Photodynamic ophthalmic treatments with amino acid-supplemented photosensitizer

Abstract

Ophthalmic compositions comprising a photosensitizer and an amino acid are disclosed. A method and a kit for using the compositions to treat ocular bacterial infections are also described herein.

Description

3724890.00383PCT APPLICATIONPhotodynamic Ophthalmic Treatments with Amino Acid-Supplemented PhotosensitizerCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 683,378, filed August 15, 2024, the entirety of which is incorporated herein.TECHNICAL FIELD

[0002] The instant disclosure relates to compositions and methods for treating infections, particularly those affecting the eye.BACKGROUND

[0003] The eye can be susceptible to infections such as ulcerative keratitis (“UK”), a general term for the group of disease processes leading to corneal ulceration, as well as the inflammation that accompanies ulceration.

[0004] UK can be caused by Pseudomonas aeruginosa (P. aeruginosa), a Gram-negative bacteria having an outer cell membrane which includes a great number of small porins and resistance-nodulation-division efflux pumps. These form a permeability barrier, thus complicating treatment with conventional antibiotic drugs.

[0005] Additionally, P. aeruginosa is able to form matrix-enclosed aggregates, known as biofilms, further reducing drug permeability. This increasing antibiotic resistance makes P. aeruginosa, infections extremely difficult to treat. Indeed, it has proved challenging to develop new antibiotics effective against Gram-negative bacteria in general.

[0006] Therefore, improved compositions and methods would address a critical clinical shortcoming.SUMMARY

[0007] Disclosed herein are compositions and methods for treating infections caused by bacterial, viral, and fugal pathogens. For example, disclosed embodiments can treat ophthalmic diseases including UK. In particular, disclosed embodiments can be used to treat bacterial infections, such as those caused by Gram-negative bacteria.3724890.00383PCT APPLICATION

[0008] Disclosed compositions and methods include novel treatments for infections such as those caused by P. aeruginosa. For example, some disclosed methods comprise combining: a) photodynamic therapy (PDT); with b) an amino acid-supplemented photosensitizer to kill bacterial cells.

[0009] Further embodiments comprise treatments under hyperoxic conditions, such as 90% O2.

[0010] Also disclosed are kits comprising disclosed compositions and methods as well as instructions for use.BRIEF DESCRIPTION OF THE FIGURES

[0011] FIG. 1 shows an experimental procedure for testing riboflavin / histidine / ultraviolet A radiation (UVA) treatment on P. aeruginosa cell death.

[0012] FIG. 2 shows agar plate colony forming units (CFUs) of P. aeruginosa cells with or without riboflavin / histidine / UVA treatment.

[0013] FIG. 3 shows the effect of histidine concentration on P. aeruginosa cell killing.

[0014] FIG. 4 shows the effect of UVA dose on P. aeruginosa cell killing.

[0015] FIG. 5A-5G shows the experimental procedure used in Example 2.

[0016] FIG. 6 shows impact of histidine concentration on P. aeruginosa cell killing.

[0017] FIG. 7 shows the effect of PDT with 0.2% w / w riboflavin / 50 mM histidine I phosphate buffered saline formulation under hyperoxic conditions on P. aeruginosa (ATCC 15692) cell killing.

[0018] FIG. 8 shows the effects of varying amounts of riboflavin and histidine in phosphate buffered saline formulations on P. aeruginosa cell killing under normoxic and hyperoxic conditions.

[0019] FIG. 9 shows the experimental procedure used in Example 4.

[0020] FIG. 10A illustrates a top view of an example system for applying ophthalmic treatments with a mask device.3724890.00383PCT APPLICATION

[0021] FIG. 10B illustrates a detailed view of the mask device from an alternate perspective.DETAILED DESCRIPTION

[0022] Antibiotics are generally used to treat P. aeruginosa infections. However, conventional antibiotic treatments show very limited efficacy because of the permeability barrier formed by the bacterium’s intrinsic structure and increasing resistance to antibiotics due to evolutionary pressure. Further, the determination of antibiotic type for treating P. aeruginosa infections is in turn based on specimen laboratory test results, which typically take a significant amount of time and can thus delay treatment.

[0023] In contrast, the present disclosure provides an efficient and timely approach to control bacterial infections, including Gram-negative bacterial infections.

[0024] The disclosed photodynamic therapy (PDT) uses ultraviolet A (UVA) light, an amino acid-supplemented photosensitizer solution and, optionally, hyperoxic conditions to efficiently kill P. aeruginosa cells. Compared to conventional treatments for P. aeruginosa infections, the present methods excelled in at least two areas - elevated levels of cell death and short treatment time. The disclosed methods do not require that the specimen be laboratory tested prior to treatment. Because the doubling time of P. aeruginosa is as short as 30 minutes, the disclosed methods (with a total UVA treatment time of 10-30 minutes) enable simple and timely intervention.

[0025] PDT is a therapeutic method that combines light energy with a photosensitizer designed to cause microbial cell death after light irradiation. UVA activated photosensitizers, such as riboflavin, generate reactive oxygen species (ROS) through mixed type I and type II photochemical reactions. ROS are a group of derivatives of molecular oxygen. Excessive levels of ROS in cells cause damage to DNA, RNA, and proteins, resulting in cell death.

[0026] UVA irradiation induces ROS and increases the intracellular level of H2O2.. However, photosensitizers, such as riboflavin, and UVA light alone do not bring about sufficient cell death in a short time period. Herein we demonstrate the synergetic role of photosensitizers and amino acids that produce significant P. aeruginosa cell death. Results presented herein demonstrate UVA irradiation under a hyperoxic atmosphere with L-histidine supplemented riboflavin can kill about 99% of P. aeruginosa cells.

[0027] It is hypothesized that the side chains of aromatic amino acids may help to increase H2O2-induced bacterial cell toxicity by promoting the formation of highly reactive hydroxyl radicals.3724890.00383PCT APPLICATIONFor example, histidine’s imidazole ring may bind to metal ions such as iron ions, which then catalyze oxidative reactions that damage cellular components. Histidine’s imidazole ring may also directly react with singlet oxygen generated from the activation of O2 by a photosensitizer and photoactivating light (e.g., riboflavin and UVA light). In turn, this can produce hydroxyl radicals and reactive oxygen species by subsequent reactions that then induce bacterial cell toxicity and subsequent cell death of the bacteria.

