Compositions and methods for treating the eye

Abstract

To provide compositions and methods for treating an eye.SOLUTION: The present invention relates to compositions comprising one or more extracts and / or compounds having retinol-like activity and properties, and to methods of using the compositions to treat an eye.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of each of the earlier-filed U.S. Provisional Patent Applications Nos. 62 / 937,450 and 62 / 937,467, filed November 19, 2019, each of which is incorporated by reference in its entirety as if fully set forth herein.

[0002] FIELD OF THE INVENTION The present invention relates to one or more extracts and / or compounds having retinol-like activity and properties, and methods of using the compositions to treat the eye. [Background technology]

[0003] "Dry eye is a multifactorial disease of the ocular surface characterized by loss of tear film homeostasis, with ocular symptoms in which tear film instability and hyperopia, ocular surface inflammation and damage, and neurosensory abnormalities play a pathogenetic role." Craig, JP et al. TFOS DEWS II definition and classification report. Ocul Surf. 2017;15:276-283. Dry eye can result from abnormal or insufficient tear formation and deficient mucin secretion (i.e., corneal inflammation). Dry eye symptoms can manifest as a result of various underlying disorders, such as rheumatoid arthritis, Sjögren's syndrome, systemic lupus erythematosus, and autoimmune disorders that damage the lacrimal (i.e., tear-producing) glands, such as systemic sclerosis and sarcoidosis. Dry eye can also be induced after ophthalmic surgery, such as LASIK™ surgery. Dry eye is estimated to affect more than 13 million individuals in the United States.

[0004] Regardless of the underlying pathology, dry eye is usually accompanied by a rapid breakdown of the preocular tear film, resulting in dehydration of the exposed outer surface. Normal tear formation is necessary to maintain moist conditions of the cornea and conjunctiva, which helps prevent ulcers on both, maintaining corneal transparency. In addition, tears facilitate eyelid movement (e.g., blinking) over the ocular surface and the removal of foreign bodies from the eye. Tears usually contain lysozyme, which is useful for preventing ocular infection. Dry eye may be associated with mild to severe discomfort in the eye. If it occurs over a long period of time, it may cause blurred vision, a foreign body sensation, and / or a burning sensation, and itching. If the condition persists without treatment, it may further lead to corneal ulcers and / or scarring.

[0005] Dry eye symptoms include eye pain or fatigue, increased blinking, and bloodshot eyes. Furthermore, bacteria can enter through scratching, causing infection and, if the scratch is deep enough, affecting a person's vision. In addition to eye strain, causes of dry eye include Sjögren's syndrome, Shuveni-Johnson syndrome, burns and eye injuries, and side effects of hypotenuse medications, psychiatric medications, eye drops to treat glaucoma, and other such medications.

[0006] Eye drops are an effective method for treating dry eye. These eye drops typically contain a dry eye treatment active, a common active ingredient in such eye drops being hyaluronic acid. Hyaluronic acid is a biologically derived polymeric substance with unique properties, such as extremely high water retention, high viscoelasticity, good thickening properties, and good thread-forming ability, and has been used as a moisturizer in topical preparations for treating various skin problems. In the case of dry eye caused by Sjögren's syndrome, where dryness is observed throughout the body, instillation of eye drops containing hyaluronic acid is effective. However, when instilled as eye drops, hyaluronic acid has a relatively short residence time on the cornea, and therefore, the effects of hyaluronic acid eye drops only last about two or three hours, meaning that patients must instill the eye drops more frequently (e.g., five to ten times per day).

[0007] Hyaluronic acid (HA) is produced by corneal epithelial cells in the eye. In particular, significantly higher hyaluronic acid concentrations have been found in the corneas of younger human populations than in older individuals. (See Pacella, E., Pascella, F., De Paolis, G., et al. Glycosaminoglycans in the human cornea: age-related changes. Ophthalmol. Eye Dis. 7:1-5, 2015.)

[0008] Additionally, the tear film is the body's natural defense against dry eye. The tear film contains ocular mucins, essential for maintaining moist ocular surface homeostasis. Mucins are produced, in particular, by corneal epithelial cells in the eye. Mucins are glycoproteins expressed by epithelial tissues of mucosal surfaces. They function as antioxidants and protect tissues by providing lubrication. Mucin genes associated with the tear film include MUC1, MUC2, MUC4, MUC5AC, MUC5B, MUC7, and MUC16.

[0009] Mucins are also useful as antibacterial agents, and both mucins and hyaluronic acid are useful in general wound healing and are essential for maintaining overall eye health.

[0010] Therefore, there is a need for ophthalmic pharmaceutical compositions that would promote and / or improve the production and / or release of hyaluronic acid and / or mucin from and / or in the cornea.

[0011] The inventors have discovered compounds and / or extracts with retinol-like properties and / or benefits that can induce, promote and / or improve the production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or in the cornea.

[0012] Accordingly, one aspect of the present invention relates to compositions comprising a safe and effective amount of one or more compounds and / or extracts having retinol-like properties and / or benefits for treating dry eye.

[0013] Another aspect of the present invention relates to microemulsion compositions comprising a safe and effective amount of one or more compounds and / or extracts with retinol-like properties and / or benefits that induce, promote and / or improve the production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or in the cornea.

[0014] Another aspect of the present invention relates to methods of preventing and treating (e.g., reducing) ocular symptoms associated with dry eye and / or resulting from reduced or low levels of production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or in the cornea by administering a composition comprising a safe and effective amount of one or more compounds and / or extracts having retinol-like properties and / or benefits.

[0015] Specifically, one aspect of the present invention is a composition comprising a safe and effective amount of one or more compounds and / or extracts having retinol-like properties and / or benefits that induce, promote, and / or improve the production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or in the cornea, which composition can be administered to a patient having a concentration of hyaluronic acid in the patient's tears of less than 10 (or about 10) nanograms per milligram of protein, optionally less than 15 (or about 15) nanograms per milligram of protein, optionally less than 20 (or about 20) nanograms per milligram of protein, or optionally less than 25 (or about 25) nanograms per milligram of protein, such that the concentration of hyaluronic acid in the patient's tears is less than 10 (or about 10) nanograms per milligram of protein, optionally less than 15 (or about 15) nanograms per milligram of protein, or optionally less than 20 (or about 20) nanograms per milligram of protein. and optionally increasing from 10 (or about 10) nanograms or more, optionally 15 (or about 15) nanograms or more, optionally 20 (or about 20) nanograms or more, optionally 25 (or about 25) nanograms or more, optionally 30 (or about 30) nanograms or more, optionally 35 (or about 35) nanograms or more, optionally 40 (or about 40) nanograms or more, or optionally 45 (or about 45) nanograms or more to 100 (or about 100) nanograms, optionally 90 (or about 90) nanograms, optionally 80 (or about 80) nanograms, optionally 70 (or about 70) nanograms, or optionally 60 (or about 60) nanograms per milligram of protein.

[0016] In certain embodiments, the above-described concentrations of hyaluronic acid in the patient's tears resulting from the compounds and / or extracts having retinol-like properties and / or benefits are maintained for at least about 2 hours, optionally about 4 hours, optionally about 6 hours, optionally about 8 hours, optionally about 10 hours, optionally about 12 hours, or optionally about 12 to about 24 hours.

[0017] The concentrations of hyaluronic acid detailed above are determined using the Dreyfuss method (described in the definitions below).

[0018] A similar aspect of the present invention relates to a composition comprising a safe and effective amount of one or more compounds and / or extracts with retinol-like properties and / or benefits that induce, promote, and / or improve the production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or in the cornea, wherein this composition can be administered to a patient having a concentration of MUC5AC in tears of less than 6 (or about 6) nanograms per milligram of protein, optionally of 8 (or about 8) nanograms per milligram of protein, thereby elevating the concentration of MUC5AC in tears to 8 (or about 8) nanograms to 15 (or about 15) nanograms per milligram of protein, optionally 9 (or about 9) nanograms to 12 (or about 12) nanograms per milligram of protein.

[0019] In certain embodiments, the above-mentioned concentration of MUC5AC in tears (i.e., 8 (or about 8) nanograms to 15 (or about 15) nanograms, optionally 9 (or about 9) nanograms to 12 (or about 12) nanograms per milligram of protein) resulting from compounds and / or extracts with retinol-like properties and / or benefits that induce, promote and / or improve hyaluronic acid and / or mucin production / release / delivery / excretion from and / or at the cornea is maintained for up to at least about 2 hours, optionally about 4 hours, optionally about 6 hours, optionally about 8 hours, optionally about 10 hours, optionally about 12 hours, or optionally about 12 to about 24 hours.

[0020] The concentration of MUC5AC in tears detailed above is determined using the Uchino method (described in the definitions below).

[0021] A further aspect of the present invention relates to methods for promoting or increasing the rate of healing of wounds in and / or on the eye (e.g., non-dry eye related, ocular trauma, post-operative surgical or non-specific wounds) in a patient by administering a composition comprising a safe and effective amount of one or more compounds and / or extracts with retinol-like properties and / or benefits that induce, promote and / or improve hyaluronic acid and / or mucin production / release / delivery / excretion from and / or at the cornea (i.e., in certain embodiments, increasing the production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or at the cornea above the concentration levels of hyaluronic acid and / or mucin, respectively, that would normally be produced by such patient without (or in the absence of) the administration of a composition comprising a safe and effective amount of one or more compounds and / or extracts with retinol-like properties and / or benefits.

[0022] A still further aspect of the present invention relates to methods of improving the antibacterial properties in the tears (or tear film of the eye) of a patient by administering a composition comprising a safe and effective amount of one or more compounds and / or extracts that induce, promote and / or improve the production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or at the cornea (i.e., in certain embodiments, increasing the production / release / delivery / excretion of mucin from and / or at the cornea above the concentration levels of mucin normally produced by such patient without (or in the absence of) administration of a composition comprising a safe and effective amount of such one or more compounds and / or extracts having retinol-like properties and / or benefits). Summary of the Invention [Means for solving the problem]

[0023] The present invention provides a microemulsion composition for treating the eye, comprising: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating the eye selected from one or more of: a plant extract or source of extract from a plant of the genus Acronychia, Licaria, Calendula and / or Retinol; a bacterial extract or source of extract from the genus Actinomyces; and a compound of formula (I),

[0024] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; and optionally 2-methyl-propa-1,3-diene; ii) an ophthalmically acceptable carrier; The present invention relates to a microemulsion composition wherein the microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0025] The present invention provides a method for preventing or treating symptoms associated with dry eye, optionally in a patient in need of such prevention or treatment, comprising administering a microemulsion composition comprising: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0026] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; and optionally 2-methyl-propa-1,3-diene; ii) an ophthalmically acceptable carrier; The microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0027] The present invention provides a method for treating a patient having reduced or low levels of production / release / delivery / excretion of hyaluronic acid from and / or in the cornea, comprising topically administering to the patient's eye a microemulsion composition, wherein the microemulsion composition: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0028] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; and optionally 2-methyl-propa-1,3-diene; ii) optionally, an ophthalmologically acceptable carrier; The microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0029] The present invention provides a method for preventing or treating symptoms associated with dry eye (optionally in the patient's need for such prevention or reduction in dry eye symptoms) comprising topically administering to a patient a microemulsion composition, wherein the microemulsion composition: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0030] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; and optionally 2-methyl-propa-1,3-diene; ii) optionally, an ophthalmologically acceptable carrier; The microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0031] The present invention provides a method for promoting or increasing the rate of healing of a wound in and / or on a patient's eye (optionally in the patient's need for healing of such eye wound) by administering a composition (i.e., in certain embodiments, increasing the production / release / delivery / excretion of hyaluronic acid from and / or at the cornea above the concentration levels of hyaluronic acid produced by such patient in the absence (or absence) of administration of an emulsion composition comprising a safe and effective amount of one or more compounds and / or extracts having retinol-like properties and / or benefits), wherein the emulsion composition: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0032] [ka] During the ceremony, the dotted lines represent single or double bonds, preferably one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 is a group having 1 to 20 carbon atoms, preferably methyl represents a linear, cyclic, or branched saturated or unsaturated carbonate chain containing a (-CH3) or methylene (=CH2) moiety, a represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; preferably 1 to 10 carbon atoms; more preferably 6 carbon atoms; preferably an aromatic moiety, preferably a phenyl moiety; preferably 2-methyl-propa-1,3-diene; and ii) optionally, an ophthalmologically acceptable carrier; The microemulsion droplets or particles have a maximum dimension of less than 1,500 Å. [Brief explanation of the drawings]

[0033] [Figure 1] 1 shows a bar graph depicting MUC1, MUC4, and MUC16 gene expression induced by lemon aspen extract in corneal epithelial cells. [Figure 2] 1 shows a bar graph illustrating mucin-1 secretion induced by lemon aspen extract in corneal epithelial cells. [Figure 3] 1 shows a bar graph illustrating HAS3 gene expression induced by lemon aspen extract in corneal epithelial cells. [Figure 4] 1 shows a bar graph illustrating HA secretion induced by lemon aspen extract in corneal epithelial cells. [Figure 5] 1 shows a bar graph illustrating HAS3 gene expression induced by lemon aspen extract in corneal epithelial cells. [Figure 6] 1 shows a bar graph illustrating HA secretion induced by lemon aspen extract in corneal epithelial cells. DETAILED DESCRIPTION OF THE INVENTION

[0034] It is believed that one skilled in the art can, based on the description herein, utilize the present invention to its fullest extent. The following specific embodiments are to be construed as merely illustrative, and not limitative of the following disclosure in any way.

[0035] The compositions of the present invention can comprise, consist of, or consist essentially of the elements, steps, and limitations of the present invention described herein, as well as additional or optional ingredients, components, or limitations described herein.

[0036] As used herein, the word "comprising" (and grammatical variations thereof) is used in the inclusive sense of "having" or "including," and not in the exclusive sense of "consisting only of." As used herein, the terms "a" and "the" are understood to include the plural as well as the singular.

[0037] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Furthermore, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety to the extent they are not inconsistent with this specification. As used herein, all percentages are by weight of the total composition unless otherwise specified.

[0038] As used herein, the terms "cornea" or "corneal" include and / or relate to the clear anterior portion of the eye covering the iris, pupil, and anterior chamber, and the layers of the clear anterior portion include the corneal epithelial layer (comprising corneal epithelial cells), Bowman's membrane (also known as the anterior limiting membrane), corneal stroma (also called the substrata), Descemet's membrane (also called the posterior limiting membrane), and the inner corneal endothelium (a simple squamous or hypocuboidal single layer, approximately 5 μm thick, with mitochondria-rich cells).

