 5 R1, R2, R3, and R4 may each independently be -C(O)R 5 R5 may be a linear, branched, or cyclic alkyl having 1 to 26 carbon atoms. R1, R2, R3, and R4 may each independently be selected from hydrogen or -C(O)R5; and at least one of R1, R2, R3, and R4 may be -C(O)R 5 or at least two of R1, R2, R3, and R4 may be -C(O)R 5 or at least three of R1, R2, R3, and R4 may be -C(O)R 5 wherein R5 can be selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms. R1, R2, R3, and R4 can each independently be -C(O)R5, wherein R5 can be selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms. The compound of Formula I, in some embodiments, can be selected from riboflavin tetraacetate, riboflavin tetrapropionate, riboflavin tetrabutyrate, riboflavin tetrapalmitate, or a combination thereof.

[0012] In these and other embodiments, the composition may optionally have the following features: The MCT may be a triglyceride of a fatty acid, where the fatty acid is selected from the group consisting of hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, and combinations thereof. The semifluorinated alkane compound may have the formula RFRH or RFRHRF; RF may be a perfluorinated hydrocarbon having 1 to 15 carbon atoms, and RH may be a non-fluorinated hydrocarbon having 1 to 15 carbon atoms. The semifluorinated alkane may be selected from perfluorobutylbutane (F4H4), perfluorobutylpentane (F4H5), perfluorobutylhexane (F4H6), perfluorohexylethane (F6H2), perfluorohexylbutane (F6H4), perfluorohexyloctane (F6H8), and perfluorohexyldecane (F6H10); preferably, perfluorohexylhexane (F6H6) and perfluorohexyloctane (F6H8). In some embodiments, the semifluorinated alkane may be perfluorohexyloctane (F6H8).

[0013] In these and other embodiments, the compound of Formula I is optionally present in an amount of about 0.001% to about 1% (w / w), about 0.01% to about 1% (w / w), about 0.05% to about 0.5% (w / w), about 0.01% (w / w), 0.02% (w / w), 0.03% (w / w), 0.04% (w / w), 0.05% (w / w), 0.06% (w / w), , 0.07% (w / w), 0.08% (w / w), 0.09% (w / w), 0.1% (w / w), 0.2% (w / w), 0.3% (w / w), 0.4% (w / w), 0.5% (w / w), 0.6% (w / w), 0.7% (w / w), 0.8% (w / w), 0.9% (w / w), or 1% (w / w).

[0014] In these and other embodiments, the MCTs are optionally present in an amount of about 0% to about 99.9% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), or about 60% to about 99.9% (w / w). / w), about 65% to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 3 0% to about 80% (w / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / (w / w), about 60% (w / w), about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w).

[0015] In these and other embodiments, the semifluorinated alkane is optionally present in an amount of from about 0% to about 99.9% (w / w), from about 0.1% to about 99.9% (w / w), from about 0.1% to about 30% (w / w), from about 1% to about 25% (w / w), from about 1% to about 20% (w / w), from about 5% to about 20% (w / w), from about 5% to about 15% (w / w), from about 10% to about 99.9% (w / w), from about 20% to about 99.9% (w / w), from about 30% to about 99.9% (w / w), from about 40% to about 99.9% (w / w), from about 50% to about 99.9% (w / w), from about 6 ... 0.9% (w / w), about 65% to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w) , about 30% to about 80% (w / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% ( (w / w), about 60% (w / w), about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w).

[0016] In these and other embodiments, the composition may optionally have the following features: MCT may be present in an amount of about 1% (w / w) and SFA may be present in an amount of about 99% (w / w); MCT may be present in an amount of about 2% (w / w) and SFA may be present in an amount of about 98% (w / w); MCT may be present in an amount of about 5% (w / w) and SFA may be present in an amount of about 95% (w / w); MCT may be present in an amount of about 10% (w / w) and SFA may be present in an amount of about 90% (w / w); MCT may be present in an amount of about 15% (w / w) and SFA may be present in an amount of about 85% (w / w); MCT may be present in an amount of about 20% (w / w) and SFA may be present in an amount of about 80% (w / w). ); MCT may be present in an amount of about 25% (w / w) and SFA may be present in an amount of about 75% (w / w); MCT may be present in an amount of about 30% (w / w) and SFA may be present in an amount of about 70% (w / w); MCT may be present in an amount of about 35% (w / w) and SFA may be present in an amount of about 65% (w / w); MCT may be present in an amount of about 40% (w / w) and SFA may be present in an amount of about 60% (w / w); MCT may be present in an amount of about 45% (w / w) and SFA may be present in an amount of about 55% (w / w); MCT may be present in an amount of about 50% (w / w), SFA may be present in an amount of about 50% (w / w); MCT may be present in an amount of about 55% (w / w) and SFA may be present in an amount of about 45% (w / w); MCT may be present in an amount of about 60% (w / w) and SFA may be present in an amount of about 40% (w / w); MCT may be present in an amount of about 65% (w / w) and SFA may be present in an amount of about 35% (w / w); MCT may be present in an amount of about 70% (w / w) and SFA may be present in an amount of about 30% (w / w); MCT may be present in an amount of about 75% (w / w) and SFA may be present in an amount of about 25% (w / w); MCT may be present in an amount of about 80% (w / w). MCT may be present in an amount of about 85% (w / w) and SFA may be present in an amount of about 15% (w / w); MCT may be present in an amount of about 90% (w / w) and SFA may be present in an amount of about 10% (w / w); MCT may be present in an amount of about 95% (w / w) and SFA may be present in an amount of about 5% (w / w); MCT may be present in an amount of about 98% (w / w) and SFA may be present in an amount of about 2% (w / w); MCT may be present in an amount of about 99% (w / w) and SFA may be present in an amount of about 1% (w / w);MCT may be the only vehicle, and MCT may be present in an amount of about 99% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.5% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.6% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.7% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.8% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.9% (w / w); SF A may be the only vehicle and SFA may be present in an amount of about 99% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.5% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.6% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.7% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.8% (w / w); or SFA may be the only vehicle and SFA may be present in an amount of about 99.9% (w / w);

[0017] In these and other embodiments, the composition may optionally have the following features: The composition may consist essentially of a compound of Formula I and a vehicle; The composition may be an organic composition with no aqueous component; The composition may be a topical composition or a topical ophthalmic composition, where the composition is a solution, suspension, or emulsion; The composition may further comprise a viscosity enhancing agent; The composition may be preservative-free or eye irritant-free.

[0018] In another aspect, described herein is a method for crosslinking collagen in a subject's cornea, comprising administering the composition described above to at least a portion of the subject's cornea; and irradiating at least a portion of the subject's cornea with ultraviolet light or blue light.

[0019] In another aspect, described herein is a method for crosslinking collagen in tissue of a subject, comprising administering the composition to at least a portion of the subject's cornea; and irradiating at least a portion of the subject's cornea with ultraviolet light or blue light.

[0020] In another aspect, described herein is a method for crosslinking a structural protein, comprising contacting the structural protein, or a biological tissue containing the structural protein, with the composition described above; and irradiating the structural protein or the tissue of the subject with ultraviolet or blue light.

[0021] In these and other embodiments of the methods described herein, the structural protein, or optionally, collagen, may be present in a biological tissue or in the body of a subject. The biological tissue may be ex vivo. The biological tissue may be present in the body of a subject. Contacting the structural protein or contacting the tissue may include administering the composition to the tissue in the body of the subject. The tissue may be selected from collagenous tissue, epidermal tissue, or keratinous tissue. The keratinous tissue may be selected from hair or nails. The tissue may be selected from corneal tissue.

[0022] In these and other embodiments, the method may optionally have the following features: the subject has a chemical burn or keratoconus, corneal thinning, keratitis, infectious keratitis including bacterial or fungal keratitis, corneal ulcer, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune disease, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., following laser in situ keratomileusis (LASIK) and laser photorefractive keratectomy (PRK), post-refractive keratomileusis, or post-refractive keratomileusis. The subject may have a disease selected from the group consisting of corneal degeneration, brittle cornea syndrome, type VI Ehlers-Danlos syndrome, corneal neovascularization, corneal melting including corneal melting in type I Boston keratoprosthesis, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid marginal corneal degeneration (PMD). The composition may be in the form of eye drops, and the method may further comprise applying the eye drops to the subject's eye. The method may further comprise de-epithelializing at least a portion of the cornea. Crosslinking may be performed without de-epithelializing any portion of the cornea.

[0023] In another aspect, The tissue of interest is treated with a compound of Formula II: TIFF2025542093000003.tif40128[in the formula, R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms. administering a compound of the formula: Illuminating tissue with ultraviolet or blue light Described herein are methods for cross-linking collagen in tissue of a subject, comprising:

[0024] In another aspect, The structural protein, or biological tissue containing the structural protein, is selected from the group consisting of: TIFF2025542093000004.tif40128[in the formula, R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms. contacting the compound of Irradiating structural proteins or tissues with ultraviolet or blue light Described herein are methods for cross-linking structural proteins, including:

[0025] In these and other embodiments, Formula II may optionally have the following features: R1, R2, R3, and R4 are each independently hydrogen or -C(O)R 5R5 may be selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms; at least one of R1, R2, R3, and R4 may be -C(O)R 5 or at least two of R1, R2, R3, and R4 may be -C(O)R 5 or at least three of R1, R2, R3, and R4 may be -C(O)R 5 R1, R2, R3, and R4 may each independently be -C(O)R 5 R5 may be selected from linear, branched, or cyclic alkyl having 1 to 26 carbon atoms. R1, R2, R3, and R4 may each independently be selected from hydrogen or -C(O)R5; and at least one of R1, R2, R3, and R4 may be -C(O)R 5 or at least two of R1, R2, R3, and R4 may be -C(O)R 5 or at least three of R1, R2, R3, and R4 may be -C(O)R 5 wherein R5 can be selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms. R1, R2, R3, and R4 can each independently be -C(O)R5, wherein R5 can be selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms. In some embodiments, the compound of Formula II can be selected from riboflavin tetraacetate, riboflavin tetrapropionate, riboflavin tetrabutyrate, riboflavin tetrapalmitate, or a combination thereof.

[0026] In these and other embodiments, the method may optionally have the following features: The structural protein may be selected from collagen, keratin, derivatives thereof, and combinations thereof. The structural protein may be present in a biological tissue or may be present in a subject's body. The biological tissue may be ex vivo. The biological tissue may be present in a subject's body. Contacting the structural protein or contacting the tissue may include administering a compound of Formula II to a tissue of the subject's body. The tissue may be selected from collagenous tissue, epidermal tissue, or keratinous tissue. The keratinous tissue may be selected from hair or nails. Administering the compound of Formula II to the tissue may include administering a composition comprising the compound of Formula II to the tissue, wherein the composition may further include a vehicle. The vehicle may be selected from medium-chain triglycerides (MCTs), semi-fluorinated alkanes, or combinations thereof. The MCTs may be triglycerides of fatty acids, wherein the fatty acids are selected from the group consisting of hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, and combinations thereof. The semifluorinated alkane compound may have the formula RFRH or RFRHRF; RF may be a perfluorinated hydrocarbon having 1 to 15 carbon atoms, and RH may be a non-fluorinated hydrocarbon having 1 to 15 carbon atoms. The semifluorinated alkane may be selected from perfluorobutylbutane (F4H4), perfluorobutylpentane (F4H5), perfluorobutylhexane (F4H6), perfluorohexylethane (F6H2), perfluorohexylbutane (F6H4), perfluorohexyloctane (F6H8), and perfluorohexyldecane (F6H10); preferably, perfluorohexylhexane (F6H6) and perfluorohexyloctane (F6H8). The semifluorinated alkane may be perfluorohexyloctane (F6H8).