[0028] Disclosed embodiments provide an effective response to antibiotic resistance.

[0029] Further embodiments comprise kits, for example kits comprising disclosed compositions and devices, for use in disclosed methods. Disclosed kits can further comprise instructions for use, for example use in performing disclosed methods.DEFINITIONS:

[0030] “A” and “an” are used herein to refer to one or to more than one ( / .e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0031] “Administration,” or “to administer” means the step of giving ( / .e. administering) a pharmaceutical composition or active ingredient to a subject. The pharmaceutical compositions disclosed herein can be administered via a number of appropriate routes including topically.

[0032] “Comprise,” “comprising,” “include,” “including,” “have,” and “having” are used in the inclusive, open sense, meaning that additional elements may be included. The terms “such as”, “e.g.”, as used herein are non-limiting and are for illustrative purposes only. “Including” and “including but not limited to” are used interchangeably.

[0033] “Or” as used herein should be understood to mean “and / or”, unless the context clearly indicates otherwise.

[0034] “Patient” means a human or non-human subject receiving medical or veterinary care.

[0035] “Pharmaceutically acceptable” or “therapeutically acceptable” refers to a substance which does not interfere with the effectiveness or the biological activity of the active ingredients and which is not toxic to a patient.3724890.00383PCT APPLICATION

[0036] “Pharmaceutically acceptable carrier” is art-recognized, and includes, for example, pharmaceutically acceptable materials, compositions or vehicles, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, involved in carrying or transporting any subject composition from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of a subject composition and not injurious to the patient. In certain embodiments, a pharmaceutically acceptable carrier is non-pyrogenic. Exemplary materials which can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0037] “Pharmaceutical composition” means a formulation comprising an active agent. The word “formulation” means that there is at least one additional ingredient (such as, for example and not limited to, an albumin [such as a human serum albumin or a recombinant human albumin] and / or sodium chloride) in the pharmaceutical composition in addition to a botulinum neurotoxin active ingredient. A pharmaceutical composition is therefore a formulation which is suitable for diagnostic, therapeutic or cosmetic administration to a subject, such as a human patient. The pharmaceutical composition can be: in a lyophilized or vacuum dried condition, a solution formed after reconstitution of the lyophilized or vacuum dried pharmaceutical composition with saline or water, for example, or; as a solution that does not require reconstitution. As stated, a pharmaceutical composition can be liquid, semi-solid, or solid. A pharmaceutical composition can be animal-protein free.

[0038] “Therapeutic formulation” means a formulation that can be used to treat and thereby alleviate a disorder or a disease and / or symptom associated thereof.3724890.00383PCT APPLICATION

[0039] “Therapeutically effective amount” means the level, amount or concentration of an agent needed to treat a disease, disorder or condition without causing significant negative or adverse side effects.

[0040] “Treat,” “treating,” or “treatment” means an alleviation or a reduction (which includes some reduction, a significant reduction a near total reduction, and a total reduction), resolution or prevention (temporarily or permanently) of a symptom, disease, disorder or condition, so as to achieve a desired therapeutic or cosmetic result, such as by healing of injured or damaged tissue, or by altering, changing, enhancing, improving, ameliorating and / or beautifying an existing or perceived symptom, disease, disorder or condition.

[0041] Disclosed compositions and methods include novel treatments for bacterial infections such as those caused by P. aeruginosa. In some embodiments, methods disclosed herein comprise combining: a) photodynamic therapy; with b) an amino acid-supplemented photosensitizer composition to kill the bacterial cells.

[0042] Further embodiments comprise treatments under hyperoxic conditions.Photodynamic Therapy

[0043] Disclosed systems and methods comprise photodynamic therapy (PDT), a treatment involving light-sensitive molecules and a light source to destroy abnormal cells. PDT can be used to treat some skin and eye conditions, as well as certain types of cancer.

[0044] Disclosed PDT embodiments can comprise three components: a photosensitizer, a light source, and tissue oxygen. In embodiments, the wavelength of the light source is appropriate for activating the photosensitizer to produce radicals and / or reactive oxygen species. These are free radicals (Type I) generated through electron abstraction or transfer from a substrate molecule and highly reactive state of oxygen known as singlet oxygen (Type II).

[0045] In some embodiments, the light source comprises an ultraviolet (UV) light source, for example a UVA light source. In some embodiments, the wavelength of the UV light supplied by the UV light source can be one that is appropriate to activate the photosensitizer. If the3724890.00383PCT APPLICATION photosensitizer is riboflavin, the UV light may have a wavelength between about 320 nm and about 390 nm.

[0046] In some embodiments, the light source includes a visible light source or a near infrared light source.

[0047] In some embodiments, the UV light source is a KXL® LED UV system, which emits UVA radiation at a wavelength of 365 nm at an intensity, or treatment power, of 3 mW / cm2to 45 mW / cm2.

[0048] In some embodiments, the intensity of the UVA light used can be about 10 mW / cm2to about 40 mW / cm2, about 10-15 mW / cm2, about 15-20 mW / cm2, about 20-25 mW / cm2, about 25-30 mW / cm2, about 30-35 mW / cm2, about 35-40 mW / cm2, or about 10 mW / cm2, about 15 mW / cm2, about 20 J / cm2, about 25 mW / cm2, about 30 mW / cm2, about 35 mW / cm2, about 40 mW / cm2, or any intensity in a range bounded by any of these values.

[0049] In some embodiments, the total energy of the UV dose can be about 10 J / cm2to about 40 J / cm2, about 10-15 J / cm2, about 15-20 J / cm2, about 20-25 J / cm2, about 25-30 J / cm2, about 30-35 J / cm2, about 35-40 J / cm2, or about 10 J / cm2, about 15 J / cm2, about 17.5 J / cm2, about 20 J / cm2, about 26.25 J / cm2, about 20 J / cm2, about 25 J / cm2, about 30 J / cm2, about 35 J / cm2, about 40 J / cm2, or any total energy in a range bounded by any of these values.

