Compositions and methods for treating the eye

文档序号：538229 发布日期：2021-06-01 浏览：201次中文

阅读说明：本技术 用于治疗眼睛的组合物和方法 (Compositions and methods for treating the eye ) 是由李丁文华 K·马哈茂德 R·帕萨拜明岐 K·T·霍列娃于 2019-07-24 设计创作，主要内容包括：本发明涉及包含一种或多种具有视黄醇样活性和特性的提取物和/或化合物的组合物以及使用所述组合物治疗眼睛的方法。(The present invention relates to compositions comprising one or more extracts and/or compounds having retinol-like activity and properties and methods of using the same to treat the eye.)

1. A method for producing/releasing/delivering/excreting mucin from and/or in the cornea, comprising the step of administering a composition comprising:

i) a safe and effective amount of a compound and/or extract having retinol-like properties and/or beneficial effects for the treatment of dry eye, said compound and/or extract being selected from one or more of the following: a plant extract or source of extract from a plant of the genus acronychia, illicium, marigold, and/or trigonella; a bacterial extract or extract source of the genus actinomycete; and a compound of formula (I):

wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (= CH)₂) A moiety; a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, optionally from 1 to 10 carbon atoms, optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; optionally 2-methylpropan-1, 3-diene; and

ii) optionally, an ophthalmically acceptable carrier.

2. The method of claim 1, wherein said compound and/or extract having retinol-like properties and/or beneficial effects is a plant extract or extract source from a plant of the genus acronychia and/or pistia.

3. The method of claim 2, wherein said compound and/or extract having retinol-like properties and/or beneficial effects is a plant extract or extract source from a plant of the genus acronychia.

4. The method of claim 3, wherein the plant extract or extract source from a acronychia plant is selected from the group consisting of:Acronychia aberrans(ii) wild citrus reticulataAcronychia acidula）、Acronychia acronychioides、Acronychia acuminate、Acronychia baeuerlenii、Acronychia chooreechillum、Acronychia crassipetala、Acronychia eungellensis、Acronychia imperforate、Acronychia laevis、Acronychia laurifolia、Acronychia littoralis、Acronychia oblongifolia、Acronychia octandra、Acronychia parviflora、Acronychia pauciflora、Acronychia pedunculata、Acronychia pubescensCitrus reticulata species (Batavia Downs),Acronychia suberosa、Acronychia vestita、Acronychia wilcoxianaAnd combinations of two or more thereof.

5. The method of claim 4, wherein the plant extract or extract source from a plant of the genus acronychia is acronychia.

6. The method of claim 2, wherein the plant extract of acronychia and/or pistia comprises from about 1% to about 20%, by weight of the extract, of the compound of formula II

(II)

Wherein:

R₁selected from the group consisting of: c₁-C₂₀Alkyl radical, C₂-C₂₀Alkenyl radical, C₂-C₂₀Alkynyl and C₃-C₈Cycloalkyl or aryl;

R₂selected from the group consisting of: hydrogen, hydroxy, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₈Cycloalkyl or aryl, -OC₁-C₆Alkyl, -OC₂-C₆Alkenyl, -OC₂-C₆Alkynyl, -OC₃-C₈Cycloalkyl or aryl, thiols, -SC₁-C₆Alkyl, -SC₂-C₆Alkenyl, -SC₂-C₆Alkynyl, -SC₃-C₈Cycloalkyl or aryl, -NR₄C₁-C₆Alkyl, -NR₄C₂-C₆Alkenyl, -NR₄C₂-C₆Alkynyl and-NR₄C₃-C₈Cycloalkyl or aryl;

R₃is selected from-CO₂H、-CO₂R₄Or isosteric equivalents of carboxyl groups wherein R₄Is C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₃-C₈Cycloalkyl or aryl; and is

Y is- (CH)₂—CH₂) -, - (CH ═ CH) -or- (C.ident.C) -.

7. The method of claim 6, wherein the plant extract of acronychia and/or pistia comprises from about 1% to about 20%, by weight of the extract, of the compound of formula II

(II)

Wherein:

R₁selected from the group consisting of: c₅-C₁₆Alkyl radical, C₅-C₁₆Alkenyl and C₅-C₁₆Alkynyl, more preferably C₅-C₁₆Alkenyl groups including, for example, farnesyl;

R₂selected from the group consisting of: hydrogen, hydroxy, -OC₁-C₆Alkyl, -OC₂-C₆Alkenyl, -OC₂-C₆Alkynyl, -OC₃-C₈Cycloalkyl, more preferably hydrogen, hydroxy, -OC₁-C₆Alkyl, even more preferably hydrogen or-OC₁-C₃An alkyl group;

R₃is selected from-CO₂H、-CO₂R₄Wherein R is₄Is C₁-C₆An isosteric equivalent of an alkyl, or carboxyl group; and is

Y is- (CH)₂-CH₂) -or- (CH ═ CH) -.

8. The method of claim 7, wherein the compound of formula (II) is in the form of an acid or alkyl ester selected from the group consisting of 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 process according to claim 8, wherein the compound of formula II useful in the present invention is 3- (4-farnesyloxyphenyl) -propionic acid and/or the ethyl ester thereof.

10. The method of claim 2, wherein said compound and/or extract having retinol-like properties and/or beneficial effects is a plant extract or extract source from plants of the genus cinnamomum.

11. The method of claim 10, wherein the plant extract from plants of the genus litseaOr the extract source is selected from the group consisting of:Licaria vernicosa、Licaria brittoniana、Licaria canella、Licaria cubensis、Licaria velutinaandLicaria triandraand combinations of two or more thereof.

12. The method of claim 11, wherein the plant extract or extract source from a plant of the genus cinnamomum isLicaria vernicosa。

13. The method of claim 1, wherein said compound and/or extract having retinol-like properties and/or beneficial effects is a bacterial extract or extract source of said actinomycete.

14. The method of claim 13, wherein said compound and/or extract having retinol-like properties and/or beneficial effects is a bacterial extract or extract source of actinomycete species a 5640.

15. The method of claim 1, wherein the compound and/or extract having retinol-like properties and/or beneficial effects comprises a compound of formula (I):

wherein-

The dotted line represents a single or double bond; optionally, one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; a linear, cyclic or branched, saturated or unsaturated carbonising chain containing from 1 to 20 carbon atoms.

16. The method of claim 15, wherein the compound of formula I is selected from the group consisting of (2E,4E,6E) -7- (1,1,2,2,3, 3-hexamethyl-2, 3-dihydro-1H-inden-5-yl) -3-methyloct-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 its derivatives that exhibit retinoid-like activity, and mixtures thereof.

17. A method for maintaining a concentration of MU5AC in tear fluid in a range of equal to or greater than 8 ng/mg protein to 15 ng/mg protein, comprising the step of administering a composition comprising:

wherein-

The dotted line represents a single or double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (= CH)₂) A moiety;

a represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; and

ii) optionally, an ophthalmically acceptable carrier.

18. A method for treating a patient suffering from reduced or low levels of production/release/delivery/excretion of mucin from and/or in the cornea, comprising the step of topically administering to the eye of the patient a composition comprising:

wherein-

The dotted line represents a single or double bond; preferably one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (= CH)₂) A moiety;

a represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, preferably from 1 to 10 carbon atoms, more preferably 6 carbon atoms; preferably an aromatic moiety, preferably a phenyl moiety; preferably 2-methylpropan-1, 3-diene,

ii) optionally, an ophthalmically acceptable carrier.

19. A method for preventing or treating symptoms associated with dry eye, comprising the step of topically administering to a patient a composition comprising:

wherein-

The dotted line represents a single or double bond; preferably one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (= CH)₂) A moiety;

ii) one or more demulcents or soothing agents; and

iii) optionally, an ophthalmically acceptable carrier.

20. A method for promoting wound healing or increasing the rate of healing in and/or on the eye of a patient comprising the step of administering a composition comprising:

wherein-

The dotted line represents a single or double bond; preferably one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (= CH)₂) A moiety;

ii) optionally, an ophthalmically acceptable carrier.

21. A method for improving antimicrobial properties in the tear fluid of a patient comprising the step of administering a composition comprising:

wherein-

The dotted line represents a single or double bond; preferably one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (= CH)₂) A moiety;

ii) optionally, an ophthalmically acceptable carrier.

Technical Field

The present invention relates to compositions comprising one or more extracts and/or compounds having retinol-like activity and properties and methods of using the same to treat the eye.

Background

"Dry eye" is a multifactorial disease of the ocular surface characterized by a loss of tear film homeostasis and is associated with ocular symptoms where tear film instability and high osmotic pressure, ocular surface inflammation and damage, and sensory nerve abnormalities play a causative role. "Craig, j.p. et al," TFOS dess II definition and classification report, "Ocul surf.2017; volume 15 page 276 and 283. Dry eye can be caused by abnormal or insufficient tear formation and insufficient mucin secretion (i.e., dry keratoconjunctivitis). Dry eye can be the result of a variety of underlying diseases, such as autoimmune diseases that damage the lacrimal glands (i.e., produce tears), such as rheumatoid arthritis, sjogren's syndrome, systemic lupus erythematosus, and systemic sclerosis and sarcoidosis. In eye surgery (such asSurgery) may also cause dry eye. Dry eye is estimated to affect more than thirty million people in the united states.

Regardless of the underlying pathology, dry eye generally involves rapid breakdown of the pre-ocular tear film, resulting in dehydration of the exposed outer surface. Normal tear formation is required to keep the cornea and conjunctiva moist, and this in turn helps to prevent ulceration of both, as well as to maintain corneal transparency. In addition, tears facilitate movement of the eyelids on the surface of the eye (e.g., blinking) and removal of foreign matter from the eye. Tears typically also contain lysozyme, which can be used to prevent ocular infections. Dry eye can be associated with mild discomfort to severe pain in the eye. When dry eye occurs for a long period of time, it can cause blurred vision, a gritty and/or burning sensation, and itching. The condition may further lead to corneal ulceration and/or scarring if allowed to persist without treatment.