[0027] In these and other embodiments, the compound of formula II is present in an amount of about 0.001% to about 1% (w / w), about 0.01% to about 1% (w / w), about 0.05% to about 0.5% (w / w), about 0.01% (w / w), 0.02% (w / w), 0.03% (w / w), 0.04% (w / w), 0.05% (w / w), 0.06% (w / w), 0.07% (w / w), 0.08% (w / w), 0.09% (w / w), 0.10% (w / w), 0.11% (w / w), 0.12% (w / w), 0.13% (w / w), 0.14% (w / w), 0.15% (w / w), 0.16% (w / w), 0.17% (w / w), 0.18% (w / w), 0.19% (w / w), 0.20% (w / w), 0.21% (w / w), 0.22% (w / w), 0.23% (w / w), 0.24% (w / w), 0.25% (w / w), 0.26% (w / w), 0.27% (w / w), 0.28% (w / w), 0.29% (w / w), 0.30% (w / w), 0.31% (w / w), 0.32% (w / w), 0.33% (w / w), 0.34% (w / w), 0.35% (w / w), 0.36% (w / w), 0.37% (w / w), 0.38% (w / w), 0.39% (w / w), 0.40% (w / w It may be present in the composition in an amount of 0.07% (w / w), 0.08% (w / w), 0.09% (w / w), 0.1% (w / w), 0.2% (w / w), 0.3% (w / w), 0.4% (w / w), 0.5% (w / w), 0.6% (w / w), 0.7% (w / w), 0.8% (w / w), 0.9% (w / w), or 1% (w / w).

[0028] In these and other embodiments, the MCTs are optionally present in an amount of about 0% to about 99.9% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), or about 60% to about 99.9% (w / w). / w), about 65% to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 3 0% to about 80% (w / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / (w / w), about 60% (w / w), about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w).

[0029] In these and other embodiments, the semifluorinated alkane is optionally present in an amount of from about 0% to about 99.9% (w / w), from about 0.1% to about 99.9% (w / w), from about 0.1% to about 30% (w / w), from about 1% to about 25% (w / w), from about 1% to about 20% (w / w), from about 5% to about 20% (w / w), from about 5% to about 15% (w / w), from about 10% to about 99.9% (w / w), from about 20% to about 99.9% (w / w), from about 30% to about 99.9% (w / w), from about 40% to about 99.9% (w / w), from about 50% to about 99.9% (w / w), from about 6 ... 0.9% (w / w), about 65% to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w) , about 30% to about 80% (w / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% ( (w / w), about 60% (w / w), about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w).

[0030] In these and other embodiments, the composition may optionally have the following features: MCT may be present in an amount of about 1% (w / w) and SFA may be present in an amount of about 99% (w / w); MCT may be present in an amount of about 2% (w / w) and SFA may be present in an amount of about 98% (w / w); MCT may be present in an amount of about 5% (w / w) and SFA may be present in an amount of about 95% (w / w); MCT may be present in an amount of about 10% (w / w) and SFA may be present in an amount of about 90% (w / w); MCT may be present in an amount of about 15% (w / w) and SFA may be present in an amount of about 85% (w / w); MCT may be present in an amount of about 20% (w / w) and SFA may be present in an amount of about 80% (w / w). ); MCT may be present in an amount of about 25% (w / w) and SFA may be present in an amount of about 75% (w / w); MCT may be present in an amount of about 30% (w / w) and SFA may be present in an amount of about 70% (w / w); MCT may be present in an amount of about 35% (w / w) and SFA may be present in an amount of about 65% (w / w); MCT may be present in an amount of about 40% (w / w) and SFA may be present in an amount of about 60% (w / w); MCT may be present in an amount of about 45% (w / w) and SFA may be present in an amount of about 55% (w / w); MCT may be present in an amount of about 50% (w / w), SFA may be present in an amount of about 50% (w / w); MCT may be present in an amount of about 55% (w / w) and SFA may be present in an amount of about 45% (w / w); MCT may be present in an amount of about 60% (w / w) and SFA may be present in an amount of about 40% (w / w); MCT may be present in an amount of about 65% (w / w) and SFA may be present in an amount of about 35% (w / w); MCT may be present in an amount of about 70% (w / w) and SFA may be present in an amount of about 30% (w / w); MCT may be present in an amount of about 75% (w / w) and SFA may be present in an amount of about 25% (w / w); MCT may be present in an amount of about 80% (w / w). MCT may be present in an amount of about 85% (w / w) and SFA may be present in an amount of about 15% (w / w); MCT may be present in an amount of about 90% (w / w) and SFA may be present in an amount of about 10% (w / w); MCT may be present in an amount of about 95% (w / w) and SFA may be present in an amount of about 5% (w / w); MCT may be present in an amount of about 98% (w / w) and SFA may be present in an amount of about 2% (w / w); MCT may be present in an amount of about 99% (w / w) and SFA may be present in an amount of about 1% (w / w);MCT may be the only vehicle, and MCT may be present in an amount of about 99% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.5% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.6% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.7% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.8% (w / w); MCT may be the only vehicle, and MCT may be present in an amount of about 99.9% (w / w); SF A may be the only vehicle and SFA may be present in an amount of about 99% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.5% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.6% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.7% (w / w); SFA may be the only vehicle and SFA may be present in an amount of about 99.8% (w / w); or SFA may be the only vehicle and SFA may be present in an amount of about 99.9% (w / w);

[0031] In these and other embodiments of the method, the composition may optionally have the following features: The composition may consist essentially of a compound of Formula II and a vehicle; The composition may be an organic composition with no aqueous component; The composition may be a topical composition or a topical ophthalmic composition; The composition may be a solution, suspension, or emulsion; The composition may further comprise a viscosity enhancing agent; The composition may be preservative-free or eye irritant-free.

[0032] In these and other aspects, the method may optionally have the following features: The tissue may be corneal tissue. The subject may have a disease selected from chemical burns, keratoconus, corneal thinning, infectious keratitis including keratitis, bacterial or fungal keratitis, corneal ulcers, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., corneal ectasia appearing after laser in situ keratomileusis (LASIK) and laser photorefractive keratectomy (PRK), postrefractive ectasia, corneal degeneration, brittle cornea syndrome, Ehlers-Danlos syndrome type VI, corneal neovascularization, corneal melting including corneal melting in type I Boston keratoprosthesis, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD). The composition may be in the form of eye drops, and the method may further comprise applying the eye drops to the subject's eye. The method may further comprise de-epithelializing at least a portion of the cornea. Crosslinking may be performed without de-epithelializing any portion of the cornea.

[0033] In some embodiments, the compositions and methods provided herein can provide several advantages.First, in some embodiments, the compositions provided herein contain riboflavin derivatives with variable hydrophobicity, thus allowing those skilled in the art to fine-tune the optimal permeability, stability and effectiveness (compared to riboflavin or riboflavin-5-phosphate) for the intended use of the composition.

[0034] Second, in some embodiments of the compositions and methods provided herein, the compound can penetrate the target tissue, such as the corneal epithelium, better. This provides some additional benefits, such as improved cross-linking in tissue, because the compound can reach the structural protein intended for cross-linking. Furthermore, increased permeability can, in some embodiments, lead to the use of a smaller amount or dosage of the compound or composition, or a smaller application time, or a shorter amount of time to treat the tissue. Furthermore, improved permeability can eliminate the need to de-epithelialize the target tissue, such as the corneal epithelium, thereby reducing pain and discomfort for the patient, and can also shorten healing time and minimize the chance of infection.

[0035] Third, in some embodiments, the riboflavin derivative compounds described herein exhibit greater stability than, for example, riboflavin or riboflavin-5-phosphate, thus reducing the likelihood of excess free radicals being present in tissues. In some embodiments, this improved stability allows for the use of compositions that do not contain preservatives or additional penetration enhancers or other irritants used in aqueous formulations to confer stability or enhance absorption into corneal tissue, thereby simplifying the composition and reducing the potential for irritation, such as eye irritation or allergic reactions, associated with the inclusion of such additives. In some embodiments of the riboflavin derivative compounds described herein, the compounds are less susceptible to photodegradation than riboflavin or riboflavin-5-phosphate, yet can still act as photosensitizers for photocrosslinking of structural proteins, such as collagen. Without being bound by theory, it is believed that in some embodiments, the compounds may act directly as photosensitizers or act through prodrug-like activity, in which the compounds enter a crosslinking environment, such as corneal tissue, and are then converted to normal riboflavin in that environment (e.g., a mammalian eye). In either case, these embodiments can provide both enhanced permeability and desired cross-linking activity.

[0036] Fourth, in some embodiments described herein, the compounds may be formulated in compositions comprising organic media, such as semifluorinated alkanes (SFAs) and / or medium-chain triglycerides (MCTs). These formulations may, in some embodiments, allow for increased stability of the compounds, and thus increased stability of the formulation, desirable dosage, and / or lack of impurities.

[0037] Other features and advantages of the present application will become apparent from the following detailed description and drawings, and from the claims. DETAILED DESCRIPTION OF THE INVENTION

[0038] Detailed Description Provided herein are compositions capable of promoting photocrosslinking of structural proteins, such as collagen, and methods of crosslinking structural proteins, such as collagen and keratin, using the compounds and compositions, as further described herein. In some embodiments, the structural protein may be present in the tissue or body of a subject, such as a mammal, for example, a human.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this application belongs.Methods and materials for use in this application are described herein, but other suitable methods and materials known in the art can also be used.Materials, methods, and examples are illustrative only and are not intended to be limiting.All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety.In the event of any discrepancy, the present specification, including definitions, shall prevail.

[0040] As used herein, the term "about" means "approximately" (eg, plus or minus approximately 10% of the indicated value).

[0041] The term "alkyl," as used herein, refers to a saturated monovalent hydrocarbon moiety having linear or branched moieties or combinations thereof and containing the specified number of carbon atoms (e.g., 1 to 26).

[0042] The term "cycloalkyl," as used herein, refers to a monovalent or divalent group derived from a saturated cyclic hydrocarbon, e.g., of 3 to 8 carbon atoms. Cycloalkyl groups can be monocyclic or polycyclic.

[0043] The formula "H," as used herein, represents a hydrogen atom.

[0044] The formula "O," as used herein, represents an oxygen atom.

[0045] The formula "P," as used herein, represents a phosphorus atom.

[0046] The term "compound," as used herein, is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure, e.g., racemic mixtures. A compound identified herein by name or structure as one particular tautomeric form is intended to include other tautomeric forms unless otherwise specified.

[0047] As used herein, the term "contacting" refers to bringing the indicated moieties together in an in vitro system or in vivo system. For example, "contacting" ocular tissue with a compound of the invention includes administering a compound of the invention to an individual or patient, e.g., a human, having ocular tissue in need of treatment, and also includes introducing a compound of the invention into a sample containing, for example, a cell preparation or purified preparation containing ocular tissue.

[0048] As used herein, the terms "individual," "patient," or "subject," used interchangeably, refer to any animal, e.g., a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, most preferably a human. In some embodiments, a human subject can be of any age, e.g., 0-100 years old. As used herein, "pediatric subject" and "child," used interchangeably, refer to a subject aged 0-18 years, e.g., 1, 2, 5, 10, 12, or 14 years old.