[0050] In embodiments, various UV light intensities / dosages can be applied. In some embodiments, the intensity / dosage can be 20 mW / cm2 / 20 J / cm2, 20 mW / cm2 / 25 J / cm2, 25 mW / cm21 25 J / cm2, 25 mW / cm2 / 30 J / cm2, 30 mW / cm21 30 J / cm2, 30 mW / cm2 / 35 J / cm2, 35 mW / cm2I 35 J / cm2, 35 mW / cm2 / 40 J / cm2, 40 mW / cm2 / 40 J / cm2, or the like. In some embodiments, the intensity / dosage can be about 20 mW / cm2 / 20 J / cm2to about 20 mW / cm2 / 25 J / cm2, about 20 mW / cm2120 J / cm2to about 25 mW / cm2 / 25 J / cm2, about 20 mW / cm2 / 20 J / cm2to about 25 mW / cm2 / 30 J / cm2, about 20 mW / cm21 20 J / cm2to about 30 mW / cm2 / 30 J / cm2, about 20 mW / cm2 / 20 J / cm2to about 30 mW / cm2 / 35 J / cm2, about 20 mW / cm2 / 20 J / cm2to about 35 mW / cm2 / 35 J / cm2, about 20 mW / cm2 / 20 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 20 mW / cm2 / 20 J / cm2to about 40 mW / cm2 / 40 J / cm2, about 20 mW / cm2 / 25 J / cm2to about 25 mW / cm2125 J / cm2, about 20 mW / cm2125 J / cm2to about 25 mW / cm2130 J / cm2, about 20 mW / cm2I 25 J / cm2to about 30 mW / cm2 / 30 J / cm2, about 20 mW / cm2 / 25 J / cm2to about 30 mW / cm2 / 35 J / cm2, about 20 mW / cm2 / 25 J / cm2to about 35 mW / cm2 / 35 J / cm2, about 20 mW / cm2 / 25 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 20 mW / cm2 / 25 J / cm2to about 40 mW / cm2 / 40 J / cm2,3724890.00383PCT APPLICATION about 25 mW / cm2 / 25 J / cm2to about 25 mW / cm2 / 30 J / cm2, about 25 mW / cm2 / 25 J / cm2to about 30 mW / cm2 / 30 J / cm2, about 25 mW / cm2 / 25 J / cm2to about 30 mW / cm2 / 35 J / cm2, about 25 mW / cm2 / 25 J / cm2to about 35 mW / cm2 / 35 J / cm2, about 25 mW / cm2 / 25 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 25 mW / cm2 / 25 J / cm2to about 40 mW / cm2 / 40 J / cm2, about 25 mW / cm2I 30 J / cm2to about 30 mW / cm2 / 30 J / cm2, about 25 mW / cm2 / 30 J / cm2to about 30 mW / cm2 / 35 J / cm2, about 25 mW / cm21 30 J / cm2to about 35 mW / cm2 / 35 J / cm2, about 25 mW / cm2 / 30 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 25 mW / cm2! 30 J / cm2to about 40 mW / cm21 40 J / cm2, about 30 mW / cm2 / 30 J / cm2to about 30 mW / cm2 / 35 J / cm2, about 30 mW / cm2 / 30 J / cm2to about 35 mW / cm2 / 35 J / cm2, about 30 mW / cm2 / 30 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 30 mW / cm2 / 30 J / cm2to about 40 mW / cm2 / 40 J / cm2, about 30 mW / cm2 / 35 J / cm2to about 35 mW / cm2 / 35 J / cm2, about 30 mW / cm2 / 35 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 30 mW / cm2I 35 J / cm2to about 40 mW / cm2 / 40 J / cm2, about 35 mW / cm2 / 35 J / cm2to about 35 mW / cm2 / 40 J / cm2, about 35 mW / cm21 35 J / cm2to about 40 mW / cm2140 J / cm2, about 35 mW / cm2 / 40 J / cm2to about 40 mW / cm2 / 40 J / cm2, or any UV light intensity / dosage in a range bounded by any of these values.

[0051] In some embodiments, the light source can be applied to a patient in need thereof for about 2 minutes to about 40 minutes or more. In some embodiments, the light source can be applied for about 2-5 minutes, about 5-10 minutes, about 10-15 minutes, about 15-20 minutes, about 20-25 minutes, 25-30 minutes, about 30-35 minutes, about 35-40 minutes, or for about 2 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, or any application time in a range bounded by any of these values.

[0052] In some embodiments, the light source can be applied to a patient in need thereof for about 40 minutes or more. In some embodiments, the light source can be applied from about 1 minute to about 60 minutes, about 2 minutes to about 40 minutes, about 5 minutes to about 30 minutes, or any total time range bounded by these values. In some embodiments, the light source can be applied for less than 1 minute.

[0053] Photosensitizers

[0054] Disclosed embodiments comprise photosensitizers. Photosensitizers are molecules which absorb light and transfer energy from the incident light to another nearby molecule. In3724890.00383PCT APPLICATION practice, this typically results in the generation of reactive oxygen species (ROS), which can be deadly to cells.

[0055] In embodiments, the photosensitizer is not active until it is activated by light. After light activation, the photosensitizer becomes reactive to the targeted tissue.

[0056] In embodiments, the photosensitizer can comprise, for example, a cyanine, a phthalocyanine, a hematoporphyrin derivative, a phenothiazines (such as methylene blue and toluidine blue), 5-ALA, a psoralen, a quinone, an anthraquinone, an anthracycline, an anthracenedione, hypericin, xanthene, eosin, a rhodamine, a kryptocyanine, rose bengal, or a combination thereof, and the like. In various embodiments, the photosensitizer, when activated by photoactivating light, can produce reactive oxygen species, such as singlet oxygen.

[0057] In some embodiments, the photosensitizer comprises riboflavin (optionally termed “Rf” herein). In some embodiments, riboflavin may be present in an amount of 100 pM, 200 pM, 300 pM, 400 pM, 500 pM, 600 pM, 700 pM, or the like. In embodiments, about 0.1% w / w to about 0.8% w / w riboflavin is used. In some embodiments, riboflavin is used in an amount of about 0.1- 0.2% w / w, about 0.1-0.3% w / w, about 0.2-0.3% w / w, about 0.3-0.5% w / w, about 0.2-0.4% w / w, about 0.3-0.4% w / w, about 0.4-0.5% w / w, about 0.5-0.8% w / w, about 0.5-0.6% w / w, about 0.6- 0.7% w / w, about 0.7-0.8% w / w, or about 0.1% w / w, about 0.2% w / w, about 0.22% w / w, 0.25% w / w, 0.3% w / w, about 0.4% w / w, about 0.5% w / w, 0.6% w / w, about 0.7% w / w, about 0.8% w / w, or about any weight percentage of riboflavin in a range bounded by any of these values.

