# Patent

## Title
Prostaglandin and vasoconstrictor pharmaceutical compositions and methods of use

## Abstract

Compositions containing a prostaglandin agent and a vasoconstrictor agent for ophthalmic applications are provided as well as methods of treating ophthalmic diseases using the same. In certain embodiments, the compositions and methods are useful for treating glaucoma without causing conjunctival hyperemia.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Patent Application Ser. No. 61/637,597, filed Apr. 24, 2012, the disclosure of which is hereby incorporated in its entirety herein by reference.


BACKGROUND OF THE INVENTION
Based on its etiology, glaucoma can be classified into primary and secondary glaucoma. Primary glaucoma, also known as congenital glaucoma, can occur in the absence of other ocular conditions and its underlying causes are not known. However, it is known that increased intraocular pressure (IOP) observed in primary glaucoma is due to the obstruction of aqueous humor flow out of the eye. Secondary glaucoma results from another pre-existing ocular disease such as, uveitis, an intraocular tumor, enlarged cataract, central retinal vein occlusion, trauma to the eye, operative procedures and intraocular hemorrhage. Generally, any interference with the outward flow of aqueous humor from the posterior chamber into the anterior chamber and subsequently, into the canal of Schlemm can lead to secondary glaucoma.
Considering all types of glaucoma together, this ocular disorder occurs in about 2% of all persons over the age of 40. Unfortunately, glaucoma can be asymptomatic for years before progressing to a rapid loss of vision. In cases where surgery is not indicated, topical β-adrenoreceptor antagonists (or β-blockers) have traditionally been the drugs of choice for treating glaucoma. Further, certain prostaglandins and their analogues have been recommended for use in glaucoma management. For example, the prostaglandin analogue bimatoprost (Lumigan®) given at a dose of 0.03%, once daily, is a highly efficacious intraocular pressure (IOP) lowering drug. It has a very high systemic safety margin in studies conducted in laboratory animals. However, bimatoprost as well as many other topical anti-glaucoma medications produce conjunctival hyperemia as a side effect. Conjunctival hyperemia (or “red eye”) refers to vasodilatation of the conjunctival blood vessels. The vasodilatation of the conjunctival blood vessels results in increased blood supply to the vessels or even rupture of the vessels, which causes redness of the eye, reduced visual acuity, severe ocular pain, and photophobia (light sensitivity).
The present invention solves these as well as other problems in the art by, inter alia, providing compositions of sub-therapeutic concentrations of a vasoconstrictor agent (e.g., a beta adrenergic antagonist) in combination with a pharmaceutically effective amount of a prostaglandin agent (e.g., is defined to include prostaglandins, prostaglandin analogs, prostaglandin derivatives or prostamides such as bimatoprost) and methods of using such compositions to treat ophthalmic diseases without causing conjunctival hyperemia.
BRIEF SUMMARY OF THE INVENTION
Presented herein are, inter alia, compositions containing a prostaglandin agent and a vasoconstrictor agent (e.g. at sub-therapeutic concentrations) for ophthalmic applications as well as methods of treating ophthalmic diseases. In certain embodiments, the compositions and methods are useful for treating the symptoms of glaucoma. The compositions and methods provided herein are particularly useful for treating increased intraocular pressure (IOP) without causing the adverse effect of conjunctival hyperemia.
In one aspect, a composition including a prostaglandin agent and a vasoconstrictor agent is provided. In another aspect, a method of reducing increased intraocular pressure (IOP) in a subject in need thereof is provided. The method includes administering to the subject a therapeutically effective amount of an ophthalmic pharmaceutical formulation including a prostaglandin agent and a vasoconstrictor agent.
Some embodiments of the invention include:
1. A composition comprising a prostaglandin agent and a vasoconstrictor agent.
2. The composition of paragraph 1, wherein said composition is an ophthalmic pharmaceutical formulation further comprising an ophthalmically acceptable excipient.
3. The composition of paragraph 1, wherein said prostaglandin agent is present in a therapeutically effective amount.
4. The composition of paragraph 1, wherein said vasoconstrictor agent is present in a sub-therapeutic amount.
5. The composition of paragraph 2, wherein said prostaglandin agent and said vasoconstrictor agent are present in a combined amount effective to treat an ophthalmic disease.
6. The composition of paragraph 2, wherein said ophthalmic pharmaceutical formulation is a gel formulation.
7. The composition of paragraph 2, wherein said ophthalmic pharmaceutical formulation is an aqueous solution.
8. The composition of paragraph 2, wherein said prostaglandin agent is bimatoprost.
9. The composition of paragraph 8, wherein said bimatoprost is present in an amount approximately equal to or less than about 0.1% w/w.
10. The composition of paragraph 8, wherein said bimatoprost is present in an amount of about 0.03% w/w.
11. The composition of paragraph 2, wherein said prostaglandin agent is travoprost.
12. The composition of paragraph 11, wherein said travoprost is present in an amount approximately equal to or less than about 0.1% w/w.
13. The composition of paragraph 11, wherein said travoprost is present in an amount of about 0.004% w/w.
14. The composition of paragraph 2, wherein said prostaglandin agent is latanoprost.
15. The composition of paragraph 14, wherein said latanoprost is present in an amount approximately equal to or less than about 0.1% w/w.
16. The composition of paragraph 14, wherein said latanoprost is present in an amount of about 0.005% w/w.
17. The composition of paragraph 2, wherein said vasoconstrictor agent is an alpha adrenergic agonist.
18. The composition of paragraph 2, wherein said vasoconstrictor agent is a beta adrenergic antagonist.
19. The composition of paragraph 18, wherein said beta adrenergic antagonist is timolol.
20. The composition of paragraph 19, wherein said timolol is present in a sub-therapeutic amount.
21. The composition of paragraphs 18 or 20, wherein said sub-therapeutic amount is an amount less than about 0.25% w/w or less than 0.1% w/w or less than 0.05% w/w or in a range from 0.0001%-0.001% w/w.
22. The composition of paragraph 19, wherein said timolol is timolol maleate.
23. The composition of paragraph 19, wherein said timolol is timolol hemihydrate.
24. The composition of paragraph 18, wherein said beta adrenergic antagonist is betaxolol.
25. The composition of paragraph 24, wherein said betaxolol is present in a sub-therapeutic amount.
26. The composition of paragraph 25, wherein said sub-therapeutic amount is an amount less than about 0.05% w/w or in an amount less than 0.005% w/w.
27. The composition of paragraph 24, wherein said betaxolol is a betaxolol salt.
28. The composition of paragraph 18, wherein said beta adrenergic antagonist is levobunolol.
29. The composition of paragraph 28, wherein said levobunolol is present in a sub-therapeutic amount.
30. The composition of paragraph 29, wherein said sub-therapeutic amount is an amount less than about 0.25% w/w or less than 0.025% w/w.
31. The composition of paragraph 28, wherein said levobunolol is a levobunolol salt.
32. The composition of paragraph 18, wherein said beta adrenergic antagonist is metipranolol.
33. The composition of paragraph 32, wherein said metipranolol is present in a sub-therapeutic amount.
34. The composition of paragraph 33, wherein said sub-therapeutic amount is an amount less than about 0.3% w/w or 0.03% w/w.
35. The composition of paragraph 32, wherein said metipranolol is a metipranolol salt.
36. The composition of paragraph 2, further comprising a buffering agent, a tonicity agent, a salt, and a preservative.
37. The composition of paragraph 2, consisting essentially of bimatoprost, timolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
38. The composition of paragraph 37, wherein said timolol is present in a sub-therapeutic amount.
39. The composition of paragraph 38, wherein said sub-therapeutic amount is an amount less than about 0.25% w/w or less than 0.05% w/w or 0.005% w/w.
40. The composition of paragraph 2, consisting essentially of bimatoprost, betaxolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
41. The composition of paragraph 40, wherein said betaxolol is present in a sub-therapeutic amount.
42. The composition of paragraph 41, wherein said sub-therapeutic amount is an amount less than about 0.25% w/w.
43. The composition of paragraph 2, consisting essentially of bimatoprost, levobunolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
44. The composition of paragraph 43, wherein said levobunolol is present in a sub-therapeutic amount.
45. The composition of paragraph 44, wherein said sub-therapeutic amount is an amount less than about 0.25% w/w.
46. The composition of paragraph 2, consisting essentially of bimatoprost, metipranolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
47. The composition of paragraph 46, wherein said metipranolol is present in a sub-therapeutic amount.
48. The composition of paragraph 47, wherein said sub-therapeutic amount is an amount less than about 0.3% w/w.
49. A method of reducing intraocular pressure (IOP) in a subject in need thereof, said method comprising:
administering to said subject a therapeutically effective amount of an ophthalmic pharmaceutical formulation comprising a prostaglandin agent and a vasoconstrictor agent.