[0049] As used herein, the phrase "effective amount" or "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal, individual, or human that is desired by a researcher, veterinarian, physician, or other clinician.

[0050] As used herein, the term "treating" or "treatment" refers to (1) inhibiting a disease; e.g., inhibiting a disease, condition, or disorder (i.e., arresting further development of the pathology and / or symptomology) in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder, or (2) ameliorating a disease; e.g., ameliorating a disease, condition, or disorder (i.e., reversing the pathology and / or symptomology) in an individual experiencing or exhibiting the pathology or symptomology of the disease, condition, or disorder.

[0051] In some embodiments, a compound of general formula (I): Compositions comprising a compound having TIFF2025542093000005.tif41128 are described herein.

[0052] In formula (I), R1, R2, and R3 are each independently hydrogen or -C(O)R 5 More selected; R4 is hydrogen, -C(O)R5 , phosphoric acid, or salts thereof; R5 is a straight, branched or cyclic alkyl having 1 to 26 carbon atoms (eg, methyl, ethyl, propyl, butyl, sec-butyl, tert-butyl, etc.).

[0053] In some embodiments of Formula (I), R, R, R, and R are each independently hydrogen or —C(O)R 5 In some embodiments of Formula (I), at least one of R1, R2, R3, and R4 is selected from -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 or all of R1, R2, R3, and R4 are -C(O)R 5 In some embodiments, each of R1, R2, R3, and R4 may be the same group or may have a different group.

[0054] In some embodiments of Formula (I), R, R, R, and R are each independently hydrogen or —C(O)R 5 at least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 In some embodiments, R1, R2, R3, and R4 are each independently -C(O)R 5 is.

[0055] In some embodiments of Formula (I), R1, R2, R3, and R4 are each independently selected from hydrogen or -C(O)R5; and at least one of R1, R2, R3, and R4 is -C(O)R 5or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 and R5 can be a straight or branched alkyl having 1 or 3 carbon atoms.

[0056] In some embodiments, R1, R2, R3, and R4 are each independently -C(O)R5, and R5 can be a straight-chain or branched alkyl having 1 or 3 carbon atoms. In some embodiments, exemplary compounds of formula (I) include riboflavin, riboflavin-5-phosphate, riboflavin tetraacetate: (2R,3S,4S)-5-(7,8-dimethyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl)pentane-1,2,3,4-tetrayltetraacetate, riboflavin tetrapropionate: (2R,3S,4S)-5-(7,8-dimethyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl)pentane-1,2,3,4-tetrayltetraacetate, and riboflavin tetrapropionate: (2R,3S,4S)-5-(7,8-dimethyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl)pentane-1,2,3,4-tetrayltetraacetate. tolyl tetrapropionate, riboflavin tetrabutylate: (2R,3S,4S)-5-(7,8-dimethyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl)pentane-1,2,3,4-tetrayltetrabutylate, riboflavin tetrapalmitate: (2R,3S,4S)-5-(7,8-dimethyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl)pentane-1,2,3,4-tetrayltetrapalmitate, or combinations thereof.

[0057] In some embodiments, the compound of formula (I) is present in an amount of from about 0.001% to about 1% (w / w), 0.001% to about 0.95% (w / w), 0.001% to about 0.9% (w / w), 0.001% to about 0.85% (w / w), 0.001% to about 0.8% (w / w), 0.001% to about 0.75% (w / w), 0.001% to about 0.7% (w / w), 0.001% to about 0.65% (w / w), 0.001% to about 0.6% (w / w), 0.001% to about 0.55% (w / w), 0.001% to about 0.5% (w / w), 0.001% to about 0.45% (w / w), 0.001% ~ about 0.4% (w / w), 0.001% to about 0.35% (w / w), 0.001% to about 0.3% (w / w), 0.001% to about 0.25% (w / w), 0.001% to about 0.2% (w / w), 0.001% to about 0.15% (w / w), 0.001% to about 0.1% (w / w), 0 0.001% to approximately 0.09% (w / w), 0.001% to approximately 0.08% (w / w), 0.001% to approximately 0.07% (w / w), 0.001% to approximately 0.06% (w / w), 0.001% to approximately 0.05% (w / w), 0.001% to approximately 0.04% (w / w), 0.001% to approximately 0.03 % (w / w), 0.001% to approximately 0.02% (w / w), 0.001% to approximately 0.01% (w / w), approximately 0.01% to approximately 1%, 0.01% to approximately 0.95% (w / w), 0.01% to approximately 0.9% (w / w), 0.01% to approximately 0.85% (w / w), 0.01% to approximately 0.8% (w / w) , 0.01% to about 0.75% (w / w), 0.01% to about 0.7% (w / w), 0.01% to about 0.65% (w / w), 0.01% to about 0.6% (w / w), 0.01% to about 0.55% (w / w), 0.01% to about 0.5% (w / w), 0.01% to about 0.45% (w / w), 0 0.01% to about 0.4% (w / w), 0.01% to about 0.35% (w / w), 0.01% to about 0.3% (w / w), 0.01% to about 0.25% (w / w), 0.01% to about 0.2% (w / w), 0.01% to about 0.15% (w / w), 0.01% to about 0.1% (w / w), 0.0 1% to approximately 0.09% (w / w), 0.01% to approximately 0.08% (w / w), 0.01% to approximately 0.07% (w / w), 0.01% to approximately 0.06% (w / w), 0.01% to approximately 0.05% (w / w), 0.01% to approximately 0.04% (w / w), 0.01% to approximately 0.03% (w / w), 0.0.1% to about 0.02% (w / w), about 0.05% to about 0.5% (w / w), about 0.05% to about 0.45% (w / w), about 0.05% to about 0.4% (w / w), about 0.05% to about 0.35% (w / w), about 0.05% to about 0.3% (w / w), about 0.05% to about 0.25% (w / w), about 0.05% to about 0.2% (w / w), about 0.05% to about 0.15% (w / w), about 0.05% to about 0.1% (w / w), about 0.05% to about 0.09% (w / w), about 0.05% to about 0.08% (w / w), about 0.05% to about 0.07% (w / w), about 0.05% to about 0.06% (w / w), about 0.06% to about 0.5% (w / w), about 0.07% to about 0.5% (w / w), about 0.08% to about 0.5% (w / w), about 0.09% to about 0.5% (w / w), about 0.1% to about 0.5% (w / w), about 0.15% to about 0.5% (w / w), about 0.2% to about 0.5% (w / w), about 0.25% to about 0 .5% (w / w), about 0.3% to about 0.5% (w / w), about 0.35% to about 0.5% (w / w), about 0.4% to about 0.5% (w / w), about 0.45% to about 0.5% (w / w), about 0.01% (w / w), 0.02% (w) / w), 0.03%(w / w), 0.04%(w / w), 0.05%(w / w), 0.06%(w / w), 0.07%(w / w), 0.08%(w / w), 0.09%(w / w), 0.1%(w / w), 0.15%(w) It may be present in the composition in an amount of 0.2% (w / w), 0.25% (w / w), 0.3% (w / w), 0.35% (w / w), 0.4% (w / w), 0.45% (w / w), 0.5% (w / w), 0.55% (w / w), 0.6% (w / w), 0.65% (w / w), 0.7% (w / w), 0.75% (w / w), 0.8% (w / w), 0.85% (w / w), 0.9% (w / w), 0.95% (w / w), or 1% (w / w).

[0058] In some embodiments, a compound represented by the general formula (II): Provided herein are compositions comprising a compound having TIFF2025542093000006.tif40128.

[0059] In formula (II), R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is a straight, branched or cyclic alkyl having 1 to 26 carbon atoms.

[0060] In some embodiments of Formula (II), R6, R7, R8, and R9, at least one of R6, R7, R8, and R9 is —C(O)R 5 or at least two of R6, R7, R8, and R9 are -C(O)R 5 or at least three of R6, R7, R8, and R9 are —C(O)R 5 or R6, R7, R8, and R9 are all -C(O)R 5 In some embodiments, each of R6, R7, R8, and R9 may have the same group or different groups.

[0061] In some embodiments of Formula (II), R, R, R, and R are each independently hydrogen or —C(O)R 5 R5 may be selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms; at least two of R6, R7, R8, and R9 are -C(O)R 5 or at least three of R6, R7, R8, and R9 are —C(O)R 5 is.

[0062] In some embodiments of Formula (II), R, R, R, and R are each independently —C(O)R 5 and R5 may be selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms.

[0063] In some embodiments of Formula (II), R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; and at least two of R6, R7, R8, and R9 are —C(O)R 5 or at least three of R6, R7, R8, and R9 are —C(O)R 5 and R5 may be selected from linear or branched alkyl having 1 or 3 carbon atoms.

[0064] In some embodiments of Formula (II), R6, R7, R8, and R9 are each independently -C(O)R5, where R5 can be selected from linear or branched alkyl having 1 or 3 carbon atoms. In some embodiments, exemplary compounds of Formula (II) can be selected from riboflavin tetraacetate, riboflavin tetrapropionate, riboflavin tetrabutyrate, riboflavin tetrapalmitate, or a combination thereof.

[0065] In some embodiments, the compound of formula (II) is present in an amount of from about 0.001% to about 1% (w / w), 0.001% to about 0.95% (w / w), 0.001% to about 0.9% (w / w), 0.001% to about 0.85% (w / w), 0.001% to about 0.8% (w / w), 0.001% to about 0.75% (w / w), 0.001% to about 0.7% (w / w), 0.001% to about 0.65% (w / w), 0.001% to about 0.6% (w / w), 0.001% to about 0.55% (w / w), 0.001% to about 0.5% (w / w), 0.001% to about 0.45% (w / w), 0.001% ~ about 0.4% (w / w), 0.001% to about 0.35% (w / w), 0.001% to about 0.3% (w / w), 0.001% to about 0.25% (w / w), 0.001% to about 0.2% (w / w), 0.001% to about 0.15% (w / w), 0.001% to about 0.1% (w / w), 0 0.001% to approximately 0.09% (w / w), 0.001% to approximately 0.08% (w / w), 0.001% to approximately 0.07% (w / w), 0.001% to approximately 0.06% (w / w), 0.001% to approximately 0.05% (w / w), 0.001% to approximately 0.04% (w / w), 0.001% to approximately 0.03 % (w / w), 0.001% to approximately 0.02% (w / w), 0.001% to approximately 0.01% (w / w), approximately 0.01% to approximately 1%, 0.01% to approximately 0.95% (w / w), 0.01% to approximately 0.9% (w / w), 0.01% to approximately 0.85% (w / w), 0.01% to approximately 0.8% (w / w) , 0.01% to about 0.75% (w / w), 0.01% to about 0.7% (w / w), 0.01% to about 0.65% (w / w), 0.01% to about 0.6% (w / w), 0.01% to about 0.55% (w / w), 0.01% to about 0.5% (w / w), 0.01% to about 0.45% (w / w), 0 0.01% to about 0.4% (w / w), 0.01% to about 0.35% (w / w), 0.01% to about 0.3% (w / w), 0.01% to about 0.25% (w / w), 0.01% to about 0.2% (w / w), 0.01% to about 0.15% (w / w), 0.01% to about 0.1% (w / w), 0.0 1% to approximately 0.09% (w / w), 0.01% to approximately 0.08% (w / w), 0.01% to approximately 0.07% (w / w), 0.01% to approximately 0.06% (w / w), 0.01% to approximately 0.05% (w / w), 0.01% to approximately 0.04% (w / w), 0.01% to approximately 0.03% (w / w), 0.0.1% to about 0.02% (w / w), about 0.05% to about 0.5% (w / w), about 0.05% to about 0.45% (w / w), about 0.05% to about 0.4% (w / w), about 0.05% to about 0.35% (w / w), about 0.05% to about 0.3% (w / w), about 0.05% to about 0.25% (w / w), about 0.05% to about 0.2% (w / w), about 0.05% to about 0.15% (w / w), about 0.05% to about 0.1% (w / w), about 0.05% to about 0.09% (w / w), about 0.05% to about 0.08% (w / w), about 0.05% to about 0.07% (w / w), about 0.05% to about 0.06% (w / w), about 0.06% to about 0.5% (w / w), about 0.07% to about 0.5% (w / w), about 0.08% to about 0.5% (w / w), about 0.09% to about 0.5% (w / w), about 0.1% to about 0.5% (w / w), about 0.15% to about 0.5% (w / w), about 0.2% to about 0.5% (w / w), about 0.25% to about 0 .5% (w / w), about 0.3% to about 0.5% (w / w), about 0.35% to about 0.5% (w / w), about 0.4% to about 0.5% (w / w), about 0.45% to about 0.5% (w / w), about 0.01% (w / w), 0.02% (w) / w), 0.03%(w / w), 0.04%(w / w), 0.05%(w / w), 0.06%(w / w), 0.07%(w / w), 0.08%(w / w), 0.09%(w / w), 0.1%(w / w), 0.15%(w) It may be present in the composition in an amount of 0.2% (w / w), 0.25% (w / w), 0.3% (w / w), 0.35% (w / w), 0.4% (w / w), 0.45% (w / w), 0.5% (w / w), 0.55% (w / w), 0.6% (w / w), 0.65% (w / w), 0.7% (w / w), 0.75% (w / w), 0.8% (w / w), 0.85% (w / w), 0.9% (w / w), 0.95% (w / w), or 1% (w / w).