[0058] Disclosed photosensitizers are activated by specific wavelengths of light energy, for example UV light, in embodiments, the photosensitizer is activated by UV light of a wavelength of between about 10 nm and about 400 nm, between about 50 nm and about 350 nm, between about 100 nm and about 300 nm, between about 150 nm and 250 nm, or the like.

[0059] In some embodiments, a photosensitizer is activated by visible light of a wavelength between about 380 nm and about 750 nm.

[0060] In some embodiments, a photosensitizer is activated by near-infrared light of a wavelength between about 750 nm to about 2500 nm.

[0061] In some embodiments, the photosensitizer is activated by UVA light of a wavelength of 365 nm, 370 nm, 375 nm, or the like. In some embodiments, an appropriate activation3724890.00383PCT APPLICATION wavelength can be about 350-400 nm, about 355-395 nm, about 360-390 nm, about 365-385 nm, about 370-380 nm, or about 366 nm, about 368 nm, about 370 nm, about 372 nm, about 374 nm, about 376 nm, or any wavelength in a range bounded by any of these values. In embodiments, an appropriate activation wavelength can be, for example, 366 nm, 368 nm, 370 nm, 372 nm, 374 nm, 376 nm, or the like.

[0062] Disclosed embodiments comprise activation of a photosensitizer with an appropriate wavelength of light, also an appropriate intensity and total energy for a given amount of time.

[0063] Amino Acids

[0064] In some embodiments, at least one amino acid is included. In some embodiments, the amino acid can comprise at least one of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine. Some embodiments include a combination of two or more amino acids.

[0065] In some embodiments, the amino acid concentration is up to about 250 mmol / L (mM). In some embodiments, the amino acid concentration is at least about 1 mM. In some embodiments, the amino acid concentration is about 1 mM to about 250 mM, about 10 mM to about 250 mM, about 25 mM to about 200 mM, or about 50 mM to about 150 mM.

[0066] In an embodiment, the amino acid of the composition can be histidine. In some embodiments, the amino acid is L-histidine. In some embodiments, the amount of histidine is up to about 150 mmol / L (mM), about 25 mM to about 150 mM, about 25 mM to about 50 mM, or about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM, about 100 mM, or about 150 mM.

[0067] In some embodiments, the amino acid of the composition can include an aromatic ring. In some embodiments, the amino acid including an aromatic ring is one or more of histidine, phenylalanine, tryptophan, and tyrosine. In some embodiments, the composition includes more than one kind of aromatic amino acid. In some embodiments, the composition includes more than one kind of amino acids including at least one aromatic amino acid.3724890.00383PCT APPLICATION

[0068] Compositions

[0069] Pharmaceutical compositions are described herein. In some embodiments, the pharmaceutical compositions are ophthalmic compositions. In some embodiments, the ophthalmic compositions comprise a photosensitizer selected from the photosensitizers described above. In some embodiments, the ophthalmic compositions comprise an amino acid selected from the amino acids described above. In some embodiments, the ophthalmic compositions comprise a photosensitizer and an amino acid, such as those described above. In some embodiments, the ophthalmic compositions comprise a photosensitizer and an aromatic amino acid. In some embodiments, the ophthalmic compositions comprise riboflavin and an aromatic amino acid. In some embodiments, the ophthalmic compositions comprise riboflavin and histidine.

[0070] Some embodiments include an antibiotic compound. In some embodiments, the antibiotic compound is included in the pharmaceutical ophthalmic compositions described herein. In embodiments wherein the antibiotic compound is provided as a separate composition, it may be administered before, concurrently with, or after the ophthalmic compositions described herein. In some embodiments, any suitable class of antibiotic may be used, such as penicillins, tetracyclines, cephalosporins, quinolones, lincomycins, macrolides, sulfonamides, glycopeptides, aminoglycosides, and carbapenems.

[0071] In some embodiments, wherein the antibiotic compound is an ophthalmic antibiotic, the antibiotic compound may include, but is not limited to, chloramphenicol, erythromycin, azithromycin, ciprofloxacin, neomycin, moxifloxacin, tobramycin, bacitracin, gentamicin, polymyxin B, gatifloxacin, besifloxacin, and levofloxacin. In some embodiments, wherein the antibiotic compound is a topical antibiotic compound, the antibiotic compound may include, but is not limited to, bacitracin, neomycin, polymyxin B, mupirocin, fusidic acid, clindamycin, erythromycin, neomycin / polymyxin B, ozenoxacin, retapamulin, silver sulfadiazine, gentamicin, gentian violet, azelaic acid, 4,4'-sulfonyldianiline or diaminodiphenyl sulfone, mafenide, metronidazole, and sulfacetamide / sulfur. In some embodiments, wherein the antibiotic compound is a systemic antibiotic compound, the systemic antibiotic compound may include, but is not limited to, amoxicillin, penicillin G, flucloxacillin, cefalexin, cefaclor, ceftriaxone, doxycycline, minocycline, tetracycline, gentamicin, tobramycin, amikacin, azithromycin, clarithromycin, erythromycin, ciprofloxacin, levofloxacin, moxifloxacin, vancomycin (glycopeptide), clindamycin (lincosamide), and linezolid (oxazolidinone).3724890.00383PCT APPLICATION

[0072] In some embodiments, the ophthalmic composition is an aqueous solution. In some embodiments, the ophthalmic compositions comprise a buffer. In some embodiments, the ophthalmic compositions are saline solutions. In some embodiments, the ophthalmic composition is a buffered saline solution. In some embodiments, the buffer is an aqueous phosphate buffer. In some embodiments, the ophthalmic composition is a phosphate buffered saline solution. In some embodiments, the phosphate buffer comprises sodium dihydrogen phosphate. In some embodiments, the buffer comprises tris(hydroxymethyl)aminomethane (Tris). In some embodiments, the buffer does not comprise Tris. In some embodiments, the ophthalmic composition is a buffered saline solution wherein the NaCI concentration is about 0.1% w / v to about 5% w / v. In some embodiments, the ophthalmic composition is a buffered saline solution wherein the NaCI concentration is about 0.1 % w / v to about 2% w / v. In some embodiments, the ophthalmic composition is a buffered saline solution wherein the NaCI concentration is about 0.85% w / v.