50. The method of paragraph 49, wherein said prostaglandin agent is present in a therapeutically effective amount and said vasoconstrictor agent is present in a sub-therapeutic amount.
51. The method of paragraph 49, wherein said prostaglandin agent is bimatoprost and wherein said vasoconstrictor agent is timolol.
52. The method of paragraph, wherein timolol is present in an amount less than about 0.25% w/w.
53. The method of paragraph 49, wherein said ophthalmic pharmaceutical formulation further comprises a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
54. The method of paragraph 49, wherein said ophthalmic pharmaceutical formulation consists essentially of bimatoprost, timolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
55. The method of paragraph 54, wherein bimatoprost is present in a therapeutically effective amount and the timolol is present in a sub-therapeutic amount.
56. The composition of paragraph 4, wherein the vasoconstrictor is selected from the group consisting of befunolol, betaxolol, carteolol, levobunolol, metipranolol, timolol, brimonidine, tetrahydrozolone hydrochloride and mepindolol.
57. The composition of paragraph 56, where the vasoconstrictor is present in an amount selected from the group consisting of 0.01 to about 0.06, from about 0.02 to about 0.06, from about 0.03 to about 0.06, from about 0.04 to about 0.06, from about 0.05 to about 0.06, from about 0.001 to about 0.04, from about 0.002 to about 0.04, from about 0.003 to about 0.04, from about 0.004 to about 0.04, from about 0.005 to about 0.04, from about 0.006 to about 0.04, from about 0.007 to about 0.04, from about 0.008 to about 0.04, from about 0.009 to about 0.04, from about 0.01 to about 0.04, from about 0.02 to about 0.04, from about 0.03 to about 0.04, from about 0.001 to about 0.02, from about 0.002 to about 0.02, from about 0.003 to about 0.02, from about 0.004 to about 0.02, from about 0.005 to about 0.02, from about 0.006 to about 0.02, from about 0.007 to about 0.02, from about 0.008 to about 0.02, from about 0.009 to about 0.02, from about 0.01 to about 0.02, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.003 to about 0.01, from about 0.004 to about 0.01, from about 0.005 to about 0.01, from about 0.006 to about 0.01, from about 0.007 to about 0.01, from about 0.008 to about 0.01, or from about 0.009 to about 0.01% (w/w).



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1. Conjunctival hyperemia scale scores at each study visit in patients treated with topically applied bimatoprost 0.03% once daily. Results are expressed as the mean hyperemia score of three vessel-bed scores from 33-39 patients with bilateral open-angle glaucoma or ocular hypertension. Adapted from Abelson M B et al., Adv. Ther. 2003; 20:1-13.
FIG. 2. (Top) Photomicrographs of conjunctival specimens stained with hematoxylin and eosin (H&E, ×200), showing vascular congestion (arrows). (Top left) Specimen from patient 3 of the control group. (Top right) Specimen from patient 4 of the bimatoprost group. (Bottom) Photomicrographs of conjunctival specimens stained with H&E (×400) showing features of acute inflammatory response. (Bottom left) Specimen from patient 1 of the control group, showing vascular congestion with polymorphonuclear leukocyte margination (long arrow), diapedesis phenomenon (short arrow), and inflammatory cell infiltrate. (Bottom right) Specimen from patient 8 of the bimatoprost group, showing vascular congestion with polymorphonuclear leukocyte margination (arrow). Reprinted from FIG. 1 of Leal B C et al, Am J Ophthalmol 2004; 138:310-313, with permission from Elsevier.
FIG. 3. Concentration-response curves for (FIG. 3A) bimatoprost and (FIG. 3B) latanoprost free acid in histamine-precontracted endothelium-intact and endothelium-denuded rabbit isolated jugular veins. Results are expressed as mean±S.E.M. of 5-6 animals. FIG. 3A adapted from Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609; FIG. 3B extends results reported in Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609; and Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda2001/21275_Lumigan.htm.
FIG. 4. Concentration-response curves for (FIG. 4A) bimatoprost and (FIG. 4B) latanoprost free acid in the presence or absence of indomethacin in endothelium-intact rabbit isolated jugular veins. Results are expressed as mean±S.E.M. of 7 animals and extend previous results reported in Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275_Lumigan.htm.
FIG. 5. Concentration-response curves for (FIG. 5A) bimatoprost and (FIG. 5B) latanoprost free acid in the presence of 100 μM L-NAME or 100 (μM D-NAME control in endothelium-intact rabbit isolated jugular veins. Results are expressed as mean±S.E.M. of 6 animals. *P≦0.05, compared to responses in D-NAME pretreated tissues at the same concentration, paired t-test.
FIG. 6. Effects of L-NAME (test eye) and D-NAME control (contralateral fellow eye) on ocular surface hyperemia elicited by (FIG. 6A) 0.1% bimatoprost, (FIG. 6B) 0.005% latanoprost, or (FIG. 6C) 0.01% PGE2, topically applied to both eyes of each dog at time 0. Values are expressed as mean±S.E.M. of 8 animals. *P≦0.05, difference from baseline (time 0), Wilcoxon signed-rank test for paired observations. Data from Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609.



DETAILED DESCRIPTION OF THE INVENTION
The terms “a,” “an,” or “the” as used herein not only include aspects with one member, but also aspects with more than one member. For example, an embodiment including “a buffer and a chelating agent” should be understood to present aspects with at least a second buffer, at least a second chelating agent, or both.
The term “or” as used herein should in general be construed non-exclusively. For example, an embodiment of “a formulation including A or B” would typically present an aspect with a formulation including both A and B. “Or” should, however, be construed to exclude those aspects presented that cannot be combined without contradiction (e.g., a formulation pH that is between 9 and 10 or between 7 and 8).
“Agent” as used herein indicates a compound or mixture of compounds that, when added to a pharmaceutical formulation, tend to produce a particular effect on the formulation's properties. For example, a formulation including a thickening agent is likely to be more viscous than an otherwise identical comparative formulation that lacks the thickening agent.
“Formulation,” “composition,” and “preparation” as used herein are equivalent terms referring to a composition of matter suitable for pharmaceutical use (i.e., producing a therapeutic effect as well as possessing acceptable pharmacokinetic and toxicological properties).
As used herein, the term “pharmaceutically” acceptable is used as equivalent to physiologically acceptable. In certain embodiments, a pharmaceutically acceptable composition or preparation will include agents for buffering and preservation in storage, and can include buffers and carriers for appropriate delivery, depending on the route of administration.
As used herein, the terms “prevent” and “treat” are not intended to be absolute terms. Treatment can refer to any delay in onset, e.g., reduction in the frequency or severity of symptoms, amelioration of symptoms, improvement in patient comfort, and the like. The effect of treatment can be compared to an individual or pool of individuals not receiving a given treatment, or to the same patient before, or after cessation of, treatment.
The terms “subject,” “patient,” “individual,” and the like as used herein are not intended to be limiting and can be generally interchanged. That is, an individual described as a “patient” does not necessarily have a given disease, but may be merely seeking medical advice.
The term “subject” as used herein includes all members of the animal kingdom prone to suffering from the indicated disorder. In some aspects, the subject is a mammal, and in some aspects, the subject is a human.
The terms “effective amount,” “therapeutically effective amount” or “pharmaceutically effective amount” as used herein refer to that amount of the therapeutic agent sufficient to ameliorate one or more aspects of the disorder. The result can be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an “effective amount” for therapeutic uses is the amount of the composition comprising an agent as set forth herein required to provide a clinically significant decrease in an ophthalmic disease. For example, for the given aspect (e.g., length of incidence), a therapeutically effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as “-fold” increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control. An appropriate “effective” amount in any individual case may be determined using techniques, such as a dose escalation study.
The term “sub-therapeutic amount” or “sub-therapeutic concentration” as provided herein refers to an amount of a therapeutic agent (e.g. drug) that is insufficient to provide a therapeutic effect in the absence of an additional active agent. In some embodiments, the sub-therapeutic amount is less than the clinically approved amount or concentration of that therapeutic agent. In some embodiments, a sub-therapeutic amount is less than the lowest clinically approved amount of a therapeutic agent. A clinically approved amount of a therapeutic agent is the amount approved by the U.S. Food and Drug Administration (FDA) or an equivalent regulatory entity, to be safe and effective when used as directed.