[0039] As used herein, the phrase "reduced or low levels of hyaluronic acid production / release / release / excretion from and / or within the cornea" refers to a concentration of hyaluronic acid that is less than the concentration of hyaluronic acid in the tear fluid of a healthy (i.e., non-diseased) person, or in certain embodiments, less than 25 (or about 25) nanograms per milligram of protein, as determined using the method described in Dreyfuss JL, Regatieri CV, Coelho B et al., "Altered hyaluronic acid content in tear fluid of patients with adenoviral conjunctivitis. An Acad Bras Cienc. 2015;87(1):455-462." This method (Dreyfuss's method) is reproduced below. ·Sample collection To collect tear fluid, Schirmer strips were placed under the temporal eyelid of each eye without topical anesthetic for 5 minutes. The strips were allowed to dry at room temperature and stored at -20°C until analysis. Tear sample preparation Tear compounds were eluted from Schirmer strips using 100 μL of distilled water and subjected to hyaluronic acid and protein content analysis. Hyaluronic acid measurement Hyaluronan content in tears was measured using a nonisotopic fluorescence assay (Martins Jr, Passerotti CC, Maciel RM, Sampaio Lo, Dietrich CP, and Nader HB. 2003. Practical determination of hyaluronan by a new noncompetitive fluorescence-based assay on serum of normal and cirrhotic patients. Anal Biochem 319:65-72). Eluted tears and standard concentrations of hyaluronan (Sigma, St. Louis, MO) were added to a 96-multiwell plate (FluoroNUNC Maxisorp-microtiterplate, Roskilde, Denmark) precoated with hyaluronan-binding protein. The plate was then sequentially incubated with biotinylated hyaluronan-binding protein and europium-labeled streptavidin (Amershan, Piscataway, NJ). The remaining europium on the solid phase was then released with the fortifying solution, and fluorescence was measured using a time-resolved fluorometer (Perkin-Elmer Life Sciences Wallac Oy, Turku, Finland). Data (counts / s) were automatically processed using the MultiCalc software program (Perkin-Elmer Life Sciences Wallac Oy), and values ​​are expressed as the amount of protein in ng / mg. Protein analysis Total tear protein concentration was determined using a colorimetric assay kit according to the manufacturer's instructions (Protein Assay Kit from Bio-Rad, Hercules, CA). Protein profiles were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) as previously described (see Laemmli, UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685). Briefly, 10 μg of protein from tear samples was applied to a 3-20% linear gradient polyacrylamide gel under reducing conditions. After electrophoresis, the gel was stained with Coomassie Blue (Bio-Rad, Hercules, CA). Each protein band was quantified by densitometry using ImageJ Version 10.2 for Mac software (National Institutes of Health, Bethesda, MD, USA). Results are expressed in arbitrary densitometric units (ADU).

[0040] As used herein, the phrase "reduced or low levels of mucin production / release / release / excretion from and / or within the cornea" refers to a concentration of MUC5AC less than the concentration of MUC5AC in tears of healthy (i.e., non-diseased) individuals, or, in certain embodiments, less than 6 nanograms, optionally less than 8 nanograms, of MUC5AC per milligram of protein, as determined using the method described in Uchino Y, Uchino M, Yokoi N, et al. Alteration of Tear Mucin 5AC in Office Workers Using Visual Display Terminals: The Osaka Study. JAMA Ophthalmol. n 2014; 132(8), 985-992. This method (Uchino's method) is reproduced below. Tear fluid MUC5AC concentration The concentration of secreted mucin MUC5AC in tear samples was quantified by enzyme-linked immunoassay (E90756Hu; USCN Life Science). (See Maker AV, Katabi N, Gonen M, et al. Pancreatic cyst fluid and serum mucin levels predict dysplasia in intraductal papillary mucinous neoplasms of the pancreas. Ann Surg Oncol. 2011;18(1):199-206.) All samples were analyzed according to the manufacturer's instructions. Absorbance was measured at 450 nm, and the standard solution in the kit was recombinant human MUC5AC. Protein concentrations in tear samples were determined using a protein assay reagent kit (BCA Protein Assay Kit; Pierce). MUC5AC concentrations were normalized to tear protein content and expressed as nanograms of MUC5AC protein per milligram of total tear protein.

[0041] As used herein, a composition that is "essentially free" of a component refers to a composition that has about 2% by weight or less of the component, based on the total weight of the composition. Preferably, a composition that is essentially free of a component has about 1% by weight or less of the component, more preferably about 0.5% by weight or less, more preferably about 0.1% by weight or less, more preferably about 0.05% by weight or less, and more preferably about 0.01% by weight or less of the component, based on the total weight of the composition. In certain more preferred embodiments, a composition that is essentially free of a component does not include the component, i.e., the component is not present in the composition.

[0042] As used herein, "microemulsion" refers to emulsions that have one or more of the following properties: i) they form spontaneously or substantially spontaneously when their components are brought into contact, i.e., substantially without the input of energy, e.g., in the absence of heating or the use of high-shear devices or other substantial agitation; ii) they exhibit thermodynamic stability; iii) they are monophasic; iv) they are substantially non-opaque, i.e., transparent or opalescent when viewed by optical microscopy; and / or v) in the undisturbed state, they are optically isotropic, although anisotropic structures may be observable, e.g., using X-ray techniques. Microemulsion particles may be spherical, although other structures, e.g., lamellar, hexagonal, or liquid crystals with isotropic symmetry, are possible. Generally, microemulsions comprise droplets or particles having a maximum dimension (e.g., diameter) of less than 1,500 Å, preferably less than 1,000 Å, and preferably less than 500 Å, but greater than 100 Å. This definition of microemulsions includes self-emulsifying drug delivery systems (SEDDS). SEDDS are isotropic mixtures of oil, surfactant (with or without co-surfactant), and co-solvent that spontaneously emulsify when exposed to aqueous media with gentle agitation. SEDDS can be used to improve the bioavailability of poorly water-soluble drugs via oral administration. The addition of a co-solvent significantly reduces interfacial tension, thereby facilitating the formation of self-emulsifying systems. In doing so, a fluid interfacial membrane is created that is flexible enough to assume the different curvatures required to form microemulsions across a wide range of compositions. Further details regarding SEDDS can be found in U.S. Patent Application Publication No. 2018 / 0036233 A1 (Shabaik et al.), which is incorporated herein by reference in its entirety.

[0043] As used herein, "ophthalmically acceptable" means that the component to which this term refers is suitable for use in contact with tissue (e.g., the soft tissue of the eye or periorbital skin tissue) without undue toxicity, incompatibility, instability, irritation, allergic response, etc. As will be recognized by those skilled in the art, ophthalmically acceptable salts are acidic / anionic or basic / cationic salts.

[0044] As used herein, the term "safe and effective amount" refers to an amount of the disclosed extracts, compounds, or compositions that is sufficient to induce, promote, and / or improve mucin production / release / delivery / excretion from and / or in one or more layers of the cornea, but low enough to avoid serious side effects. A safe and effective amount of a compound, extract, or composition will vary depending, for example, on the age, health, and environmental exposure of the end user, the duration and nature of the treatment, the specific extract, ingredient, or composition used, the specific pharmaceutically acceptable carrier used, and similar factors.

[0045] The term "retinol-like properties and / or benefits" means properties and / or benefits induced by retinol.

[0046] In certain embodiments, the invention disclosed herein can be practiced in the absence of any compound or element (or group of compounds or elements) not specifically disclosed herein.

[0047] Generally, IUPAC nomenclature is used herein in accordance with the following definitions of terms.

[0048] The term "Ci_8 alkyl," whether used alone or as part of a substituent, refers to a saturated aliphatic branched or straight-chain monovalent hydrocarbon radical having 1 to 8 carbon atoms. For example, "Ci_8 alkyl" specifically includes the radicals methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, tert-butyl, 1-butyl, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 1-octyl, 2-octyl, 3-octyl, and the like. The term may also refer to the corresponding alkyldiyl radical. Alkyl and alkyldiyl radicals may be attached to a core molecule through a terminal carbon atom or through an atom within the chain. Similarly, any number of substituent variables may be attached to an alkyl or alkyldiyl radical, if available valences permit.

[0049] The term “C 1~4 "Alkyl," whether used alone or as part of a substituent group, refers to a saturated aliphatic branched or straight-chain monovalent hydrocarbon radical or alkyldiyl linking group having the specified number of carbon atoms, where the radical is derived by removing one hydrogen atom from a carbon atom and the alkyldiyl linking group is derived by removing one hydrogen atom from each of two carbon atoms in the chain. 1~4 "Alkyl" refers to a radical having from 1 to 4 carbon atoms in a straight or branched arrangement. For example, "C 1~4 "Alkyl" specifically includes radicals such as methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, tert-butyl, 1-butyl, etc. Alkyl and alkyldiyl radicals can be attached to the core molecule through a terminal carbon atom or through an atom within the chain. Similarly, any number of substituent variables can be attached to an alkyl or alkyldiyl radical, available valences permitting.

[0050] The term “C 2~4"Alkenyl" refers to an alkenyl radical having 2 to 4 carbon atoms. For example, it specifically includes the radicals ethenyl, propenyl, allyl (2-propenyl), butenyl, and the like. As noted above, alkenyl radicals may also be attached to a core molecule and may be further substituted, if specified.

[0051] The term "halo," by itself or in combination with other terms, means a halogen atom, such as fluoro, chloro, bromo, or iodo.

[0052] The term "substituted" refers to a core molecule in which one or more hydrogen atoms have been replaced with a substituent, in the amount permitted by available valences. Substitution is not limited to the core molecule, but can also occur on substituent radicals, thereby rendering the radical a linking group.

[0053] The term "independently selected" refers to two or more substituents that may be selected from a group of substituent variables, where the selected substituents may be the same or different.

[0054] The term "dependently selected" refers to one or more substituent variables that are specified in a specified combination for substitution within a core molecule (e.g., a variable that refers to a group of substituents listed in a table of compounds).

[0055] Acceptable salts derived from inorganic bases include, for example, sodium or potassium salts, etc. Acceptable salts derived from organic bases include, for example, salts formed with primary, secondary, or tertiary amines, etc.

[0056] Compounds / extracts that exhibit retinol-like properties and / or benefits The present invention provides a safe and effective amount of a compound and / or extract having retinol-like properties and / or benefits for use in treating dry eye, selected from one or more of a plant extract or source of an extract from a plant of the genus Acronychia, Licaria, Calendula and / or Retinol, a bacterial extract or source of an extract from the genus Actinomyces, and a compound of formula (I):

[0057] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain having 1 to 20 carbon atoms; optionally, a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain having 1 to 20 carbon atoms; optionally, 1 to 10 carbon atoms; optionally, 6 carbon atoms; optionally, an aromatic moiety, optionally, a phenyl moiety; and optionally, 2-methyl-propa-1,3-diene.

[0058] Plant extracts of Acronychia species, Licoricea species, Calendula species and / or Reilly-wood species In certain embodiments, the compound / extract exhibiting retinol-like properties and / or benefits comprises an extract or source of extracts of the plants Acronychia, Licaria, Calendula, and / or Reilly-Ossop. The extract of Acronychia, Licaria, Calendula, or Reilly-Ossop, or a source of such extract, is obtained from a plant of the genus Acronychia, Licaria, Calendula, or Reilly-Ossop.

[0059] Plants of the genus Acronichia from which extracts useful in the present invention can be obtained include, for example, Acronichia averans, Acronichia acidula (also referred to herein as lemon aspen), Acronichia acronychioides, Acronichia acuminate, Acronichia bauerennii, Acronichia coolichilum, Acronichia crassipetala, Acronichia eugelensis, Acronichia imperforate, Acronichia laevis, Acronichia spp. Examples of suitable Acronychia species include Acronychia laurifolia, Acronychia litoralis, Acronychia oblongifolia, Acronychia octanara, Acronychia parviflora, Acronychia pausiflora, Acronychia pedunculata, Acronychia pubescens, Acronychia species (Batavia dauns), Acronychia suberosa, Acronychia vestita, Acronychia wilcoxinia, and combinations of two or more thereof. In one embodiment, the extract used in the present invention is obtained from Acronychia acidula.

[0060] Plants of the genus Licaria from which extracts useful in the present invention can be obtained include, for example, Licaria vernicosa, Licaria brittoniana, Licaria canella, Licaria cubensis, Licaria velutina, and Licaria triandra, as well as combinations of two or more thereof. There are approximately 40 reported species of Licaria, all of which are endemic to Central and South America. In one embodiment, the extract used in the present invention is obtained from Licaria vernicosa.

[0061] There are approximately 15 to 20 reported species of Calendula, found in Southwest Asia, Western Europe, Macaronesia, and the Mediterranean basin. Plants of the genus Calendula from which extracts useful in the present invention can be obtained include, for example, Calendula arvensis (field marigold), Calendula maderensis (field marigold), and pot marigold, as well as combinations of two or more thereof. In one embodiment, the extract used in the present invention is obtained from Calendula.

[0062] The genus Trigonella includes 36 known species of Trigonella from which extracts useful in the present invention can be obtained, including, for example, Trigonella fenugreek, Trigonella balansae, Trigonella corniculata, Trigonella maritima, Trigonella spicata, Trigonella caerulea, Trigonella occulta, Trigonella polycerata, Trigonella calyceras, Trigonella cretica, and combinations of two or more thereof. Fenugreek, or herb fenugreek, is the most well-known Trigonella species. In one embodiment, the extract used in the present invention is obtained from fenugreek.

[0063] In a particular embodiment, the extract used in the present invention is a mixture of extracts obtained from plants of the Acronychia, Licaria, Calendula or Marshmallow plant genera.

[0064] Extracts of Licaria vernicosa useful in the present invention can be obtained from the Baruch S. Blumer Institute, Doylestown, PA (formerly known as IVHR). In a specific embodiment, one extract was obtained from the wood of the plant (E2) and the second extract was derived from the roots of the plant (E3). Within the nomenclature of the IHVR collection, the two extracts are labeled IHVR_40256_G10=X-005348-001E002 and IHVR_40256_E10=X-005346-001M002, respectively. The wood and roots of Licaria vernicosa (Mez) costelum can be collected in Guyana. 504.3 g of dried ground woody plant material can be extracted with enough methanol to dry in a vacuum to yield 10.54 g of crude methanol extract (E2) of X-005348-001E002. 403.8 g of dried ground root material can be extracted with enough methanol to dry in a vacuum to yield 18.11 g of crude methanol extract (E3) of X-005346-001M002.