[0073] In some embodiments, the ophthalmic compositions can comprise topically-applied compositions. In some embodiments, the ophthalmic compositions can be in the form of a cream, a gel, a lotion, a serum, an ointment, or a solution that is applied directly on affected ocular tissue. In some embodiments, the ophthalmic composition is in the form of a cream, a gel, a lotion, a serum, an ointment, or a solution that is applied directed to infected cornea. In some embodiments, the ophthalmic cream, gel, lotion, serum, ointment or solution comprises riboflavin, histidine, and a pharmaceutically-acceptable carrier.

[0074] In some embodiments, the ophthalmic compositions described herein have a pH of about 6.9 to about 9, about 7-9, about 7-8, about 8-9, about 7-7.2, about 7.2-7.4, about 7.4- 7.6, about 7.6-7.8, about 7.8-8, or about 7, about 7.5, about 7.8, about 7.9, or any pH in a range bounded by any of these values.

[0075] In some embodiments, the ophthalmic compositions described herein have a pH less than about 7 (e.g., pH less than or equal to about 6, pH less than or equal to about 5, pH less than or equal to about 4, or pH less than or equal to about 3.5). A pH less than 7 may help to solubilize and / or stabilize components of the ophthalmic compositions (e.g., a photosensitizer or amino acid supplement).3724890.00383PCT APPLICATION

[0076] In some embodiments, the ophthalmic compositions described herein have an osmolarity (mosm / L) of about 200 to about 700, about 200-500, about 200-400, about 300-400, or about any osmolarity in a range bounded by any of these values.

[0077] Methods of Use

[0078] In some embodiments, a method of use is disclosed. In some embodiments, a method of using includes administering an ophthalmic composition described herein directly to a Gramnegative infection. In some embodiments, a method of using includes administering an ophthalmic composition described herein directly to a Gram-positive infection. In some embodiments, a method of using includes administering an ophthalmic composition described herein directly to antimicrobial-resistant bacteria or fungi.

[0079] In some examples, the ophthalmic composition is administered topically. In some embodiments, the ophthalmic composition may be administered directly into an open lesion resulting from a Gram-negative bacteria infection.

[0080] In some embodiments, the ophthalmic composition is administered directly onto a biofilm that is on ocular tissue. The biofilm may include P. aeruginosa encased in a polymeric extracellular matrix. In some embodiments, the biofilm includes other micro-organisms.

[0081] In some embodiments, the ophthalmic composition that is administered to the biofilm is activated with a photoactivating light. The activated composition may be effective in disrupting the biofilm.

[0082] In some embodiments, the ophthalmic composition is administered over a time period of a few seconds to several minutes. In some embodiments, the ophthalmic composition is administered over less than about 60 seconds, less than 30 about seconds, less than about 15 seconds, or less than about 5 seconds. In some embodiments, the ophthalmic composition is administered over about 1-5 minutes, about 5-10 minutes, about 10-15 minutes, about 15-20 minutes, about 20-25 minutes, or about 25-30 minutes.

[0083] In some embodiments, the ophthalmic composition is administered in a hyperoxic environment, for example about 80% O2 to about 95% O2. In some embodiments, the compositions may be administered under normoxic conditions ( / .e., 21% O2). In some3724890.00383PCT APPLICATION embodiments, a hyperoxic environment may be maintained by using goggles or a mask designed for such use.

[0084] In some embodiments, an example mask device for delivering concentrations of oxygen as well as photoactivating light in ophthalmic composition treatments is described in U.S. Patent No. 10,646,372, the contents of which are incorporated entirely herein by reference. For instance, a mask may be placed over the eye(s) to produce a consistent and known oxygen concentration above the surface.

[0085] In some embodiments, as shown in FIG. 10A and FIG. 10B, a mask treatment system 900 includes a mask device 901. The mask device is defined by a body having an anterior side 301a and a posterior side 301b, where the anterior side 301a is positioned distally from the face 3 and the posterior side 301b is positioned proximate to (e.g., against) the face 3. The posterior side 301b may define an opening for receiving the face 3. The body includes an outer wall 301c extending at least between the anterior side 301a and the posterior side 301 b. The wall 301c defines a chamber 302 that extends across the right and left eyes, 1a and 1 b. The chamber 302 includes a right section 302a that is positioned over the right eye 1a and a left section 302b that is positioned over the left eye 1b. Each section, 302a and 302b, facilitates the delivery or application of an ophthalmic composition to the cornea in the respective eye 1a and 1 b. The sections 302a and 302b may be physically divided by an internal wall 303 to reduce any likelihood that treatment of one eye will affect treatment of the other eye. For oxygen delivery or administration to the surface of an eye, the mask device 901 includes a right port 908a leading to the right section 302a and a left port 908b leading to the left section 302b. The right port 908a and the left port 908b may be coupled to the same external oxygen source or different oxygen sources via right and left tubes 909a, 909b, respectively, to deliver the oxygen gas to the chamber 302. If the right section 302a and the left section 302b are not separated by an interior wall 303, the delivered oxygen gas fills the entire chamber 302. Alternatively, if the right section 302a and the left section 302b are separated by an interior wall 303, the oxygen gas delivered from the oxygen source 940 via the port 908a, 908b fills the respective section 302a or 302b. In some embodiments, the oxygen gas is introduced into the chambers with minimal turbulence and backpressure by directing the oxygen gas toward the temples 4a and 4b of the face 3. In particular, the right port 908a leads to a right diffuser 911a that directs the oxygen gas toward the right temple 4a and the left port 908b leads to a left diffuser 911 b to direct the oxygen gas toward the left temple 4b. In addition to minimizing turbulence and backpressure, the oxygen gas is directed so that it does not flow directly over the eyes 1a and 1b, which minimizes undesired3724890.00383PCT APPLICATION drying of the eyes. Once the oxygen gas reaches the temples 4a and 4b, the oxygen gas flows over the eyes 1a and 1 b. Some embodiments include an illumination device 306, which may be positioned relative to the mask device 300 to deliver a first photoactivating light to the right eye 1a or a second photoactivating light to the left eye 1b. In some embodiments, the illumination device 306 may be separately supported, e.g., by a stand, over the right transmission region 304a or the left transmission region 304b. In some embodiments, the illumination device 306 may be supported over both the right transmission region 304a and the left transmission region 304b. Aspects of the mask device 901 may be formed from plastic and / or other suitable materials.