[0066] The compounds of formula (I) or formula (II) and their salts may be in the form of solvates, which are included within the scope of the present invention. Such solvates include, for example, hydrates, alcoholates (e.g., ethanol solvates), etc. The compounds according to the present invention may exist in various polymorphic forms. Although not explicitly shown in the formula above, such forms are intended to be included within the scope of the present disclosure.

[0067] In some embodiments, a composition comprising Formula (I), Formula (II), or a combination thereof may be a pharmaceutical composition. In some embodiments of a pharmaceutical composition, a compound disclosed herein (e.g., a compound of Formula (I)) may be present in an effective amount (e.g., a therapeutically effective amount). The effective dose may vary depending on the disease being treated, the severity of the disease, the route of administration, the subject's sex, age, and general health, the use of excipients, the possibility of co-administration with other therapeutic treatments, e.g., the use of other drugs, and the judgment of the treating physician. For example, in some embodiments, when the compositions described herein are used to treat a patient's eye disease, such as keratoconus, the treating physician may adjust the dosage and administration schedule based on the thickness or condition of the patient's cornea.

[0068] In some embodiments, an effective amount (e.g., in a systemic formulation) of a compound (e.g., Formula (I) or a pharmaceutically acceptable salt thereof) can be, for example, about 0.001 mg / kg to about 500 mg / kg (e.g., about 0.001 mg / kg to about 200 mg / kg; about 0.01 mg / kg to about 200 mg / kg; about 0.01 mg / kg to about 150 mg / kg; about 0.01 mg / kg to about 100 mg / kg; about 0.01 mg / kg to about 50 mg / kg; about 0.01 mg / kg to about 10 mg / kg; about 0.01 mg / kg to about 5 mg / kg; about 0.01 mg / kg to about 1 mg / kg). The effective dose of the compound of Formula (I) can be in the range of about 0.1 mg / kg to about 1 mg / kg; about 0.01 mg / kg to about 0.5 mg / kg; about 0.01 mg / kg to about 0.1 mg / kg; about 0.1 mg / kg to about 200 mg / kg; about 0.1 mg / kg to about 150 mg / kg; about 0.1 mg / kg to about 100 mg / kg; about 0.1 mg / kg to about 50 mg / kg; about 0.1 mg / kg to about 10 mg / kg; about 0.1 mg / kg to about 5 mg / kg; about 0.1 mg / kg to about 2 mg / kg; about 0.1 mg / kg to about 1 mg / kg; or about 0.1 mg / kg to about 0.5 mg / kg. In some embodiments, the effective amount of the compound of Formula (I) is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, or about 5 mg / kg.

[0069] In some embodiments, dosages may be administered daily (e.g., as a single dose or as two or more divided doses, e.g., once daily, twice daily, three times daily, or four times daily) or on a basis other than daily (e.g., every other day, every two days, every three days, once weekly, twice weekly, once every two weeks, once monthly).

[0070] In some embodiments, the composition may include a vehicle. The vehicle may act as a carrier or solvent for one or more compounds of formula (I), one or more compounds of formula (II), or a combination thereof. In some embodiments, the composition may be in the form of a solution, suspension, or emulsion. In some embodiments, the composition provided herein may consist essentially of one or more compounds of formula (I), one or more compounds of formula (II), or a combination thereof. In some embodiments, the composition may be an organic composition without an aqueous component.

[0071] In some embodiments, the medium may be a medium-chain triglyceride (MCT). Medium-chain triglycerides (MCT) are triglycerides of fatty acids. The fatty acids have an aliphatic chain of 6 to 12 carbon atoms, such as hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid. MCT may be a single triglyceride or a mixture of triglycerides. A representative chemical structure of MCT is shown below. TIFF2025542093000007.tif79128

[0072] In some embodiments, the MCTs can be triglycerides of fatty acids, and the fatty acids can be selected from the group consisting of hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, and combinations thereof.

[0073] In some embodiments, the MCT is from about 0% to about 99.9% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), about 60% to about 99.9% (w / w), about 65% to about 99.9% (w / w), or about 70% to about 99.9% (w / w). % to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 30% to About 80% (w / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / w), about 60% (w / w), about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w).

[0074] In some embodiments, the medium can be a semi-fluorinated alkane. A semi-fluorinated alkane is an amphiphilic liquid in which two mutually immiscible portions (a hydrocarbon segment and a fully fluorinated segment) are covalently bonded. In one example, the semi-fluorinated alkane compound useful in the formulation of the present disclosure has the formula CF3(CF2) m (CH2) n In some embodiments, m is an integer from 1 to 10, and n is an integer from 1 to 10. For example, m can be 1, 2, 3, 4, 5, or 6. In another example, n can be 1, 2, 3, 4, 5, or 6. Examples of semifluorinated alkanes include perfluorobutylbutane (F4H4), perfluorobutylpentane (F4H5), perfluorobutylhexane (F4H6), perfluorohexylethane (F6H2), perfluorohexylbutane (F6H4), perfluorohexyloctane (F6H8), and perfluorohexyldecane (F6H10), preferably perfluorohexylhexane (F6H6) and perfluorohexyloctane (F6H8).

[0075] As an example, the structure of F6H8 is: TIFF2025542093000008.tif27128.

[0076] In some embodiments, the semifluorinated alkane compound may have the formula RFRH or RFRHRF; where RF is a perfluorinated hydrocarbon having 1 to 15 carbon atoms and RH is a non-fluorinated hydrocarbon having 1 to 15 carbon atoms.

[0077] In some embodiments, the concentration of the compound of Formula (I) or Formula (II) in the pharmaceutical composition comprising a semifluorinated alkane liquid vehicle can be about 1 mg / mL to about 10 mg / mL. For example, the concentration can be about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, or about 7 mg / mL. In one example, the composition comprising a semifluorinated alkane liquid as a pharmaceutically acceptable vehicle can be a topical ophthalmic composition.

[0078] In some embodiments, the semifluorinated alkane is from about 0% to about 99.9% (w / w), from about 0.1% to about 99.9% (w / w), from about 0.1% to about 30% (w / w), from about 1% to about 25% (w / w), from about 1% to about 20% (w / w), from about 5% to about 20% (w / w), from about 5% to about 15% (w / w), from about 10% to about 99.9% (w / w), from about 20% to about 99.9% (w / w), from about 30% to about 99.9% (w / w), from about 40% to about 99.9% (w / w), from about 50% to about 99.9% (w / w), or from about 60% to about 99.9% (w / w). w), about 65% to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 30% to about 80% (w / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / w), about 60% (w / w), about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w).

[0079] In some embodiments, the vehicle can be a combination of one or more MCTs described herein and one or more SFAs described herein. In some embodiments, MCT may be present in an amount of about 1% (w / w) and SFA may be present in an amount of about 99% (w / w); MCT may be present in an amount of about 2% (w / w) and SFA may be present in an amount of about 98% (w / w); MCT may be present in an amount of about 5% (w / w) and SFA may be present in an amount of about 95% (w / w); MCT may be present in an amount of about 10% (w / w) and SFA may be present in an amount of about 90% (w / w); MCT may be present in an amount of about 15% (w / w) and SFA may be present in an amount of about 85% (w / w); MCT may be present in an amount of about 20% (w / w) and SFA may be present in an amount of about 80% (w / w); MCT may be present in an amount of about 25% (w / w) and SFA may be present in an amount of about 75% (w / w); MCT may be present in an amount of about 30% (w / w) and SFA may be present in an amount of about 70% (w / w); MCT may be present in an amount of about 35% (w / w) and SFA may be present in an amount of about 65% (w / w); MCT may be present in an amount of about 40% (w / w) and SFA may be present in an amount of about 60% (w / w); MCT may be present in an amount of about 45% (w / w) and SFA may be present in an amount of about 55% (w / w); MCT may be present in an amount of about 50% (w / w) and SFA may be present in an amount of about 50% (w / w); MCT may be present in an amount of about 55% (w / w) and SFA may be present in an amount of about 45% (w / w); MCT may be present in an amount of about 60% (w / w) and SFA may be present in an amount of about 40% (w / w). MCT may be present in an amount of about 65% (w / w) and SFA may be present in an amount of about 35% (w / w); MCT may be present in an amount of about 70% (w / w) and SFA may be present in an amount of about 30% (w / w); MCT may be present in an amount of about 75% (w / w) and SFA may be present in an amount of about 25% (w / w); MCT may be present in an amount of about 80% (w / w) and SFA may be present in an amount of about 20% (w / w); MCT may be present in an amount of about 85% (w / w) and SFA may be present in an amount of about 15% (w / w); MCT may be present in an amount of about 90% (w / w) and SFA may be present in an amount of about 10% (w / w); MCT may be present in an amount of about 95% (w / w) and SFA may be present in an amount of about 5% (w / w);MCT may be present in an amount of about 98% (w / w) and SFA may be present in an amount of about 2% (w / w); MCT may be present in an amount of about 99% (w / w) and SFA may be present in an amount of about 1% (w / w); MCT may be the only vehicle and MCT may be present in an amount of about 99% (w / w); MCT may be the only vehicle and MCT may be present in an amount of about 99.5% (w / w); MCT may be the only vehicle and MCT may be present in an amount of about 99.6% (w / w); MCT may be the only vehicle and MCT may be present in an amount of about 99.7% (w / w); MCT may be the only vehicle and MCT may be present in an amount of about 99.8% (w / w). MCT may be the only vehicle, and MCT may be present in an amount of about 99.9% (w / w); SFA may be the only vehicle, and SFA may be present in an amount of about 99% (w / w); SFA may be the only vehicle, and SFA may be present in an amount of about 99.5% (w / w); SFA may be the only vehicle, and SFA may be present in an amount of about 99.6% (w / w); SFA may be the only vehicle, and SFA may be present in an amount of about 99.7% (w / w); SFA may be the only vehicle, and SFA may be present in an amount of about 99.8% (w / w); or SFA may be the only vehicle, and SFA may be present in an amount of about 99.9% (w / w);

[0080] The compositions disclosed herein may be pharmaceutical compositions comprising an effective amount of a compound of Formula (I), Formula (II), or a pharmaceutically acceptable salt thereof, or a combination thereof, and a pharmaceutically acceptable carrier or vehicle as described herein. In some embodiments, the composition may be a topical composition. In some embodiments, the composition may be a topical ophthalmic composition.