[0065] In certain embodiments, the Acronychia and / or Likaria extracts useful in the present invention comprise a compound having formula II:

[0066] [ka] During the ceremony, R1 is C1~C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 selected from the group consisting of alkynyl, and C3-C8 cycloalkyl or aryl; R2 is selected from the group consisting of hydrogen, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl or aryl, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC2-C6 alkynyl, -OC3-C8 cycloalkyl or aryl, thiol, -SC1-C6 alkyl, -SC2-C6 alkenyl, -SC2-C6 alkynyl, SC3-C8 cycloalkyl or aryl, -NR4C1-C6 alkyl, -NR4C2-C6 alkenyl, -NR4C2-C6 alkynyl, and -NR4C3-C8 cycloalkyl or aryl; R3 is selected from -CO2H, -CO2R4 or an isosteric equivalent of a carboxy group, and R4 is C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl or aryl; Y is -(CH2-CH2)-, -(CH=CH)-, or -(C=C)-, or a pharmaceutically acceptable salt thereof.

[0067] In certain embodiments, the Acronychia and / or Likaria extracts useful in the present invention comprise a compound having formula II:

[0068] [ka] During the ceremony, R1 is C5~C 16 Alkyl, C5-C 16 Alkenyl, C5-C 16Alkynyl, more preferably C5-C, including, for example, farnesyl 16 alkenyl, R2 is selected from the group consisting of hydrogen, hydroxyl, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC2-C6 alkynyl, -OC3-C8 cycloalkyl, more preferably hydrogen, hydroxyl, -OC1-C6 alkyl, even more preferably hydrogen or -OC1-C3 alkyl; R3 is selected from -CO2H, -CO2R4, and R4 is selected from C1-C6 alkyl or an isosteric equivalent of a carboxy group; Y is -(CH2-CH2)- or -(CH=CH)-, or an ophthalmologically acceptable salt thereof.

[0069] In certain embodiments, at least one of the compounds of formula II is present in the extract of Acronychia and / or Likaria at a concentration of greater than or equal to 1% (or about 1%) to about 20%, or optionally about 7% (or about 7%) to about 10% (or about 10%) by weight of the extract of Acronychia and / or Likaria.

[0070] In certain embodiments, the compound of formula II useful in the present invention is in the form of an acid or alkyl ester selected from 3-(4-farnesyloxyphenyl)-propionic acid, 3-(4-farnesioxy-3-hydroxyphenyl)-propionic acid, 3-(4-farnesioxy-3-methoxyphenyl)-propionic acid, alkyl esters thereof, particularly ethyl esters thereof, and combinations of two or more thereof.

[0071] In certain embodiments, the compound of formula II useful in the present invention is 3-(4-farnesyloxyphenyl)-propionic acid and / or its ethyl ester.

[0072] In certain embodiments, the compound of formula II useful in the present invention is 3-(4-farnesyloxy-3-hydroxyphenyl)-propionic acid and / or its ethyl ester.

[0073] In certain embodiments, the compound of formula II useful in the present invention is 3-(4-farnesyloxy-3-methoxyphenyl)-propionic acid and / or its ethyl ester.

[0074] Compounds and extracts derived from Acronychia species are described in US Pat. No. 9,220,928, which is incorporated herein by reference in its entirety.

[0075] In certain embodiments, 3-(4-farnesyloxyphenyl)-propionic acid and / or its ethyl ester is present in the extract of Acronychia and / or Likaria at a concentration of greater than or equal to 1% (or about 1%) to about 20%, or optionally from about 7% (or about 7%) to about 10% (or about 10%) by weight of the extract of Acronychia and / or Likaria.

[0076] For embodiments in which the method includes applying such an extract, any of a variety of extracts of Acronychia and / or Likaria may be used. The extract may be obtained from any part of the plant, including the fruit, seeds, bark, leaves, flowers, roots, and wood.

[0077] In certain embodiments, the extract is obtained from the fruit of the plant. Suitable extracts of the fruits, seeds, bark, leaves, flowers, roots, and wood of Acronychia and / or Likaria may be obtained using conventional methods, including, but not limited to, crushing, maceration, pressing, squeezing, grinding, centrifugation, and / or direct extraction of materials from the biomass, including cold irrigation, stirring / distillation, microwave-assisted extraction, supercritical / subcritical CO2 compressed gas extraction, pressurized solvent extraction, accelerated solvent extraction, pressurized or regular hot water extraction, surfactant-assisted heated hot water extraction, oil extraction, membrane extraction, Soxhlet extraction, by the Kinzaji distillation / extraction and / or process disclosed in, for example, U.S. Patent Nos. 7,442,391, 7,473,435, and 7,537,791 (Integrated Botanical Technologies, LLC), which are incorporated herein by reference, or by other methods such as solvent extraction. Any of a variety of solvents may be used in methods involving solvent extraction, including polar solvents, non-polar solvents, or combinations of two or more thereof.

[0078] Suitable polar solvents include polar inorganic solvents such as water, polar organic solvents such as alcohols and corresponding organic acids, e.g., C1-C8 alcohols including methanol, ethanol, propanol, butanol, etc., and organic acids including acetic acid, formic acid, propanoic acid, etc., polyols and glycols including C1-C8 polyols / glycols, etc., and combinations of two or more thereof. Suitable nonpolar solvents include nonpolar organic solvents such as alkanes including C1-C8 alkanes, cycloalkanes including C1-C8 alkanes, alkyl ethers including C1-C8 alkyl ethers, petroleum ethers, ketones including C1-C8 ketones, ketones including methylene chloride, ethyl acetate, xylene, toluene, chloroform, vegetable oil, mineral oil, etc. In another embodiment, extraction can be obtained by supercritical fluid extraction with or without the above nonpolar solvents or polar modifiers such as C1-C8 alcohols, water, C1-C8 polyols / glycols, or C1-C8 organic acids.

[0079] In one embodiment, the extract comprises an extract of Acronychia acidula. In another embodiment, the extract of the present invention comprises a combination of polar and non-polar extracts from the fruit of Acronychia acidula. In another embodiment, the extract of the present invention comprises an alcoholic or glycolic extract of the fruit of Acronychia acidula.

[0080] In one embodiment, the extract comprises an extract of Licaria vernicosa. In another embodiment, the extract of the present invention comprises a combination of polar and non-polar extracts from the Licaria vernicosa tree or the root of Licaria vernicosa. In another embodiment, the extract of the present invention comprises an alcoholic extract of the Licaria vernicosa tree or the root of Licaria vernicosa.

[0081] In another embodiment, the extract of the present invention comprises a polar extract prepared by extracting the fruit of Acronychia acidula, the tree of Licaria vernicosa, or the root of Licaria vernicosa using a polar solvent including water, a C1-C8 alcohol, a C1-C8 polyol, a C1-C8 glycol, and a combination of two or more thereof. In certain embodiments, the extract is extracted using one or more C1-C4 alcohols, C1-C4 polyols, and / or C1-C4 glycols. In certain embodiments, the extract is prepared using a solvent including methanol, ethanol, or a combination thereof with or without water. In another embodiment, the extract is a polar extract extracted from the fruit of Acronychia acidula using a combination of alcohol and water. In yet another embodiment, the extract is a polar extract extracted from the crushed tree of Licaria vernicosa or the crushed root of Licaria vernicosa using methanol.

[0082] In yet another embodiment, the extract comprises a non-polar extract prepared by extraction from the fruit of Acronychia acidula, the tree of Licaria vernicosa, or the root of Licaria vernicosa using a non-polar solvent comprising one or more C1-C8 alkanes, C1-C8 cycloalkanes, C1-C8 alkyl ethers, C1-C8 alkyl esters, and / or chloroform, more preferably one or more C1-C8 alkanes, C1-C8 alkyl esters, and / or chloroform. In yet another embodiment, the non-polar extract is extracted from the fruit of Acronychia acidula, the tree of Licaria vernicosa, or the root of Licaria vernicosa using hexane, ethyl acetate, chloroform, or a mixture of two or more thereof. In yet another embodiment, the non-polar extract is extracted from the fruit of Acronychia acidula using ethyl acetate.

[0083] In one embodiment, the extract comprises an extract of calendula. In another embodiment, the extract of the present invention comprises a combination of a polar extract and a non-polar extract of calendula petals. In another embodiment, the extract of the present invention comprises a non-polar extract of calendula petals.

[0084] In one embodiment, the extract comprises an extract of fenugreek. In another embodiment, the extract of the present invention comprises a combination of polar and non-polar extracts of fenugreek leaves. In another embodiment, the extract of the present invention comprises a non-polar extract of fenugreek leaves.

[0085] In yet another embodiment, the calendula and / or Reishi extract is a non-polar extract prepared using a non-polar solvent comprising one or more C1-C8 alkanes, C1-C8 cycloalkanes, C1-C8 alkyl ethers, C1-C8 alkyl esters and / or chloroform, more preferably one or more C1-C8 alkanes, C1-C8 alkyl esters and / or chloroform.

[0086] In yet another embodiment, the calendula and / or Reitang Root extract is a non-polar extract prepared using hexane, ethyl acetate, chloroform, or a mixture of two or more thereof, hi yet another embodiment, the extract is a non-polar extract prepared using ethyl acetate.

[0087] In one embodiment, the plant extract may be obtained by extracting cell cultures of various plants, including cell cultures of the genera Acronychia, Licaria, Calendula, and / or Trellis. The cell cultures extracted to obtain the plant extracts used in the present invention may be in any form, including suspension cell cultures, etc.

[0088] Calendula and fenugreek extracts are available from Caithness Biotechnologies Ltd, UK (http: / / www.caithnessbiotechnes.com). These extracts are part of the Phytotitre Natural Product Library, available for purchase. Alternatively, the extracts can be obtained using a preparation method described by Caithness Biotechnologies Ltd. as non-polar and prepared with a mixture of methanol and methylene chloride. For a detailed description, see the webpage http: / / caithnessbiotechnlogies.com / contact_t.html. In a typical extraction, a pre-weighed amount of dried biomass powder is suspended and stirred in a mixture of methanol / methylene chloride (1:1) overnight at ambient temperature. The suspension is then filtered, and the filtrate is dried under reduced pressure to a solvent-free residue.

[0089] In certain embodiments, extracts of Acronychia, Licoricea, Calendula, and / or Retinol are present in the compositions of the present invention in an amount of from about 0.001% to about 10%, optionally from about 0.001% to about 5%, or optionally from about 0.01% to about 1% by weight of the composition.

[0090] Actinomyces bacterial extracts exhibiting retinol-like properties and / or benefits In certain embodiments, the compound / extract exhibiting retinol-like properties and / or benefits is or includes a bacterial extract of the genus Actinomyces. Actinomyces bacteria include many fully characterized species, some of which are less well characterized, such as those collected from the United States and labeled as species A5640. Samples of this bacterium are collected, grown in culture, and extracted. The extract is currently part of a natural product collection under the stewardship of the Baruch S. Blumer Institute, Doylestown, Pennsylvania (formerly known as the Institute of Heabc Virus Research Labs (IHVR)). Within the nomenclature of the IHVR collection, the extract is labeled as IHVR_39565_F7.

[0091] In one embodiment, the extract used in the present invention is obtained from an Actinomyces species capable of producing a chemical composition similar to that produced by extract A5640. In another embodiment, the bacterium is collected in the United States and the strain is identical to the previously assigned species A5640.

[0092] In certain embodiments, the extract of Actinomyces is present in the composition in an amount of from about 0.001% to about 10%, optionally from about 0.001% to about 5%, or optionally from about 0.01% to about 1% by weight of the composition.

[0093] The compound(s) of formula (I) exhibit retinoic properties and / or benefits. In certain embodiments, the compound / extract exhibiting retinol-like properties and / or effects is or comprises a compound of formula (I):

[0094] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; R2 is a linear, cyclic, or branched, saturated or unsaturated carbonate chain having 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain having 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; preferably 2-methyl-propa-1,3-diene.

[0095] In certain embodiments, the compound of formula (I) is Li + , Na + , K. + , Ca 2+ , or Mg 2+ This includes, but is not limited to, the corresponding salts of metal ions such as:

[0096] In certain embodiments, the compound of formula I is

[0097] [ka] (2E,4E,6E)-7-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)-3-methylocta-2,4,6-trienoic acid or

[0098] [ka] 4-(1-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)vinyl)benzoic acid, and in each case derivatives thereof that exhibit retinoid-like activity. These compounds are referred to below in Examples 8 and 9 as Compound 1 and Compound 2, respectively.

[0099] The present invention relates to compounds of formula I, such as (2E,4E,6E)-7-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)-3-methylocta-2,4,6-trienoic acid and 4-(1-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)vinyl)benzoic acid, derivatives thereof, and mixtures thereof, which exhibit retinoid-like activity.

[0100] Any mixture of compounds / extracts / extract sources that exhibit retinol-like properties and / or benefits may also be used.

[0101] The compounds and extracts of Formula I are described in U.S. Patent Publication No. 2019 / 0091122, which is incorporated herein by reference in its entirety.

[0102] In certain embodiments, the compound(s) of Formula I are present in the compositions of the invention in an amount of from about 0.0001% to about 20%, optionally from about 0.001% to about 10%, optionally from about 0.01% to about 5%, or optionally from about 0.2 to about 2% by weight of the composition. In yet other embodiments, the compound(s) of Formula I are present in the compositions of the invention in an amount of from about 0.0001% to about 1%, optionally from about 0.001% to about 1%, or optionally from about 0.01 to about 1% by weight of the composition.

[0103] The present inventors have discovered that compounds and / or extracts with retinol-like properties and / or benefits can enhance and / or improve the delivery / excretion of hyaluronic acid from corneal epithelial cells.

[0104] Permeation enhancers In certain embodiments, the compositions of the present invention optionally include a permeation enhancer.

[0105] Suitable permeation enhancers include saponin, polyoxyethylene, polyoxyethylene ethers of fatty acids such as polyoxyethylene 4-, 9-, 10-, and 23-lauryl ether, polyoxyethylene 10- and 20-cetyl ether, polyoxyethylene 10- and 20-stearyl ether, polyoxyethylene sorbitans such as sorbitan monooleate, sorbitan monolaurate, polyoxyethylene monolaurate, polyoxyethylene sorbitan monolaurate, surfactants such as decamethonium, decamethonium bromide, and dodecyltrimethylammonium bromide; natural polyacids (e.g., citric acid), phosphates (e.g., disodium pyrophosphate), phosphonates, bisphosphonates (e.g., etridronic acid), aminocarboxylic acids (e.g., ethylenediaminetetraacetic acid (EDTA) and disodium EDTA), and ethylenediamine-N , N'-disuccinic acid (EDDS); bile salts and acids such as cholic acid, deoxycholic acid, glycocholic acid, glycodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, sodium cholate, sodium glycocholate, glycocholate, sodium deoxycholate, sodium taurocholate, sodium glycodeoxycholate, sodium taurodeoxycholate, chenodeoxycholic acid, and ursodeoxycholic acid; fusidic acid derivatives, including saponin EDTA, fusidic acid, polyoxyethylene 9-lauryl ether, polyoxyethylene 20-stearyl ether, glycocholate, or mixtures of any of the foregoing, glycyrrhizic acid, and ammonium glycyrrhizinate, either alone or in combination.