[0086] In some embodiments, a UV light source is applied as described herein, introducing UVA light to the infected area of a patient that has been administered the ophthalmic composition described herein.

[0087] In some embodiments, the composition and treatment described herein is applied to one eye. In some embodiments, the composition and treatment described herein is applied to both eyes. In some embodiments, the composition and treatment described herein is applied to both eyes simultaneously or sequentially.

[0088] Disclosed methods can comprise use in ophthalmic oncology applications, or in conjunction with other anti-microbial therapies, for example nanoparticle adjuvants such as silver salts or titanium oxide.

[0089] An effective method to employ disclosed treatments is to apply the ophthalmic compositions described herein to an open lesion, such as ulcerative keratitis caused by P. aeruginosa. In some embodiments, the treatment is done under a hyperoxic atmosphere of at least 90% O2, with a riboflavin formulation containing 0.1-0.3% w / w riboflavin and an amino acid or acids such as 25-50 mM histidine in phosphate buffered saline (0.85%). This is instilled topically to the cornea over a 10 minute period, followed by administration of UVA light at 365- 375 nm (preferably 372 nm) at an irradiance of 10-40 mW / cm2for a period of 10-30 min.

[0090] An effective method to employ disclosed treatments is to apply disclosed compositions to a treatment area. Typically, the treatment is done under a hyperoxic atmosphere of at least 95% O2, with a riboflavin formulation containing 0.05-0.5% w / w riboflavin and an amino acid or acids such as 15-60 mM amino acid in saline (0.85%). This is applied topically to the treatment area over a 20 minute period, followed by administration of UVA light at 355-385 nm at an irradiance of 10-40 mW / cm2for a period of 10-30 min.3724890.00383PCT APPLICATION

[0091] An effective method to employ disclosed treatments is to apply disclosed compositions to a treatment area. Typically, the treatment is done under a hyperoxic atmosphere of at least 80% O2, with a riboflavin formulation containing 0.1-0.9% w / w riboflavin and an amino acid or acids such as 15-60 mM histidine in saline (0.85%). This is applied topically to the treatment area over a 20 minute period, followed by administration of UVA light at 355-385 nm at an irradiance of 15-45 mW / cm2for a period of 10-30 min.

[0092] An effective method to employ disclosed treatments is to apply disclosed compositions to a treatment area. Typically, the treatment is done under a hyperoxic atmosphere of at least 80% O2, with a formulation containing 0.1-0.6% w / w photosensitizer and an amino acid or acids such as 25-50 mM histidine in saline (0.85%). This is instilled topically to the cornea over a 10 minute period, followed by administration of UVA light at an irradiance of 10-40 mW / cm2for a period of 5-40 min.

[0093] An effective method to employ disclosed treatments is to apply disclosed compositions to a treatment area. Typically, the treatment is done under a hyperoxic atmosphere of at least 80% O2, with a formulation containing 0.1-0.8% photosensitizer and an amino acid or acids such as 20-100 mM amino acid in saline (0.85%). This is applied topically to the treatment area over a 10 minute period, followed by administration of UVA light for a period of 5-50 min.

[0094] An effective method to employ disclosed treatments is to apply disclosed compositions to an open lesion in the eye, such as ulcerative keratitis caused by P. aeruginosa. Typically, the treatment is done under a hyperoxic atmosphere of at least 90% O2, with a riboflavin formulation containing 0.2% w / w riboflavin and 25-50 mM histidine in saline (0.85%). This is instilled topically to the cornea over a 5-10 minute period, followed by administration of UVA light at 365 nm at an intensity / dosage of 10-40 mW / cm2 / 10-40 J / cm2for a period of 5-40 min.

[0095] In some embodiments, UVA irradiation (372 nm) in combination with L-histidine- supplemented riboflavin under a hyperoxic atmosphere causes about 99% P. aeruginosa cell death (see FIG. 2).

[0096] In some embodiments, when P. aeruginosa cell death is studied under normoxic and hyperoxic conditions, a hyperoxic atmosphere provides an additional effect (see FIG. 3).

[0097] In some embodiments, P. aeruginosa cell kill generally increases with UVA dose, with a plateau of cell kill, in some conditions, close to a 20 mW / cm2 / 20 J / cm2dosage (see FIG. 4).3724890.00383PCT APPLICATION

[0098] In some embodiments, P. aeruginosa cell killing under hyperoxic conditions was maximal at >25 mM of histidine (see FIG. 6).

[0099] In some embodiments, extending the UVA irradiation time enhances P. aeruginosa cell killing using 0.2% w / w riboflavin, 50 mM histidine in phosphate buffered saline under hyperoxic conditions. In these embodiments, treatment with a UVA dose of 35 J / cm2with 20 mW / cm2eradicated the cells, while treatment with UVA dose of 20 J / cm2with 10m W / cm2demonstrated 98.9% death (see FIG. 7).