[0081] Pharmaceutical compositions may also include one or more additional therapeutic agents, excipients, or diluents, such as, but not limited to, absorbents, anti-irritants, preservatives, antioxidants, colorants / dyes, emollients (moisturizers), emulsifiers, film formers / retentive agents, formulating agents, surfactants / detergents, cleaning agents, penetration enhancers, viscosity enhancers, and thickeners.

[0082] The pharmaceutical compositions of the present application may be suitable for any acceptable route of administration. Suitable examples of acceptable routes of administration include topical, ocular, topical ocular, injection, and the like.

[0083] The compositions and formulations described herein can be conveniently presented in unit or multiple dosage forms. In some embodiments, the compositions and formulations described herein can be conveniently presented in dosage forms (unit or multiple) that can be used by or in automated administration devices or schedules.

[0084] In some embodiments, the compositions are prepared by uniformly and intimately admixing a compound of Formula (I) or Formula (II) disclosed herein with a liquid medium disclosed herein.

[0085] In some embodiments, the topical composition of the present disclosure can be prepared and used in the form of an aerosol spray, cream, emulsion, solid, liquid, dispersion, foam, oil, gel, hydrogel, lotion, mousse, ointment, pomade, or solution. In some embodiments, the composition is in any form commonly used in the fields of cosmetic and skin care formulations. In some embodiments, the topical composition can be in the form of an emulsion. In some embodiments, the topical composition can be an organic solution. In some embodiments, the composition can be in the form of an organic composition without an aqueous component.

[0086] In some embodiments, the topical composition is an ophthalmic composition, for example, for intraocular administration. Suitable examples of ophthalmic compositions include eye drops, suspensions, ointments, emulsions, nanoemulsions, oils, gels, hydrogels, and solutions. Any of these ophthalmic compositions can be administered directly to the cornea, for example, using a plastic or latex applicator (e.g., a disposable applicator), an eye dropper, a glass pipette, or a rubber bulb. These compositions can include, for example, a semifluorinated alkane liquid medium, or an MCT liquid medium, or a combination thereof, as described above.

[0087] In some embodiments, the topical composition is, for example, a cosmetic composition for topical administration to epidermal tissue, nails, or hair. Suitable examples of cosmetic compositions include suspensions, ointments, emulsions, nanoemulsions, oils, gels, creams, mousses, sprays, hydrogels, and solutions. Any of these cosmetic compositions can be administered directly to skin, hair, or nails using, for example, a plastic or latex applicator (e.g., a disposable applicator), a dropper, a cotton or fabric applicator, a sponge, a glass pipette, or a rubber bulb. These compositions can include, for example, a semifluorinated alkane liquid medium, or an MCT liquid medium, or a combination thereof, as described above.

[0088] The present disclosure also includes pharmaceutical kits, such as those useful in treating disorders, diseases, and conditions mentioned herein, comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present disclosure. Such kits may optionally further comprise one or more of various conventional pharmaceutical kit components, such as a container containing one or more pharmaceutically acceptable carriers, additional containers, etc. Instructions indicating the amount of components to be administered, directions for administration, and / or directions for mixing components may also be included in the kit, either as an insert or label. The kit may optionally include additional therapeutic agents as described herein.

[0089] In some embodiments, the kits may include dispensing devices pre-loaded with the compositions described herein.

[0090] The present disclosure also provides methods for crosslinking structural proteins using the compounds and compositions described herein. Structural proteins that can be photocrosslinked using the compounds, compositions, or methods described herein can include any structural protein that can be photocrosslinked in the presence of an active oxygen promoter. Exemplary structural proteins include, but are not limited to, collagen and collagen derivatives, keratin and keratin derivatives, elastin and elastin derivatives, etc. As used herein, "collagen" includes, but is not limited to, collagen fibers or collagen fibrils. As used herein, "keratin" includes, but is not limited to, keratin fibers or keratin fibrils.

[0091] In some embodiments, the method may include contacting the structural protein, or a biological tissue containing the structural protein, with an effective amount of a compound of Formula (I) or Formula (II) or a combination thereof, or a composition containing an effective amount of Formula (I) or Formula (II) or a combination thereof; and irradiating the structural protein or the tissue containing the structural protein.

[0092] In some embodiments, provided is a method for cross-linking collagen in the tissue of a target.In some embodiments, the method can include: administering to the tissue of the target an effective amount of the compound of formula (I) or formula (II) or a combination thereof, or a composition comprising an effective amount of formula (I) or formula (II) or a combination thereof; and irradiating the tissue.

[0093] In some embodiments, irradiating may include irradiating with ultraviolet light (e.g., UVA light) or blue light. In some embodiments, contacting the structural protein or biological tissue comprising the structural protein may include, but is not limited to, topically applying the compound or composition to the protein or tissue, immersing the protein or tissue in the composition, spraying the compound or composition onto the protein or tissue, injecting the compound or composition into the tissue or matrix comprising the structural protein, etc. In some embodiments, contacting the structural protein or biological tissue comprising the structural protein may include administering to the tissue of the subject's body an effective amount of a compound of Formula (I) or Formula (II) or a combination thereof, or a composition comprising an effective amount of Formula (I) or Formula (II) or a combination thereof.

[0094] An effective amount of a compound of Formula (I) or Formula (II) or a combination thereof, or a composition comprising an effective amount of a compound of Formula (I) or Formula (II) or a combination thereof, can be administered to various bodily or biological tissues. In some embodiments, exemplary bodily or biological tissues can include, but are not limited to, collagenous tissue, keratinous tissue, corneal tissue, epithelial tissue, epidermal tissue, pericardial tissue, skin, hair, or nails. In some embodiments, the tissue can be present in a subject's body. In some embodiments, the tissue can be ex vivo or in vitro. In some embodiments, the tissue can be bovine or porcine tissue.

[0095] Also provided herein are methods of treating ocular diseases or conditions using the compounds, compositions, and methods described herein. In some embodiments, the ocular disease or condition can be associated with the cornea or corneal tissue. Exemplary, non-limiting ocular diseases that can be treated by the compounds, compositions, and methods described herein include chemical burns, or keratoconus, corneal thinning, keratitis, infectious keratitis, including bacterial or fungal keratitis, corneal ulcers, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases, including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., leukemia, leukemia, and the like. This includes corneal ectasia after Laser In Situ Keratectomy (LASIK) and Laser Photorefractive Keratectomy (PRK), postrefractive ectasia, corneal degeneration, brittle cornea syndrome, Ehlers-Danlos syndrome type VI, corneal neovascularization, corneal melting including corneal melting in Boston keratoprosthesis type I, hyperopia, hyperopic astigmatism, myopic astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD).

[0096] In some embodiments, a method is provided for administering to a subject's eye and / or corneal tissue an effective amount of a compound of Formula (I) or Formula (II) or a combination thereof, or a composition comprising an effective amount of Formula (I) or Formula (II) or a combination thereof; and irradiating at least a portion of the eye and / or corneal tissue with, for example, ultraviolet light or blue light. In some embodiments, the compound of Formula (I) or Formula (II) or a combination thereof, or the composition comprising Formula (I) or Formula (II) or a combination thereof, can be in the form of eye drops. In some embodiments, the method can further include applying the eye drops to the subject's eye. In some embodiments, the method can further include de-epithelializing at least a portion of the cornea. In some embodiments, the crosslinking method can be performed without de-epithelializing any portion of the cornea.

[0097] The compounds, compositions, and methods described herein may be useful in some embodiments for crosslinking structural proteins in ex vivo biological tissues or other formats. Exemplary ex vivo applications include, but are not limited to, compositions, biomaterials, biological matrices, and biological tissues for experimental or in vitro use, or biomaterials, tissues, hydrogels, and matrices for wound healing, tissue regeneration, tissue engineering, or drug delivery applications, as well as compositions, biological matrices, and biological tissues for use in, for example, hydrogels, biological scaffolds, surgical implants, bioprostheses, grafts, and transplants. Exemplary biomaterials include, but are not limited to, natural or synthetic heart valve replacements, corneal transplants, arterial prostheses, arterial grafts, wound healing matrices or grafts, skin grafts, vascular grafts, transdermal drug delivery devices, or patches. In some embodiments, these biomaterials can also undergo photocrosslinking in a subject's body using the compounds, compositions, and methods described herein, for example, during or after delivery of the biomaterial to the subject via surgery or other delivery methods.

[0098] The present invention is further described in the following examples, which are not intended to limit the scope of the invention.

[0099] While the present invention has been described in conjunction with the detailed description thereof, it should be understood that the foregoing description is intended to be illustrative, but not limiting, of the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. [Example]

[0100] Example 1a - Solubility of Riboflavin in Organic Solvents The solubility of riboflavin in medium-chain triglycerides (MCT), F6H8, or a mixture of MCT / F6H8 was tested at 100° C. Solubility was determined by HPLC according to the conditions outlined in Table 1.

[0101] (Table 1) Chromatography conditions TIFF2025542093000009.tif100149

[0102] The results are shown in Table 1a below.

[0103] (Table 1a) Solubility of riboflavin in organic solvents TIFF2025542093000010.tif38137

[0104] Example 1b - Solubility of Riboflavin Tetrabutyrate in MCT, F6H8, or Mixed Solvents The solubility of riboflavin tetrabutyrate (RTB) was tested in medium-chain triglycerides (MCT), F6H8, or a mixture of MCT / F6H8 at room temperature, and in F6H8 at 100° C. Solubility was analyzed as in Example 1a.

[0105] The results are shown in Table 1b below.

[0106] (Table 1b) Solubility of RTB in organic solvents TIFF2025542093000011.tif53137

[0107] Example 1c - Water solubility of riboflavin tetrabutyrate The solubility of riboflavin tetrabutyrate (RTB) in PBS solution was tested at various pH levels with and without 10% hydroxypropyl-β-cyclodextrin (HP-β-CD) as a solubilizer. For each formulation, 5 mg of RTB was added to 5 mL of pH-adjusted PBS containing 10% HP-β-CD at 25°C and mixed thoroughly. The solubility was analyzed as in Example 1a. The results are shown in Table 1c below.

[0108] (Table 1c) Saturated water solubility of RTB TIFF2025542093000012.tif52142

[0109] Example 2 – Stability of Riboflavin Tetrabutyrate in Organic Solvents The stability of riboflavin tetrabutylate (RTB) in various solvents was investigated. Using the same sample analysis method as described in Examples 1a-1c, the stability of RTB in MCT, F6H8, and combination formulations was monitored at various temperatures. The results are summarized in Table 2 below.