[0106] The concentration of permeation enhancer administered should be the minimum amount necessary to sufficiently increase absorption of the compound and / or extract through the ocular mucosa or other barrier membrane.Generally from 0.01% (or about 0.01%), optionally from 0.05% (or about 0.05%), optionally from 0.1% (or about 0.1%), optionally from 0.15% (or about 0.15%), optionally from 0.2% (or about 0.2%), optionally from 0.25% (or about 0.25%) to 2% (or about 2%), optionally to 2.5% (or about 2.5%), optionally to 3% (or about 3%), optionally to 3.5% (or about 3.5%), optionally to 4% (or about 4%) of the total composition (w / v) up to 4.5% (or about 4.5%), optionally up to 5% (or about 5%), optionally up to 5.5% (or about 5.5%), optionally up to 6% (or about 6%), optionally up to 6.5% (or about 6.5%), optionally up to 7% (or about 7%), optionally up to 7.5% (or about 7.5%), optionally up to 8% (or about 8%), optionally up to 8.5% (or about 8.5%), optionally up to 9% (or about 9%), optionally up to 9.5% (or about 9.5%), optionally up to 10% (or is up to about 10%), optionally up to 10.5% (or about 10.5%), optionally up to 11% (or about 11%), optionally up to 11.5% (or about 11.5%), optionally up to 12% (or about 12%), optionally up to 12.5% ​​(or about 12.5%), optionally up to 13% (or about 13%), optionally up to 13.5% (or about 13.5%), optionally up to 14% (or about 14%), optionally up to 14.5% (or about 14.5%), optionally up to 15% (or about 15%), optionally up to 1 Concentrations ranging from up to 5.5% (or about 15.5%), optionally up to 16% (or about 16%), optionally up to 16.5% (or about 16.5%), optionally up to 17% (or about 17%), optionally up to 17.5% (or about 17.5%), optionally up to 18% (or about 18%), optionally up to 18.5% (or about 18.5%), optionally up to 19% (or about 19%), optionally up to 19.5% (or about 19.5%), optionally up to 20% (or about 20%) are useful in the compositions of the present invention.

[0107] Ophthalmically acceptable carrier The compositions of the present invention may also comprise aqueous, oil-in-water emulsion, water-in-oil emulsion carrier, oil-in-water microemulsion, or water-in-oil microemulsion carrier. The carrier is ophthalmically acceptable. Useful oil-in-water carriers and water-oil carriers can be found in U.S. Patent Publication No. 20030165545A1 and U.S. Patent Nos. 9,480,645, 8,828,412, and 8,496,976, each of which is incorporated herein by reference in its entirety.

[0108] In certain embodiments, the compositions of the present invention comprise an emulsion, optionally a self-emulsifying emulsion comprising an oil component, such as, for example, one or more oils, and, without limitation, mineral oil and / or one or more conventionally known and / or commercially available oils suitable for use in the present invention; a surfactant component comprising three or more surfactants; and an aqueous component comprising an aqueous phase. In addition, multiple additional components may be included in the composition. The compositions of the present invention are substantially non-toxic and / or non-irritating and / or non-damaging to the eye and may provide protective functions for ocular cells and tissues.

[0109] One or more oils or oily substances are used to form the composition. Any suitable oil or oily substance or combination of oils or oily substances may be used, as long as such oil and / or oily substance is effective in the composition, and the combination does not cause any substantial or significant adverse effects on the human or animal to which the composition is administered, or on the treated contact lens, or on the wearer of the treated contact lens. The oily component may be, for example, without limitation, a higher fatty acid glyceride, such as castor oil, corn oil, sunflower oil, etc., and mixtures thereof. The oily component may also include one or more non-polar oils, such as mineral oil, silicone oil, etc., and mixtures thereof.

[0110] The ophthalmically acceptable carrier (or composition of the present invention) may optionally include one or more additional excipients and / or one or more additional active ingredients, examples of which are described below.

[0111] Excipients used in ophthalmic compositions include, but are not limited to, demulcents, tonicity agents, preservatives, chelating agents, buffers (other than and in addition to the organic acids of the present invention), and surfactants. Other excipients include solubilizers, stabilizers, comfort enhancers, polymers, emollients, pH adjusters (other than and in addition to the organic acids of the present invention), and / or lubricants. Any of a variety of excipients may be used in the compositions of the present invention, including water, mixtures of water and water-miscible solvents, e.g., vegetable or mineral oils containing 0.5% to 5% of a non-toxic, water-soluble polymer, natural products such as agar and acacia, starch derivatives such as starch acetate and hydroxypropyl starch, and other synthetic products such as polyvinyl alcohol, polyvinylpyrrolidone, polyvinylmethyl ether, polyethylene oxide, and preferably cross-linked polyacrylic acid, as well as combinations thereof.

[0112] Demulcents or smoothing agents for use with embodiments of the present invention include, but are not limited to, cellulose derivatives (such as hydroxyethyl cellulose, methyl cellulose, hypromellose, or mixtures thereof), hyaluronic acid or its salts (such as sodium hyaluronate), tamarind seed extract, glycerin, polyvinylpyrrolidone, polyethylene oxide, polyethylene glycol, propylene glycol, and polyacrylic acid, and mixtures thereof. In certain embodiments, one or more of hyaluronic acid, propylene glycol, tamarind seed extract, glycerin, and / or polyethylene glycol 400 are the demulcents or smoothing agents. In certain embodiments, the demulcent or smoothing agent is selected from hyaluronic acid, tamarind seed extract, or mixtures thereof.

[0113] The compositions of the present invention are ophthalmologically suitable for application to a subject's eyes. The term "water-soluble" typically refers to an aqueous formulation in which the vehicle is greater than about 50% by weight, more preferably greater than about 75% by weight, and particularly greater than about 90% by weight water. In certain embodiments, the compositions of the present invention are also essentially free of compounds that are irritating to the eye. In certain embodiments, the compositions of the present invention are essentially free of free fatty acids and C1-C4 alcohols. In certain embodiments, the compositions of the present invention comprise less than 40% by weight (or about 40%) of the total composition, optionally less than 35% by weight (or about 35%), optionally less than 30% by weight (or about 30%), optionally less than 25% by weight (or about 25%), optionally less than 20% by weight (or about 20%), optionally less than 15% by weight (or about 15%), optionally less than 10% by weight (or about 10%), or optionally less than 5% by weight (or about 5%) of non-alcohol, organic excipients or solvents. These drops may be delivered from a single-dose ampoule, which may preferably be sterile and thus obviate the need for a bacteriostatic component of the formulation. Alternatively, the drops may be delivered from a multi-dose bottle, which may preferably be equipped with a device that extracts any preservatives from the composition as it is delivered, such as devices well known in the art.

[0114] In certain embodiments, the compositions of the present invention are isotonic or slightly hypotonic to combat any tear hypertonicity caused by evaporation and / or disease. This may require a tonicity agent to bring the osmolality of the formulation closer to levels at or near 210-320 milliosmoles per kilogram (mOsm / kg). Compositions of the present invention generally have an osmolality in the range of 220-320 mOsm / kg, or optionally, an osmolality in the range of 235-300 mOsm / kg. Ophthalmic compositions can generally be formulated as sterile aqueous solutions.

[0115] The osmolality of the compositions of the present invention can be adjusted to a value compatible with the intended use of the composition using an isotonicity agent.For example, the osmolality of the composition can be adjusted to approximate the osmolality of normal tears, which corresponds to about 0.9 w / v% sodium chloride in water.Examples of suitable isotonicity adjusting agents include, but are not limited to, sodium, potassium, calcium, and magnesium chloride, dextrose, glycerin, propylene glycol, mannitol, sorbitol, etc., and mixtures thereof.In one embodiment, a combination of sodium chloride and potassium chloride is used to adjust the tonicity of the composition.

[0116] The compositions of the present invention can also be used to administer pharmaceutically active compounds. Such compounds include, but are not limited to, glaucoma medications, analgesics, anti-inflammatory and anti-allergic drugs, and antibacterial drugs. More specific examples of pharmaceutically active compounds include betaxolol, timolol, pilocarpine, carbonic anhydrase inhibitors and prostaglandins, dopaminergic antagonists, postoperative antihypertensive drugs, anti-infective drugs such as para-aminoclonidine (apraclonidine) salts, ciprofloxacin, moxifloxacin, and tobramycin, non-steroidal and steroidal anti-inflammatory drugs, such as naproxen, diclofenac, nepafenac, suprofen, ketorolac, tetrahydrocortisol, and dexamethasone, dry eye medications such as PDE4 inhibitors, and anti-allergic drugs such as H1 / H4 inhibitors, H4 inhibitors, olopadine, or mixtures thereof.

[0117] It is also contemplated that the concentrations of the components comprising the formulations of the present invention may vary, and one of skill in the art will understand that concentrations may vary depending on the addition, substitution, and / or omission of components in a given formulation.

[0118] In certain embodiments, the compositions of the present invention may have a pH compatible with their intended use, often in the range of 4 (or about 4) to 10 (or about 10), optionally 6 (or about 6) to 8 (or about 8), optionally 6.5 (or about 6.5) to 7.5 (or about 7.5), or optionally 6.8 (or about 6.8) to 7.2 (or about 7.2).

[0119] In certain embodiments, various conventional buffers may be used, such as phosphate, borate, citrate, acetate, histidine, Tris, Bis-Tris, etc., and mixtures thereof. Borate buffers include boric acid and its salts, such as sodium borate or potassium borate. Potassium tetraborate or potassium metaborate, which form boric acid or a salt of boric acid in solution, may also be used. Hydrated salts, such as sodium borate decahydrate, may also be used. Phosphate buffers include phosphoric acid and its salts, such as M2HPO4 and MH2PO4, where M is an alkali metal, such as sodium or potassium. Hydrated salts may also be used. In one embodiment of the present invention, Na2HPO4.7H2O and NaH2PO2.H2O are used as buffers. The term phosphate also includes compounds that form phosphoric acid or a salt of phosphoric acid in solution. In addition, organic counterions for the above buffers may also be used. The concentration of the buffering agent generally varies from about 0.01 to 2.5 w / v %, more preferably from about 0.05 to about 0.5 w / v %.

[0120] In certain embodiments, the viscosity of the compositions of the present invention ranges from about 1 to about 500 cps, optionally from about 10 to about 200 cps, or optionally from about 10 to about 100 cps, as measured using a TA Instrument AR 2000 rheometer. The TA Instrument AR 2000 rheometer should be used with the AR2000 flow test method in the TA Rheological Advantage software using a 40 mm steel plate geometry, and the viscosity range is from 0 sec -1 ~200 seconds -1 should be obtained by measuring the steady-state flow rate controlling the shear rate.

[0121] In certain embodiments, the compositions of the present invention are useful as and in the form of eye drop solutions, eyewash solutions, contact lens lubricating and / or rewetting solutions, sprays, mists, or any other mode of administering the composition to the eye.

[0122] The compositions of the present invention may also be useful as and in the form of a packing solution for contact lenses. In certain embodiments, as a packing solution, the compositions of the present invention may be sealed in a blister package and may be suitable for sterilization.

[0123] Examples of blister packaging and sterilization techniques are disclosed in the following references, which are incorporated herein by reference in their entireties: U.S. Patent Nos. D435,966, D4,691,820, D5,467,868, D5,704,468, D5,823,327, D6,050,398, D5,696,686, D6,018,931, D5,577,367, and D5,488,815. This part of the manufacturing process presents an alternative method for treating the ophthalmic device with an antiallergic agent, i.e., adding the antiallergic agent to a solution before sealing the package, followed by sterilization of the package. This is a preferred method for treating the ophthalmic device with an antiallergic agent.

[0124] Sterilization can be carried out at different temperatures and times. Preferred sterilization conditions range from about 100°C for about 8 hours to about 150°C for about 0.5 minutes. More preferred sterilization conditions range from about 115°C for about 2.5 hours to about 130°C for about 5.0 minutes. The most preferred sterilization conditions are about 124°C for about 18 minutes.

[0125] When used as a loading solution, the compositions of the present invention may be aqueous solutions. Typical solutions include, but are not limited to, saline, other buffer solutions, and deionized water. In certain embodiments, the loading solution is an aqueous solution of deionized water or saline containing salts, including, but not limited to, sodium chloride, sodium borate, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, or the corresponding potassium salts of the same acids. These components generally combine to form a buffer containing an acid and its conjugate base, so that the addition of an acid and a base results in relatively little change in pH. In certain embodiments, the pH of the loading solution is as described above. The buffer may further include 2-(N-morpholino)ethanesulfonic acid (MES), sodium hydroxide, 2,2-bis(hydroxymethyl)-2,2',2''-nitrilotriethanol, n-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid, citric acid, sodium citrate, sodium carbonate, sodium bicarbonate, acetic acid, sodium acetate, ethylenediaminetetraacetic acid, and the like, and combinations thereof. Preferably, the solution is phosphate buffered saline or deionized water. Particularly preferred solutions contain from about 500 ppm to about 18,500 ppm sodium borate, most particularly preferably about 1000 ppm sodium borate.