Dry eye includes eye pain or fatigue, increased frequency of blinking, and congestion of the eyes. Furthermore, bacteria can enter through the scratch and cause infection, and if the scratch is deep enough, it can even affect a person's vision. In addition to eye fatigue, causes of dry eye include sjogren's syndrome, stevens-johnson syndrome, burns and eye injuries, as well as side effects of antihypertensive drugs, tranquilizers, eye drops for the treatment of glaucoma, and other such drugs.

The tear film is the body's natural defense against dry eye. The tear film contains ocular mucin and is essential for maintaining homeostasis of the wet ocular surface. Mucins are produced by, among other things, corneal epithelial cells in the eye. Mucins are glycoproteins expressed by epithelial tissue of mucosal surfaces. They protect tissue by acting as antioxidants and providing lubrication. Mucin genes associated with tear film include MUC1, MUC2, MUC4, MUC5AC, MUC5B, MUC7, and MUC 16.

Mucins are also useful as antimicrobial agents for general wound healing and are essential for overall eye health maintenance.

Accordingly, there is a need for ophthalmic pharmaceutical compositions that will facilitate and/or improve the production and/or release of mucin from and/or in the cornea.

The present inventors have discovered compounds and/or extracts having retinol-like properties and/or beneficial effects that can induce, promote and/or improve the production/release/delivery/excretion of mucin from and/or in the cornea.

Accordingly, aspects of the present invention relate to compositions comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects to induce, promote and/or improve the production/release/delivery/excretion of mucin from and/or in the cornea.

Another 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 beneficial effects, the compounds and/or extracts induce, promote and/or improve the production/release/delivery/excretion of mucin from and/or in the cornea, the composition can be administered to a patient having a concentration of MUC5AC in tear fluid of less than 6 (or about 6) ng/mg protein, optionally 8 (or about 8) ng/mg protein, such that the concentration of MUC5AC in the tear fluid is increased to (or brought to) equal to or greater than 8 (or about 8) ng/mg protein to 15 (or about 15) ng/mg protein, optionally 9 (or about 9) ng/mg protein to 12 (or about 12) ng/mg protein.

In certain embodiments, the above-described concentration of MU5AC in tear fluid (i.e., equal to or greater than 8 (or about 8) ng/mg protein to 15 (or about 15) ng/mg protein, optionally 9 (or about 9) ng/mg protein to 12 (or about 12) ng/mg protein) is produced by a compound and/or extract having retinol-like properties and/or beneficial effects to induce, promote and/or improve mucin production/release/delivery/excretion from and/or in the cornea for a period of time of 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.

The concentration of MUC5AC in tear fluid detailed above was determined using the Uchino method (described below in the definition).

Another aspect of the invention relates to methods of preventing and/or treating (e.g., reducing) ocular symptoms associated with dry eye and/or caused by reduced or low level production/release/delivery/excretion of 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 beneficial effects which induce, promote and/or improve the production/release/delivery/excretion of mucin from and/or in the cornea.

Another aspect of the invention relates to methods of promoting healing or increasing the rate of healing of wounds (e.g., ocular wounds not associated with dry eye, post-operative surgery, or non-specific wounds) in and/or on the eye of a patient by: administering a composition comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects that induce, promote and/or improve production/release/delivery/excretion of mucin from and/or in the cornea (i.e., increase production/release/delivery/excretion of mucin from and/or in the cornea, in certain embodiments, over the concentration level of mucin produced by such patients without administration (or absence) of a composition comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects that induce, promote and/or improve production/release/delivery of mucin from and/or in the cornea Excretion).

Another aspect of the invention relates to a method of improving antimicrobial properties in a tear fluid of a patient (or tear film of the eye) by: administering a composition comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects that induce, promote and/or improve production/release/delivery/excretion of mucin from and/or in the cornea (i.e., increase production/release/delivery/excretion of mucin from and/or in the cornea, in certain embodiments, over the concentration level of mucin produced by such patients without administration (or absence) of a composition comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects that induce, promote and/or improve production/release/delivery of mucin from and/or in the cornea Excretion).

Disclosure of Invention

The present invention relates to a method for producing/releasing/delivering/excreting mucin from and/or in the cornea (optionally, in a patient requiring such production/release/delivery/excretion of mucin), the method comprising the step of administering a composition comprising:

wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (═ CH)₂) A moiety; containing 1 to 20 carbon atoms, optionally 1 to 10 carbon atoms, optionally 6 carbon atomsLinear, cyclic or branched, saturated or unsaturated, carbonised chains of subunits; optionally an aromatic moiety, optionally a phenyl moiety; optionally 2-methylpropan-1, 3-diene; and

ii) optionally, an ophthalmically acceptable carrier.

The present invention relates to a method for maintaining a concentration of MU5AC in tear fluid in the range of equal to or greater than 8 ng/mg protein to 15 ng/mg protein, comprising the step of administering a composition comprising:

wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (═ CH)₂) A moiety; a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, optionally from 1 to 10 carbon atoms, optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; optionally 2-methylpropan-1, 3-diene; and

ii) optionally, an ophthalmically acceptable carrier.

The present invention relates to a method for treating a patient suffering from reduced or low levels of production/release/delivery/excretion of mucin from and/or in the cornea, the method comprising the step of topically administering to the eye of the patient a composition comprising:

wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (═ CH)₂) A moiety; a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, optionally from 1 to 10 carbon atoms, optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; optionally 2-methylpropan-1, 3-diene; and

ii) optionally, an ophthalmically acceptable carrier.

The present invention relates to methods for preventing or treating symptoms associated with dry eye, comprising the step of topically administering to a patient (optionally, in a patient in need of such prevention or reduction of dry eye symptoms) a composition comprising:

wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (═ CH)₂) A moiety; a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, optionally from 1 to 10 carbon atoms, optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; optionally 2-methylpropan-1, 3-diene;

ii) one or more demulcents or soothing agents; and

iii) optionally, an ophthalmically acceptable carrier.

The present invention relates to a method for promoting wound healing or increasing the rate of healing in and/or on the eye of a patient (optionally, in a patient in need of such eye wound healing), the method comprising the step of administering a composition (i.e., increasing production/release/delivery/excretion of mucin from and/or in the cornea, in certain embodiments, above the concentration level of mucin produced by such a patient in the absence of administration (or absence) of a composition comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects) comprising:

wherein-

The dotted line represents a single or double bond; preferably one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (═ CH)₂) A moiety;

ii) optionally, an ophthalmically acceptable carrier.

The present invention relates to a method for improving antimicrobial properties in the tear fluid of a patient (or tear film of the eye), optionally in a patient in need of such antimicrobial properties, the method comprising the step of administering a composition (i.e., increasing production/release/delivery/excretion of mucin from and/or in the cornea, in certain embodiments, above the concentration level of mucin produced by such a patient in the absence of administration (or lack thereof) of a composition comprising a safe and effective amount of one or more compounds and/or extracts having retinol-like properties and/or beneficial effects) comprising:

wherein-

The dotted line represents a single or double bond; preferably one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; preferably methyl (-CH)₃) Or methylene (═ CH)₂) A moiety;

ii) optionally, an ophthalmically acceptable carrier.

Drawings

Figure 1 depicts a bar graph showing MUC1, MUC4, and MUC16 gene expression induced in corneal epithelial cells by the extract of lemon aspen (lemon aspen).

Figure 2 depicts a bar graph showing the induction of Mucin-1 secretion by aspen lemon extract in corneal epithelial cells.

Detailed Description

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 remainder of the disclosure in any way whatsoever.

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

As used herein, the term "comprising (and grammatical variations thereof)" is used in an inclusive sense of "having" or "including" and not in an exclusive sense of "including only". As used herein, the terms "a" and "an" and "the" should be understood to encompass the plural as well as the singular.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety to the extent not inconsistent with this specification. As used herein, all percentages are by weight of the total composition, unless otherwise specified.

As used herein, the term "cornea" or "corneal" is, includes and/or relates to the transparent anterior portion of an eye covering the iris, pupil and anterior chamber, the layers of which transparent anterior portion include the corneal epithelial layer (including corneal epithelial cells), bowman's layer (also known as the anterior limiting membrane), corneal stroma (also the proper stroma), the posterior elastic layer (also the posterior limiting membrane) and the corneal endothelium (a simple squamous or low cubic monolayer of mitochondria-rich cells, about 5 μm thick).

As used herein, the phrase "reduced or low level production/release/delivery/excretion of mucin from and/or in the cornea" refers to a concentration of MUC5AC that is less than the concentration of MUC5AC in normal human (i.e., non-diseased) tears, or in certain embodiments less than 6 nanograms per milligram of protein, optionally less than 8 nanograms per milligram of protein, as determined using the methods described in Uchino Y, Uchino M, Yokoi N, et al. Change in office tear mucin 5AC using visual display terminal: the Osaka study, jama ophthalmol, n 2014; 132(8):985-992. The method (Uchino method) is reproduced as follows:

concentration of MUC5AC in lacrimal fluid

By enzyme-linked immunoassay (E90756 Hu; USCN Life Science). (see Maker AV, Katabi N, Gonen M et al, pancreatic cyst fluid and serum mucin levels predict abnormal proliferation of papillary mucinous tumors in pancreatic ducts, Ann Surg Oncol.2011; 18(1): 199-. All samples were analyzed according to the manufacturer's instructions. The absorbance was measured at 450nm and the standard solution in the kit was recombinant human MUC5 AC. A protein assay kit (BCA protein assay kit; Pierce) was used to determine the protein concentration in the tear sample. The MUC5AC concentration was normalized to the tear protein content and expressed as MUC5AC protein (nanograms)/total tear protein (milligrams).