“Treating” or “treatment” as used herein (and as well-understood in the art) also broadly includes any approach for obtaining beneficial or desired results in a subject's condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, prevention of a disease's transmission or spread, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. In other words, “treatment” as used herein includes any cure, amelioration, or prevention of a disease. Treatment may prevent the disease from occurring; inhibit the disease's spread; relieve the disease's symptoms (e.g., ocular pain, seeing halos around lights, red eye, very high intraocular pressure), fully or partially remove the disease's underlying cause, shorten a disease's duration, or do a combination of these things.
“Treating” and “treatment” as used herein include prophylactic treatment. Treatment methods include administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may include a series of administrations. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the age of the patient, the concentration of active agent, the activity of the compositions used in the treatment, or a combination thereof. It will also be appreciated that the effective dosage of an agent used for the treatment or prophylaxis may increase or decrease over the course of a particular treatment or prophylaxis regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. For example, the compositions are administered to the subject in an amount and for a duration sufficient to treat the patient.
The term “disease” refers to any deviation from the normal health of a mammal and includes a state when disease symptoms are present, as well as conditions in which a deviation (e.g., infection, gene mutation, genetic defect, etc.) has occurred, but symptoms are not yet manifested. According to the present invention, the methods disclosed herein are suitable for use in a patient that is a member of the Vertebrate class, Mammalia, including, without limitation, primates, livestock and domestic pets (e.g., a companion animal). Typically, a patient will be a human patient.
As used herein, “topical application,” “topical administration,” and “topically administering” are used interchangeably herein and include the administration of a composition to the eye, the mucosal or dermal area proximal to the eye. Topical application or administering may result in the delivery of an active agent to the eye or skin, a localized region of the body, a localized volume of the body, or the systemic circulation.
“Topical formulation” and “topical pharmaceutical composition” are used interchangeably herein and include a formulation that is suitable for topical application to the eye or dermal area proximal to the eye, or other localized region of the body. A topical formulation may, for example, be used to confer a therapeutic benefit to its user. Specific topical formulations can be used for topical, local, regional, or transdermal application of substances.
As used herein, the terms “application,” “apply,” and “applying” used in reference to a topical composition product or method of using a composition or a product, refer to any manner of administering a topical composition or a product to the eye, the mucosal or dermal area proximal to the eye of a patient which, in medical or cosmetology practice, delivers the composition or the product to patient's eye, the mucosal or dermal area proximal to the eye. Smearing, rubbing, spreading, spraying a topical composition, with or without the aid of suitable devices, on a patient's skin are all included within the scope of the term “application,” as used herein. The term “topical” or “topically” in reference to administration or application of a composition or a product refers to epicutaneous administration or application, or administration onto skin. The term “topically active agent” as used herein refers to a compound that is effective in a treatment of a skin condition when administered topically. It is to be understood that a topically active agent can have a local or a systemic effect, or both, when administered topically. The term “topical,” when used in reference to a composition or a product refers to a composition or a product formulated for topical application.
The terms “adverse effect” or “side effects” as used herein refer to a harmful and undesired effect associated with the use of a medication (i.e. drug). An adverse effect may be termed a side effect, when judged to be secondary to a main or therapeutic effect. Adverse effects or side effects may occur when starting treatment, increasing treatment dosage or discontinuing treatment with a drug. Adverse effects may also be caused by drug interaction when more than one drug is administered. In some embodiments, the side effect of administering therapeutically effective amounts
The abbreviations used herein have their conventional meaning within the chemical, biological or pharmaceutical arts.
The terms “about” and “approximately equal” are used herein to modify a numerical value and indicate a defined range around that value. If “X” were the value, “about X” or “approximately equal to X” would generally indicate a value from 0.90X to 1.10X. Any reference to “about X” minimally indicates at least the values X, 0.90X, 0.91X, 0.92X, 0.93X, 0.94X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, 1.05X, 1.06X, 1.07X, 1.08X, 1.09X, and 1.10X. Thus, “about X” is intended to disclose, e.g., “0.98X.” When “about” is applied to the beginning of a numerical range, it applies to both ends of the range. Thus, “from about 6 to 8.5” is equivalent to “from about 6 to about 8.5.” When “about” is applied to the first value of a set of values, it applies to all values in that set. Thus, “about 7, 9, or 11%” is equivalent to “about 7%, about 9%, or about 11%.”
As used herein, the phrase “pharmaceutically acceptable salts” refers to salts of the active compound(s) which possess the same pharmacological activity as the active compound(s) and which are neither biologically nor otherwise undesirable. A salt can be formed with, for example, organic or inorganic acids. Non-limiting examples of suitable acids include acetic acid, acetylsalicylic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzoic acid, benzenesulfonic acid, bisulfic acid, boric acid, butyric acid, camphoric acid, camphorsulfonic acid, carbonic acid, citric acid, cyclopentanepropionic acid, digluconic acid, dodecylsulfic acid, ethanesulfonic acid, formic acid, fumaric acid, glyceric acid, glycerophosphoric acid, glycine, glucoheptanoic acid, gluconic acid, glutamic acid, glutaric acid, glycolic acid, hemisulfic acid, heptanoic acid, hexanoic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthylanesulfonic acid, naphthylic acid, nicotinic acid, nitrous acid, oxalic acid, pelargonic, phosphoric acid, propionic acid, saccharin, salicylic acid, sorbic acid, succinic acid, sulfuric acid, tartaric acid, thiocyanic acid, thioglycolic acid, thiosulfuric acid, tosylic acid, undecylenic acid, naturally and synthetically derived amino acids. Non-limiting examples of base salts include ammonium salts; alkali metal salts, such as sodium and potassium salts; alkaline earth metal salts, such as calcium and magnesium salts; salts with organic bases, such as dicyclohexylamine salts; methyl-D-glucamine; and salts with amino acids, such as arginine, lysine, and so forth. Also, the basic nitrogen-containing groups can be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as dimethyl, diethyl, dibutyl, and diamyl sulfates; long chain halides, such as decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; asthma halides, such as benzyl and phenethyl bromides; and others.
In formulations including an “additional,” “further,” or “second” component, the second component as used herein is chemically different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.
The term “hydrophobic” is used herein in accordance with its plain ordinary meaning and refers to a chemical group having a tendency to attract non-polar or uncharged chemical groups, e.g. hexane, and to repel polar or charged chemical groups, e.g. water.
“Conjunctival hyperemia” or “red eye” refers to an ophthalmic condition caused by vasodilatation of the conjunctival blood vessels. The vasodilatation of the conjunctival blood vessels results in increased blood supply to the vessels or even rupture of the vessels, which causes redness of the eye, reduced visual acuity, severe ocular pain, and photophobia (light sensitivity). Conjunctival hyperemia is a common side effect caused by many topical glaucoma medications (e.g., bimatoprost or Lumigan®).
I. COMPOSITIONS
The present invention provides pharmaceutical compositions including a pharmaceutically active ingredient (e.g., two pharmaceutically active ingredients) and an ophthalmically acceptable excipient. In some embodiments, the pharmaceutical composition includes a prostaglandin agent and a vasoconstrictor agent. In some embodiments, the vasoconstrictor agent is present in a sub-therapeutic amount. The compositions provided herein may be used for the treatment of ophthalmic diseases. In some embodiments, the compositions are used for the treatment of increased intraocular pressure (IOP). Gels, aqueous solutions, emulsions, creams or ointments are contemplated as useful pharmaceutical formulations including the compositions provided herein.
In one aspect, a composition including a prostaglandin agent and a vasoconstrictor agent is provided. In some embodiments, the composition is an ophthalmic pharmaceutical formulation further including an ophthalmically acceptable excipient. Generally, ophthalmically acceptable excipients commonly known in the fields of ophthalmology and cosmetology as useful in topical compositions, and any non-toxic, inert, and effective topical carriers, are contemplated as useful in the compositions and products according to the embodiments of the present invention.
In some embodiments, the prostaglandin agent is present in a therapeutically effective amount. In other embodiments, the vasoconstrictor agent is present in a sub-therapeutic amount. As described above a sub-therapeutic amount is an amount of a therapeutic agent (e.g. drug), which is different to the clinically approved amount or concentration of that therapeutic agent. Thus, when a therapeutic agent (e.g. drug) is present at a sub-therapeutic amount it may be present at an amount that is less than the clinically approved amount for that therapeutic agent. In some embodiments, a sub-therapeutic amount of a therapeutic agent is less than the lowest clinically approved amount of that therapeutic agent.