[0126] If any of the components incorporated into the loading solution are susceptible to oxidative degradation, an agent may be added to stabilize the loading solution containing such components. Examples of such "oxidative stabilizers" include, but are not limited to, EDTA, Dequest, Desferal, silica, chitin derivatives such as chitosan, cellulose, and its derivatives, N,N,N',N',N'',N''-hexane(2-pyridyl)-1,3,5-tris(aminomethyl)benzene, and certain macrocyclic ligands such as crown ether, knot, and catenane-containing ligands. See David A. Leigh et al., Angew. Chem Int. Ed., 2001, 40, No. 8, pp. 1538-1542 and Jean-Claude Chambron et al., Pure & Appl. Chem, 1990, Vol. 62, No. 6, pp. 1027-1034. Oxidation stabilizers include 2,2',2'',6,6',6''-hexane-(1,1-dimethylethyl)4,4',4''-[(2,4,6-trimethyl-1,3,5-benzenetriyl)-trimethylene]-triphenol (Irganox 1330), 1,3,5-tris[3,5-di(1,1-dimethylethyl)4-hydroxybenzyl]-1H,3H,5H-1,3,5-triazine-2,4,6 trione, pentaerythrityl tetrakis[3-[3,5-di(1,1-dimethylethyl)-4-hydroxyphenyl]-propionate], octadecyl-3-[3,5-di(1,1-dimethylethyl)-4-hydroxyphenyl]-propionate, tris[2,4-di(1,1-dimethylethyl)-phenyl]-phosphine Other compounds that inhibit oxidation may be included, such as a compound selected from the group consisting of thiophene, 2,2'-di(octadecyloxy)-5,5'-spirobi(1,3,2-dioxaphosphorinane), dioctadecyl disulfide, dioctadecyl-3,3'-thiodipropionate, dioctadecyl-3,3'-thiodipropionate, butylhydroxytoluene, ethylene bis[3,3-di[3-(1,1-dimethylethyl)-4-hydroxyphenyl]butyrate], and mixtures thereof.A preferred oxidative stabilizer is diethylenetriaminepentaacetic acid ("DTPA") or a salt of DTPA, such as CaNaDTPA, ZnNaDTPA, and CaDTPA. See U.S. Patent Application Publication No. 60 / 783,557, filed March 17, 2006, entitled "Methods for Stabilizing Oxidatively Unstable Pharmaceutical Compositions," and its corresponding nonprovisional application, which are incorporated herein by reference in their entireties. In certain embodiments, the concentration of the oxidative stabilizer in the solution is from about 2.5 μmol / L to about 5000 μmol / L, optionally from about 20 μmol / L to about 1000 μmol / L, optionally from about 100 μmol / L to about 1000 μmol / L, or optionally from about 100 μmol / L to about 500 μmol / L.

[0127] In certain embodiments, the compositions of the present invention are formulated for administration at any administration frequency, including once a week, once every 5 days, once every 3 days, once every 2 days, twice a day, three times a day, four times a day, five times a day, six times a day, eight times a day, hourly, or more frequently.Such administration frequencies are also maintained for varying durations depending on the user's treatment needs.The duration of a particular treatment regimen can vary from a single administration to a regimen that lasts for months or years.Those skilled in the art will be familiar with determining the treatment regimen for a particular indication.

[0128] Compositions and products containing such compositions of the present invention can be prepared using methods well known to those skilled in the art. [Example]

[0129] Any compositions of the present invention as illustrated in the following examples are intended to illustrate specific embodiments of the compositions of the present invention and are not intended to limit the invention. Other modifications can be made by those skilled in the art without departing from the spirit and scope of the present invention.

[0130] The following test methods were used in the examples. Example 1 The composition of lemon aspen extract showed an increase in MUC1, MUC4 and MUC16 gene expression in human epicorneal 3D tissue when treated with culture medium.

[0131] EpiCorneal 3D human tissue was purchased from MatTek Company (Ashland, Massachusetts, USA). Upon receipt, the epicorneal 3D human tissues were incubated overnight in MatTek assay medium according to the manufacturer's instructions. The epicorneal 3D human tissues were divided into three treatment groups with at least three tissues per group. Lemon aspen extract was added to the culture medium containing two human epicorneal tissues from each treatment group to produce a medium concentration of 0.001% or 0.01% (w / v), respectively. The epicorneal tissues in all four treatment groups were allowed to incubate for two days. Lemon aspen extract was extracted and supplied by Southern Cross Botanicals (Knockroe, New South Wales, Australia). After two days of incubation, gene expression of mucin 1 (MUC1), mucin 4 (MUC4), and mucin 16 (MUC16) was analyzed. After 2 days of incubation, the human epicorneal 3D tissue was cut in half, and one half of the tissue was lysed in 350 μL of lysis buffer consisting of 100 parts RLT buffer (RNeasy Mini Kit, Qiagen, Valencia, CA) to 1 part 2-mercaptoethanol. RNA was extracted from the solution using the RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions, and the RNA was eluted in 25 μL of RNase-free water.

[0132] Reverse transcription (RT) was performed using the Applied Biosystems High Capacity Reverse Transcription Kit (ThermoFisher Scientific, Bridgewater, NJ). Gene expression assays sold under the trade name TAQMAN for mucin-1 (MUC1), mucin-4 (MUC4), and mucin-16 (MUC16) polymerase (RNA) II polypeptide A (POLR2A), and Master Mix were purchased from ThermoFisher Scientific (Bridgewater, NJ). qPCR analysis was performed using TaqMan® Master Mix (ThermoFisher Scientific, Bridgewater, NJ) and run on a real-time PCR system sold under the trade name QUANTSTUDIO 7 Flex System (ThermoFisher Scientific, Bridgewater, NJ). Expression of the MUC1, MUC4, and MUC16 genes was normalized to expression of the human POLR2A housekeeping gene. Fold changes were calculated relative to the untreated control (UT) and a two-step, two-sample Student's t test (Microsoft Office Excel 2007; Microsoft, Redmond, WA, USA) was performed. The results are shown in Figure 1.

[0133] Example 2 The composition of lemon aspen extract showed increased mucin-1 secretion in human epicorneal 3D tissue when treated in culture medium.

[0134] EpiCorneal 3D human tissue was purchased from MatTek Company (Ashland, Massachusetts, USA). Upon receipt, the epicorneal 3D human tissues were incubated overnight in MatTek assay medium according to the manufacturer's instructions. The epicorneal 3D human tissues were divided into three treatment groups with at least three tissues per group. Lemon aspen extract was added to the culture medium containing two human epicorneal tissues from each treatment group to produce a medium concentration of 0.001% or 0.01% (w / v), respectively. The epicorneal tissues in all four treatment groups were allowed to incubate for two days. After two days, the culture medium was collected to measure mucin 1 secretion using a human mucin-1 (CA15-3) enzyme-linked immunosorbent assay (ELISA) kit (EHMUC1, ThermoFisher Scientific, Bridgewater, NJ) according to the manufacturer's protocol. To assess activity, colorimetric changes were measured using a microplate reader (SpectraMax M2E, Molecular Devices, Sunnyvale, CA, USA). The assay employed a standard enzyme-linked immunosorbent assay, and a linear correlation existed between the mucin-1 concentration in the sample and the colorimetric change. A standard curve was generated to plot the corresponding mucin-1 concentration, with the mucin-1 concentration on the x-axis and the absorbance on the y-axis. The results are shown in Figure 2.

[0135] Example 3 Extracts of lemon aspen (Acronychia acidula) showed increased induction of HAS3 gene expression in human epicorneal 3D tissue when applied topically.

[0136] EpiCorneal 3D human tissue was purchased from MatTek Company (Ashland, Massachusetts, USA). Upon receiving the epicorneal 3D human tissue, it was incubated overnight in MatTek assay medium according to the manufacturer's instructions. The epicorneal 3D human tissue was divided into two treatment groups with at least three tissues per group. A solution of lemon aspen extract (0.1% w / v extract in a 30 / 70 (v / v) polypropylene glycol / ethanol vehicle) was topically applied to the human epicorneal tissue in one of the treatment groups. The epicorneal tissues in both treatment groups were incubated for two days. After two days, gene expression of hyaluronan synthase 3 (HAS3) was analyzed. HAS3 is an enzyme involved in the synthesis of hyaluronic acid. Lemon aspen extract was extracted and supplied by Southern Cross Botanicals (Knockroe, New South Wales, Australia). After 2 days of incubation, the human epicorneal 3D tissue was cut in half, and one half of the tissue was lysed in 350 μL of lysis buffer consisting of 100 parts RLT buffer (RNeasy Mini Kit, Qiagen, Valencia, CA) to 1 part 2-mercaptoethanol. RNA was extracted from the solution using the RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions, and the RNA was eluted in 25 μL of RNase-free water.

[0137] Reverse transcription (RT) was performed using the Applied Biosystems High Capacity Reverse Transcription Kit (ThermoFisher Scientific, Bridgewater, NJ). Gene expression assays sold under the trade name TAQMAN for hyaluronan synthase 3 (HAS3), polymerase (RNA) II polypeptide A (POLR2A), and Master Mix were purchased from ThermoFisher Scientific (Bridgewater, NJ). qPCR analysis was performed using TaqMan® Master Mix (ThermoFisher Scientific, Bridgewater, NJ) and run on a real-time PCR system sold under the trade name QUANTSTUDIO 7 Flex System (ThermoFisher Scientific, Bridgewater, NJ). Expression of the HAS3 gene was normalized to the expression of the human POLR2A housekeeping gene. Fold changes were calculated relative to the untreated control (UT) using a two-step, two-sample Student's t test (Microsoft Office Excel 2007; Microsoft, Redmond, WA, USA). The results are shown in Figure 3.

[0138] Example 4 Lemon aspin extract, when applied topically, showed increased bioactivity of hyaluronic acid in human epicorneal 3D tissue.

[0139] The epicorneal 3D human tissue was purchased from MatTek Company (Ashland, Massachusetts, USA). Upon receiving the epicorneal 3D human tissue, it was incubated overnight in MatTek assay medium according to the manufacturer's instructions. The epicorneal 3D human tissue was divided into two treatment groups with at least three tissues per group. A solution of lemon aspen extract (0.1% w / v extract in a 30 / 70 (v / v) polypropylene glycol / ethanol vehicle) was topically applied to the human epicorneal tissue in one of the treatment groups. The epicorneal tissues in both treatment groups were allowed to incubate for two days. After two days, the culture medium was collected to measure hyaluronic acid (HA) secretion using an HA enzyme-linked immunosorbent assay (ELISA) kit (K-1200, Echelon, Salt Lake City, Utah, USA) according to the manufacturer's protocol. The lemon aspen extract used in this Example 4 is the same as the lemon aspen extract used in Example 3. To assess activity, the colorimetric change was measured at 405 nm using a microplate reader (SpectraMax M2E, Molecular Devices, Sunnyvale, CA, USA). Because this assay employs a competitive enzyme-linked immunosorbent assay, there is an inverse correlation between the HA concentration in the sample and the colorimetric change. A standard curve was generated to plot the corresponding HA concentration, with HA concentration on the x-axis and absorbance on the y-axis. The results are shown in Figure 4.

[0140] Example 5 Lemon aspen extract showed increased induction of HAS3 gene expression in human epicorneal 3D tissues when treated in culture medium.

[0141] EpiCorneal 3D human tissue was purchased from MatTek Company (Ashland, Massachusetts, USA). Upon receiving the epicorneal 3D human tissues, they were incubated overnight in MatTek assay medium according to the manufacturer's instructions. The epicorneal 3D human tissues were divided into four treatment groups with at least three tissues per group. Lemon aspen extract was added to the culture medium containing three human epicorneal tissues from each treatment group to produce medium concentrations of 0.0001%, 0.001%, or 0.01% (w / v), respectively. The epicorneal tissues in all four treatment groups were allowed to incubate for two days. The lemon aspen extract used in this Example 5 was extracted and supplied by Southern Cross Botanicals (Knockroe, New South Wales, Australia), but using a different extraction process than that used to extract the extract used in Example 3. After 2 days of incubation, hyaluronan synthase 3 (HAS3) gene expression was analyzed as follows: Human epicorneal 3D tissue was cut in half, and one half of the tissue was lysed in 350 μL of lysis buffer consisting of 100 parts RLT buffer (RNeasy Mini Kit, Qiagen, Valencia, CA) and 1 part 2-mercaptoethanol. RNA was extracted from the solution using the RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions, and the RNA was eluted in 25 μL of RNase-free water.

[0142] Reverse transcription (RT) was performed using the Applied Biosystems High Capacity Reverse Transcription Kit (ThermoFisher Scientific, Bridgewater, NJ). Gene expression assays sold under the trade name TAQMAN for hyaluronan synthase 3 (HAS3), polymerase (RNA) II polypeptide A (POLR2A), and Master Mix were purchased from ThermoFisher Scientific (Bridgewater, NJ). qPCR analysis was performed using TaqMan® Master Mix (ThermoFisher Scientific, Bridgewater, NJ) and run on a real-time PCR system sold under the trade name QUANTSTUDIO 7 Flex System (ThermoFisher Scientific, Bridgewater, NJ). Expression of the HAS3 gene was normalized to the expression of the human POLR2A housekeeping gene. Fold changes were calculated relative to the untreated control (UT) and a two-step, two-sample Student's t-test (Microsoft Office Excel 2007; Microsoft, Redmond, WA, USA) was performed. The results are shown in Figure 5.

[0143] Example 6 lemon a vinegar P The extract of hyaluronic acid showed increased bioactivity of hyaluronic acid in human epicorneal 3D tissues when treated with culture medium.

[0144] The epicorneal 3D human tissues were purchased from MatTek Company (Ashland, Massachusetts, USA). Upon receiving the epicorneal 3D human tissues, they were incubated overnight in MatTek assay medium according to the manufacturer's instructions. The epicorneal 3D human tissues were divided into four treatment groups with at least three tissues per group. Lemon aspen extract was added to the culture medium containing three human epicorneal tissues from each treatment group to produce medium concentrations of 0.0001%, 0.001%, or 0.01% (w / v), respectively. The epicorneal tissues in all four treatment groups were allowed to incubate for two days. The lemon aspen extract used in this Example 6 is the same as the lemon aspen extract used in Example 5. After 2 days, the culture medium was collected to measure hyaluronic acid (HA) secretion using an HA enzyme-linked immunosorbent assay (ELISA) kit (K-1200, Echelon, Salt Lake City, UT, USA) according to the manufacturer's protocol. To assess activity, the colorimetric change was measured using a microplate reader (SpectraMax M2E, Molecular Devices, Sunnyvale, CA, USA). Because this assay employs a competitive enzyme-linked immunosorbent assay method, there is an inverse correlation between the HA concentration in the sample and the colorimetric change. A standard curve was generated to plot the corresponding HA concentration, with HA concentration on the x-axis and absorbance on the y-axis. The results are shown in Figure 6.

[0145] Example 7 Table 1 illustrates the components of the formulations (as shown in Formulations 7A-7D), ie, which components may be combined as described below using conventional mixing techniques.

[0146] [Table 1] * Tonicity can be adjusted to 280~290mOsm / Kg ** Can be adjusted to pH 7.2 ***Optionally, in amounts up to 100% w / w

[0147] For Examples 7A-7D, sodium hyaluronate may be supplied by CONTIPRO AS (Dolný, Dobrócz, Czech Republic).

[0148] For Examples 7A-7D, lemon aspen extract (Abacross™ Acronychia acidula fruit extract) may be supplied by SOUTHERN CROSS BOTANICALS (Knockroe, New South Wales, Australia).

[0149] For Examples 7A-7D, polysorbate 20 may be supplied by Merck KGaA (Darmstadt, Germany).

[0150] For Examples 7A-7D, Polysorbate 80 may be supplied by Merck KGaA (Darmstadt, Germany).