As used herein, a composition that is "substantially free" of an ingredient refers to a composition having about 2% by weight or less of the ingredient (based on the total weight of the composition). Preferably, a composition that is substantially free of an ingredient has about 1% by weight or less of the ingredient (based on the total weight of the composition), 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, more preferably about 0.01% by weight or less of the ingredient. In certain more preferred embodiments, a composition that is substantially free of an ingredient is free of that ingredient, that is, the composition is completely free of that ingredient.

As used herein, "ophthalmically acceptable" means that the ingredients described by the term are suitable for use in contact with tissue (e.g., soft eye tissue or periorbital skin tissue) without causing undue toxicity, incompatibility, instability, irritation, allergic response, and the like. As will be appreciated by those skilled in the art, ophthalmically acceptable salts are acidic/anionic or basic/cationic salts.

As used herein, the term "safe and effective amount" means an amount of the disclosed extract, compound, or composition that is up to sufficient to induce, promote, and/or improve production/release/delivery/excretion of mucin from and/or in the cornea, but is as little as sufficient to avoid serious side effects. The safe and effective amount of the compound, extract or composition will vary with such factors as the age, health and environmental exposure of the end user, the duration and nature of the treatment, the particular extract, ingredient or composition employed, and the particular pharmaceutically acceptable carrier employed.

The term "retinol-like properties and/or benefits" refers to the properties and/or benefits induced by retinol-like.

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

Generally, IUPAC nomenclature is used herein and is defined in accordance with the following terms.

The term "C1-8 alkyl" whether used alone or as part of a substituent group, refers to a saturated aliphatic branched or straight chain monovalent hydrocarbon radical having from 1 to 8 carbon atoms. For example, "C1-8 alkyl" specifically includes the groups methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, t-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 alkanediyl. Alkyl and alkanediyl groups may be attached to the core molecule via a terminal carbon atom or via a carbon atom in the chain. Similarly, any number of substituent variables may be attached to an alkyl or alkanediyl group as the available valences permit.

The term "C_1-4Alkyl "whether used alone or as part of a substituent group, refers to a saturated aliphatic branched or straight chain monovalent hydrocarbon or alkanediyl linking group having the specified number of carbon atoms, wherein the group is derived by removal of one hydrogen atom from a carbon atom and the alkanediyl linking group is derived by removal of one hydrogen atom from each of two carbon atoms in the chain. The term "C_1-4Alkyl "refers to a straight or branched chain arrangement of groups having 1 to 4 carbon atoms. For example, "C_1-4Alkyl "specifically includes the groups methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-butyl, t-butyl, 1-butyl, and the like. Alkyl and alkanediyl groups may be attached to the core molecule via a terminal carbon atom or via a carbon atom in the chain. Similarly, anyThe number of substituent variables may be attached to an alkyl or alkanediyl group as the available valences permit.

The term "C_2-4Alkenyl "means alkenyl having 2 to 4 carbon atoms. For example, specifically included are the groups vinyl, propenyl, allyl (2-propenyl), butenyl, and the like. As described above, alkenyl groups may be similarly attached to the core molecule and further substituted as indicated.

The term "halogen" by itself or in combination with other terms refers to a halogen atom, such as fluorine, chlorine, bromine, or iodine.

The term "substituted" means that one or more hydrogen atoms on the core molecule have been replaced with a substituent that is available in a valence-permitting amount. The substitution is not limited to the core molecule, but may be carried out on a substituent such that the group becomes a linking group.

The term "independently selected" refers to two or more substituents that may be selected from a variable group of substituents, wherein the selected substituents may be the same or different.

The term "dependently selected" means that one or more substituent variables designated in the indicated combination for substitution in the core molecule (e.g., variables relating to the group of substituents appearing in a tabular listing of compounds).

Acceptable salts from inorganic bases include, for example, sodium or potassium salts and the like. Acceptable salts from organic bases include, for example, salts with primary, secondary, tertiary amines, and the like.

Compounds/extracts exhibiting retinol-like biological activity and/or properties

The present invention includes compounds and/or extracts having retinol-like properties and/or beneficial effects for the treatment of dry eye, comprising a compound and/or extract selected from or consisting of: a plant extract or extract source from the genus acronychia, the genus illicium, the genus marigold, and/or the genus trigonella; a bacterial extract or extract source of the genus actinomycete; and a compound of formula (I):

wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (═ CH)₂) A moiety; a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, optionally from 1 to 10 carbon atoms, optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; optionally 2-methylpropan-1, 3-diene.

Plant extracts of acronychia tangutica, cinnamomum kohlii, calendula and/or trigonella foenum graecum

In certain embodiments, the compound/extract exhibiting retinol-like properties and/or beneficial effects is or comprises an extract or extract source of the plants acronychia, pistia cinnamomea, marigold, and/or trigonella foenum graecum. The acronychia, illicium, marigold and/or trigonella extract or the source of such extract is obtained from a plant of the acronychia, illicium, marigold and/or trigonella.

The Acronychia plants from which the extracts useful in the present invention are obtained include species such as Acronychia berrans, Acronychia olea (also referred to herein as aspen lemon), Acronychia acronoides, Acronychia acuminata, Acronychia baueritenii, Acronychia choorechalium, Acronychia brachycanthum, Acronychia crassimum (Acronychia crassipes), Acronychia euglensis, Acronychia immunoproformate, Acronychia laevis, Acronychia laurifolia, Acronychia toruloides, Acronychia oblongifolia, Acronychia octandra, Acronychia parvira, Acronychia papyrifera, Acronychia acerolata, Acronychia cinerea, or a combination of two or more species thereof. In one embodiment, the extract used in the present invention is obtained from Acronychia acidula.

Plants of the genus litsea from which extracts useful in the present invention are obtained include, for example, Licaria verniciosa, Licaria brittoniana, Licaria canella, Licaria cubensis, Licaria velutina, and Licaria triandra, and combinations of two or more thereof. About 40 species of illicium are reported to be unique to central and south america. In one embodiment, the extract used in the present invention is obtained from Licaria vernicosa.

About 15 to 20 species of marigold are reported to be present in southwestern asia, western asia, macaroni and mediterranean. Plants of the genus Calendula from which extracts useful in the present invention are obtained include, for example, Calendula (field marigold); calendaula maderaensis (Madeiran marigold); and calendiula officinalis (pot marigold) and combinations of two or more thereof. In one embodiment, the extract used in the present invention is obtained from calendiula officinalis.

The plant of the genus Trigonella from which the extract useful in the present invention is obtained includes 36 known species of plants of the genus Trigonella, including, for example, Trigonella foenum-graecum, Trigonella balansae, Trigonella corniculata, Trigonella maritima, Trigonella spicata, blue Trigonella (Trigonella calerulea), Trigonella occulta, Trigonella polytera, Trigonella calliera, Trigonella Cretica, and combinations of two or more thereof. Trigonella foenum-graecum or fenugreek (herb fenugreek) is known as the best member of the genus Trigonella. In one embodiment, the extract used in the present invention is obtained from Trigonella foenum-graecum.

In certain embodiments, the extract used in the present invention is a mixture of extracts from plants of the acronychia, illicium, calendula and/or trigonella genus.

The extract of Licaria vernicosa useful in the present invention is available from the bouch s. In certain embodiments, one extract is from the woody part of the plant (E2) and a second extract is from the roots of the plant (E3). In the nomenclature of the IHVR collection, the two extracts were labeled IHVR _40256_ G10 ═ X-005348-. The xylem and roots of Licaria vernicosa (Mez) Kosterm can be collected from Guiana. 504.3g of dried ground woody plant material can be extracted with sufficient methanol, which can be dried under vacuum to provide 10.54g of crude methanol extract labeled X-005348-001E002 (E2). 403.8g of dried ground root material can be extracted with sufficient methanol that it can be dried under vacuum to provide 18.11g of crude methanol extract labeled X-005346-001M002 (E3).

In certain embodiments, the acronychia and/or pistia extracts useful in the invention comprise a compound having formula II:

wherein:

R₁selected from the group consisting of: c₁-C₂₀Alkyl radical, C₂-C₂₀Alkenyl radical, C₂-C₂₀Alkynyl and C₃-C₈Cycloalkyl or aryl;

R₃is selected from-CO₂H、-CO₂R₄Or isosteric equivalents of carboxyl groups wherein R₄Is C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₃-C₈Cycloalkyl or aryl; and is

Y is- (CH)₂-CH₂) -, - (CH ═ CH) -or- (C ≡ C) -;

or an ophthalmically acceptable salt thereof.

In certain embodiments, the acronychia and/or pistia extracts useful in the invention comprise a compound having formula II:

wherein:

R₁selected from the group consisting of: c₅-C₁₆Alkyl radical, C₅-C₁₆Alkenyl and C₅-C₁₆Alkynyl, more preferably C₅-C₁₆Alkenyl groups including, for example, farnesyl;

R₃is selected from-CO₂H、-CO₂R₄Wherein R is₄Is C₁-C₆An isosteric equivalent of an alkyl, or carboxyl group; and is

Y is- (CH)₂-CH₂) -or- (CH ═ CH) -;

or an ophthalmically acceptable salt thereof.

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

In certain embodiments, the compounds of formula II useful in the present invention are in the form of an acid or alkyl ester selected from (or selected from the group consisting of) 3- (4-farnesyloxyphenyl) -propionic acid, 3- (4-farnesyloxy-3-hydroxyphenyl) -propionic acid, 3- (4-farnesyloxy-3-methoxyphenyl) -propionic acid, alkyl esters thereof (specifically, ethyl esters thereof), and combinations of two or more thereof.

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

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

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

Compounds and extracts derived from acronychia species are described in US 9,220,928, which is incorporated herein by reference in its entirety.