Where the ophthalmic compositions provided herein include more than one active pharmaceutical ingredient, the active pharmaceutical ingredients are present in a combined amount effective to treat ophthalmic diseases. A combined effective amount is the total amount of active pharmaceutical ingredients that, in combination, provide an effective amount. For example, a combined effective amount may include a first amount of a first active pharmaceutical ingredient (e.g. a prostaglandin agent) and a second amount of a second active pharmaceutical ingredient (e.g. a vasoconstrictor agent). The first amount of the first active pharmaceutical ingredient may be less than an effective amount of the first active pharmaceutical ingredient when the first active pharmaceutical ingredient is administered without the second active pharmaceutical ingredient. On the other hand, the first amount of the first active pharmaceutical ingredient may be more than an effective amount of the first active pharmaceutical ingredient when the first active pharmaceutical ingredient is administered without the second active pharmaceutical ingredient. Correspondingly, the second amount of the second active pharmaceutical ingredient may be less than an effective amount of the second active pharmaceutical ingredient when the second active pharmaceutical ingredient is administered without the first active pharmaceutical ingredient. Equally, the second amount of the second active pharmaceutical ingredient may be more than an effective amount of the second active pharmaceutical ingredient when the second active pharmaceutical ingredient is administered without the first active pharmaceutical ingredient. Thus, in some embodiments, the prostaglandin agent and the vasoconstrictor agent are present in a combined amount effective to treat ophthalmic diseases.
The ophthalmic pharmaceutical compositions provided herein may be administered in various ways e.g., a foam, a gel, a cream, jelly, solution, suspension, a spray (e.g., a solution), an ointment, ointment films, occlusive films, sustained release films, fast drying films, slow drying films, patches, semi solids or stick formulation comprising a semi-solid vehicle with a melting point near physiological temperature. Topical compositions and products according to embodiments of the present invention can be formulated as creams, which can be semi-solid emulsions of oil and water, and lotions, including suspensions of powdered material in water or alcohol base and water-based emulsions. Topical compositions and products according to embodiments of the present invention can also be formulated as ointments, which are oleaginous and contain little if any water. In some embodiments, the ophthalmic pharmaceutical formulation is a gel formulation. In other embodiments, the ophthalmic pharmaceutical formulation is an aqueous formulation.
The compositions provided herein may include a prostaglandin agent and a vasoconstrictor agent as active pharmaceutical ingredients. It is also to be understood that pharmaceutically acceptable salts of the active pharmaceutical ingredients may be included in the compositions provided herein. A prostaglandin agent is a compound capable of binding a prostaglandin E2 receptor. A prostaglandin E2 receptor as defined herein is a G-protein coupled receptor that is being bound by prostaglandin E2. Prostaglandin E2 is a lipid mediator that is derived enzymatically from fatty acids and has important functions in the animal body. E2 prostaglandins have a variety of strong physiological effects, such as regulating the contraction and relaxation of smooth muscle tissue. Non-limiting examples of prostaglandin agents contemplated for the compositions and methods provided herein are travoprost, latanoprost, unoprostone, tafluprost and bimatoprost. In some embodiments, the prostaglandin agent is bimatoprost. Bimatoprost refers, in the customary sense, to CAS Registry No. 155206-00-1. In some embodiments, bimatoprost is present in an amount approximately equal to or less than about 0.1% w/w. In some embodiments, bimatoprost is present from about 0.001 to about 0.1, from about 0.002 to about 0.1, from about 0.003 to about 0.1, from about 0.004 to about 0.1, from about 0.005 to about 0.1, from about 0.006 to about 0.1, from about 0.007 to about 0.1, from about 0.008 to about 0.1, from about 0.009 to about 0.1, from about 0.01 to about 0.1, from about 0.02 to about 0.1, from about 0.03 to about 0.1, from about 0.04 to about 0.1, from about 0.05 to about 0.1, from about 0.06 to about 0.1, from about 0.07 to about 0.1, from about 0.08 to about 0.1, from about 0.09 to about 0.1, from about 0.001 to about 0.08, from about 0.002 to about 0.08, from about 0.003 to about 0.08, from about 0.004 to about 0.08, from about 0.005 to about 0.08, from about 0.006 to about 0.08, from about 0.007 to about 0.08, from about 0.008 to about 0.08, from about 0.009 to about 0.08, from about 0.01 to about 0.08, from about 0.02 to about 0.08, from about 0.03 to about 0.08, from about 0.04 to about 0.08, from about 0.05 to about 0.08, from about 0.06 to about 0.08, from about 0.07 to about 0.08, from about 0.001 to about 0.06, from about 0.002 to about 0.06, from about 0.003 to about 0.06, from about 0.004 to about 0.06, from about 0.005 to about 0.06, from about 0.006 to about 0.06, from about 0.007 to about 0.06, from about 0.008 to about 0.06, from about 0.009 to about 0.06, from about 0.01 to about 0.06, from about 0.02 to about 0.06, from about 0.03 to about 0.06, from about 0.04 to about 0.06, from about 0.05 to about 0.06, from about 0.001 to about 0.04, from about 0.002 to about 0.04, from about 0.003 to about 0.04, from about 0.004 to about 0.04, from about 0.005 to about 0.04, from about 0.006 to about 0.04, from about 0.007 to about 0.04, from about 0.008 to about 0.04, from about 0.009 to about 0.04, from about 0.01 to about 0.04, from about 0.02 to about 0.04, from about 0.03 to about 0.04, from about 0.001 to about 0.02, from about 0.002 to about 0.02, from about 0.003 to about 0.02, from about 0.004 to about 0.02, from about 0.005 to about 0.02, from about 0.006 to about 0.02, from about 0.007 to about 0.02, from about 0.008 to about 0.02, from about 0.009 to about 0.02, from about 0.01 to about 0.02, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.003 to about 0.01, from about 0.004 to about 0.01, from about 0.005 to about 0.01, from about 0.006 to about 0.01, from about 0.007 to about 0.01, from about 0.008 to about 0.01, or from about 0.009 to about 0.01% (w/w). In some embodiments, bimatoprost is present at about 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.1% (w/w). In some embodiments, bimatoprost is present in an amount of about 0.03% w/w.
In other embodiments, the prostaglandin agent is travoprost. Travoprost refers, in the customary sense, to CAS Registry No. 157283-68-6. In some embodiments, the travoprost is present in an amount approximately equal to or less than about 0.1% w/w. In some embodiments, the travoprost is present in an amount from about 0.0002 to about 0.1, from about 0.0004 to about 0.1, from about 0.0006 to about 0.1, from about 0.0008 to about 0.1, from about 0.001 to about 0.1, from about 0.002 to about 0.1, from about 0.004 to about 0.1, from about 0.006 to about 0.1, from about 0.008 to about 0.1, from about 0.01 to about 0.1, from about 0.02 to about 0.1, from about 0.04 to about 0.1, from about 0.06 to about 0.1, from about 0.08 to about 0.1, from about 0.0002 to about 0.08, from about 0.0004 to about 0.08, from about 0.0006 to about 0.08, from about 0.0008 to about 0.08, from about 0.001 to about 0.08, from about 0.002 to about 0.08, from about 0.004 to about 0.08, from about 0.006 to about 0.08, from about 0.008 to about 0.08, from about 0.01 to about 0.08, from about 0.02 to about 0.08, from about 0.04 to about 0.08, from about 0.06 to about 0.08, from about 0.0002 to about 0.06, from about 0.0004 to about 0.06, from about 0.0006 to about 0.06, from about 0.0008 to about 0.06, from about 0.001 to about 0.06, from about 0.002 to about 0.06, from about 0.004 to about 0.06, from about 0.006 to about 0.06, from about 0.008 to about 0.06, from about 0.01 to about 0.06, from about 0.02 to about 0.06, from about 0.04 to about 0.06, from about 0.0002 to about 0.04, from about 0.0004 to about 0.04, from about 0.0006 to about 0.04, from about 0.0008 to about 0.04, from about 0.001 to about 0.04, from about 0.002 to about 0.04, from about 0.004 to about 0.04, from about 0.006 to about 0.04, from about 0.008 to about 0.04, from about 0.01 to about 0.04, from about 0.02 to about 0.04, from about 0.0002 to about 0.02, from about 0.0004 to about 0.02, from about 0.0006 to about 0.02, from about 0.0008 to about 0.02, from about 0.001 to about 0.02, from about 0.002 to about 0.02, from about 0.004 to about 0.02, from about 0.006 to about 0.02, from about 0.008 to about 0.02, from about 0.01 to about 0.02, from about 0.0002 to about 0.01, from about 0.0004 to about 0.01, from about 0.0006 to about 0.01, from about 0.0008 to about 0.01, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.004 to about 0.01, from about 0.006 to about 0.01, from about 0.008 to about 0.01, from about 0.0002 to about 0.008, from about 0.0004 to about 0.008, from about 0.0006 to about 0.008, from about 0.0008 to about 0.008, from about 0.001 to about 0.008, from about 0.002 to about 0.008, from about 0.004 to about 0.008, from about 0.006 to about 0.008, from about 0.0002 to about 0.006, from about 0.0004 to about 0.006, from about 0.0006 to about 0.006, from about 0.0008 to about 0.006, from about 0.001 to about 0.006, from about 0.002 to about 0.006, from about 0.004 to about 0.006, from about 0.0002 to about 0.004, from about 0.0004 to about 0.004, from about 0.0006 to about 0.004, from about 0.0008 to about 0.004, from about 0.001 to about 0.004, from about 0.002 to about 0.004, from about 0.0002 to about 0.002, from about 0.0004 to about 0.002, from about 0.0006 to about 0.002, from about 0.0008 to about 0.002, from about 0.001 to about 0.002, from about 0.0002 to about 0.001, from about 0.0004 to about 0.001, from about 0.0006 to about 0.001, from about 0.0008 to about 0.001, from about 0.0002 to about 0.0008, from about 0.0004 to about 0.0008, from about 0.0006 to about 0.0008, from about 0.0002 to about 0.0006, from about 0.0004 to about 0.0006, or from about 0.0002 to about 0.0004% (w/w). In some embodiments, the travoprost is present at about 0.1, 0.08, 0.06, 0.04, 0.02, 0.01, 0.008, 0.006, 0.004, 0.002, 0.001, 0.0008, 0.0006, 0.0004, or 0.0002% (w/w). In some embodiments, travoprost is present in an amount of about 0.004% w/w.