[0110] Table 2: Stability of riboflavin tetrabutylate in organic solvents TIFF2025542093000013.tif151149

[0111] Example 3a - Impurities from riboflavin tetrabutylate in aqueous solution The impurities present in PBS solutions containing riboflavin tetrabutyrate (RTB) were investigated at various pH levels, with and without 10% hydroxypropyl-β-cyclodextrin (HP-β-CD) as a solubilizing agent. The impurities were determined using the same sample analysis method as described in Examples 1a-1c. The results are summarized in Table 3a below.

[0112] Table 3a. Impurities from riboflavin tetrabutylate in aqueous solution TIFF2025542093000014.tif59149

[0113] Example 3b - Impurities from riboflavin tetrabutylate in organic solvents The impurities present in MCT, F6H8, or a combination containing riboflavin tetrabutylate (RTB) were investigated at 25°C. The impurities were determined using sample analysis methods similar to those described in Examples 1a-1c. The results are summarized in Table 3b below.

[0114] Table 3b. Impurities from riboflavin tetrabutylate in organic solvents TIFF2025542093000015.tif65159

[0115] Example 3 - Ex vivo studies (Dresden method) Based on Gregor Wollensak, Eberhard Spoerl, and Theo Seiler. Riboflavin / ultraviolet-induced collagen crosslinking for the treatment of keratoconus. Am J Ophthalmol. 2003 May;135(5):620-7, fresh cadaveric pig eyes containing intact epithelium and clear corneas were collected from a local slaughterhouse within 2-5 hours of death. The eyes were mechanically de-epithelialized, and the corneal limbus was removed with scissors. Using a homemade double-blade scalpel, a single corneal slice measuring 5.0 mm wide, 850 μm central corneal thickness, and 14.0 mm long, including 1.0 mm of sclera on each side, was cut superiorly and inferiorly from the 12 o'clock position of the cornea, identified by its oval shape. Due to the natural thickness of the porcine cornea, only one corneal slice with clearly cut vertical edges could be adequately prepared from each eye. Half of the corneas are treated with RTB-UVA irradiation and half serve as untreated controls.Ultrasound pachymetry (Pachette, Technomed) is used to determine the central corneal thickness of the eyes.

[0116] treatment Five minutes before treatment, 0.1% riboflavin tetrabutyrate (RTB) photosensitizer solution (10 mL of MCT, F6H8, or their combination containing 10 mg of riboflavin tetrabutyrate) was instilled into the treated section at 5-minute intervals, and MCT, F6H8, or the combined solution was instilled into the control section. Long-wave ultraviolet irradiation (370 nm) was applied for 30 minutes at a distance of 1.0 cm from the cornea using two dual UVA diodes (Roithner Lasertechnik) at an irradiance of 3 mW / cm². This is equivalent to a dose of 5.4 J / cm². Exposure parameters were selected depending on the treatment technique used in keratoconus patients. Three 1.3 V accumulators were used as generators. Before treatment, the desired irradiance of 3 mW / cm2 is adjusted by a calibrated UVA meter (LaserMate-Q, Laser2000) at a distance of 1.0 cm and, if necessary, controlled by a potentiometer.

[0117] Static stress and strain measurement The porcine corneal slices are clamped horizontally at a distance of 8.0 mm between the clamps of a commercially available microcomputer-controlled biomaterial tester (Minimat, Rheometric Scientific GmbH). To cover the physiological stress range, a 5 × 10 force is applied, which requires a force of 20 mN for the porcine cornea. 3 A prestress of 1 Pa (1 Pa = 1 N / m2) is used. Then, the strain is increased to 1.5 mm min -1 The stress is increased linearly at a rate of 2×10 5 The stress-strain values ​​are fitted by an exponential function σ = A exp(B × ε) using the SPSS calculation program (SPSS GmbH Software, Munich). Young's modulus (E) is calculated for strains of 4%, 6%, and 8% as the slope of the stress-strain graph (E = dσ / dε = A × B exp(B × ε).

[0118] Statistical evaluation The stress data required for 4%, 6%, and 8% strain in treated and untreated corneas are compared using a Student's t-test.

[0119] result Stress-strain curve The stress-strain curves showed the typical exponential increase of a biological viscoelastic solid. In the porcine cornea, the stress using strain was greater in the treated cornea and less in the untreated cornea, corresponding to an increase in the treated cornea.

[0120] No epithelial removal The same experiment is repeated for intact pig eyes, except that the corneas are not de-epithelialized. MCT or F6H8 RTB formulations are applied to half of the intact eyes, which are then treated with UVA rays. After application and treatment, one corneal section is collected from each eye (treated and untreated) according to the procedure described above. Corneal thickness and stress / strain measurements are obtained according to the procedure described above. In pig corneas, stress using strain is greater in treated corneas and less in untreated corneas, which corresponds to an increase in treated corneas.

[0121] Other Aspects Although the present application has been described in conjunction with its detailed description, it should be understood that the foregoing description is intended to be illustrative, but not limiting, of the scope of the application, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0122] Aspect 1 is Formula I: TIFF2025542093000016.tif41128[in the formula, R1, R2, and R3 are each independently hydrogen or -C(O)R 5 More selected; R4 is hydrogen, -C(O)R 5 , phosphoric acid, or salts thereof; R5 is a linear, branched or cyclic alkyl having 1 to 26 carbon atoms. with compounds of; a medium selected from medium-chain triglycerides (MCTs), semi-fluorinated alkanes, or a combination thereof; a composition comprising is.

[0123] Aspect 2 is The composition of embodiment 1, wherein the MCTs are triglycerides of fatty acids, and the fatty acids are selected from the group consisting of hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, and combinations thereof. is.

[0124] Aspect 3 is The composition of any of embodiments 1 or 2, wherein the semifluorinated alkane compound has the formula RFRH or RFRHRF, where RF is a perfluorinated hydrocarbon having 1 to 15 carbon atoms and RH is a non-fluorinated hydrocarbon having 1 to 15 carbon atoms. is.

[0125] Aspect 4 is The composition of any of embodiments 1-3, wherein the semifluorinated alkane is selected from perfluorobutylbutane (F4H4), perfluorobutylpentane (F4H5), perfluorobutylhexane (F4H6), perfluorohexylethane (F6H2), perfluorohexylbutane (F6H4), perfluorohexyloctane (F6H8), and perfluorohexyldecane (F6H10), preferably perfluorohexylhexane (F6H6) and perfluorohexyloctane (F6H8). is.

[0126] Aspect 5 is The composition of any of embodiments 1-4, wherein the semifluorinated alkane is perfluorohexyl octane (F6H8). is.

[0127] Aspect 6 is The compound of formula I is about 0.001% to about 1% (w / w), about 0.01% to about 1% (w / w), about 0.05% to about 0.5% (w / w), about 0.01% (w / w), 0.02% (w / w), 0.03% (w / w), 0.04% (w / w), 0.05% (w / w), 0.06% (w / w), 0.07% (w / w), 0.08% (w / w), 6. The composition of any of embodiments 1-5, wherein the soluble ... is.

[0128] Aspect 7 is MCT is about 0% to about 99.9% (w / w), about 0% to about 80% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), about 60% to about 99.9% (w / w), about 65% to Approximately 99.9% (w / w), approximately 70% to approximately 99.9% (w / w), approximately 75% to approximately 99.9% (w / w), approximately 80% to approximately 99.9% (w / w), approximately 85% to approximately 99.9% (w / w), approximately 90% to approximately 99.9% (w / w), approximately 95% to approximately 99.9% (w / w), approximately 98% to approximately 99.9% (w / w), approximately 20% to approximately 80% (w / w), approximately 20% to approximately 70% (w / w), approximately 20% to approximately 60% (w / w), approximately 20% to approximately 50% (w / w), approximately 20% to approximately 40% (w / w), approximately 20% to approximately 30% (w / w), approximately 30% to approximately 80% (w / w) / w), about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / w), about 60% ( 7. The composition of any of embodiments 1-6, wherein the hydroxybenzoate is present in an amount of about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w). is.

[0129] Aspect 8 is The concentration of semi-fluorinated alkanes is about 0% to about 99.9% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), about 60% to about 99.9% (w / w), about 65% to about 99.9 % (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 30% to about 80% (w / w) , about 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / w), about 60% (w / 8. The composition of any of embodiments 1-7, wherein the soluble ... is.

[0130] Aspect 9 is MCTs are present in an amount of about 1% (w / w) and SFAs are present in an amount of about 99% (w / w), MCTs are present in an amount of about 2% (w / w) and SFAs are present in an amount of about 98% (w / w), MCTs are present in an amount of about 5% (w / w) and SFAs are present in an amount of about 95% (w / w), MCTs are present in an amount of about 10% (w / w) and SFAs are present in an amount of about 90% (w / w), MCTs are present in an amount of about 15% (w / w) and SFAs are present in an amount of about 85% (w / w), or MCTs are present in an amount of about 20% (w / w) and SFAs are present in an amount of about 80% (w / w). , MCT is present in an amount of about 25% (w / w) and SFA is present in an amount of about 75% (w / w), or MCT is present in an amount of about 30% (w / w) and SFA is present in an amount of about 70% (w / w), or MCT is present in an amount of about 35% (w / w) and SFA is present in an amount of about 65% (w / w), or MCT is present in an amount of about 40% (w / w) and SFA is present in an amount of about 60% (w / w), or MCT is present in an amount of about 45% (w / w) and SFA is present in an amount of about 55% (w / w), or MCT is present in an amount of about 50% (w / w) and SFA is present in an amount of about 50% (w / w). or MCTs are present in an amount of about 55% (w / w) and SFAs are present in an amount of about 45% (w / w); MCTs are present in an amount of about 60% (w / w) and SFAs are present in an amount of about 40% (w / w); MCTs are present in an amount of about 65% (w / w) and SFAs are present in an amount of about 35% (w / w); MCTs are present in an amount of about 70% (w / w) and SFAs are present in an amount of about 30% (w / w); MCTs are present in an amount of about 75% (w / w) and SFAs are present in an amount of about 25% (w / w); or MCTs are present in an amount of about 80% (w / w) and SFAs are present in an amount of about 20% (w / w). or MCT is present in an amount of about 85% (w / w) and SFA is present in an amount of about 15% (w / w), or MCT is present in an amount of about 90% (w / w) and SFA is present in an amount of about 10% (w / w), or MCT is present in an amount of about 95% (w / w) and SFA is present in an amount of about 5% (w / w), or MCT is present in an amount of about 98% (w / w) and SFA is present in an amount of about 2% (w / w), or MCT is present in an amount of about 99% (w / w) and SFA is present in an amount of about 1% (w / w), or MCT is the only vehicle and MCT is present in an amount of about 99% (w / w),MCT is the only vehicle and MCT is present in an amount of about 99.5% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.6% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.7% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.8% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.9% (w / w); or SFA is the only vehicle and SFA is present in an amount of about 99% (w / w). 99.5% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.6% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.7% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.8% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.9% (w / w). is.

[0131] Aspect 10 is R1, R2, R3, and R4 are each independently hydrogen or -C(O)R 5 More selected; R5 is a linear, branched or cyclic alkyl having 1 to 26 carbon atoms; At least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 That is, The composition of any one of aspects 1 to 9 is.