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

Any of a variety of extracts of acronychia and/or pistia may be used in embodiments where the method includes applying such an extract. The extract may be obtained from any part of the plant, such as fruit, seeds, bark, leaves, flowers, roots and wood.

In certain embodiments, the extract is obtained from the fruit of a plant. Suitable extracts of the fruit, seeds, bark, leaves, flowers, roots and wood of acronychia or pistia can be obtained using conventional methods including, but not limited to, by milling, macerating, pressing, squeezing, mashing, centrifugation and/or processes such as cold percolation, stirring/distillation, microwave assisted extraction, supercritical/subcritical CO with or without polar modifiers₂Compressed gas extraction, pressurized solvent extraction, accelerated solvent extraction, pressurized or normal hot water extraction, surfactant assisted pressurized hot water extraction, oil extraction, membrane extraction, soxhlet extraction, gold finger distillation/extraction, and/or the like as disclosed, for example, in U.S. patents 7442391, 7473435, and 7537791 to Integrated botanicals Technologies, LLC, all incorporated herein by reference, or by other methods such as solvent extraction, or the like, to extract material directly from biomass. Any of a variety of solvents, including polar solvents, non-polar solvents, or combinations of two or more thereof, can be used in the process including solvent extraction.

Suitable polar solvents include polar inorganic solvents such as water and the like; polar organic solvents such as alcohols and corresponding organic acids, e.g. C including methanol, ethanol, propanol, butanol, etc₁-C₈Alcohols, organic acids including acetic, formic, propionic, and the like, including C₁-C₈Polyols and diols such as polyols/diols, and combinations of two or more thereof. Suitable non-polar solvents include non-polar organic solvents such as alkanes (including C)₁-C₈Alkanes), cycloalkanes (including C)₁-C₈Alkanes), alkyl ethers (including C)₁-C₈Alkyl ethers), petroleum ethers, ketones (including C)₁-C₈Ketones), methylene chloride, ethyl acetate, xylene, toluene, chloroform, vegetable oils, mineral oils, and the like. In another embodiment, extraction may be obtained by the above-described non-polar solvent or supercritical fluid extraction with or without a polar modifier, such as C₁-C₈Alcohol, water, C₁-C₈Polyol/diol or C₁-C₈An organic acid.

In one embodiment, the extract comprises an extract of acronychia. In another embodiment, the extract of the present invention comprises a combination of a polar extract and a non-polar extract from the fruit of acronychia tangutorum. In another embodiment, the extract of the present invention comprises an ethanol or glycolic acid extract of the fruit of acronychia tangutorum.

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 wood of, or root of, Licaria vernicosa. In another embodiment, the extract of the present invention comprises an ethanol extract from lica vernicosa wood or lica vernicosa root.

In another embodiment, the extract of the invention comprises by using a composition comprising water, C₁-C₈Alcohol, C₁-C₈Polyol, C₁-C₈The polar solvent of the glycols and combinations of two or more thereof is a polar extract prepared by extraction from the fruit of acronychia, the wood of Licaria vernicosa, or the root of Licaria vernicosa. In certain embodiments, one or more C's are used₁-C₄Alcohol, C₁-C₄Polyol and/or C₁-C₄Glycol to extract the extract. In certain embodiments, the extract is prepared using a solvent comprising methanol, ethanol, or a combination thereof, in the presence or absence of water. In another embodiment, the extract is a polar extract extracted from the fruit of satsuma aurantiaca using a combination of alcohol and water. In another embodiment, the extract is a polar extract extracted from ground wood of lica vernicosa or ground roots of lica vernicosa using methanol.

In another embodiment, the extract comprises a non-polar extract prepared by extracting from the fruit of Phyllanthus Amarus, Licaria veronica wood, or Licaria veronica root using a non-polar solvent comprising one or more ofSeed C₁-C₈Alkane, C₁-C₈Cycloalkanes, C₁-C₈Alkyl ethers, C₁-C₈Alkyl esters and/or chloroform, more preferably one or more C₁-C₈Alkane, C₁-C₈Alkyl esters and/or chloroform. In another embodiment, the non-polar extract is extracted from the fruit of acronychia tangutorum, Licaria verniciosa wood, or Licaria verniciosa root using hexane, ethyl acetate, chloroform, or a mixture of two or more thereof. In another embodiment, the non-polar extract is extracted from the satsuma aurantifolia fruit using ethyl acetate.

In one embodiment, the extract comprises an extract of marigold. In another embodiment, the extract of the invention comprises a combination of polar and non-polar extracts of marigold petals. In another embodiment, the extract of the invention comprises a non-polar extract of marigold petals.

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.

In another embodiment, the calendula and/or trigonella extract is a non-polar extract prepared using a non-polar solvent comprising one or more C₁-C₈Alkane, C₁-C₈Cycloalkanes, C₁-C₈Alkyl ethers, C₁-C₈Alkyl esters and/or chloroform, more preferably one or more C₁-C₈Alkane, C₁-C₈Alkyl esters and/or chloroform.

In another embodiment, the calendula and/or trigonella extract is a non-polar extract prepared using hexane, ethyl acetate, chloroform or a mixture of two or more thereof. In another embodiment, the extract is a non-polar extract prepared using ethyl acetate.

In one embodiment, the plant extract may be obtained by extracting a cell culture of a variety of plants, including cell cultures of acronychia, illicium, marigold, and/or trigonella. The cell culture extracted to obtain the plant extract used in the present invention may be in any form, including suspension cell cultures and the like.

Extracts of marigold and fenugreek are available from Caithress Biotechnologies Ltd, UK (http:// www.caithnessbiotechnologies.com /). These extracts are part of the Phytottre Natural Product Library, which is available to everyone. Alternatively, the extract may be obtained using a preparation method described as non-polar by caittress biotechnology ltd. and prepared with a mixture of methanol and dichloromethane. For details, please refer to the web page http:// caithtechnitology. In a typical extraction, a pre-weighed amount of dry powdered biomass was suspended in a mixture of methanol/dichloromethane (1:1) at ambient temperature and stirred overnight. The suspension was then filtered and the filtrate was dried under reduced pressure to a solvent-free residue.

In certain embodiments, the extract of acronychia, pistia stratiotes, marigold, and/or fenugreek is present in the compositions of the 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.

Bacterial extracts of actinomycetes exhibiting retinol-like properties and/or beneficial effects

In certain embodiments, the compound/extract exhibiting retinol-like properties and/or beneficial effects is or comprises a bacterial extract of the genus actinomycete. Bacteria of the genus actinomycete include many well characterized species, as well as some insufficiently characterized species, such as one collected from the united states and labeled as species a 5640. Samples of this bacteria were collected, grown in culture, and made into extracts. Extracts are part of the natural products collection (natural products collection) and are now under the control of the Barukenburg institute (Doylestrown, PA), formerly known as the hepatitis and Virus Institute (IHVR). Within the nomenclature of the IHVR collection, extracts are labeled IHVR _39565_ F7.

In one embodiment, the extract used in the present invention is obtained from an actinomycete species having the ability to produce a chemical composition similar to that produced by extract a 5640. In another embodiment, the bacterium is deposited in the united states and the strain is the same as previously designated species a 5640.

In certain embodiments, the extract of actinomyces is present in the compositions of the invention in an amount of from 0.001% to 10%, optionally from 0.001% to 5%, or optionally from 0.01% to 1%, by weight of the composition.

A compound of formula (I) exhibiting retinol-like biological activity and/or properties.

In certain embodiments, the compound/extract exhibiting retinol-like properties and/or beneficial effects is or comprises a compound of formula (I).

Wherein-

The dotted line represents a single or double bond; optionally one of the dotted lines is a double bond;

R₁represents H, a linear, cyclic or branched, saturated or unsaturated, carbonising chain comprising from 1 to 20 carbon atoms;

R₂represents a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms; optionally methyl (-CH)₃) Or methylene (═ CH)₂) A moiety; a linear, cyclic or branched, saturated or unsaturated carbonising chain comprising from 1 to 20 carbon atoms, optionally from 1 to 10 carbon atoms, optionally 6 carbon atoms; optionally an aromatic moiety, optionally a phenyl moiety; preferably 2-methylpropan-1, 3-diene.

In certain embodiments, the compounds of formula (I) include the corresponding salts of such metal ions, such as, but not limited to, Li⁺、Na⁺、K⁺、Ca²⁺Or Mg²⁺。

In certain embodiments, the compound of formula I is:

(2E,4E,6E) -7- (1,1,2,2,3, 3-hexamethyl-2, 3-dihydro-1H-inden-5-yl) -3-methyloctane-2, 4, 6-trienoic acid

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 which exhibit retinoid-like activity. These compounds are referred to as compound 1 and compound 2, respectively, in examples 6 and 7 below.

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-methyloct-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 its derivatives exhibiting retinoid-like activity, and mixtures thereof.

Mixtures of any of the above compound/extract sources that exhibit retinol-like properties and/or beneficial effects may also be used.

Compounds and extracts of formula I are described in U.S. patent publication 2019/0091122, which is incorporated herein by reference in its entirety.

In certain embodiments, the compound of formula I is present in the compositions of the present invention in an amount 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 another embodiment, the compound of formula I is present in the compositions of the present invention in an amount 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.

Compositions the present inventors have found that compounds and/or extracts having retinol-like properties and/or beneficial effects can facilitate and/or improve the delivery/excretion of mucins from corneal epithelial cells.

Penetration enhancer

In certain embodiments, the compositions of the present invention optionally comprise a penetration enhancer.