In some embodiments, the prostaglandin agent is latanoprost. Latanoprost refers, in the customary sense, to CAS Registry No. 130209-82-4. In some embodiments, the latanoprost is present in an amount approximately equal to or less than about 0.1% w/w. In some embodiments, latanoprost is present from about 0.0003 to about 0.1, from about 0.0005 to about 0.1, from about 0.0007 to about 0.1, from about 0.0009 to about 0.1, from about 0.001 to about 0.1, from about 0.003 to about 0.1, from about 0.005 to about 0.1, from about 0.007 to about 0.1, from about 0.009 to about 0.1, from about 0.01 to about 0.1, from about 0.03 to about 0.1, from about 0.05 to about 0.1, from about 0.07 to about 0.1, from about 0.09 to about 0.1, from about 0.0003 to about 0.09, from about 0.0005 to about 0.09, from about 0.0007 to about 0.09, from about 0.0009 to about 0.09, from about 0.001 to about 0.09, from about 0.003 to about 0.09, from about 0.005 to about 0.09, from about 0.007 to about 0.09, from about 0.009 to about 0.09, from about 0.01 to about 0.09, from about 0.03 to about 0.09, from about 0.05 to about 0.09, from about 0.07 to about 0.09, from about 0.0003 to about 0.07, from about 0.0005 to about 0.07, from about 0.0007 to about 0.07, from about 0.0009 to about 0.07, from about 0.001 to about 0.07, from about 0.003 to about 0.07, from about 0.005 to about 0.07, from about 0.007 to about 0.07, from about 0.009 to about 0.07, from about 0.01 to about 0.07, from about 0.03 to about 0.07, from about 0.05 to about 0.07, from about 0.0003 to about 0.05, from about 0.0005 to about 0.05, from about 0.0007 to about 0.05, from about 0.0009 to about 0.05, from about 0.001 to about 0.05, from about 0.003 to about 0.05, from about 0.005 to about 0.05, from about 0.007 to about 0.05, from about 0.009 to about 0.05, from about 0.01 to about 0.05, from about 0.03 to about 0.05, from about 0.0003 to about 0.03, from about 0.0005 to about 0.03, from about 0.0007 to about 0.03, from about 0.0009 to about 0.03, from about 0.001 to about 0.03, from about 0.003 to about 0.03, from about 0.005 to about 0.03, from about 0.007 to about 0.03, from about 0.009 to about 0.03, from about 0.01 to about 0.03, from about 0.0003 to about 0.01, from about 0.0005 to about 0.01, from about 0.0007 to about 0.01, from about 0.0009 to about 0.01, from about 0.001 to about 0.01, from about 0.003 to about 0.01, from about 0.005 to about 0.01, from about 0.007 to about 0.01, from about 0.009 to about 0.01, from about 0.0003 to about 0.009, from about 0.0005 to about 0.009, from about 0.0007 to about 0.009, from about 0.0009 to about 0.009, from about 0.001 to about 0.009, from about 0.003 to about 0.009, from about 0.005 to about 0.009, from about 0.007 to about 0.009, from about 0.0003 to about 0.007, from about 0.0005 to about 0.007, from about 0.0007 to about 0.007, from about 0.0009 to about 0.007, from about 0.001 to about 0.007, from about 0.003 to about 0.007, from about 0.005 to about 0.007, from about 0.0003 to about 0.005, from about 0.0005 to about 0.005, from about 0.0007 to about 0.005, from about 0.0009 to about 0.005, from about 0.001 to about 0.005, from about 0.003 to about 0.005, from about 0.0003 to about 0.003, from about 0.0005 to about 0.003, from about 0.0007 to about 0.003, from about 0.0009 to about 0.003, from about 0.001 to about 0.003, from about 0.0003 to about 0.001, from about 0.0005 to about 0.001, from about 0.0007 to about 0.001, from about 0.0009 to about 0.001, from about 0.0003 to about 0.0009, from about 0.0005 to about 0.0009, from about 0.0007 to about 0.0009, from about 0.0003 to about 0.0007, from about 0.0005 to about 0.0007, or from about 0.0003 to about 0.0005% (w/w). In some embodiments, the latanoprost is present at about 0.1, 0.09, 0.07, 0.05, 0.03, 0.01, 0.009, 0.007, 0.005, 0.003, 0.001, 0.0009, 0.0007, 0.0005, or 0.0003% (w/w). In some embodiments, the latanoprost is present in an amount of about 0.005% w/w.
As mentioned above the ophthalmic pharmaceutical formulations provided herein may include a vasoconstrictor agent. A vasoconstrictor agent is an agent having a vasoconstriction effect on blood vessels. Vasoconstriction is the narrowing of blood vessels resulting from contraction of the muscular wall of the vessels. Vasoconstriction is a mechanism by which the body regulates and maintains mean arterial pressure. Therefore, vasoconstrictors or vasoconstrictor agents are agents causing a general increase in systemic blood pressure, but at the same time may cause a localized reduction in blood flow. In some embodiments, the vasoconstrictor agent is an alpha adrenergic agonist. An alpha adrenergic agonist is an agent (e.g., drug, compound), which selectively stimulates alpha adrenergic receptors. Alpha adrenergic receptors are G protein-coupled receptors that are bound by noradrenalin and adrenaline. Binding of an agonist to an alpha adrenergic receptor leads to vasoconstriction, which causes a sympathetic response, where the heart rate increases, the pupils dilate and blood flow is being diverted from non-essential organs to the skeletal muscle. A non-limiting example of an alpha adrenergic agonist is brimonidine. Brimonidine refers, in the customary sense, to CAS Registry No. 59803-98-4. In some embodiments, the alpha adrenergic agonist is brimonidine.
In other embodiments, the vasoconstrictor agent is a beta adrenergic antagonist. A beta adrenergic antagonist is an agent (e.g., drug, compound), which blocks the stimulation of beta adrenergic receptors. Stimulation of beta adrenergic receptors induces smooth muscle relaxation, whereas blocking beta adrenergic receptors causes contraction of smooth muscles. Therefore, beta adrenergic antagonists can cause vasoconstriction. Examples of beta adrenergic antagonists are without limitation befunolol, betaxolol, carteolol, levobunolol, metipranolol, timolol, and mepindolol.