[0151] For Examples 7A and 7B, polyethylene glycol 400 may be supplied by Clariant Produkte (Burgkirchen, Germany).

[0152] For Examples 7A-7D, boric acid may be supplied by Merck KGaA (Darmstadt, Germany).

[0153] For Examples 7A-7D, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0154] For Examples 7A-7D, sodium chloride may be supplied by Caldic (Düsseldorf, Germany).

[0155] For Examples 7A-7D, potassium chloride may be supplied by KGaA (Darmstadt, Germany).

[0156] For Examples 7A-7D, calcium chloride dihydrate may be supplied by Merck KGaA (Darmstadt, Germany).

[0157] For Examples 7A-7D, magnesium chloride may be supplied by KGaA (Darmstadt, Germany).

[0158] For Examples 7A-7D, Polyquaternium-42 (33% aqueous solution) may be supplied by DSM BIOMEDICAL (Berkeley, Calif., USA).

[0159] For Examples 7A-7D, sodium chlorite dihydrate may be supplied by Oxychem (Wichita, Kansas, USA).

[0160] For Examples 7A-7D, 1N sodium hydroxide can be supplied by VWR (Radnor, PA, USA).

[0161] For Examples 7A-7D, 1N hydrochloric acid can be supplied by VWR (Radnor, PA, USA).

[0162] Solution 7A can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 50 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 1.0g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 2.0 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.5 grams of polyethylene glycol 400, 6.0 grams of boric acid, 0.05 grams of sodium borate, 1.0 grams of potassium chloride, 0.06 grams of calcium chloride dihydrate, 0.06 grams of magnesium chloride, and 0.0015 grams of Polyquaternium 42 (aqueous solution). 7. While continuing to mix, add 0.14 grams of sodium chlorite dihydrate and mix to dissolve. 8. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. The pH of the formulation is adjusted to 7.2 using 91N sodium hydroxide and / or 1N hydrochloric acid. 10. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure complete homogeneity. 11. Filter the solution using a 0.22 micrometer filter.

[0163] Solution 7B can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 2g of Polysorbate 80 and 10g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 0.1g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 3.0 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.5 grams of polyethylene glycol 400, 6.0 grams of boric acid, 0.05 grams of sodium borate, 1.0 grams of potassium chloride, 0.06 grams of calcium chloride dihydrate, 0.06 grams of magnesium chloride, and 0.0015 grams of Polyquaternium 42 (aqueous solution). 7. While continuing to mix, add 0.14 grams of sodium chlorite dihydrate and mix to dissolve. 8. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. Adjust the pH of the formulation to 7.2 using 9.1 N sodium hydroxide and / or 1 N hydrochloric acid. 10. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure complete homogeneity. 11. Filter the solution using a 0.22 micrometer filter.

[0164] Solution 7C can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 20 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 1.0g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.5 grams of sodium citrate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 6.0 grams boric acid, 0.05 grams sodium borate, 1.0 gram potassium chloride, 0.06 grams calcium chloride dihydrate, 0.06 grams magnesium chloride, and 0.0015 grams Polyquaternium 42 (aqueous solution). 7. While continuing to mix, add 0.14 grams of sodium chloride dihydrate and mix to dissolve. 8. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. Adjust the pH of the formulation to 7.2 using 9.1 N sodium hydroxide and / or 1 N hydrochloric acid. 10. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure complete homogeneity. 11. Filter the solution using a 0.22 micrometer filter.

[0165] Solution 7D can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 2g of Polysorbate 10 and 50g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 0.1g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.5 grams of sodium citrate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 6.0 grams boric acid, 0.05 grams sodium borate, 1.0 gram potassium chloride, 0.06 grams calcium chloride dihydrate, 0.06 grams magnesium chloride, and 0.0015 grams Polyquaternium 42 (aqueous). 7. While continuing to mix, add 0.14 grams of sodium chloride dihydrate and mix to dissolve. 8. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. Adjust the pH of the formulation to 7.2 using 9.1 N sodium hydroxide and / or 1 N hydrochloric acid. 10. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure complete homogeneity. 11. Filter the solution using a 0.22 micrometer filter.

[0166] Example 8 Table 2 shows the ingredients of formulations of the present invention (as shown in Formulations 8A-8D), which can be combined using conventional mixing techniques as described below.

[0167] [Table 2] * Tonicity can be adjusted to 280~290mOsm / Kg ** Can be adjusted to pH 7.2 *** Optionally, the amount to be added is 100% w / w

[0168] For Examples 8C and 8D, sodium hyaluronate may be supplied by CONTIPRO AS (Dolný, Dobrócz, Czech Republic).

[0169] For Examples 8C and 8D, tamarind seed polysaccharide may be supplied by INDENA (Milan, Italy).

[0170] For Examples 8C and 8D, lemon aspen extract (Abacross™ Acronychia acidula fruit extract) may be supplied by SOUTHERN CROSS BOTANICALS (Knockroe, New South Wales, Australia).

[0171] For Examples 8A-8D, polysorbate 20 may be supplied by Merck KGaA (Darmstadt, Germany).

[0172] For Examples 8A-8D, Polysorbate 80 may be supplied by Merck KGaA (Darmstadt, Germany).

[0173] For Examples 8C-8D, polyethylene glycol 400 may be supplied by Clariant Produkte (Burgkirchen, Germany).

[0174] For Examples 8A-8D, boric acid may be supplied by Merck KGaA (Darmstadt, Germany).

[0175] For Examples 8A-8D, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0176] For Examples 8A-8D, sodium chloride may be supplied by Caldic (Düsseldorf, Germany).

[0177] For Examples 8A-8D, potassium chloride may be supplied by KGaA (Darmstadt, Germany).

[0178] For Examples 8A-8D, Hypromellose E3 2910 may be supplied by DOW CHEMICAL (Plaquemine, Louisiana, USA).

[0179] For Examples 8A-8D, glycerin may be supplied by Emery Oleochemicals GmbH (Düsseldorf, Germany).

[0180] For Examples 8A-8D, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0181] For Examples 8A-8D, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0182] For Examples 8A-8D, sodium lactate may be supplied by Merck KGaA (Darmstadt, Germany) as sodium lactate (50% aqueous solution).

[0183] For Examples 8A-8D, glucose may be supplied by Roquette Freres (Restron, France).

[0184] For Examples 8A-8D, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0185] For Examples 8A-8D, ascorbic acid may be supplied by DSM NUTRITIONAL Products (DRAKEMYRE, Scotland, UK).

[0186] For Examples 8A-8D, Polyquaternium 42 can be supplied by DSM BIOMEDICAL (Berkeley, Calif.) as Polyquaternium 42 (33% aqueous solution).

[0187] For Examples 8A-8D, diisodium edate may be supplied by Merck NV / SA (Overijse, Belgium).

[0188] For Examples 8A-8D, 1N sodium hydroxide can be supplied by VWR (Radnor, PA, USA).

[0189] For Examples 8A-8D, 1N hydrochloric acid can be supplied by VWR (Radnor, PA, USA).

[0190] For Examples 8A-8D, sodium chlorite dihydrate may be supplied by Oxychem (Wichita, Kansas, USA).

[0191] Solution 8A can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 50 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 1.0g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.98g of Hypromellose E3 Premium to the above. Mix the solution until the Hypromellose E3 Premium is dissolved. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.50 grams glycerin, 4.0 grams boric acid, 0.22 grams sodium borate, 0.27 grams disodium phosphate, 4.00 grams sodium citrate dihydrate, 1 gram potassium chloride, 0.57 grams sodium lactate (50% solution in water), 0.13 grams magnesium chloride, 0.036 grams glucose, 0.0002 grams glycine, 0.0001 grams ascorbic acid, 0.10 grams edetate disodium, 0.030 grams polyquaternium-42 (33% solution in water), and 0.14 grams sodium chlorite. 7. Determine the tonicity of the solution and adjust to 280 mOsm with sodium chloride. 8. Measure the pH of the solution and adjust to 7.2 with 1N sodium hydroxide and / or 1N hydrochloric acid. 9. Bring the solution to a volume of 1,000.00 grams with purified water and mix for 10 minutes. 10. Filter the solution using a 0.22 micrometer filter.

[0192] Solution 8B can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 50 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 1.0g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.98g of Hypromellose E3 Premium to the above. Mix the solution until the Hypromellose E3 Premium is dissolved. 6. The following ingredients are then added in order, allowing each to dissolve before adding the next: 2.50 grams glycerin, 4.0 grams boric acid, 0.22 grams sodium borate, 0.27 grams disodium phosphate, 4.00 grams sodium citrate dihydrate, 1 gram potassium chloride, 0.57 grams sodium lactate (50% solution in water), 0.13 grams magnesium chloride, 0.036 grams glucose, 0.0002 grams glycine, 0.0001 grams ascorbic acid, 0.05 grams edetate disodium, 0.015 grams polyquaternium-42 (33% solution in water), and 0.14 grams sodium chlorite. 7. Determine the tonicity of the solution and adjust to 280 mOsm with sodium chloride. 8. Measure the pH of the solution and adjust to 7.2 with 1N sodium hydroxide and / or 1N hydrochloric acid. 9. Bring the solution to a volume of 1,000.00 grams with purified water and mix for 10 minutes. 10. Filter the solution using a 0.22 micrometer filter.

[0193] Solution 8C can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 2g of Polysorbate 80 and 10g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 0.1g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.2 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Add 2.0 grams of tamarind seed polysaccharide to the above and mix the solution to completely dissolve the tamarind seed polysaccharide. 7. Add 1.98g of Hypromellose E3 Premium to the above. Mix the solution until the Hypromellose E3 Premium is dissolved. 8. The following ingredients are then added in order, allowing each to dissolve before adding the next: 2.50 grams polyethylene glycol 400, 2.50 grams glycerin, 4.0 grams boric acid, 0.22 grams sodium borate, 0.27 grams disodium phosphate, 4.00 grams sodium citrate dihydrate, 1 gram potassium chloride, 0.57 grams sodium lactate (50% solution in water), 0.13 grams magnesium chloride, 0.036 grams glucose, 0.0002 grams glycine, 0.0001 grams ascorbic acid, 0.10 grams edetate disodium, 0.030 grams polyquaternium-42 (33% solution in water), and 0.14 grams sodium chlorite. 9. Determine the tonicity of the solution and adjust to 280 mOsm with sodium chloride. 10. Measure the pH of the solution and adjust to 7.2 with 1N sodium hydroxide and / or 1N hydrochloric acid. 11. Bring the solution to a volume of 1,000.00 grams with purified water and mix for 10 minutes. 12. Filter the solution using a 0.22 micrometer filter.

[0194] Solution 8D can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 2g of Polysorbate 80 and 10g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 0.1g of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.2 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Add 2.0 grams of tamarind seed polysaccharide to the above and mix the solution to completely dissolve the tamarind seed polysaccharide. 7. Add 1.98g of Hypromellose E3 Premium to the above. Mix the solution until the Hypromellose E3 Premium is dissolved. 8. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.50 grams polyethylene glycol 400, 2.50 grams glycerin, 4.0 grams boric acid, 0.22 grams sodium borate, 0.27 grams disodium phosphate, 4.00 grams sodium citrate dihydrate, 1 gram potassium chloride, 0.57 grams sodium lactate (50% solution in water), 0.13 grams magnesium chloride, 0.036 grams glucose, 0.0002 grams glycine, 0.0001 grams ascorbic acid, 0.10 grams edetate disodium, 0.015 grams polyquaternium-42 (33% solution in water), and 0.14 grams sodium chlorite. 9. Determine the tonicity of the solution and adjust to 280 mOsm with sodium chloride. 10. Measure the pH of the solution and adjust to 7.2 with 1N sodium hydroxide and / or 1N hydrochloric acid. 11. Bring the solution to a volume of 1,000.00 grams with purified water and mix for 10 minutes. 12. Filter the solution using a 0.22 micrometer hydrophilic filter.

[0195] Example 9 Table 3 shows the ingredients of formulations of the present invention (as shown in Formulations 9A and 9B) which can be combined using conventional mixing techniques as described below.

[0196] [Table 3] * Tonicity can be adjusted to 280~290mOsm / Kg ** Can be adjusted to pH 7.2 *** Optionally, the amount to be added is 100% w / w

[0197] For Examples 9A and 9B, sodium hyaluronate may be supplied by CONTIPRO AS (Dolny, Dobrócz, Czech Republic).

[0198] For Examples 9A and 9B, lemon aspen extract (Abacross™ Acronychia acidula fruit extract) may be supplied by SOUTHERN CROSS BOTANICALS (Knockroe, New South Wales, Australia).

[0199] For Examples 9A and 9B, polysorbate 20 may be supplied by Merck KGaA (Darmstadt, Germany).

[0200] For Examples 9A and 9B, Polysorbate 80 may be supplied by Merck KGaA (Darmstadt, Germany).

[0201] For Examples 9A and 9B, polyethylene glycol 400 may be supplied by Clariant Produkte (Burgkirchen, Germany).

[0202] For Examples 9A and 9B, boric acid may be supplied by Merck KGaA (Darmstadt, Germany).

[0203] For Examples 9A and 9B, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0204] For Examples 9A and 9B, sodium chloride may be supplied by Caldic (Dusseldorf, Germany).

[0205] For Examples 9A and 9B, 1N sodium hydroxide can be supplied by VWR (Radnor, PA, USA).

[0206] For Examples 9A and 9B, 1N hydrochloric acid can be supplied by VWR (Radnor, PA, USA).

[0207] For Examples 9A and 9B, Lumuluse GRH-40 may be supplied by VANTAGE (Gurnee, Illinois, USA).

[0208] For Examples 9A and 9B, ultra-refined castor oil may be supplied by CRODA (Edison, NJ, USA).

[0209] For Examples 9A and 9B, ethyl linolenate may be supplied by SIGMA-ALDRICH (St. Louis, MO, USA).

[0210] For Examples 9A and 9B, retinol palmitate may be supplied by SIGMA-ALDRICH (St. Louis, MO, USA).

[0211] For Examples 9A and 9B, Polyquaternium-42 (33% aqueous solution) may be supplied by DSM BIOMEDICAL (Berkeley, Calif., USA).

[0212] For Example 9B, tamarind seed extract may be supplied by INDENA (Milan, Italy).