Suitable penetration enhancers include (selected from or selected from the group consisting of: surfactants such as saponin, 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), sorbitan monooleate, sorbitan monolaurate, polyoxyethylene sorbitan (such as polyoxyethylene sorbitan monolaurate), quaternary ammonium decahydrocarbide, and dodecyltrimethylammonium bromide; chelating agents such as natural polyacids (e.g., citric acid), phosphates (e.g., disodium pyrophosphate), phosphonates, bisphosphonates (e.g., hydroxyethylidene diphosphonic acid), aminocarboxylic acids (e.g., ethylenediaminetetraacetic acid (EDTA) and disodium edetate) 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, sodium deoxycholate, sodium taurodeoxycholate, chenodeoxycholic acid, and ursodeoxycholic acid; fusidic acid derivatives, glycyrrhizic acid and ammonium glycyrrhizinate, and saponin EDTA, fusidic acid, polyoxyethylene 9-lauryl ether, polyoxyethylene 20-stearyl ether, glycocholate or a mixture of any of the above.

The concentration of the permeation enhancer applied should be the minimum amount necessary to increase absorption of the compound and/or extract through the mucus or other barrier membrane of the eye. Generally, ranges 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%), optionally to 4.5% (or about 4.5%), optionally to 5% (or about 5%), optionally to 5.5% (or about 5.5%), optionally to 6% (or about 6%), optionally to 6.5% (or about 6.5%), optionally to 7% (or about 7%), optionally to 7.5% (or about 7.5%), optionally to 8% (or about 8.5%), optionally to 8.5% (or about 8.5%), optionally to 9% (or about 9%), optionally to 9.5% (or about 9.5%), optionally to 10% (or about 10%), optionally to 10.5% (or about 10.5%), optionally to 11% (or about 11%), optionally to 11.5% (or about 11.5%), optionally to 12% (or about 12%), optionally to 12.5% (or about 12.5%), optionally to 13% (or about 13%), optionally to 13.5% (or about 13.5%), optionally to 14% (or about 14%), optionally to 14.5% (or about 14.5%), optionally to 15% (or about 15%), optionally to 15.5% (or about 15.5%), optionally to 16% (or about 16%), optionally to 16.5% (or about 16.5%), optionally to 17% (or about 17%), optionally to 17.5% (or about 17.5%), optionally to 18% (or about 18), optionally to 18.5% (or about 18.5%), optionally to 11%, optionally to 18.5%), optionally to 12%, optionally to 12.5%, optionally to 12%, optionally to 12.5%, or to 12%, optionally to 12., optionally, a concentration (w/v) of up to 19% (or about 19%), optionally up to 19.5% (or about 19.5%), optionally up to 20% (or about 20%) of the total composition may be used in the compositions of the invention.

Ophthalmologically acceptable carrier

The compositions of the present invention also comprise an aqueous, oil-in-water, or water-in-oil emulsion carrier. The carrier is ophthalmically acceptable. Useful oil-in-water and water-in-oil carriers can be found in U.S. patent publication 20030165545a1 and U.S. patents 9480645, 8828412, and 8496976, each of which is incorporated herein by reference in its entirety.

The ophthalmically acceptable carrier (or composition of the present invention) may optionally comprise one or more additional excipients and/or one or more additional active ingredients. Examples of such optional components are described below.

Excipients commonly used in ophthalmic compositions include, but are not limited to, demulcents, tonicity agents, preservatives, chelating agents, buffering agents (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 (not 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 invention, including mixtures of water, water and water-miscible solvents, such as vegetable or mineral oils containing from 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, polyvinyl pyrrolidone, polyvinyl methyl ether, polyethylene oxide, and preferably cross-linked polyacrylic acid and mixtures thereof.

Demulcents or soothing agents for use with embodiments of the present invention include, but are not limited to, cellulose derivatives (such as hydroxyethylcellulose, methylcellulose, hypromellose, or mixtures thereof), hyaluronic acid, 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 is a soothing or soothing agent. In certain embodiments, the soothing or soothing agent is selected from hyaluronic acid, tamarind seed extract, or mixtures thereof.

The compositions of the present invention are ophthalmically suitable for administration to the eye of a subject. The term "aqueous" generally refers to an aqueous formulation wherein the excipient is greater than about 50%, more preferably greater than about 75%, and specifically greater than about 90% water by weight. In certain embodiments, the compositions of the present invention are substantially free of compounds that irritate the eye. In certain embodiments, the compositions of the present invention are substantially free of free fatty acids and C₁To C₄An alcohol. In certain embodiments, the compositions of the present invention compriseLess than 40% (or about 40%), optionally less than 35% (or about 35%), optionally less than 30% (or about 30%), optionally less than 25% (or about 25%), optionally less than 20% (or about 20%), optionally less than 15% (or about 15%), optionally less than 10% (or about 10%) or optionally less than 5% (or about 5%) by weight of the total composition of non-alcohol, organic excipient or solvent. These drops may be delivered in single dose ampoules, preferably sterile, so that no bacteriostatic component is required in the formulation. Alternatively, the drops may be delivered in a multi-dose bottle which may preferably include means to extract any preservatives from the composition as it is delivered, such means being known in the art.

In certain embodiments, the compositions of the present invention are isotonic or slightly hypotonic in order to combat the hypertonicity of tears caused by evaporation and/or disease. This may require a tonicity agent to bring the osmolality of the formulation to a level of or near 210-320 millimoles per kilogram (mOsm/kg). The compositions of the present invention typically have an osmolarity in the range of 220-320mOsm/kg, or optionally in the range of 235-300 mOsm/kg. Ophthalmic compositions will generally be formulated as sterile aqueous solutions.

The degree of penetration of the compositions of the present invention may be adjusted with tonicity agents to a value compatible with the intended use of the composition. For example, the permeability of the composition can be adjusted to approximate the permeability of normal tears, which is equivalent to about 0.9 w/v% sodium chloride in water. Examples of suitable tonicity adjusting agents include, but are not limited to, sodium chloride, potassium chloride, calcium chloride, and magnesium chloride; dextrose; glycerol; propylene glycol; mannitol; sorbitol, and the like, and mixtures thereof. In one embodiment, a combination of sodium chloride and potassium chloride is used to adjust the tonicity of the composition.

The compositions of the present invention may also be used to administer pharmaceutically active compounds. Such compounds include, but are not limited to, glaucoma therapeutics, analgesics, anti-inflammatory and anti-allergic drugs, and antimicrobial agents. More specific examples of pharmaceutically active compounds include betaxolol, timolol, pilocarpine, carbonic anhydrase inhibitors, and prostaglandins; a dopaminergic antagonist; postoperative antihypertensive agents such as p-aminocclonidine (alaclonidine); anti-infective agents such as ciprofloxacin, moxifloxacin, and tobramycin; non-steroidal and steroidal anti-inflammatory agents such as naproxen, diclofenac, nepafenac, suprofen, ketorolac, tetrahydrocortisol, and dexamethasone; dry eye therapeutic agents, for example, PDE4 inhibitors; and antiallergic drugs such as H1/H4 inhibitors, H4 inhibitors, olopatadine, or mixtures thereof.

It is also contemplated that the concentration of the ingredients comprising the formulations of the present invention may vary. One of ordinary skill in the art will appreciate that the concentration may vary depending on the addition, substitution, and/or subtraction of ingredients in a given formulation.

In certain embodiments, the compositions of the present invention may have a pH compatible with the intended use, and typically range from 4 (or about 4) to 10 (or about 10), optionally between 6 (or about 6) to 8 (to about 8), optionally between 6.5 (or about 6.5) to 7.5 (or about 7.5), or optionally between 6.8 (or about 6.8) to 7.2 (or about 7.2).

In certain embodiments, various conventional buffering agents may be employed, such as phosphates, borates, citrates, acetates, histidines, tris, bis-tris, and the like, and mixtures thereof. Borate buffers include boric acid and salts thereof, such as sodium or potassium borate. Potassium tetraborate or metaborate may also be used, which in solution produces boric acid or a borate salt. Hydrated salts, such as sodium borate decahydrate, may also be used. Phosphate buffers include phosphoric acid and its salts; for example, M₂HPO₄And MH₂PO4, where M is an alkali metal such as sodium and potassium. Hydrated salts may also be used. In one embodiment of the invention, Na is used₂HPO₄.7H₂O and NaH₂PO₂.H₂O as a buffer. The term phosphate also includes compounds that produce phosphoric acid or phosphate in solution. In addition, organic counterions to the above-mentioned buffers can also be used. The concentration of the buffer typically varies between about 0.01 w/v% and 2.5 w/v%, more preferably between about 0.05 w/v% and about 0.5 w/v%.

In certain embodiments, the compositions of the present invention have a viscosity ranging from about 1cps to about 500cps, optionally from about 10cps to about 200cps, or optionally from about 10cps to about 100cps when measured using a TA Instrument AR2000 rheometer. The TA Instrument AR2000 rheometer should be used with the AR2000 flow test method of the TA rheologic Advantage software, the rheometer having a 40mm steel plate geometry; the viscosity range should be obtained by measuring the steady-state flow rate, which is controlled at a shear rate of 0 sec-1 to 200 sec-1.

In certain embodiments, the compositions of the present invention may be used as an eye drop solution, an eye wash solution, a contact lens lubricating and/or rewetting solution, a spray, a mist, or any other form of applying the composition to the eye.

The compositions of the present invention may also be used in the form of a wetting solution for contact lenses. In certain embodiments, the compositions of the present invention are sealable in blister packages as a wetting solution, and are also suitable for undergoing a sterilization process.

Examples of blister packages and sterilization techniques are disclosed in the following references, which are incorporated herein by reference in their entirety: U.S. patent D435,966; 4,691,820; 5,467,868, respectively; 5,704,468, respectively; 5,823,327, respectively; 6,050,398, 5,696,686; 6,018,931, respectively; 5,577,367, respectively; and 5,488,815. This part of the manufacturing process provides another method of treating an ophthalmic device with an anti-allergic agent by adding the anti-allergic agent to the solution prior to sealing the package, followed by sterilization of the package. This is the preferred method of treating ophthalmic devices with anti-allergic agents.