In some embodiments, the beta adrenergic antagonist is timolol. In some embodiments, the timolol is timolol maleate. Timolol maleate refers, in the customary sense, to CAS Registry No. 26839-75-8. The chemical name of timolol maleate is (−)-1-tert-butylamino-3-[(4-morpholino-1,2,5-thiodiazol-3yl)oxy]-2-propanol maleate. Timolol maleate has a molecular weight of 432.50 g/mol and is commercially available from Merck as TIMOPTIC®. In other embodiments, the timolol is timolol hemihydrate. In some embodiments, the timolol is present in a sub-therapeutic amount. In some embodiments, the sub-therapeutic amount is an amount less than about 0.25% w/w. In some embodiments, the sub-therapeutic amount is an amount from about 0.0002 to about 0.25, from about 0.0004 to about 0.25, from about 0.0006 to about 0.25, from about 0.0008 to about 0.25, from about 0.001 to about 0.25, from about 0.002 to about 0.25, from about 0.004 to about 0.25, from about 0.006 to about 0.25, from about 0.008 to about 0.25, from about 0.01 to about 0.25, from about 0.02 to about 0.25, from about 0.04 to about 0.25, from about 0.06 to about 0.25, from about 0.08 to about 0.25, from about 0.1 to about 0.25, from about 0.15 to about 0.25, from about 0.20 to about 0.25, from about 0.0002 to about 0.20, from about 0.0004 to about 0.20, from about 0.0006 to about 0.20, from about 0.0008 to about 0.20, from about 0.001 to about 0.20, from about 0.002 to about 0.20, from about 0.004 to about 0.20, from about 0.006 to about 0.20, from about 0.008 to about 0.20, from about 0.01 to about 0.20, from about 0.02 to about 0.20, from about 0.04 to about 0.20, from about 0.06 to about 0.20, from about 0.08 to about 0.20, from about 0.1 to about 0.20, from about 0.15 to about 0.20, from about 0.0002 to about 0.015, from about 0.0004 to about 0.015, from about 0.0006 to about 0.015, from about 0.0008 to about 0.015, from about 0.001 to about 0.015, from about 0.002 to about 0.015, from about 0.004 to about 0.015, from about 0.006 to about 0.015, from about 0.008 to about 0.015, from about 0.01 to about 0.15, from about 0.02 to about 0.15, from about 0.04 to about 0.15, from about 0.06 to about 0.15, from about 0.08 to about 0.15, from about 0.1 to about 0.15, from about 0.0002 to about 0.001, from about 0.0004 to about 0.001, from about 0.0006 to about 0.001, from about 0.0008 to about 0.001, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.004 to about 0.01, from about 0.006 to about 0.01, from about 0.008 to about 0.01, from about 0.01 to about 0.1, from about 0.02 to about 0.1, from about 0.04 to about 0.1, from about 0.06 to about 0.1, from about 0.08 to about 0.1, from about 0.0002 to about 0.08, from about 0.0004 to about 0.08, from about 0.0006 to about 0.08, from about 0.0008 to about 0.08, from about 0.001 to about 0.08, from about 0.002 to about 0.08, from about 0.004 to about 0.08, from about 0.006 to about 0.08, from about 0.008 to about 0.08, from about 0.01 to about 0.08, from about 0.02 to about 0.08, from about 0.04 to about 0.08, from about 0.06 to about 0.08, from about 0.0002 to about 0.06, from about 0.0004 to about 0.06, from about 0.0006 to about 0.06, from about 0.0008 to about 0.06, from about 0.001 to about 0.06, from about 0.002 to about 0.06, from about 0.004 to about 0.06, from about 0.006 to about 0.06, from about 0.008 to about 0.06, from about 0.01 to about 0.06, from about 0.02 to about 0.06, from about 0.04 to about 0.06, from about 0.0002 to about 0.04, from about 0.0004 to about 0.04, from about 0.0006 to about 0.04, from about 0.0008 to about 0.04, from about 0.001 to about 0.04, from about 0.002 to about 0.04, from about 0.004 to about 0.04, from about 0.006 to about 0.04, from about 0.008 to about 0.04, from about 0.01 to about 0.04, from about 0.02 to about 0.04, from about 0.0002 to about 0.02, from about 0.0004 to about 0.02, from about 0.0006 to about 0.02, from about 0.0008 to about 0.02, from about 0.001 to about 0.02, from about 0.002 to about 0.02, from about 0.004 to about 0.02, from about 0.006 to about 0.02, from about 0.008 to about 0.02, from about 0.01 to about 0.02, from about 0.0002 to about 0.01, from about 0.0004 to about 0.01, from about 0.0006 to about 0.01, from about 0.0008 to about 0.01, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.004 to about 0.01, from about 0.006 to about 0.01, from about 0.008 to about 0.01, from about 0.0002 to about 0.008, from about 0.0004 to about 0.008, from about 0.0006 to about 0.008, from about 0.0008 to about 0.008, from about 0.001 to about 0.008, from about 0.002 to about 0.008, from about 0.004 to about 0.008, from about 0.006 to about 0.008, from about 0.0002 to about 0.006, from about 0.0004 to about 0.006, from about 0.0006 to about 0.006, from about 0.0008 to about 0.006, from about 0.001 to about 0.006, from about 0.002 to about 0.006, from about 0.004 to about 0.006, from about 0.0002 to about 0.004, from about 0.0004 to about 0.004, from about 0.0006 to about 0.004, from about 0.0008 to about 0.004, from about 0.001 to about 0.004, from about 0.002 to about 0.004, from about 0.0002 to about 0.002, from about 0.0004 to about 0.002, from about 0.0006 to about 0.002, from about 0.0008 to about 0.002, from about 0.001 to about 0.002, from about 0.0002 to about 0.001, from about 0.0004 to about 0.001, from about 0.0006 to about 0.001, from about 0.0008 to about 0.001, from about 0.0002 to about 0.0008, from about 0.0004 to about 0.0008, from about 0.0006 to about 0.0008, from about 0.0002 to about 0.0006, from about 0.0004 to about 0.0006, or from about 0.0002 to about 0.0004% (w/w). In other embodiments, the sub-therapeutic amount is an amount of about 0.0002, 0.0004, 0.0006, 0.0008, 0.001, 0.002, 0.004, 0.006, 0.008, 0.01, 0.02, 0.04, 0.06, 0.08, 0.1, 0.15, 0.20, or 0.25% (w/w).
In other embodiments, beta adrenergic antagonist is betaxolol. In some embodiments, the betaxolol is present in a sub-therapeutic amount. Betaxolol refers, in the customary sense, to CAS Registry No. 63659-18-7. In some embodiments, the sub-therapeutic amount is an amount less than about 0.25% w/w. In some embodiments, the sub-therapeutic amount is an amount from about 0.0002 to about 0.25, from about 0.0004 to about 0.25, from about 0.0006 to about 0.25, from about 0.0008 to about 0.25, from about 0.001 to about 0.25, from about 0.002 to about 0.25, from about 0.004 to about 0.25, from about 0.006 to about 0.25, from about 0.008 to about 0.25, from about 0.01 to about 0.25, from about 0.02 to about 0.25, from about 0.04 to about 0.25, from about 0.06 to about 0.25, from about 0.08 to about 0.25, from about 0.1 to about 0.25, from about 0.15 to about 0.25, from about 0.20 to about 0.25, from about 0.0002 to about 0.20, from about 0.0004 to about 0.20, from about 0.0006 to about 0.20, from about 0.0008 to about 0.20, from about 0.001 to about 0.20, from about 0.002 to about 0.20, from about 0.004 to about 0.20, from about 0.006 to about 0.20, from about 0.008 to about 0.20, from about 0.01 to about 0.20, from about 0.02 to about 0.20, from about 0.04 to about 0.20, from about 0.06 to about 0.20, from about 0.08 to about 0.20, from about 0.1 to about 0.20, from about 0.15 to about 0.20, from about 0.0002 to about 0.15, from about 0.0004 to about 0.15, from about 0.0006 to about 0.15, from about 0.0008 to about 0.15, from about 0.001 to about 0.15, from about 0.002 to about 0.15, from about 0.004 to about 0.15, from about 0.006 to about 0.15, from about 0.008 to about 0.15, from about 0.01 to about 0.15, from about 0.02 to about 0.15, from about 0.04 to about 0.15, from about 0.06 to about 0.15, from about 0.08 to about 0.15, from about 0.1 to about 0.15, from about 0.0002 to about 0.1, from about 0.0004 to about 0.1, from about 0.0006 to about 0.1, from about 0.0008 to about 0.1, from about 0.001 to about 0.1, from about 0.002 to about 0.1, from about 0.004 to about 0.1, from about 0.006 to about 0.1, from about 0.008 to about 0.1, from about 0.01 to about 0.1, from about 0.02 to about 0.1, from about 0.04 to about 0.1, from about 0.06 to about 0.1, from about 0.08 to about 0.1, from about 0.0002 to about 0.08, from about 0.0004 to about 0.08, from about 0.0006 to about 0.08, from about 0.0008 to about 0.08, from about 0.001 to about 0.08, from about 0.002 to about 0.08, from about 0.004 to about 0.08, from about 0.006 to about 0.08, from about 0.008 to about 0.08, from about 0.01 to about 0.08, from