[0213] Solution 9A can be prepared as follows. To a 1.50 mL beaker, add 5.0 grams of Lumuluse GRH-40. 2. While mixing, add 6.25 grams of ultra refined castor oil. 3. Next, add 1 gram of ethyl linolenate and 0.5 grams of retinol palmitate, Mix until uniform. 4. To a separate 1500 mL beaker, add 500 grams of purified water. 5. To the above, add 2 g of Polysorbate 80 and 10 g of Polysorbate 20. Mix the solution until both are fully dissolved. 6. Add 1.0 gram of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 7. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 8. Add 2.0 grams of sodium hyaluronate to the solution from step 7. Mix the solution to completely dissolve the sodium hyaluronate. 9. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.5 grams of polyethylene glycol 400, 6.0 grams of boric acid, 0.06 grams of sodium borate, and 1 gram of polyquaternium-42 (33% aqueous solution). 10. Add the contents of step 3 and mix with a homogenizer until uniform. 11. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. Adjust the pH of the formulation to a pH of 7.2 using 12.1 N sodium hydroxide and / or 1 N hydrochloric acid. 13. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure thorough homogeneity. 14. Filter the solution using a 0.22 micrometer filter.

[0214] Solution 9B can be prepared as follows. To a 1.50 mL beaker, add 5.0 grams of Lumuluse GRH-40. 2. While mixing, add 6.25 grams of ultra refined castor oil. 3. Retain the homogeneous solution for future use. 4. To a separate 1500 mL beaker, add 500 grams of purified water. 5. To the above, add 10 g of Polysorbate 80 and 50 g of Polysorbate 20. Mix the solution until both are dissolved. 6. Add 1.0 gram of lemon aspen extract to the above. Mix the solution until the lemon aspen extract is dissolved. 7. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 8. Add 1.0 gram of sodium hyaluronate to the solution from step 7. Mix the solution to completely dissolve the sodium hyaluronate. 9. Next, add 2.0 grams of tamarind seed polysaccharides. Mix the solution to completely dissolve the tamarind seed polysaccharides. 10. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.5 grams of polyethylene glycol 400, 6.0 grams of boric acid, 0.06 grams of sodium borate, and 0.045 grams of polyquaternium-42 (33% solution in water). 11. Add the contents of step 3 and mix with a homogenizer until uniform. 12. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. The pH of the formulation is adjusted to a pH of 7.2 using 13.1 N sodium hydroxide and / or 1 N hydrochloric acid. 14. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure uniformity. 15. Filter the solution using a 0.22 micrometer filter.

[0215] Example 10 Table 4 shows the ingredients of formulations of the present invention (as shown in Formulations 10A and 10B), which can be combined using conventional mixing techniques as described below.

[0216] [Table 4] * Adjust to a tonicity of 280-290mOsm / Kg ** Adjust to pH 7.2 *** Dosage up to 100% w / w **** (2E,4E,6E)-7-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)-3-methylocta-2,4,6-trienoic acid

[0217] For Examples 10A and 10B, sodium hyaluronate may be supplied by CONTIPRO AS (Dolny, Dobrócz, Czech Republic).

[0218] For Example 10A, Actinomyces sp. A5640 extract (IHVR collection bacterial extract labeled IHVR_39565_F7 in the IHVR collection nomenclature) may be supplied by the Baruch S. Blumberg Institute (Doylestown, PA).

[0219] For Example 10B, Compound 1 may be supplied by Sigma-Aldrich.

[0220] For Examples 10A and 10B, polysorbate 20 may be supplied by Merck KGaA (Darmstadt, Germany).

[0221] For Examples 10A and 10B, Polysorbate 80 may be supplied by Merck KGaA (Darmstadt, Germany).

[0222] For Example 10A, polyethylene glycol 400 may be supplied by Clariant Produkte (Burgkirchen, Germany).

[0223] For Examples 10A and 10B, boric acid may be supplied by Merck KGaA (Darmstadt, Germany).

[0224] For Examples 10A and 10B, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0225] For Examples 10A and 10B, sodium chloride may be supplied by Caldic (Dusseldorf, Germany).

[0226] For Examples 10A and 10B, potassium chloride may be supplied by KGaA (Darmstadt, Germany).

[0227] For Examples 10A and 10B, calcium chloride dihydrate may be supplied by Merck KGaA (Darmstadt, Germany).

[0228] For Examples 10A and 10B, magnesium chloride may be supplied by KGaA (Darmstadt, Germany).

[0229] For Examples 10A and 10B, Polyquaternium-42 (33% aqueous solution) can be supplied by DSM BIOMEDICAL (Berkeley, Calif., USA).

[0230] For Examples 10A and 10B, sodium chlorite dihydrate may be supplied by Oxychem (Wichita, Kansas, USA).

[0231] For Examples 10A and 10B, 1N sodium hydroxide can be supplied by VWR (Radnor, Pennsylvania, USA).

[0232] For Examples 10A and 10B, 1N hydrochloric acid can be supplied by VWR (Radnor, PA, USA).

[0233] Solution 10A can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 100 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. Add 50.0 g of Actinomyces extract to the above. Mix the solution until the Actinomyces extract is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 3.0 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.5 grams of polyethylene glycol 400, 6.0 grams of boric acid, 0.05 grams of sodium borate, 1.0 grams of potassium chloride, 0.06 grams of calcium chloride dihydrate, 0.06 grams of magnesium chloride, and 0.0015 grams of Polyquaternium 42 (aqueous solution). 7. While continuing to mix, add 0.14 grams of sodium chloride dihydrate and mix to dissolve. 8. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. The pH of the formulation is adjusted to a pH of 7.2 using 9.1 N sodium hydroxide and / or 1 N hydrochloric acid. 10. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure complete homogeneity. 11. Filter the solution using a 0.22 micrometer filter.

[0234] Solution 10B can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 75 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. To the above, add 50.0 g of Compound I. Mix the solution until Compound I is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.5 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 6.0 grams boric acid, 0.05 grams sodium borate, 1.0 gram potassium chloride, 0.06 grams calcium chloride dihydrate, 0.06 grams magnesium chloride, and 0.0015 grams Polyquaternium 42 (aqueous solution). 7. While continuing to mix, add 0.14 grams of sodium chloride dihydrate and mix to dissolve. 8. Determine tonicity of formulation and adjust to 280 mOsm / Kg with sodium chloride. The pH of the formulation is adjusted to a pH of 7.2 using 9.1 N sodium hydroxide and / or 1 N hydrochloric acid. 10. Bring the solution to 1000.0 grams with purified water USP and mix for 10 minutes to ensure complete homogeneity. 11. Filter the solution using a 0.22 micrometer filter.

[0235] Example 11 Table 5 shows the ingredients of formulations of the present invention (as shown in Formulations 11A and 11B) which can be combined using conventional mixing techniques as described below.

[0236] [Table 5] * Adjust to a tonicity of 280-290mOsm / Kg ** Adjust to pH 7.2 *** Adequate amount up to 100.00% by volume ****4-(1-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)vinyl)benzoic acid

[0237] For Examples 11A and 11B, sodium hyaluronate may be supplied by CONTIPRO AS (Dolný, Dobrócz, Czech Republic).

[0238] For Examples 11A and 11B, tamarind seed polysaccharide may be supplied by INDENA (Milan, Italy).

[0239] For Example 11A, Actinomyces sp. A5640 extract (IHVR collection bacterial extract labeled IHVR_39565_F7 in the IHVR collection nomenclature) may be supplied by the Baruch S. Blumberg Institute (Doylestown, PA).

[0240] For Example 11B, compound 2 may be supplied by Sigma-Aldrich.

[0241] For Examples 11A and 11B, polysorbate 20 may be supplied by Merck KGaA (Darmstadt, Germany).

[0242] For Examples 11A and 11B, Polysorbate 80 may be supplied by Merck KGaA (Darmstadt, Germany).

[0243] For Examples 11A and 11B, polyethylene glycol 400 may be supplied by Clariant Produkte (Burgkirchen, Germany).

[0244] For Examples 11A and 11B, boric acid may be supplied by Merck KGaA (Darmstadt, Germany).

[0245] For Examples 11A and 11B, sodium borate may be supplied by Merck KGaA (Darmstadt, Germany).

[0246] For Examples 11A and 11B, sodium chloride may be supplied by Caldic (Dusseldorf, Germany).

[0247] For Examples 11A and 11B, potassium chloride may be supplied by KGaA (Darmstadt, Germany).

[0248] For Examples 11A and 11B, Hypromellose E3 2910 may be supplied by DOW CHEMICAL (Plaquemine, Louisiana, USA).

[0249] For Examples 11A and 11B, glycerin may be supplied by Emery Oleochemicals GmbH (Dusseldorf, Germany).

[0250] For Examples 11A and 11B, disodium phosphate may be supplied by Merck KGaA (Darmstadt, Germany).

[0251] For Examples 11A and 11B, sodium citrate may be supplied by Merck KGaA (Darmstadt, Germany).

[0252] For Examples 11A and 11B, sodium lactate may be supplied by Merck KGaA (Darmstadt, Germany) as sodium lactate (50% aqueous solution).

[0253] For Examples 11A and 11B, glucose may be supplied by Roquette Freres (Restron, France).

[0254] For Examples 11A and 11B, glycine may be supplied by Merck KGaA (Darmstadt, Germany).

[0255] For Examples 11A and 11B, ascorbic acid may be supplied by DSM NUTRITIONAL Products (DRAKEMYRE, Scotland, UK).

[0256] For Examples 11A and 11B, Polyquaternium 42 can be supplied by DSM BIOMEDICAL (Berkeley, Calif.) as Polyquaternium 42 (33% aqueous solution).

[0257] For Examples 11A and 11B, edetate disodium may be supplied by Merck NV / SA (Overijse, Belgium).

[0258] For Examples 11A and 11B, 1N sodium hydroxide can be supplied by VWR (Radnor, Pennsylvania, USA).

[0259] For Examples 11A and 11B, 1N hydrochloric acid can be supplied by VWR (Radnor, PA, USA). For Examples 11A and 11B, sodium chlorite dihydrate can be supplied by Oxychem (Wichita, KS, USA).

[0260] Solution 11Acan can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 100 g of Polysorbate 20. Mix the solution until both are thoroughly mixed and dissolved. 3. Add 50g of milk extract to the above and mix the solution until the sodium hyaluronate is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.2 grams of sodium citrate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Add 2.0 grams of tamarind seed polysaccharide to the above and mix the solution to completely dissolve the tamarind seed polysaccharide. 7. Add 1.98g of Hypromellose E3 Premium to the above. Mix the solution until the Hypromellose E3 Premium is dissolved. 8. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.50 grams polyethylene glycol 400, 2.50 grams glycerin, 4.0 grams boric acid, 0.22 grams sodium borate, 0.27 grams disodium phosphate, 4.00 grams sodium citrate dihydrate, 1 gram potassium chloride, 0.57 grams sodium lactate (50% solution in water), 0.13 grams magnesium chloride, 0.036 grams glucose, 0.0002 grams glycine, 0.0001 grams ascorbic acid, 0.10 grams edetate disodium, 0.030 grams polyquaternium-42 (33% solution in water), and 0.14 grams sodium chloride. 9. Determine the tonicity of the solution and adjust to 280 mOsm with sodium chloride. 10. Measure the pH of the solution and adjust to 7.2 with 1N sodium hydroxide and / or 1N hydrochloric acid. 11. Bring the solution to a volume of 1,000.00 grams with purified water and mix for 10 minutes. 12. Filter the solution using a 0.22 micrometer filter.

[0261] Solution 11B can be prepared as follows. 1. To a 1500 mL beaker, add 800 grams of purified water USP. 2. To the above, add 10 g of Polysorbate 80 and 30 g of Polysorbate 20. Mix the solution until both are well mixed and dissolved. 3. To the above, add 5.0 g of Compound I. Mix the solution until Compound I is dissolved. 4. Filter the solution through a 0.45 micrometer filter and return it to the 1500 mL beaker. 5. Add 1.2 grams of sodium hyaluronate to the solution from step 4. Mix the solution to completely dissolve the sodium hyaluronate. 6. Add 2.0 grams of tamarind seed polysaccharide to the above and mix the solution to completely dissolve the tamarind seed polysaccharide. 7. Add 1.98g of Hypromellose E3 Premium to the above. Mix the solution until the Hypromellose E3 Premium is dissolved. 8. Next, add the following ingredients in order, allowing each to dissolve before adding the next: 2.50 grams polyethylene glycol 400, 2.50 grams glycerin, 4.0 grams boric acid, 0.22 grams sodium borate, 0.27 grams disodium phosphate, 4.00 grams sodium citrate dihydrate, 1 gram potassium chloride, 0.57 grams sodium lactate (50% solution in water), 0.13 grams magnesium chloride, 0.036 grams glucose, 0.0002 grams glycine, 0.0001 grams ascorbic acid, 0.10 grams edetate disodium, 0.015 grams polyquaternium-42 (33% solution in water), and 0.14 grams sodium chlorite. 9. Determine the tonicity of the solution and adjust to 280 mOsm with sodium chloride. 10. Measure the pH of the solution and adjust to 7.2 with 1N sodium hydroxide and / or 1N hydrochloric acid. 11. Bring the solution to a volume of 1,000.00 grams with purified water and mix for 10 minutes. 12. Filter the solution using a 0.22 micrometer hydrophilic filter.

[0262] Embodiments of the present invention 1. A microemulsion composition for treating the eye, comprising: i) a safe and effective amount of a compound and / or extract having retinol-like properties and / or benefits for use in treating the eye selected from one or more of a plant extract or source of an extract from a plant of the genus Acronychia, Licaria, Calendula and / or Trellis, a bacterial extract or source of an extract of the genus Actinomyces, and a compound of formula (I),

[0263] [ka] During the ceremony, the dotted lines represent single or double bonds, optionally one of the dotted lines being a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; and optionally 2-methyl-propa-1,3-diene; ii) an ophthalmically acceptable carrier; A microemulsion composition wherein the microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0264] 2. A composition according to any one of embodiments 1 (or any of the following embodiments), wherein the compound and / or extract having retinol-like properties and / or benefits is a plant extract or source of extract from a plant of the genus Acronychia and / or Likaria.

[0265] 3. A composition according to any one of embodiments 1 and / or 2 (or any of the following embodiments), wherein the compound and / or extract having retinol-like properties and / or benefits is a plant extract or source of extract from a plant of the genus Acronychia.

[0266] 4. A composition according to any one of embodiments 1 to 3 (or any of the following embodiments), wherein the plant extract or source of extract from a plant of the genus Acronychia is selected from the group consisting of Acronychia averans, Acronychia acidula, Acronychia acronychioides, Acronychia acuminate, Acronychia bauerennii, Acronychia coolichirum, Acronychia crassipetala, Acronychia eugelensis, Acronychia imperforate, Acronychia laevis, 1. The composition, selected from the group consisting of Acronychia laurifolia, Acronychia litoralis, Acronychia oblongifolia, Acronychia octanara, Acronychia parviflora, Acronychia pauciflora, Acronychia pedunculata, Acronychia pubescens, Acronychia species (Batavia dauns), Acronychia suberosa, Acronychia vestiata, Acronychia wilcoxinia, and combinations of two or more thereof.