[0132] Aspect 11 is R1, R2, R3, and R4 are each independently —C(O)R 5 and; R5 is a linear, branched or cyclic alkyl having 1 to 26 carbon atoms; The composition of any one of aspects 1 to 10 is.

[0133] Aspect 12 is R1, R2, R3, and R4 are each independently selected from hydrogen or —C(O)R5; At least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 and; R5 is selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms; The composition of any one of embodiments 1 to 11 is.

[0134] Aspect 13 is The composition of any of embodiments 1-12, wherein R1, R2, R3, and R4 are each independently —C(O)R5, and R5 is selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms. is.

[0135] Aspect 14 is 13. The composition of any of embodiments 1-12, wherein the compound of formula I is selected from riboflavin tetraacetate, riboflavin tetrapropionate, riboflavin tetrabutyrate, riboflavin tetrapalmitate, or a combination thereof. is.

[0136] Aspect 15 is The composition of any of embodiments 1 to 14, consisting essentially of a compound of formula I and a vehicle. is.

[0137] Aspect 16 is The composition of any one of embodiments 1 to 15, which is an organic composition with no aqueous component. is.

[0138] Aspect 17 is 17. The composition of any of embodiments 1 to 16, which is a topical composition or a topical ophthalmic composition and is a solution, suspension, or emulsion. is.

[0139] Aspect 18 is 18. The composition of any of embodiments 1-17, further comprising a viscosity enhancing agent. is.

[0140] Aspect 19 is The tissue of interest is treated with a compound of Formula II: TIFF2025542093000017.tif40128[in the formula, R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms. administering a compound of the formula: Illuminating tissue with ultraviolet or blue light a method for cross-linking collagen in tissue of a subject, comprising: is.

[0141] Aspect 20 is The structural protein, or biological tissue containing the structural protein, is represented by Formula II: TIFF2025542093000018.tif40128[in the formula, R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms. contacting the compound of Irradiating structural proteins or tissues with ultraviolet or blue light A method for crosslinking structural proteins, comprising: is.

[0142] Aspect 21 is With respect to Formula II: R1, R2, R3, and R4 are each independently hydrogen or -C(O)R 5 More selected; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms; At least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 That is, The method of any of embodiments 19 or 20 is.

[0143] Aspect 22 is R1, R2, R3, and R4 are each independently —C(O)R 5 and; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms; The method of any one of aspects 19 to 21 is.

[0144] Aspect 23 is R1, R2, R3, and R4 are each independently selected from hydrogen or —C(O)R5; At least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 and; R5 is selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms; The method of any one of aspects 19 to 21 is.

[0145] Aspect 24 is The method of any of embodiments 19-23, wherein R1, R2, R3, and R4 are each independently —C(O)R5, and R5 is selected from linear or branched alkyl having 1 to 3 or 15 carbon atoms. is.

[0146] Aspect 25 is The method of any of embodiments 19-23, wherein the compound of formula II is selected from riboflavin tetraacetate, riboflavin tetrapropionate, riboflavin tetrabutyrate, riboflavin tetrapalmitate, or a combination thereof. is.

[0147] Aspect 26 is 26. The method of any one of embodiments 19 to 25, wherein the structural protein is selected from collagen, keratin, derivatives thereof, and combinations thereof. is.

[0148] Aspect 27 is 27. The method of any of embodiments 19 to 26, wherein the structural protein is present in a biological tissue or in the body of a subject. is.

[0149] Aspect 28 is The method of any one of embodiments 19 to 27, wherein the biological tissue is ex vivo. is.

[0150] Aspect 29 is The method of any of embodiments 19-27, wherein the biological tissue is present in the body of a subject. is.

[0151] Aspect 30 is 30. The method of any of embodiments 19-29, wherein contacting the structural protein or contacting the tissue comprises administering a compound of formula II to the tissue of the subject's body. is.

[0152] Aspect 31 is The method of any of embodiments 19-30, wherein the tissue is selected from collagenous tissue, epidermal tissue, keratinous tissue, or a combination thereof. is.

[0153] Aspect 32 is 32. The method of claim 31, wherein the keratinous tissue is selected from hair or nails. is.

[0154] Aspect 33 is The method of any of embodiments 19-32, wherein administering the compound of formula II to the tissue comprises administering a composition comprising the compound of formula II to the tissue, wherein the composition further comprises a vehicle. is.

[0155] Aspect 34 is 34. The method of claim 33, wherein the vehicle is selected from medium chain triglycerides (MCTs), semifluorinated alkanes, or combinations thereof. is.

[0156] Aspect 35 is 35. The method of claim 34, wherein the MCTs are triglycerides of fatty acids, and the fatty acids are selected from the group consisting of hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, and combinations thereof. is.

[0157] Aspect 36 is The method of any of embodiments 34 or 35, wherein the semifluorinated alkane compound has a formula of RFRH or RFRHRF, where RF is a perfluorinated hydrocarbon having 1 to 15 carbon atoms and RH is a non-fluorinated hydrocarbon having 1 to 15 carbon atoms. is.

[0158] Aspect 37 is The method of any of embodiments 34-36, wherein the semifluorinated alkane is selected from perfluorobutylbutane (F4H4), perfluorobutylpentane (F4H5), perfluorobutylhexane (F4H6), perfluorohexylethane (F6H2), perfluorohexylbutane (F6H4), perfluorohexyloctane (F6H8), and perfluorohexyldecane (F6H10), preferably perfluorohexylhexane (F6H6) and perfluorohexyloctane (F6H8). is.

[0159] Aspect 38 is The composition of any of embodiments 34-37, wherein the semifluorinated alkane is perfluorohexyl octane (F6H8). is.

[0160] Aspect 39 is The compound of formula II is about 0.001% to about 1% (w / w), about 0.01% to about 1% (w / w), about 0.05% to about 0.5% (w / w), about 0.01% (w / w), 0.02% (w / w), 0.03% (w / w), 0.04% (w / w), 0.05% (w / w), 0.06% (w / w), 0.07% (w / w), 0.08% (w / w), % (w / w), 0.09% (w / w), 0.1% (w / w), 0.2% (w / w), 0.3% (w / w), 0.4% (w / w), 0.5% (w / w), 0.6% (w / w), 0.7% (w / w), 0.8% (w / w), 0.9% (w / w), or 1% (w / w). is.

[0161] Aspect 40 is MCT: about 0% to about 99.9% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), about 60% to about 99.9% (w / w), about 65% to about 99.9% (w / w) , about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 30% to about 80% (w / w), about 40 % to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / w), about 60% (w / w) 40. The method of any of embodiments 34-39, wherein the soluble fraction is present in an amount of about 55% (w / w), about 50% (w / w), about 45% (w / w), about 40% (w / w), about 35% (w / w), about 30% (w / w), about 25% (w / w), about 20% (w / w), about 15% (w / w), about 10% (w / w), about 9% (w / w), about 8% (w / w), about 7% (w / w), about 6% (w / w), about 5% (w / w), about 4% (w / w), about 3% (w / w), about 2% (w / w), about 1% (w / w), about 0.5% (w / w), or about 0.1% (w / w). is.

[0162] Aspect 41 is Semifluorinated alkanes are about 0% to about 99.9% (w / w), about 0.1% to about 99.9% (w / w), about 0.1% to about 30% (w / w), about 1% to about 25% (w / w), about 1% to about 20% (w / w), about 5% to about 20% (w / w), about 5% to about 15% (w / w), about 10% to about 99.9% (w / w), about 20% to about 99.9% (w / w), about 30% to about 99.9% (w / w), about 40% to about 99.9% (w / w), about 50% to about 99.9% (w / w), about 60% to about 99.9% (w / w), and about 65% to about 99.9% (w / w), about 70% to about 99.9% (w / w), about 75% to about 99.9% (w / w), about 80% to about 99.9% (w / w), about 85% to about 99.9% (w / w), about 90% to about 99.9% (w / w), about 95% to about 99.9% (w / w), about 98% to about 99.9% (w / w), about 20% to about 80% (w / w), about 20% to about 70% (w / w), about 20% to about 60% (w / w), about 20% to about 50% (w / w), about 20% to about 40% (w / w), about 20% to about 30% (w / w), about 30% to about 80% (w / w), About 40% to about 80% (w / w), about 50% to about 80% (w / w), about 60% to about 80% (w / w), about 70% to about 80% (w / w), about 99.9% (w / w), about 99% (w / w), about 98% (w / w), about 97% (w / w), about 96% (w / w), about 95% (w / w), about 94% (w / w), about 93% (w / w), about 92% (w / w), about 91% (w / w), about 90% (w / w), about 85% (w / w), about 80% (w / w), about 75% (w / w), about 70% (w / w), about 65% (w / w), about 60% (w / w) 41. The method of any of embodiments 34-40, wherein the soluble ... is.

[0163] Aspect 42 is MCTs are present in an amount of about 1% (w / w) and SFAs are present in an amount of about 99% (w / w), MCTs are present in an amount of about 2% (w / w) and SFAs are present in an amount of about 98% (w / w), MCTs are present in an amount of about 5% (w / w) and SFAs are present in an amount of about 95% (w / w), MCTs are present in an amount of about 10% (w / w) and SFAs are present in an amount of about 90% (w / w), MCTs are present in an amount of about 15% (w / w) and SFAs are present in an amount of about 85% (w / w), or MCTs are present in an amount of about 20% (w / w) and SFAs are present in an amount of about 80% (w / w). , MCT is present in an amount of about 25% (w / w) and SFA is present in an amount of about 75% (w / w), or MCT is present in an amount of about 30% (w / w) and SFA is present in an amount of about 70% (w / w), or MCT is present in an amount of about 35% (w / w) and SFA is present in an amount of about 65% (w / w), or MCT is present in an amount of about 40% (w / w) and SFA is present in an amount of about 60% (w / w), or MCT is present in an amount of about 45% (w / w) and SFA is present in an amount of about 55% (w / w), or MCT is present in an amount of about 50% (w / w) and SFA is present in an amount of about 50% (w / w). or MCTs are present in an amount of about 55% (w / w) and SFAs are present in an amount of about 45% (w / w); MCTs are present in an amount of about 60% (w / w) and SFAs are present in an amount of about 40% (w / w); MCTs are present in an amount of about 65% (w / w) and SFAs are present in an amount of about 35% (w / w); MCTs are present in an amount of about 70% (w / w) and SFAs are present in an amount of about 30% (w / w); MCTs are present in an amount of about 75% (w / w) and SFAs are present in an amount of about 25% (w / w); or MCTs are present in an amount of about 80% (w / w) and SFAs are present in an amount of about 20% (w / w). or MCT is present in an amount of about 85% (w / w) and SFA is present in an amount of about 15% (w / w), or MCT is present in an amount of about 90% (w / w) and SFA is present in an amount of about 10% (w / w), or MCT is present in an amount of about 95% (w / w) and SFA is present in an amount of about 5% (w / w), or MCT is present in an amount of about 98% (w / w) and SFA is present in an amount of about 2% (w / w), or MCT is present in an amount of about 99% (w / w) and SFA is present in an amount of about 1% (w / w), or MCT is the only vehicle and MCT is present in an amount of about 99% (w / w),MCT is the only vehicle and MCT is present in an amount of about 99.5% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.6% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.7% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.8% (w / w); MCT is the only vehicle and MCT is present in an amount of about 99.9% (w / w); or SFA is the only vehicle and SFA is present in an amount of about 99% (w / w). 99.5% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.6% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.7% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.8% (w / w), or SFA is the only vehicle and SFA is present in an amount of about 99.9% (w / w). is.