about 0.02 to about 0.08, from about 0.04 to about 0.08, from about 0.06 to about 0.08, from about 0.0002 to about 0.06, from about 0.0004 to about 0.06, from about 0.0006 to about 0.06, from about 0.0008 to about 0.06, from about 0.001 to about 0.06, from about 0.002 to about 0.06, from about 0.004 to about 0.06, from about 0.006 to about 0.06, from about 0.008 to about 0.06, from about 0.01 to about 0.06, from about 0.02 to about 0.06, from about 0.04 to about 0.06, from about 0.0002 to about 0.04, from about 0.0004 to about 0.04, from about 0.0006 to about 0.04, from about 0.0008 to about 0.04, from about 0.001 to about 0.04, from about 0.002 to about 0.04, from about 0.004 to about 0.04, from about 0.006 to about 0.04, from about 0.008 to about 0.04, from about 0.01 to about 0.04, from about 0.02 to about 0.04, from about 0.0002 to about 0.02, from about 0.0004 to about 0.02, from about 0.0006 to about 0.02, from about 0.0008 to about 0.02, from about 0.001 to about 0.02, from about 0.002 to about 0.02, from about 0.004 to about 0.02, from about 0.006 to about 0.02, from about 0.008 to about 0.02, from about 0.01 to about 0.02, from about 0.0002 to about 0.01, from about 0.0004 to about 0.01, from about 0.0006 to about 0.01, from about 0.0008 to about 0.01, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.004 to about 0.01, from about 0.006 to about 0.01, from about 0.008 to about 0.01, from about 0.0002 to about 0.008, from about 0.0004 to about 0.008, from about 0.0006 to about 0.008, from about 0.0008 to about 0.008, from about 0.001 to about 0.008, from about 0.002 to about 0.008, from about 0.004 to about 0.008, from about 0.006 to about 0.008, from about 0.0002 to about 0.006, from about 0.0004 to about 0.006, from about 0.0006 to about 0.006, from about 0.0008 to about 0.006, from about 0.001 to about 0.006, from about 0.002 to about 0.006, from about 0.004 to about 0.006, from about 0.0002 to about 0.004, from about 0.0004 to about 0.004, from about 0.0006 to about 0.004, from about 0.0008 to about 0.004, from about 0.001 to about 0.004, from about 0.002 to about 0.004, from about 0.0002 to about 0.002, from about 0.0004 to about 0.002, from about 0.0006 to about 0.002, from about 0.0008 to about 0.002, from about 0.001 to about 0.002, from about 0.0002 to about 0.001, from about 0.0004 to about 0.001, from about 0.0006 to about 0.001, from about 0.0008 to about 0.001, from about 0.0002 to about 0.0008, from about 0.0004 to about 0.0008, from about 0.0006 to about 0.0008, from about 0.0002 to about 0.0006, from about 0.0004 to about 0.0006, or from about 0.0002 to about 0.0004% (w/w). In other embodiments, the sub-therapeutic amount is an amount of about 0.0002, 0.0004, 0.0006, 0.0008, 0.001, 0.002, 0.004, 0.006, 0.008, 0.01, 0.02, 0.04, 0.06, 0.08, 0.1, 0.15, 0.20, or 0.25% (w/w). In some embodiments, the betaxolol is a betaxolol salt.
In other embodiments, the beta adrenergic antagonist is levobunolol. Levobunolol refers, in the customary sense, to CAS Registry No. 47141-42-4. In some embodiments, the levobunolol is present in a sub-therapeutic amount. In some embodiments, the sub-therapeutic amount is an amount less than about 0.25% w/w. In some embodiments, the sub-therapeutic amount is an amount from about 0.0002 to about 0.25, from about 0.0004 to about 0.25, from about 0.0006 to about 0.25, from about 0.0008 to about 0.25, from about 0.001 to about 0.25, from about 0.002 to about 0.25, from about 0.004 to about 0.25, from about 0.006 to about 0.25, from about 0.008 to about 0.25, from about 0.01 to about 0.25, from about 0.02 to about 0.25, from about 0.04 to about 0.25, from about 0.06 to about 0.25, from about 0.08 to about 0.25, from about 0.1 to about 0.25, from about 0.15 to about 0.25, from about 0.20 to about 0.25, from about 0.0002 to about 0.20, from about 0.0004 to about 0.20, from about 0.0006 to about 0.20, from about 0.0008 to about 0.20, from about 0.001 to about 0.20, from about 0.002 to about 0.20, from about 0.004 to about 0.20, from about 0.006 to about 0.20, from about 0.008 to about 0.20, from about 0.01 to about 0.20, from about 0.02 to about 0.20, from about 0.04 to about 0.20, from about 0.06 to about 0.20, from about 0.08 to about 0.20, from about 0.1 to about 0.20, from about 0.15 to about 0.20, from about 0.0002 to about 0.15, from about 0.0004 to about 0.15, from about 0.0006 to about 0.15, from about 0.0008 to about 0.15, from about 0.001 to about 0.15, from about 0.002 to about 0.15, from about 0.004 to about 0.15, from about 0.006 to about 0.15, from about 0.008 to about 0.15, from about 0.01 to about 0.15, from about 0.02 to about 0.15, from about 0.04 to about 0.15, from about 0.06 to about 0.15, from about 0.08 to about 0.15, from about 0.1 to about 0.15, from about 0.0002 to about 0.1, from about 0.0004 to about 0.1, from about 0.0006 to about 0.1, from about 0.0008 to about 0.1, from about 0.001 to about 0.1, from about 0.002 to about 0.1, from about 0.004 to about 0.1, from about 0.006 to about 0.1, from about 0.008 to about 0.1, from about 0.01 to about 0.1, from about 0.02 to about 0.1, from about 0.04 to about 0.1, from about 0.06 to about 0.1, from about 0.08 to about 0.1, from about 0.0002 to about 0.08, from about 0.0004 to about 0.08, from about 0.0006 to about 0.08, from about 0.0008 to about 0.08, from about 0.001 to about 0.08, from about 0.002 to about 0.08, from about 0.004 to about 0.08, from about 0.006 to about 0.08, from about 0.008 to about 0.08, from about 0.01 to about 0.08, from about 0.02 to about 0.08, from about 0.04 to about 0.08, from about 0.06 to about 0.08, from about 0.0002 to about 0.06, from about 0.0004 to about 0.06, from about 0.0006 to about 0.06, from about 0.0008 to about 0.06, from about 0.001 to about 0.06, from about 0.002 to about 0.06, from about 0.004 to about 0.06, from about 0.006 to about 0.06, from about 0.008 to about 0.06, from about 0.01 to about 0.06, from about 0.02 to about 0.06, from about 0.04 to about 0.06, from about 0.0002 to about 0.04, from about 0.0004 to about 0.04, from about 0.0006 to about 0.04, from about 0.0008 to about 0.04, from about 0.001 to about 0.04, from about 0.002 to about 0.04, from about 0.004 to about 0.04, from about 0.006 to about 0.04, from about 0.008 to about 0.04, from about 0.01 to about 0.04, from about 0.02 to about 0.04, from about 0.0002 to about 0.02, from about 0.0004 to about 0.02, from about 0.0006 to about 0.02, from about 0.0008 to about 0.02, from about 0.001 to about 0.02, from about 0.002 to about 0.02, from about 0.004 to about 0.02, from about 0.006 to about 0.02, from about 0.008 to about 0.02, from about 0.01 to about 0.02, from about 0.0002 to about 0.01, from about 0.0004 to about 0.01, from about 0.0006 to about 0.01, from about 0.0008 to about 0.01, from about 0.001 to about 0.01, from about 0.002 to about 0.01, from about 0.004 to about 0.01, from about 0.006 to about 0.01, from about 0.008 to about 0.01, from about 0.0002 to about 0.008, from about 0.0004 to about 0.008, from about 0.0006 to about 0.008, from about 0.0008 to about 0.008, from about 0.001 to about 0.008, from about 0.002 to about 0.008, from about 0.004 to about 0.008, from about 0.006 to about 0.008, from about 0.0002 to about 0.006, from about 0.0004 to about 0.006, from about 0.0006 to about 0.006, from about 0.0008 to about 0.006, from about 0.001 to about 0.006, from about 0.002 to about 0.006, from about 0.004 to about 0.006, from about 0.0002 to about 0.004, from about 0.0004 to about 0.004, from about 0.0006 to about 0.004, from about 0.0008 to about 0.004, from about 0.001 to about 0.004, from about 0.002 to about 0.004, from about 0.0002 to about 0.002, from about 0.0004 to about 0.002, from about 0.0006 to about 0.002, from about 0.0008 to about 0.002, from about 0.001 to about 0.002, from about 0.0002 to about 0.001, from about 0.0004 to about 0.001, from about 0.0006 to about 0.001, from about 0.0008 to about 0.001, from about 0.0002 to about 0.0008, from about 0.0004 to about 0.0008, from about 0.0006 to about 0.0008, from about 0.0002 to about 0.0006, from about 0.0004 to about 0.0006, or from about 0.0002 to about 0.0004% (w/w). In other embodiments, the sub-therapeutic amount is an amount of about 0.0002, 0.0004, 0.0006, 0.0008, 0.001, 0.002, 0.004, 0.006, 0.008, 0.01, 0.02, 0.04, 0.06, 0.08, 0.1, 0.15, 0.20, or 0.25% (w/w). In some embodiments, the levobunolol is a levobunolol salt.