[0100] Some embodiments include a kit for treating an ocular bacterial infection. In some embodiments, the kit comprises ophthalmic compositions comprising a phosphate buffered saline solution of riboflavin and histidine. In some embodiments, the kit further includes an ultraviolet or other photoactivating light source. In other embodiments, the kit may include a mask configured to deliver oxygen to a surface of an eye. In some embodiments, the kit further includes an applicator for applying or administrating the ophthalmic composition. Some embodiments of the kit include instructions for use.EMBODIMENTSEMBODIMENT 1. An ophthalmic composition, comprising: a photosensitizer; and an amino acid, wherein the amino acid comprises alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, or a combination thereof.EMBODIMENT 2. The ophthalmic composition of embodiment 1 , wherein the photosensitizer comprises a cyanine, a phthalocyanine, a hematoporphyrin, a phenothiazine, methylene blue, toluidine blue, 5-ALA, a psoralen, a quinone, an anthraquinone, an anthracycline, an anthracenedione, hypericin, xanthene, eosin, a rhodamine, a kryptocyanine, rose bengal, or a combination thereof.EMBODIMENT 3. The ophthalmic composition of embodiment 1 , wherein the photosensitizer comprises riboflavin.3724890.00383PCT APPLICATIONEMBODIMENT 4. The ophthalmic composition of embodiment 3, wherein the riboflavin is at a concentration of about 0.1 % w / w to about 0.3% w / w.EMBODIMENT S. The ophthalmic composition of embodiment 1 , wherein the amino acid comprises an aromatic side chain.EMBODIMENT 6. The ophthalmic composition of embodiment 1 , wherein the amino acid comprises histidine.EMBODIMENT 7. The ophthalmic composition of embodiment 6, wherein the histidine is at a concentration of between about 25 mM and about 50 mM.EMBODIMENT S. The ophthalmic composition of embodiment 1 , wherein the composition further comprises an antibiotic.EMBODIMENT 9. A method for treating an ophthalmic bacterial infection, comprising: activating an ophthalmic composition comprising a photosensitizer and an amino acid on a surface of an eye of a patient with a photoactivating light.EMBODIMENT 10. The method of embodiment 9, wherein the ophthalmic bacterial infection comprises a Gram-negative bacterial infection.EMBODIMENT 11. The method of embodiment 10, wherein the Gram-negative bacterial infection is caused by P. aeruginosa.EMBODIMENT 12. The method of embodiment 9, wherein the photosensitizer comprises a cyanine, a phthalocyanine, a hematoporphyrin, a phenothiazine, methylene blue, toluidine blue, 5-ALA, a psoralen, a quinone, an anthraquinone, an anthracycline, an anthracenedione, hypericin, xanthene, eosin, a rhodamine, a kryptocyanine, rose bengal, or a combination thereof.EMBODIMENT 13. The method of embodiment 9, wherein the photosensitizer comprises riboflavin.EMBODIMENT 14. The method of embodiment 13, wherein the riboflavin is at a concentration of about 0.1 % w / w to about 0.3% w / w.3724890.00383PCT APPLICATIONEMBODIMENT 15. The method of embodiment 9, wherein the amino acid comprises alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, or a combination thereof.EMBODIMENT 16. The method of embodiment 9, wherein the amino acid comprises histidine.EMBODIMENT 17. The method of embodiment 16, wherein the histidine is at a concentration of between 25 and 50 mM.EMBODIMENT 18. The method of embodiment 9, wherein the ophthalmic bacterial infection comprises ulcerative keratitis.EMBODIMENT 19. The method of embodiment 9, wherein the method is performed under hyperoxic conditions.EMBODIMENT 20. The method of embodiment 9, further comprising administering oxygen to the surface of the eye to be treated.EMBODIMENT 21. The method of embodiment 20, wherein a mask is configured to deliver oxygen to the surface of the eye to be treated.EMBODIMENT 22. The method of embodiment 19, wherein the hyperoxic conditions comprise an oxygen level greater than about 80%.EMBODIMENT 23. The method of embodiment 19, wherein the hyperoxic conditions comprise an oxygen level greater than about 95%.EMBODIMENT 24. A kit for treating an ophthalmic bacterial infection, comprising: an ultraviolet light source; an ophthalmic composition comprising a phosphate buffered saline solution of riboflavin and histidine; and instructions for use.EMBODIMENT 25. A kit for treating an ophthalmic bacterial infection, comprising:3724890.00383PCT APPLICATION a mask configured to deliver oxygen to a surface of an eye; an ophthalmic composition comprising a phosphate buffered saline solution of riboflavin and histidine; and instructions for use.EMBODIMENT 26. A kit for treating an ophthalmic bacterial infection, comprising: a mask configured to deliver oxygen to a surface of an eye; an ultraviolet light source; an ophthalmic composition comprising a phosphate buffered saline solution of riboflavin and histidine; and instructions for use.EMBODIMENT 27. The kit of any one of embodiments 24 to 26, further comprising an applicator for applying the ophthalmic composition.EMBODIMENT 28. A method for treating a biofilm on an ocular surface, the method comprising: activating an ophthalmic composition comprising a photosensitizer and an amino acid on the biofilm with a photoactivating light.EMBODIMENT 29. The method of embodiment 28, the method is performed under hyperoxic conditions.EMBODIMENT 30. The method of embodiment 29, further comprising administering oxygen to the surface of the eye to be treated.EMBODIMENT 31. The method of embodiment 30, wherein a mask is configured to deliver oxygen to the surface of the eye to be treated.3724890.00383PCT APPLICATIONEXAMPLESExample 1

[0101] UVA Treatment with L-Histidine Supplemented Riboflavin on P. aeruginosa Cell Death Assay.

[0102] The effect of UVA-treatment with L-histidine supplemented riboflavin on P. aeruginosa cell death was investigated. P. aeruginosa bacterial strain was purchased from ATCC (27853) and incubated at 37°C. The cells mixed with riboflavin / histidine phosphate buffered saline solution were loaded into a 96 well plate followed by UVA irradiation under a hyperoxic atmosphere (> 90% O2). A sample from the plate well was subsequently spread onto an agar plate and cultured overnight at 37°C to count the colony-forming units (CFU), a readout of surviving bacteria. These experimental procedures are shown in FIG. 1.

[0103] FIG. 2 shows the results of the assay as shown on an agar plate. The assay readout compares the number of colony forming units (CFU). The cell death rate is the CFU of the cells with PDT treatment relative to that of those without treatment. Results showed that UVA irradiation (372 nm) in combination with L-histidine-supplemented riboflavin under a hyperoxic atmosphere causes about 99% P. aeruginosa cell death.

[0104] FIG. 3 shows the effect of varying riboflavin and histidine concentrations on P. aeruginosa cell killing under normoxic and hyperoxic conditions. 25 mM histidine boosts - P. aeruginosa cell death, while the hyperoxic atmosphere provides an additional effect.

[0105] FIG. 4 shows the effect of the UVA overall dose on P. aeruginosa cell killing under hyperoxic conditions. Cell kill declines dramatically with decreased UVA dose, with a plateau of cell kill close to a 20 mW / cm2 / 20 J / cm2dosage.

[0106] FIG. 6 shows the effect of varying histidine concentrations on P. aeruginosa cell killing under hyperoxic conditions. Cell kill was maximal at >25 mM of histidine.