Sterilization may be performed at different temperatures and time periods. Preferred sterilization conditions range from about 8 hours at about 100 ℃ to about 0.5 minutes at about 150 ℃. More preferred sterilization conditions range from about 2.5 hours at about 115 ℃ to about 5.0 minutes at about 130 ℃. The most preferred sterilization conditions are at about 124 ℃ for about 18 minutes.

When used as a wetting solution, the composition of the present invention may be a water-based solution. Typical wetting solutions include, but are not limited to, saline solutions, other buffers, and deionized water. In certain embodiments, the wetting solution is an aqueous solution of deionized water or an aqueous salt solution containing salts including, but not limited to, sodium chloride, sodium borate, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, or their corresponding potassium salts. These ingredients are typically combined to form a buffer solution comprising an acid and its conjugate base, such that the addition of the acid and base causes only a relatively minor change in pH. In certain embodiments, the pH of the wetting solution is as described above. The buffer may additionally comprise 2- (N-morpholino) ethanesulfonic acid (MES), sodium hydroxide, 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 a borate buffered saline solution or a phosphate buffered saline solution or deionized water. Particularly preferred solutions contain about 500ppm to about 18,500ppm sodium borate, most particularly preferably about 1000ppm sodium borate.

If any ingredients incorporated into the wetting solution undergo oxidative degradation, agents may be added to stabilize the wetting solution containing such ingredients. Such "oxidative stabilizers" include, but are not limited to, chelating agents (such as EDTA, Dequest, Desferal, silica), chitin derivatives (such as chitosan, cellulose and its derivatives, and N, N', N "-hexa (2-pyridyl) -1,3, 5-tris (aminomethyl) benzene), and certain macrocyclic ligands (such as crown ethers, ligands containing junctions and chains). See, David A.Leigh et al, Angew.Chem int. eds., 2001, Vol.40, No. 8, p.1538-1542, and Jean-Claude chamboron et al, Pure & appl.chem., 1990, Vol.62, No. 6, p.1027-1034. The oxidation stabilizer may include other compounds that inhibit oxidation, such as selected from the group consisting of: 2,2', 6,6' -hexa (1, 1-dimethylethyl) 4,4' - [ (2,4, 6-trimethyl-1, 3, 5-benzenetriyl) -trimethylene ] -trisphenol (Irganox 1330), 1,3, 5-tris [3, 5-bis (1, 1-dimethylethyl) 4-hydroxybenzyl ] -1H,3H,5H-1,3, 5-triazine-2, 4, 6-trione, pentaerythritol tetrakis [3- [3, 5-bis (1, 1-dimethylethyl) -4-hydroxyphenyl ] -propionate ], octadecyl-3- [3, 5-bis (1, 1-dimethylethyl) -4-hydroxyphenyl ] -propionate, Tris [2, 4-bis (1, 1-dimethylethyl) -phenyl ] -phosphite, 2 '-dioctadecyloxy) -5,5' -spirocyclic bis (1,3, 2-dioxolane), dioctadecyl disulfide, didodecyl-3, 3 '-thiodipropionate, dioctadecyl-3, 3' -thiodipropionate, butylhydroxytoluene, ethylenebis [3, 3-bis [3- (1, 1-dimethylethyl) -4-hydroxyphenyl ] butyrate ], and mixtures thereof. Preferred oxidation stabilizers are diethylenetriaminepentaacetic acid ("DTPA"), or salts of DTPA such as CaNa3DTPA, ZnNa3DTPA, and Ca2 DTPA. See U.S. patent application 60/783,557 entitled "Methods for Stabilizing oxygen reactive volatile Pharmaceutical Compositions" filed 2006, month 3, and day 17, and its corresponding non-provisional filed application, which are incorporated herein by reference in their entirety. In certain embodiments, the concentration of the oxidation stabilizer in the solution is from about 2.5 micromoles/liter to about 5000 micromoles/liter, optionally from about 20 micromoles/liter to about 1000 micromoles/liter, optionally from about 100 micromoles/liter to about 1000 micromoles/liter, or optionally from about 100 micromoles/liter to about 500 micromoles/liter.

In particular embodiments, the compositions of the present invention are formulated for administration at any frequency of administration, including weekly, once every five days, once every three days, once every two days, twice daily, three times daily, four times daily, five times daily, six times daily, eight times daily, hourly or more. This dosing frequency is also maintained for a variable duration of time, depending on the therapeutic needs of the user. The duration of a particular treatment regimen may vary from a single administration to a regimen that lasts for months or years. One of ordinary skill in the art will be familiar with determining a particular indicated treatment regimen.

Compositions and products comprising such compositions of the present invention can be prepared using methods well known to those of ordinary skill in the art.

Examples

Any compositions of the present invention described in the following examples illustrate specific embodiments of the compositions of the present invention, but are not intended to be limiting. Other modifications may be made by those skilled in the art without departing from the spirit and scope of the invention.

The following test methods were used in the examples:

example 1

The fractions of the lemon extract in aspen showed increased expression of MUC1, MUC4 and MUC16 genes in human corneal epithelial 3D tissue when treated in culture.

Human corneal epithelial 3D tissue was purchased from MatTek corporation (Ashland, MA, USA). After receiving the human corneal epithelial 3D tissue, the tissue was incubated overnight in MatTek assay medium according to the manufacturer's instructions. Human corneal epithelial 3D tissue was divided into three treatment groups of at least three tissues per group. The lemon extract in aspen was added to the medium containing human corneal epithelial tissue of two of the treatment groups, respectively, to produce a medium concentration of 0.001% (w/v) or 0.01% (w/v), respectively. Corneal epithelial tissue in all four treatment groups was incubated for two days. The lemon extract of aspen used was supplied and extracted from south cross plant, Knockrow Nsw, Australia. After two days of incubation, mucin 1(MUC1), mucin 4(MUC4), and mucin 16(MUC16) were analyzed for gene expression. After two days of incubation, human corneal epithelial 3D tissue was cut in half and half of the tissue was lysed in 350 μ L of lysis buffer consisting of 100 parts RLT buffer (RNeasy Mini kit, Qiagen, Valencia, CA) to one part 2-mercaptoethanol. RNA was extracted from the solution using the RNeasy Mini Kit (Qiagen, Valencia, Calif.) according to the manufacturer's instructions and eluted in 25. mu.L of RNase-free water.

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

Example 2

The fractions of the lemon extract in aspen showed increased mucin-1 secretion in human corneal epithelial 3D tissue when treated in culture.

Human corneal epithelial 3D tissue was purchased from MatTek corporation (Ashland, MA, USA). After receiving the human corneal epithelial 3D tissue, the tissue was incubated overnight in MatTek assay medium according to the manufacturer's instructions. Human corneal epithelial 3D tissue was divided into three treatment groups of at least three tissues per group. The lemon extract in aspen was added to the medium containing human corneal epithelial tissue of two of the treatment groups, respectively, to produce a medium concentration of 0.001% (w/v) or 0.01% (w/v), respectively. Corneal epithelial tissue in all four treatment groups was incubated for two days. Two days later, media was collected using human mucin-1 (CA15-3) enzyme-linked immunosorbent assay (ELISA) kit (EHMUC1, thermo fisher Scientific, Bridgewater, NJ) for determination of mucin 1 secretion following the manufacturer's protocol. To assess activity, colorimetric changes were measured using a microplate reader (SpectraMax M2E, Molecular Devices, Sunnyvale, Calif., USA). The assay employs standard enzyme-linked immunoassay techniques, so that there is a linear correlation between mucin-1 concentration in the sample and colorimetric changes. A standard curve is generated with mucin-1 concentration as the x-axis and absorbance as the y-axis, indicating the corresponding mucin-1 concentration. The results are shown in figure 2 below.

Example 3

Solutions can be prepared comprising one or more compounds and/or extracts of the invention having retinol-like properties and/or beneficial effects, as shown in examples 3-7.

Table 1 shows the components of such formulations (as shown in formulations 3A-3D), which can be incorporated as described below using conventional mixing techniques.

TABLE 1

Can be adjusted to osmotic pressure of 280mOsm/Kg-290mOsm/Kg

Can be adjusted to pH 7.2

Optionally, an amount up to 100% w/w%

For examples 3A-3D: sodium hyaluronate may be provided by contiro A.S (DOLNI, DOBROUC, CZECH REPUBLIC).

For examples 3A-3D: aspen lemon extract (Abacross)^TMExtract of the fruit of satsuma aurantiaca) can be provided by southern cross (Knockrow Nsw, Australia).

For examples 3A-3D: polysorbate 20 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 3A-3D: polysorbate 80 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 5A-5B: polyethylene glycol 400 can be provided by Clariant Produkte (Burgkirchen, GERMANY).

For examples 3A-3D: boric acid may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 3A-3D: sodium borate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 3A-3D: sodium chloride may be supplied by Caldic (DusseldoRF, GERMANY).

For examples 3A-3D: potassium chloride may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 3A-3D: calcium chloride dihydrate may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 3A-3D: magnesium chloride may be provided by KGaA (DARMSTADT, GERMANY).

For examples 3A-3D: polyquaternium-42 (33% aqueous solution) may be supplied by DSM BIOMEDICAL (BERKELEY, CA, USA).

For examples 3A-3D: sodium chlorite dihydrate can be supplied by Oxychem (WICHITA, KS, USA).

For examples 3A-3D: the 1N sodium hydroxide may be supplied by VWR (RADPER, PA, USA).

For examples 3A-3D: the 1N hydrochloric acid may be supplied by VWR (RANDER, PA, USA).

Solution 3A can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 50g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 1.0g of lemon extract in Aspongopus. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution obtained in step 4 was added 2.0 g of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

6. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 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 mixing was continued, 0.14 grams of sodium chlorite dihydrate was added and mixed until dissolved.

8. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

9. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

10. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes to completely homogenize.

11. The solution was filtered using a 0.22 micron filter.

Solution 3B can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 2g of polysorbate 80 and 10g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. 0.1g of lemon extract in Aspongopus was added to the above mixture. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 3.0 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

7. While mixing was continued, 0.14 grams of sodium chlorite dihydrate was added and mixed until dissolved.

8. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

9. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

10. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes to completely homogenize.

11. The solution was filtered using a 0.22 micron filter.

Solution 3C can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 20g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 1.0g of lemon extract in Aspongopus. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 1.5 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

6. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 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 mixing was continued, 0.14 grams of sodium chlorite dihydrate was added and mixed until dissolved.

8. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

9. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

10. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes to completely homogenize.

11. The solution was filtered using a 0.22 micron filter.

Solution 3D can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 2g of polysorbate 10 and 50g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. 0.1g of lemon extract in Aspongopus was added to the above mixture. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 1.5 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

7. While mixing was continued, 0.14 grams of sodium chlorite dihydrate was added and mixed until dissolved.

8. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

9. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

10. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes to completely homogenize.

11. The solution was filtered using a 0.22 micron filter.

Example 4

Table 2 shows the components of the formulations of the present invention (as shown in formulations 4A-4D), which can be incorporated as described below using conventional mixing techniques.

TABLE 2

Can be adjusted to osmotic pressure of 280mOsm/Kg-290mOsm/Kg

Can be adjusted to pH 7.2

Optionally, an amount up to 100% w/w%

For examples 4C-4D: sodium hyaluronate may be provided by contiro A.S (DOLNI, DOBROUC, CZECH REPUBLIC).

For examples 4C-4D: the tamarind seed polysaccharide can be provided by INDENA (mirano, ITALY).

For examples 4A-4D: aspen lemon extract (Abacross)^TMExtract of the fruit of satsuma aurantiaca) can be provided by southern cross (Knockrow Nsw, Australia).

For examples 4A-4D: polysorbate 20 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: polysorbate 80 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 6C-6D: polyethylene glycol 400 can be provided by Clariant Produkte (Burgkirchen, GERMANY).

For examples 4A-4D: boric acid may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: sodium borate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: sodium chloride may be supplied by Caldic (DusseldoRF, GERMANY).

For examples 4A-4D: potassium chloride may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: hypromellose E32910 may be supplied by DOW CHEMICAL (PLAQUEMINE, LOUISIANA, USA).

For examples 4A-4D: glycerol can be supplied by Emery Oleochemicals GmbH (DUSSELDORF, GERMANY).

For examples 4A-4D: disodium phosphate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: sodium citrate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: sodium lactate may be provided as sodium lactate (50% aqueous solution) by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: glucose can be supplied by Roquette Freres (lastem, FRANCE).

For examples 4A-4D: glycine may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 4A-4D: ascorbic acid can be obtained from DSM Nutritional Products (DRAKEMYRE, SCOTLAND, UK).

For examples 4A-4D: polyquaternium 42 may be provided by DSM BIOMEDICAL (BERKELEY, CA) as Polyquaternium 42 (33% aqueous solution).

For examples 4A-4D: disodium edetate may be supplied by Merck NV/SA (OVERIJSE, BELGIUM).

For examples 4A-4D: the 1N sodium hydroxide may be supplied by VWR (RADPER, PA, USA).

For examples 4A-4D: the 1N hydrochloric acid may be supplied by VWR (RANDER, PA, USA).

For examples 6A-6D: sodium chlorite dihydrate can be supplied by Oxychem (WICHITA, KS, USA).

Solution 4A can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 50g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 1.0g of lemon extract in Aspongopus. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the above solution was added 1.98 grams of hypromellose E3 Premium. The solution was mixed until hypromellose E3 Premium was dissolved.

6. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 2.50 grams of glycerol, 4.0 grams of boric acid, 0.22 grams of sodium borate, 0.27 grams of disodium phosphate, 4.00 grams of sodium citrate dihydrate, 1 gram of potassium chloride, 0.57 grams of sodium lactate (50% aqueous solution), 0.13 grams of magnesium chloride, 0.036 grams of glucose, 0.0002 grams of glycine, 0.0001 grams of ascorbic acid, 0.10 grams of disodium edetate, 0.030 grams of polyquaternium-42 (33% aqueous solution), and 0.14 grams of sodium chlorite.

7. The osmolality of the solution was measured and adjusted to 280mOsm with sodium chloride.

8. The pH of the solution was measured and adjusted to 7.2 with 1N sodium hydroxide and/or 1N hydrochloric acid.

9. The solution was brought to 1,000.00 g with purified water and mixed for 10 minutes.

10. The solution was filtered using a 0.22 micron filter.

Solution 4B can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 50g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 1.0g of lemon extract in Aspongopus. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the above solution was added 1.98 grams of hypromellose E3 Premium. The solution was mixed until hypromellose E3 Premium was dissolved.

6. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 2.50 grams of glycerol, 4.0 grams of boric acid, 0.22 grams of sodium borate, 0.27 grams of disodium phosphate, 4.00 grams of sodium citrate dihydrate, 1 gram of potassium chloride, 0.57 grams of sodium lactate (50% aqueous solution), 0.13 grams of magnesium chloride, 0.036 grams of glucose, 0.0002 grams of glycine, 0.0001 grams of ascorbic acid, 0.05 grams of disodium edetate, 0.015 grams of polyquaternium-42 (33% aqueous solution), and 0.14 grams of sodium chlorite.

7. The osmolality of the solution was measured and adjusted to 280mOsm with sodium chloride.

8. The pH of the solution was measured and adjusted to 7.2 with 1N sodium hydroxide and/or 1N hydrochloric acid.

9. The solution was brought to 1,000.00 g with purified water and mixed for 10 minutes.

10. The solution was filtered using a 0.22 micron filter.

Solution 4C can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 2g of polysorbate 80 and 10g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. 0.1g of lemon extract in Aspongopus was added to the above mixture. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 1.2 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

6. To the above solution was added 2.0 kr tamarind kernel polysaccharide. The solution was mixed to completely dissolve the tamarind seed polysaccharide.

7. To the above solution was added 1.98 grams of hypromellose E3 Premium. The solution was mixed until hypromellose E3 Premium was dissolved.

8. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 2.50 grams of polyethylene glycol 400, 2.50 grams of glycerol, 4.0 grams of boric acid, 0.22 grams of sodium borate, 0.27 grams of disodium phosphate, 4.00 grams of sodium citrate dihydrate, 1 gram of potassium chloride, 0.57 grams of sodium lactate (50% in water), 0.13 grams of magnesium chloride, 0.036 grams of glucose, 0.0002 grams of glycine, 0.0001 grams of ascorbic acid, 0.10 grams of disodium edetate, 0.030 grams of polyquaternium-42 (33% in water), and 0.14 grams of sodium chlorite.

9. The osmolality of the solution was measured and adjusted to 280mOsm with sodium chloride.

10. The pH of the solution was measured and adjusted to 7.2 with 1N sodium hydroxide and/or 1N hydrochloric acid.

11. The solution was brought to 1,000.00 g with purified water and mixed for 10 minutes.

12. The solution was filtered using a 0.22 micron filter.

Solution 4D can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 2g of polysorbate 80 and 10g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. 0.1g of lemon extract in Aspongopus was added to the above mixture. The solution was mixed until the aspen lemon extract dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 1.2 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

6. To the above solution was added 2.0 kr tamarind kernel polysaccharide. The solution was mixed to completely dissolve the tamarind seed polysaccharide.

7. To the above solution was added 1.98 grams of hypromellose E3 Premium. The solution was mixed until hypromellose E3 Premium was dissolved.

8. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 2.50 grams of polyethylene glycol 400, 2.50 grams of glycerol, 4.0 grams of boric acid, 0.22 grams of sodium borate, 0.27 grams of disodium phosphate, 4.00 grams of sodium citrate dihydrate, 1 gram of potassium chloride, 0.57 grams of sodium lactate (50% aqueous solution), 0.13 grams of magnesium chloride, 0.036 grams of glucose, 0.0002 grams of glycine, 0.0001 grams of ascorbic acid, 0.10 grams of disodium edetate, 0.015 grams of polyquaternium-42 (33% aqueous solution), and 0.14 grams of sodium chlorite.

9. The osmolality of the solution was measured and adjusted to 280mOsm with sodium chloride.

10. The pH of the solution was measured and adjusted to 7.2 with 1N sodium hydroxide and/or 1N hydrochloric acid.

11. The solution was brought to 1,000.00 g with purified water and mixed for 10 minutes.

12. The solution was filtered using a 0.22 micron hydrophilic filter.

Example 5

Table 3 shows the components of the formulations of the present invention (as shown in formulations 5A and 5B), which can be incorporated as described below using conventional mixing techniques.

TABLE 3

Can be adjusted to osmotic pressure of 280mOsm/Kg-290mOsm/Kg

Can be adjusted to pH 7.2

Optionally, an amount up to 100% w/w%

For examples 5A-5B: sodium hyaluronate may be provided by contiro A.S (DOLNI, DOBROUC, CZECH REPUBLIC).

For examples 5A-5B: aspen lemon extract (Abacross)^TMExtract of the fruit of satsuma aurantiaca) can be provided by southern cross (Knockrow Nsw, Australia).

For examples 5A-5B: polysorbate 20 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 5A-5B: polysorbate 80 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 5A-5B: polyethylene glycol 400 can be provided by Clariant Produkte (Burgkirchen, GERMANY).

For examples 5A-5B: boric acid may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 5A-5B: sodium borate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 5A-5B: sodium chloride may be supplied by Caldic (DusseldoRF, GERMANY).

For examples 5A-5B: the 1N sodium hydroxide may be supplied by VWR (RADPER, PA, USA).