[0164] Aspect 43 is The method of any of embodiments 33-42, wherein the composition consists essentially of the compound of formula II and a vehicle. is.

[0165] Aspect 44 is The method of any one of embodiments 33 to 43, wherein the composition is an organic composition with no aqueous component. is.

[0166] Aspect 45 is The method of any of embodiments 43-44, wherein the composition is a topical composition or a topical ophthalmic composition, and the composition is a solution, suspension, or emulsion. is.

[0167] Aspect 46 is The method of any one of embodiments 19-31 or 33-45, wherein the tissue is corneal tissue. is.

[0168] Aspect 47 is If the subject has a chemical burn or infectious keratitis including keratoconus, corneal thinning, keratitis, bacterial or fungal keratitis, corneal ulcer, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., laser in situ keratomileusis (LASIK) and laser refractive surgery 47. The method of claim 46, wherein the patient has a disease selected from corneal ectasia appearing after corrective keratectomy (PRK), postrefractive ectasia, corneal degeneration, brittle cornea syndrome, Ehlers-Danlos syndrome type VI, corneal neovascularization, corneal melt including corneal melt in Boston keratoprosthesis type I, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD). is.

[0169] Aspect 48 is The method of any one of embodiments 46-47, wherein the composition is in the form of eye drops and the method further comprises applying the eye drops to an eye of the subject. is.

[0170] Aspect 49 is 49. The method of any of embodiments 46-48, further comprising de-epithelializing at least a portion of the cornea. is.

[0171] Aspect 50 is The method of any of embodiments 46-48, wherein the crosslinking is performed without de-epithelializing any portion of the cornea. is.

[0172] Aspect 51 is administering to a tissue of a subject a composition of any of embodiments 1-18; and Illuminating the target tissue with ultraviolet or blue light a method for cross-linking collagen in tissue of a subject, comprising: is.

[0173] Aspect 52 is contacting a structural protein, or a biological tissue containing a structural protein, with the composition of any of embodiments 1-18; and Illuminating the structural proteins or tissue of interest with ultraviolet or blue light A method for crosslinking structural proteins, comprising: is.

[0174] Aspect 53 is 53. The method of any of embodiments 51-52, wherein the structural protein is present in a biological tissue or in the body of a subject. is.

[0175] Aspect 54 is 54. The method of any one of embodiments 51 to 53, wherein the biological tissue is ex vivo. is.

[0176] Aspect 55 is 54. The method of any of embodiments 51-53, wherein the biological tissue is present in the body of a subject. is.

[0177] Aspect 56 is 56. The method of any of embodiments 51-55, wherein contacting the structural protein or contacting the tissue comprises administering the composition to the tissue of the subject's body. is.

[0178] Aspect 57 is 57. The method of any of embodiments 51-56, wherein the tissue is selected from collagenous tissue, epidermal tissue, or keratinous tissue. is.

[0179] Aspect 58 is 58. The method of claim 57, wherein the keratinous tissue is selected from hair or nails. is.

[0180] Aspect 59 is 57. The method of any one of embodiments 51 to 56, wherein the tissue is corneal tissue. is.

[0181] Aspect 60 is If the subject has a chemical burn or infectious keratitis including keratoconus, corneal thinning, keratitis, bacterial or fungal keratitis, corneal ulcer, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., laser in situ keratomileusis (LASIK) and laser refractive surgery 60. The method of claim 59, wherein the patient has a disorder selected from corneal ectasia appearing after corrective keratectomy (PRK), postrefractive ectasia, corneal degeneration, brittle cornea syndrome, Ehlers-Danlos syndrome type VI, corneal neovascularization, corneal melt including corneal melt in Boston keratoprosthesis type I, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD). is.

[0182] Aspect 61 is The method of any one of embodiments 59-60, wherein the composition is in the form of eye drops and the method further comprises applying the eye drops to an eye of the subject. is.

[0183] Aspect 62 is 62. The method of any of embodiments 59-61, further comprising de-epithelializing at least a portion of the cornea. is.

[0184] Aspect 63 is 62. The method of any of embodiments 59-61, wherein the crosslinking is performed without de-epithelializing any portion of the cornea. is.

[0185] Aspect 64 is administering the composition of any of embodiments 1-18 to at least a portion of the cornea of ​​a subject; and irradiating at least a portion of the subject's cornea with ultraviolet or blue light; a method for crosslinking collagen in a subject's cornea, comprising: is.

[0186] Aspect 65 is If the subject has a chemical burn or infectious keratitis including keratoconus, corneal thinning, keratitis, bacterial or fungal keratitis, corneal ulcer, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., laser in situ keratomileusis (LASIK) and laser refractive surgery 65. The method of claim 64, wherein the patient has a disease selected from corneal ectasia appearing after corrective keratectomy (PRK), postrefractive ectasia, corneal degeneration, brittle cornea syndrome, Ehlers-Danlos syndrome type VI, corneal neovascularization, corneal melting including corneal melting in Boston keratoprosthesis type I, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD). is.

[0187] Aspect 66 is The method of any one of embodiments 64-65, wherein the composition is in the form of eye drops and the method further comprises applying the eye drops to an eye of the subject. is.

[0188] Aspect 67 is 67. The method of any of embodiments 64-66, further comprising de-epithelializing at least a portion of the cornea. is.

[0189] Aspect 68 is 67. The method of any of embodiments 64-66, wherein the crosslinking is performed without de-epithelializing any portion of the cornea. is.

[0190] Aspect 69 is The composition of any one of embodiments 1 to 18 or the method of any one of embodiments 33 to 63 or 64 to 69, wherein the composition is preservative-free or eye irritant-free. is.

Claims

1. Formula I: [In the formula, R1, R2, and R3 are each independently hydrogen or -C(O)R 5 Selected from: R4 is hydrogen, -C(O)R 5 , phosphoric acid, or salts thereof; R5 is a linear, branched or cyclic alkyl having 1 to 26 carbon atoms. and a compound of a medium selected from medium-chain triglycerides (MCTs), semi-fluorinated alkanes, or a combination thereof; A composition comprising:

2. 10. The composition of claim 1, wherein the MCTs are triglycerides of fatty acids, and the fatty acids are selected from the group consisting of hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, and combinations thereof.

3. 3. The composition according to any one of claims 1 to 2, wherein the semifluorinated alkane is selected from perfluorobutylbutane (F4H4), perfluorobutylpentane (F4H5), perfluorobutylhexane (F4H6), perfluorohexylethane (F6H2), perfluorohexylbutane (F6H4), perfluorohexyloctane (F6H8), and perfluorohexyldecane (F6H10), preferably perfluorohexylhexane (F6H6) and perfluorohexyloctane (F6H8).

4. 4. The composition of any one of claims 1 to 3, wherein the compound of formula I is present in an amount of about 0.001% to about 1% (w / w), the MCT is present in an amount of about 0.1% to about 99.9% (w / w), and the semifluorinated alkane is present in an amount of about 0.1% to about 99.9% (w / w).

5. R1, R2, R3, and R4 are each independently hydrogen or -C(O)R 5 Selected from: R5 is a linear, branched or cyclic alkyl having 1 to 26 carbon atoms; At least one of R1, R2, R3, and R4 is -C(O)R 5 or at least two of R1, R2, R3, and R4 are -C(O)R 5 or at least three of R1, R2, R3, and R4 are -C(O)R 5 That is, The composition of any one of claims 1 to 4.

6. 6. The composition of any one of claims 1 to 5, wherein the compound of formula I is selected from riboflavin tetraacetate, riboflavin tetrapropionate, riboflavin tetrabutyrate, riboflavin tetrapalmitate, or a combination thereof.

7. 7. The composition of any one of claims 1 to 6, consisting essentially of a compound of formula I and said vehicle.

8. 8. The composition of any one of claims 1 to 7, which is an organic composition with no aqueous components.

9. Administering to a tissue of a subject a composition according to any one of claims 1 to 8, or administering to a tissue of a subject a compound of formula II: [In the formula, R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms. administering a compound of formula (I) to a subject; and irradiating the tissue with ultraviolet light or blue light; 1. A method for cross-linking collagen in tissue of a subject, comprising:

10. Contacting a structural protein or a biological tissue containing a structural protein with a composition according to any one of claims 1 to 8, or contacting a structural protein or a biological tissue containing a structural protein with a compound of formula II: [In the formula, R6, R7, R8, and R9 are each independently selected from hydrogen or —C(O)R5; At least one of R6, R7, R8, and R9 is —C(O)R5; R5 is selected from linear, branched or cyclic alkyl having 1 to 26 carbon atoms. contacting the compound of formula (I) with irradiating the structural protein or the tissue with ultraviolet light or blue light; 1. A method for crosslinking structural proteins, comprising:

11. 11. The method of claim 10, wherein the structural protein is selected from collagen, keratin, derivatives thereof, and combinations thereof.

12. 11. The method of any one of claims 9 or 10, wherein the tissue is selected from collagenous tissue, epidermal tissue, or keratinous tissue.

13. The method of any one of claims 9 to 12, wherein the tissue is corneal tissue.

14. If the subject has a chemical burn or infectious keratitis including keratoconus, corneal thinning, keratitis, bacterial or fungal keratitis, corneal ulcer, bullous keratopathy and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, for example, after laser in situ keratomileusis (LASIK) and laser photorefractive keratectomy (PRK), post-refractive ectasia ectasia), corneal degeneration, brittle cornea syndrome, Type VI Ehlers-Danlos syndrome, corneal neovascularization, corneal melt including corneal melt in Type I Boston keratoprosthesis, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD).

15. 15. The method of any one of claims 13 to 14, further comprising de-epithelializing at least a portion of the cornea.

16. 15. The method of any one of claims 13-14, wherein the crosslinking is performed without de-epithelializing any portion of the cornea.

17. administering to at least a portion of the cornea of ​​a subject a composition according to any one of claims 1 to 8; and irradiating at least a portion of the cornea of ​​the subject with ultraviolet or blue light.

1. A method for crosslinking collagen in a subject's cornea, comprising:

18. If the subject has a chemical burn or keratoconus, corneal thinning, keratitis, infectious keratitis including bacterial or fungal keratitis, corneal ulcer, bullous keratopathy, and other forms of corneal edema, keratolysis, autoimmune diseases, scarring diseases including ocular cicatricial pemphigoid, Stevens-Johnson syndrome, lupus erythematosus, rheumatoid arthritis, corneal ectasia, e.g., laser in situ keratomileusis (LASIK) and laser refractive surgery, 18. The method of claim 17, wherein the patient has a disease selected from corneal ectasia appearing after orthorectal keratomileusis (PRK), postrefractive ectasia, corneal degeneration, brittle cornea syndrome, type VI Ehlers-Danlos syndrome, corneal neovascularization, corneal melting including corneal melting in type I Boston keratoprosthesis, hyperopia, hyperopic astigmatism, myopia, astigmatism, myopic astigmatism, remyopia, keratopathy, corneal astigmatism, unstable cornea, or pellucid limbal corneal degeneration (PMD).

19. 19. The method of any one of claims 17-18, further comprising de-epithelializing at least a portion of the cornea.

20. 19. The method of any one of claims 17-18, wherein the crosslinking is performed without de-epithelializing any portion of the cornea.

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