[0107] FIG. 7 shows the effects of different UVA dosages and irradiation times on P. aeruginosa cell killing using 0.2% w / w riboflavin, 50 mM histidine in phosphate buffered saline under hyperoxic conditions. Extending the UVA irradiation time enhances cell killing. The treatment with UVA dose of 35 J / cm2with 20 mW / cm2eradicated the cells, while treatment with UVA dose of 20 J / cm2with 10 mW / cm2demonstrated 98.9% death.3724890.00383PCT APPLICATION

[0108] FIG. 8 shows the effects of varying amounts of riboflavin and histidine in phosphate buffered saline on P. aeruginosa cell killing under normoxic and hyperoxic conditions.Example 2

[0109] Central epithelium was removed (debrided) from New Zealand white rabbit ex vivo eye and soaked in a 0.2% w / w riboflavin, 50 mM histidine phosphate buffered saline composition for 10 min, followed by 20mW / cm2 / 20J / cm2UVA treatment. The eye was then observed for opacity, haze, epithelial damage, etc. Next, the epithelium, stroma, endothelium and lens tissues were dissected. No adverse effects of the riboflavin / histidine formulations were observed. This example is shown in detail in FIGs. 5A-5G.Example 3

[0110] A patient suffering from UK is treated as described herein. A composition comprising histidine and riboflavin in phosphate buffered saline is applied to the affected area, then UVA is applied for 30 minutes under an 80% O2 atmosphere.Example 4

[0111] A patient suffering from a viral infection is treated as described herein. A composition comprising 50 mM histidine and 0.3% w / w riboflavin in phosphate buffered saline is applied to an affected area, then UVA is applied for 20 minutes under a 90% O2 atmosphere. FIG. 9 shows a depiction of this example.

[0112] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical3724890.00383PCT APPLICATION value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0113] The terms “a,” “an,” “the” and similar referents used in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0114] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0115] Certain embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.3724890.00383PCT APPLICATION

[0116] Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term “consisting of” excludes any element, step, or ingredient not specified in the claims. The transition term “consisting essentially of’ limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s). Embodiments of the invention so claimed are inherently or expressly described and enabled herein.

[0117] In closing, it is to be understood that the embodiments of the invention disclosed herein are illustrative of the principles of the present invention. Other modifications that may be employed are within the scope of the invention. Thus, by way of example, but not of limitation, alternative configurations of the present invention may be utilized in accordance with the teachings herein. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

3724890.00383PCT APPLICATIONWhat is claimed is:

1. An ophthalmic composition, comprising: a photosensitizer; and an amino acid, wherein the amino acid comprises alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, or a combination thereof.

2. The ophthalmic composition of claim 1 , wherein the photosensitizer comprises a cyanine, a phthalocyanine, a hematoporphyrin, a phenothiazine, methylene blue, toluidine blue, 5-ALA, a psoralen, a quinone, an anthraquinone, an anthracycline, an anthracenedione, hypericin, xanthene, eosin, a rhodamine, a kryptocyanine, rose bengal, or a combination thereof.

3. The ophthalmic composition of claim 1 , wherein the photosensitizer comprises riboflavin.

4. The ophthalmic composition of claim 3, wherein the riboflavin is at a concentration of about 0.1 % w / w to about 0.3% w / w.

5. The ophthalmic composition of claim 1 , wherein the amino acid comprises an aromatic side chain.

6. The ophthalmic composition of claim 1 , wherein the amino acid comprises histidine.

7. The ophthalmic composition of claim 6, wherein the histidine is at a concentration of between about 25 mM and about 50 mM.

8. The ophthalmic composition of claim 1, wherein the composition further comprises an antibiotic.

9. A method for treating an ophthalmic bacterial infection, comprising: activating an ophthalmic composition comprising a photosensitizer and an amino acid on a surface of an eye of a patient with a photoactivating light.3724890.00383PCT APPLICATION10. The method of claim 9, wherein the ophthalmic bacterial infection comprises a Gramnegative bacterial infection.

11. The method of claim 10, wherein the Gram-negative bacterial infection is caused by Pseudomonas aeruginosa.

12. The method of claim 9, wherein the photosensitizer comprises a cyanine, a phthalocyanine, a hematoporphyrin, a phenothiazine, methylene blue, toluidine blue, 5-ALA, a psoralen, a quinone, an anthraquinone, an anthracycline, an anthracenedione, hypericin, xanthene, eosin, a rhodamine, a kryptocyanine, rose bengal, or a combination thereof.

13. The method of claim 9, wherein the photosensitizer comprises riboflavin.

14. The method of claim 13, wherein the riboflavin is at a concentration of about 0.1% w / w to about 0.3% w / w.

15. The method of claim 9, wherein the amino acid comprises alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, or a combination thereof.

16. The method of claim 9, wherein the amino acid comprises histidine.

17. The method of claim 16, wherein the histidine is at a concentration of between 25 and 50 mM.

18. The method of claim 9, wherein the ophthalmic bacterial infection comprises ulcerative keratitis.

19. The method of claim 9, wherein the method is performed under hyperoxic conditions.

20. The method of claim 9, further comprising administering oxygen to the surface of the eye to be treated.

21. The method of claim 20, wherein a mask is configured to deliver oxygen to the surface of the eye to be treated.3724890.00383 PCT APPLICATION22. The method of claim 19, wherein the hyperoxic conditions comprise an oxygen level greater than about 80%.

23. The method of claim 19, wherein the hyperoxic conditions comprise an oxygen level greater than about 95%.

24. A kit for treating an ophthalmic bacterial infection, comprising: an ultraviolet light source; an ophthalmic composition comprising a phosphate buffered saline solution of riboflavin and histidine; and instructions for use.

25. A kit for treating an ophthalmic bacterial infection, comprising: a mask configured to deliver oxygen to a surface of an eye; an ophthalmic composition comprising a phosphate buffered saline solution of riboflavin and histidine; and instructions for use.

26. A kit for treating an ophthalmic bacterial infection, comprising: a mask configured to deliver oxygen to a surface of an eye; an ultraviolet light source; an ophthalmic composition comprising a phosphate buffered saline solution of riboflavin and histidine; and instructions for use.

27. The kit of any one of claims 24 to 26, further comprising an applicator for applying the ophthalmic composition.

28. A method for treating a biofilm on an ocular surface, the method comprising:3724890.00383PCT APPLICATION activating an ophthalmic composition comprising a photosensitizer and an amino acid on the biofilm with a photoactivating light.

29. The method of claim 28, the method is performed under hyperoxic conditions.

30. The method of claim 29, further comprising administering oxygen to the surface of the eye to be treated.

31. The method of claim 30, wherein a mask is configured to deliver oxygen to the surface of the eye to be treated.

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