For examples 5A-5B: the 1N hydrochloric acid may be supplied by VWR (RANDER, PA, USA).

For examples 5A-5B: lumuluse GRH-40 can be supplied by VANTAGE (GURNEE, IL, USA).

For examples 5A-5B: ultra refined castor oil may be provided by CRODA (EDISON, NJ, USA).

For examples 5A-5B: ethyl linoleate can be supplied by SIGMA-ALDRICH (st. louis, MO, USA).

For examples 5A-5B: retinyl palmitate may be supplied from SIGMA-ALDRICH (ST. LOUIS, MO, USA).

For examples 5A-5B: polyquaternium-42 (33% aqueous solution) may be supplied by DSM BIOMEDICAL (BERKELEY, CA, USA).

For example 5B: tamarind seed polysaccharides can be provided by INDENA (MILAN, ITALY).

Solution 5A can be prepared as follows：

1. To a 50ml beaker was added 5.0 grams of Lumulus GRH-40.

2. While mixing, 6.25 grams of ultra-refined castor oil was added.

3. Then 1 gram of ethyl linolenate and 0.5 gram of retinyl palmitate were added and mixed until homogeneous.

4. To a separate 1500ml beaker was added 500 grams of purified water.

5. To the above material were added 2g of polysorbate 80 and 10g of polysorbate 20. The solution was mixed until both were completely dissolved.

6. To the above material was added 0.1g of lemon extract in aspen. The solution was mixed until the aspen lemon extract dissolved.

7. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

8. To the solution obtained in step 7 was added 2.0 g of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

9. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 2.5 grams of polyethylene glycol 400, 6.0 grams of boric acid, 0.06 grams of sodium borate, and grams of polyquaternium-42 (33% aqueous solution).

10. Add the contents of step 3 and mix using a homogenizer until homogeneous.

11. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

12. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

13. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes until completely homogeneous.

14. The solution was filtered using a 0.22 micron filter.

Solution 5B can be prepared as follows：

1. To a 50ml beaker was added 5.0 grams of Lumulus GRH-40.

2. While mixing, 6.25 grams of ultra-refined castor oil was added.

3. The homogeneous solution was set aside for future use.

4. To a separate 1500ml beaker was added 500 grams of purified water.

5. To the above material were added 10g of polysorbate 80 and 50g of polysorbate 20. The solution was mixed until both were completely dissolved.

6. To the above material was added 1.0g of lemon extract in aspen. The solution was mixed until the aspen lemon extract dissolved.

7. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

8. To the solution of step 7 was added 1.0g of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

9. Next, 2.0 kr tamarind kernel polysaccharide was added. The solution was mixed to completely dissolve the tamarind seed polysaccharide.

10. The following ingredients were then added in order, dissolving each ingredient and then adding the latter: 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% aqueous solution).

11. Add the contents of step 3 and mix using a homogenizer until homogeneous.

12. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

13. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

14. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes until completely homogeneous.

15. The solution was filtered using a 0.22 micron filter.

Example 6

Table 4 shows the components of the formulations of the present invention (as shown in formulations 6A and 6B), which can be incorporated as described below using conventional mixing techniques.

TABLE 4

Adjusting to osmotic pressure of 280mOsm/Kg-290mOsm/Kg

Adjusting to pH 7.2

Moderate amount to 100 wt/wt%

(2E,4E,6E) -7- (1,1,2,2,3, 3-hexamethyl-2, 3-dihydro-1H-inden-5-yl) -3-methyloctane-2, 4, 6-trienoic acid

For examples 6A-6B: sodium hyaluronate may be provided by contiro A.S (DOLNI, DOBROUC, CZECH REPUBLIC).

For example 6A: extracts of actinomycete species a5640 (IHVR harvested bacterial extracts labelled IHVR _39565_ F7 according to IHVR harvested nomenclature) can be provided by barcuch s.

For example 6B: compound 1 can be supplied by Sigma-Aldrich.

For examples 6A-6B: polysorbate 20 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 6A-6B: polysorbate 80 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For example 8A: polyethylene glycol 400 can be provided by Clariant Produkte (Burgkirchen, GERMANY).

For examples 6A-6B: boric acid may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 6A-6B: sodium borate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 6A-6B: sodium chloride may be supplied by Caldic (DusseldoRF, GERMANY).

For examples 6A-6B: potassium chloride may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 6A-6B: calcium chloride dihydrate may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 6A-6B: magnesium chloride may be provided by KGaA (DARMSTADT, GERMANY).

For examples 6A-6B: polyquaternium-42 (33% aqueous solution) may be supplied by DSM BIOMEDICAL (BERKELEY, CA, USA).

For examples 6A-6B: sodium chlorite dihydrate can be supplied by Oxychem (WICHITA, KS, USA).

For examples 6A-6B: the 1N sodium hydroxide may be supplied by VWR (RADPER, PA, USA).

For examples 6A-6B: the 1N hydrochloric acid may be supplied by VWR (RANDER, PA, USA).

Solution 6A can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 100g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 50.0g of actinomycete extract. The solution was mixed until the actinomycete extract was dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 3.0 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

7. While mixing was continued, 0.14 grams of sodium chlorite dihydrate was added and mixed until dissolved.

8. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

9. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

10. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes to completely homogenize.

11. The solution was filtered using a 0.22 micron filter.

Solution 6B can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 75g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 50.0g of Compound I. The solution was mixed until compound I was dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 1.5 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

7. While mixing was continued, 0.14 grams of sodium chlorite dihydrate was added and mixed until dissolved.

8. The osmotic pressure of the formulation was measured and adjusted to 280mOsm/Kg with sodium chloride.

9. The pH of the formulation was adjusted to pH 7.2 using 1N sodium hydroxide and/or 1N hydrochloric acid.

10. The solution was adjusted to 1000.0 grams using purified water USP and mixed for 10 minutes to completely homogenize.

11. The solution was filtered using a 0.22 micron filter.

Example 7

Table 5 shows the components of the formulations of the present invention (as shown in formulations 7A-7B), which can be incorporated as described below using conventional mixing techniques.

TABLE 5

Adjusting to osmotic pressure of 280mOsm/Kg-290mOsm/Kg

Adjusting to pH 7.2

Moderate amount to 100.00% volume

4- (1- (1,1,2,2,3, 3-hexamethyl-2, 3-dihydro-1H-inden-5-yl) vinyl) benzoic acid

For examples 7A-7B: sodium hyaluronate may be provided by contiro A.S (DOLNI, DOBROUC, CZECH REPUBLIC).

For examples 7A-7B: the tamarind seed polysaccharide can be provided by INDENA (mirano, ITALY).

For example 7A: extracts of actinomycete species a5640 (IHVR harvested bacterial extracts labelled IHVR _39565_ F7 according to IHVR harvested nomenclature) can be provided by barcuch s.

For example 7B: compound 2 can be supplied by Sigma-Aldrich.

For examples 7A-7B: polysorbate 20 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: polysorbate 80 may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: polyethylene glycol 400 can be provided by Clariant Produkte (Burgkirchen, GERMANY).

For examples 7A-7B: boric acid may be provided by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: sodium borate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: sodium chloride may be supplied by Caldic (DusseldoRF, GERMANY).

For examples 7A-7B: potassium chloride may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: hypromellose E32910 may be supplied by DOW CHEMICAL (PLAQUEMINE, LOUISIANA, USA).

For examples 7A-7B: glycerol can be supplied by Emery Oleochemicals GmbH (DUSSELDORF, GERMANY).

For examples 7A-7B: disodium phosphate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: sodium citrate may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: sodium lactate may be provided as sodium lactate (50% aqueous solution) by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: glucose can be supplied by Roquette Freres (lastem, FRANCE).

For examples 7A-7B: glycine may be supplied by Merck KGaA (DARMSTADT, GERMANY).

For examples 7A-7B: ascorbic acid can be obtained from DSM Nutritional Products (DRAKEMYRE, SCOTLAND, UK).

For examples 7A-7B: polyquaternium 42 may be provided by DSM BIOMEDICAL (BERKELEY, CA) as Polyquaternium 42 (33% aqueous solution).

For examples 7A-7B: disodium edetate may be supplied by Merck NV/SA (OVERIJSE, BELGIUM).

For examples 7A-7B: the 1N sodium hydroxide may be supplied by VWR (RADPER, PA, USA).

For examples 7A-7B: the 1N hydrochloric acid may be supplied by VWR (RANDER, PA, USA).

For examples 7A-7B: sodium chlorite dihydrate can be supplied by Oxychem (WICHITA, KS, USA).

Solution 7A can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 100g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 50g of actinomycete extract. The solution was mixed until the actinomycete extract was dissolved.

4. The solution was filtered through a 0.45 micron filter and poured back into a 1500ml beaker.

5. To the solution of step 4 was added 1.2 grams of sodium hyaluronate. The solution was mixed to completely dissolve the sodium hyaluronate.

6. To the above solution was added 2.0 kr tamarind kernel polysaccharide. The solution was mixed to completely dissolve the tamarind seed polysaccharide.

7. To the above solution was added 1.98 grams of hypromellose E3 Premium. The solution was mixed until hypromellose E3 Premium was dissolved.

9. The osmolality of the solution was measured and adjusted to 280mOsm with sodium chloride.

10. The pH of the solution was measured and adjusted to 7.2 with 1N sodium hydroxide and/or 1N hydrochloric acid.

11. The solution was brought to 1,000.00 g with purified water and mixed for 10 minutes.

12. The solution was filtered using a 0.22 micron filter.

Solution 7B can be prepared as follows：

1. To a 1500ml beaker was added 800 grams of purified water USP.

2. To the above material were added 10g of polysorbate 80 and 30g of polysorbate 20. The solution was mixed until both were completely mixed and dissolved.

3. To the above material was added 5.0g of Compound I. The solution was mixed until compound I was dissolved.