[0267] 5. A composition according to any one of embodiments 1-4 (or any of the following embodiments), wherein the plant extract or source of the extract derived from a plant of the genus Acronychia is Acronychia acidula.

[0268] 6. A composition according to any one of embodiments 1-5 (or any of the following embodiments), wherein the plant extract of Acronychia and / or Likaria comprises from about 1% to about 20% of a compound of formula II by weight of the extract;

[0269] [ka] During the ceremony, R1 is C1~C 20 Alkyl, C2-C20 Alkenyl, C2-C 20 selected from the group consisting of alkynyl, and C3-C8 cycloalkyl or aryl; R2 is selected from the group consisting of hydrogen, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl or aryl, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC2-C6 alkynyl, -OC3-C8 cycloalkyl or aryl, thiol, -SC1-C6 alkyl, -SC2-C6 alkenyl, -SC2-C6 alkynyl, SC3-C8 cycloalkyl or aryl, -NR4C1-C6 alkyl, -NR4C2-C6 alkenyl, -NR4C2-C6 alkynyl, and -NR4C3-C8 cycloalkyl or aryl; R3 is selected from -CO2H, -CO2R4 or an isosteric equivalent of a carboxy group, and R4 is C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl or aryl; Y is -(CH2-CH2)-, -(CH=CH)-, or

[0270] [ka] The composition.

[0271] 7. A composition according to any one of embodiments 1-6 (or any of the following embodiments), wherein the plant extract of Acronychia and / or Likaria comprises from about 1% to about 20% of a compound of formula II by weight of the extract;

[0272] [ka] During the ceremony, R1 is C5~C 16 Alkyl, C5-C 16 Alkenyl and C5-C 16 Alkynyl, more preferably C5-C, including, for example, farnesyl 16 alkenyl, R2 is selected from the group consisting of hydrogen, hydroxyl, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC2-C6 alkynyl, -OC3-C8 cycloalkyl, more preferably hydrogen, hydroxyl, -OC1-C6 alkyl, even more preferably hydrogen or -OC1-C3 alkyl; R3 is selected from -CO2H, -CO2R4, and R4 is a C1-C6 alkyl or isosteric equivalent of a carboxy group; The composition wherein Y is —(CH2—CH2)— or —(CH═CH)—.

[0273] 8. A composition according to any one of embodiments 1-7 (or any of the following embodiments), wherein the compound of formula (II) is in the form of an acid or alkyl ester selected from 3-(4-farnesyloxyphenyl)-propionic acid, 3-(4-farnesyloxy-3-hydroxyphenyl)-propionic acid, 3-(4-farnesyloxy-3-methoxyphenyl)-propionic acid, alkyl esters thereof, and combinations of two or more thereof.

[0274] 9. A composition according to any one of embodiments 1-8 (or any of the following embodiments), wherein the compound of formula II useful in the invention is 3-(4-farnesyloxyphenyl)-propionic acid and / or its ethyl ester.

[0275] 10. A composition according to embodiments 1-9 (or any of the following embodiments), wherein the compound and / or extract having retinol-like properties and / or benefits is a plant extract or source of extract from a plant of the genus Likaria.

[0276] 11. A composition according to any one of embodiments 1-10 (or any of the following embodiments), wherein the plant extract or source of the extract from a plant of the genus Licaria is selected from the group consisting of Licaria vernicosa, Licaria brittoniana, Licaria canella, Licaria cubensis, Licaria velutina, and Licaria triandra, and combinations of two or more thereof.

[0277] 12. A composition according to any one of embodiments 1-11 (or any of the following embodiments), wherein the plant extract or source of the extract from a plant of the genus Licaria is Licaria vernicosa.

[0278] 13. A composition according to any one of embodiments 1-12 (or any of the following embodiments), wherein the compound and / or extract having retinol-like properties and / or benefits is a bacterial extract or source of extract of the genus Actinomyces.

[0279] 14. A composition according to any one of embodiments 1-13 (or any of the following embodiments), wherein the compound and / or extract having retinol-like properties and / or benefits is a bacterial extract or source of an extract of Actinomyces sp. A5640.

[0280] 15. A composition according to any one of embodiments 1-14 (or any of the following embodiments), wherein the compound and / or extract having retinol-like properties and / or benefits comprises a compound of formula (I):

[0281] [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, A composition wherein R2 represents a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms.

[0282] 16. A composition according to any one of embodiments 1 to 15 (or any of the following embodiments), wherein the compound of formula I is selected from (2E,4E,6E)-7-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)-3-methylocta-2,4,6-trienoic acid and 4-(1-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)vinyl)benzoic acid and derivatives thereof exhibiting retinoid-like activity, and mixtures thereof.

[0283] 17. A method for preventing or treating symptoms associated with dry eye, optionally in a patient in need of such prevention or treatment, comprising administering a microemulsion composition, wherein the microemulsion composition: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0284] [ka] During the ceremony, The dotted lines represent single or double bonds; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; preferably containing a methyl (—CH3) or methylene (═CH2) moiety; a compound and / or extract, wherein a represents a linear, cyclic or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; ii) optionally, an ophthalmologically acceptable carrier; The method wherein the microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0285] 18. A method for treating a patient having reduced or low levels of production / release / delivery / excretion of hyaluronic acid and / or mucin from and / or within the cornea, comprising topically administering to the patient's eye a microemulsion composition, wherein the microemulsion composition: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0286] [ka] During the ceremony, the dotted lines represent single or double bonds, preferably one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; preferably containing a methyl (—CH3) or methylene (═CH2) moiety; a compound and / or extract, wherein a represents 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms; more preferably 6 carbon atoms; preferably a linear, cyclic or branched, saturated or unsaturated carbonate chain containing an aromatic moiety, preferably a phenyl moiety; preferably 2-methyl-propa-1,3-diene; ii) optionally, an ophthalmologically acceptable carrier; The method wherein the microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0287] 19. A method for treating dry eye, comprising topically administering to a patient a microemulsion composition, wherein the microemulsion composition comprises: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0288] [ka] During the ceremony, the dotted lines represent single or double bonds, preferably one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; preferably containing a methyl (—CH3) or methylene (═CH2) moiety; a represents a linear, cyclic or branched, saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; preferably 1 to 10 carbon atoms; more preferably 6 carbon atoms; preferably an aromatic moiety, preferably a phenyl moiety; preferably 2-methyl-propa-1,3-diene; and ii) one or more viscosity reducing or soothing agents; and iii) optionally, an ophthalmically acceptable carrier. The method wherein the microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0289] 20. A method for promoting or increasing the rate of healing of wounds in and / or on the eye (in a patient in need of such promoted or increased healing rate) by administering a microemulsion composition, the microemulsion composition comprising: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating dry eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licoricea, Calendula and / or Retinol; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I):

[0290] [ka] During the ceremony, the dotted lines represent single or double bonds, preferably one of the dotted lines is a double bond; R1 represents H, a linear, cyclic or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; preferably containing a methyl (—CH3) or methylene (═CH2) moiety; a represents a linear, cyclic or branched, saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; preferably 1 to 10 carbon atoms; more preferably 6 carbon atoms; preferably an aromatic moiety, preferably a phenyl moiety; preferably 2-methyl-propa-1,3-diene; and ii) optionally, an ophthalmologically acceptable carrier; The method wherein the microemulsion droplets or particles have a maximum dimension of less than 1,500 Å.

[0291] [Embodiment] (1) A microemulsion composition for treating the eye, comprising: i) a safe and effective amount of compounds and / or extracts having retinol-like properties and / or benefits for use in treating the eye selected from one or more of: a plant extract or source of an extract from a plant of the genus Acronychia, Licaria, Calendula and / or Tapioca; a bacterial extract or source of an extract from the genus Actinomyces; and a compound of formula (I), [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally containing a methyl (-CH3) or methylene (=CH2) moiety; a linear, cyclic, or branched, saturated or unsaturated carbonate chain of 1 to 20 carbon atoms; optionally 1 to 10 carbon atoms; optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; and optionally 2-methyl-propa-1,3-diene; ii) an ophthalmically acceptable carrier; A composition wherein the microemulsion droplets or particles have a maximum dimension of less than 150 nm (1,500 Å). (2) The composition of embodiment 1, wherein the compound and / or extract having retinol-like properties and / or benefits is a plant extract or source of extract from a plant of the genus Acronychia and / or Likaria. (3) The composition of embodiment 2, wherein the compound and / or extract having retinol-like properties and / or benefits is a plant extract or source of extract from a plant of the genus Acronychia. (4) The plant extract or extract source derived from a plant of the genus Acronychia is selected from the group consisting of Acronychia aberrans, Acronychia acidula, Acronychia acronychioides, Acronychia acuminate, Acronychia baeuerlenii, Acronychia chooreechillum, Acronychia crassipetala, Acronychia eungellensis, Acronychia imperforate, Acronychia laevis, Acronychia laevis, Acronychia laurifolia, Acronychia littoralis, Acronychia oblongifolia, Acronychia octandra, Acronychia parviflora, Acronychia pauciflora, Acronychia pedunculata, Acronychia pubescens, Acronychia species (Batavia Downs), Acronychia suberosa, Acronychia vestiata vestita, Acronychia wilcoxiana, and combinations of two or more thereof. (5) The composition of embodiment 4, wherein the plant extract or extract source derived from a plant of the genus Acronychia is Acronychia acidula.

[0292] (6) the plant extract of Acronychia and / or Likaria contains from about 1% to about 20% by weight of the extract of the compound of formula II; [ka] During the ceremony, R1 is C1~C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 selected from the group consisting of alkynyl, and C3-C8 cycloalkyl or aryl; R2 is selected from the group consisting of hydrogen, hydroxyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl or aryl, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC2-C6 alkynyl, -OC3-C8 cycloalkyl or aryl, thiol, -SC1-C6 alkyl, -SC2-C6 alkenyl, -SC2-C6 alkynyl, SC3-C8 cycloalkyl or aryl, -NR4C1-C6 alkyl, -NR4C2-C6 alkenyl, -NR4C2-C6 alkynyl, and -NR4C3-C8 cycloalkyl or aryl; R3 is selected from -CO2H, -CO2R4 or an isosteric equivalent of a carboxy group, and R4 is C1-C6 alkyl, C2-C6 alkenyl, C3-C8 cycloalkyl or aryl; The composition of embodiment 2, wherein Y is —(CH—CH)—, —(CH═CH)—, or —(C═C)—. (7) The plant extract of Acronychia and / or Likaria contains about 1% to about 20% by weight of the extract of the compound of formula II; [ka] During the ceremony, R1 is C5~C 16 Alkyl, C5-C 16 Alkenyl and C5-C 16 Alkynyl, more preferably C5-C, including, for example, farnesyl 16 alkenyl, R2 is selected from the group consisting of hydrogen, hydroxyl, -OC1-C6 alkyl, -OC2-C6 alkenyl, -OC2-C6 alkynyl, -OC3-C8 cycloalkyl, more preferably hydrogen, hydroxyl, -OC1-C6 alkyl, even more preferably hydrogen or -OC1-C3 alkyl; R3 is selected from -CO2H, -CO2R4, and R4 is a C1-C6 alkyl or isosteric equivalent of a carboxy group; The composition of embodiment 6, wherein Y is —(CH—CH)— or —(CH═CH)—. (8) The composition of embodiment 7, wherein the compound of formula (II) is in the form of an acid or alkyl ester selected from 3-(4-farnesyloxyphenyl)-propionic acid, 3-(4-farnesyloxy-3-hydroxyphenyl)-propionic acid, 3-(4-farnesyloxy-3-methoxyphenyl)-propionic acid, alkyl esters thereof, and combinations of two or more thereof. (9) The composition of embodiment 8, wherein the compound of formula II useful in the present invention is 3-(4-farnesyloxyphenyl)-propionic acid and / or its ethyl ester. (10) The composition of embodiment 2, wherein the compound and / or extract having retinol-like properties and / or benefits is a plant extract or source of extract from a plant of the genus Licaria.

[0293] (11) The composition of embodiment 10, wherein the plant extract or source of the extract from a plant of the genus Licaria is selected from the group consisting of Licaria vernicosa, Licaria brittoniana, Licaria canella, Licaria cubensis, Licaria velutina, and Licaria triandra, and combinations of two or more thereof. (12) The composition of embodiment 11, wherein the plant extract or extract source derived from a plant of the genus Licaria is Licaria vernicosa. (13) The composition of embodiment 1, wherein the compound and / or extract having retinol-like properties and / or benefits is a bacterial extract or source of extract of the genus Actinomyces. (14) The composition of embodiment 13, wherein the compound and / or extract having retinol-like properties and / or benefits is a bacterial extract or source of extract of Actinomyces sp. A5640. (15) The compound and / or extract having retinol-like properties and / or benefits comprises a compound of formula (I), [ka] During the ceremony, the dotted lines represent single or double bonds, and optionally one of the dotted lines is a double bond; R1 represents H, a linear, cyclic, or branched saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms, 2. The composition of embodiment 1, wherein R2 represents a linear, cyclic, or branched, saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms; a linear, cyclic, or branched, saturated or unsaturated carbonate chain containing 1 to 20 carbon atoms.

[0294] (16) The composition according to embodiment 15, wherein the compound of formula (I) is selected from (2E,4E,6E)-7-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)-3-methylocta-2,4,6-trienoic acid and 4-(1-(1,1,2,2,3,3-hexamethyl-2,3-dihydro-1H-inden-5-yl)vinyl)benzoic acid, and derivatives thereof exhibiting retinoid-like activity, and mixtures thereof.

Claims

1. In the eyes, to increase MUC1, MUC4, and / or MUC16 gene expression; and / or to increase secretion of mucin-1, and / or To increase the induction of HAS3 gene expression, and / or To increase hyaluronic acid concentration, 1. A microemulsion composition, said composition comprising: i) a safe and effective amount of an Acronychia acidula plant extract having retinol-like properties and / or benefits for use in treating the eye; ii) an ophthalmically acceptable carrier; A composition wherein the droplets or particles of said microemulsion have a maximum dimension of less than 150 nm (1,500 Å).

2. The composition of claim 1 for increasing MUC1, MUC4, and / or MUC16 gene expression in the eye.

3. The composition of claim 1 for increasing secretion of mucin-1 in the eye.

4. The composition of claim 1 for increasing the induction of HAS3 gene expression in the eye.

5. The composition of claim 1 for increasing hyaluronic acid concentration in the eye.

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