In other embodiments, the beta adrenergic antagonist is metipranolol. Metipranolol refers, in the customary sense, to CAS Registry No. 22664-55-7. In some embodiments, the metipranolol is present in a sub-therapeutic amount. In some embodiments, the sub-therapeutic amount is an amount less than about 0.3% w/w. In some embodiments, the sub-therapeutic amount is an amount from about 0.003 to about 0.3, from about 0.004 to about 0.3, from about 0.006 to about 0.3, from about 0.008 to about 0.3, from about 0.01 to about 0.3, from about 0.03 to about 0.3, from about 0.04 to about 0.3, from about 0.06 to about 0.3, from about 0.08 to about 0.3, from about 0.1 to about 0.3, from about 0.15 to about 0.3, from about 0.20 to about 0.3, from about 0.25 to about 0.3, from about 0.003 to about 0.25, from about 0.004 to about 0.25, from about 0.006 to about 0.25, from about 0.008 to about 0.25, from about 0.01 to about 0.25, from about 0.03 to about 0.25, from about 0.04 to about 0.25, from about 0.06 to about 0.25, from about 0.08 to about 0.25, from about 0.1 to about 0.25, from about 0.15 to about 0.25, from about 0.20 to about 0.25, from about 0.003 to about 0.20, from about 0.004 to about 0.20, from about 0.006 to about 0.20, from about 0.008 to about 0.20, from about 0.01 to about 0.20, from about 0.03 to about 0.20, from about 0.04 to about 0.20, from about 0.06 to about 0.20, from about 0.08 to about 0.20, from about 0.1 to about 0.20, from about 0.15 to about 0.20, from about 0.003 to about 0.15, from about 0.004 to about 0.15, from about 0.006 to about 0.15, from about 0.008 to about 0.15, from about 0.01 to about 0.15, from about 0.03 to about 0.15, from about 0.04 to about 0.15, from about 0.06 to about 0.15, from about 0.08 to about 0.15, from about 0.1 to about 0.15, from about 0.003 to about 0.1, from about 0.004 to about 0.1, from about 0.006 to about 0.1, from about 0.008 to about 0.1, from about 0.01 to about 0.1, from about 0.03 to about 0.1, from about 0.04 to about 0.1, from about 0.06 to about 0.1, from about 0.08 to about 0.1, from about 0.003 to about 0.08, from about 0.004 to about 0.08, from about 0.006 to about 0.08, from about 0.008 to about 0.08, from about 0.01 to about 0.08, from about 0.03 to about 0.08, from about 0.04 to about 0.08, from about 0.06 to about 0.08, from about 0.003 to about 0.06, from about 0.004 to about 0.06, from about 0.006 to about 0.06, from about 0.008 to about 0.06, from about 0.01 to about 0.06, from about 0.03 to about 0.06, from about 0.04 to about 0.06, from about 0.003 to about 0.04, from about 0.004 to about 0.04, from about 0.006 to about 0.04, from about 0.008 to about 0.04, from about 0.01 to about 0.04, from about 0.03 to about 0.04, from about 0.003 to about 0.03, from about 0.004 to about 0.03, from about 0.006 to about 0.03, from about 0.008 to about 0.03, from about 0.01 to about 0.03, from about 0.003 to about 0.01, from about 0.004 to about 0.01, from about 0.006 to about 0.01, from about 0.008 to about 0.01, from about 0.003 to about 0.008, from about 0.004 to about 0.008, from about 0.006 to about 0.008, from about 0.003 to about 0.006, from about 0.004 to about 0.006, or from about 0.003 to about 0.004% (w/w). In other embodiments, the sub-therapeutic amount is an amount of about 0.3, 0.25, 0.20, 0.15, 0.1, 0.08, 0.06, 0.04, 0.03, 0.01, 0.008, 0.006, 0.004, or 0.003% (w/w). In some embodiments, the metipranolol is a metipranolol salt.
The compositions and products according to the embodiments of the present invention may include a buffering agent, a tonicity agent, a salt, a thickening agent or a preservative. In some embodiments, the composition includes a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative.
A buffering agents are means for adjusting the pH of the compositions provided to be an ophthalmically acceptable pH (e.g. neutral pH). Buffers appropriate to for the compositions provided include acetate buffers, citrate buffers, phosphate buffers and borate buffers. Acids or bases may also be used to adjust the pH of the ophthalmic formulations as needed. The pH of the disclosed compositions should preferably be maintained between 6.5 and 7.2 with an appropriate buffering agent.
The term “tonicity agent” as used herein refers to a compound or ion useful for adjusting the osmotic pressure or tension of a solution, often relative to that of blood. Examples for tonicity agents are without limitation, glycerin, erythritol, mannitol, potassium, chloride, and sodium chloride. In some embodiments, the tonicity agent is glycerin. In some further embodiments, the tonicity agent is present from about 0.5% w/w to about 6% w/w.
The formulation's viscosity is a factor that determines how well the formulation sticks to the skin or ophthalmic tissue or does not run off the skin or ophthalmic tissue when applied. The viscosity of the formulation can be optimized using one or more pharmaceutically acceptable thickening agents that do not significantly interact with the components of the formulation, do not significantly reduce flux of the formulation, and do not cause stinging or irritation. Non-limiting examples of suitable thickeners useful herein include cellulosic polymers, such as gum arabic, gum acacia, gum tragacanth, locust bean gum, guar gum, hydroxypropyl guar, xanthan gum, talc, cellulose gum, sclerotium gum, carageenan gum, karaya gum, cellulose gum, rosin, methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxymethylcellulosf, hydroxypropylmethylcellulose, methylhydroxyethylcellulose, cetyl hydroxyethylcellulose, carboxymethylcellulose, corn starch, hydroxypropyl starch phosphate, distarch phosphate, distarch dimethylene urea, aluminum starch octenyl succinate, maltodextrin, dextran, poly(acrylamide), PEG-150 distearate, PEG-150/decyl alcohol/SMDI copolymer, PEG-150/stearyl alcohol/SMDI copolymer, PEG-180/Laureth-50/TMMG copolymer, Polyether 1, acrylic acid/acrylamidomethyl propane sulfonic acid copolymer, acrylate/C10-30 alkyl acrylate cross polymer, acrylate/beheneth-25 methacrylate copolymer, acrylate/steareth-20 methacrylate copolymer, acrylate/steareth-20 copolymer, acrylate/VA cross polymer, acrylic acid/acrylonitrogen copolymer, ammonium acryloyldimethyltaurate/beheneth-25 methacrylate copolymer, ammonium acryloyldimethyltaurate/VP copolymer, caprylic/capric triglyceride (and) sodium acrylate copolymer, PVM/MA decadiene cross polymer, alginic acid, propylene glycol alginate, dimethicone, silica dimethyl silylate, a dimethylacrylamide/acrylic acid/polystyrene ethyl methacrylate copolymer, derivatives thereof, and mixtures thereof.
Preservatives that can be used in the ophthalmic pharmaceutical formulations of the present embodiments include, but are not limited to, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetate and phenylmercuric nitrate. Preservative-free compositions can be considered, for example for patients experiencing hypersensitivity reactions with the above listed preservatives or other preservatives not listed. Thus, in some embodiments, the composition further includes a buffering agent, a tonicity agent, a salt and a thickening agent.
In some embodiments, the composition consists essentially of bimatoprost, timolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative. Where the composition consists essentially of bimatoprost, timolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative, the composition consists of bimatoprost and timolol, and any appropriate buffering agent, tonicity agent, salt, thickening agent and preservative. In some embodiments, the timolol is present in a sub-therapeutic amount. As described earlier the sub-therapeutic amount may be an amount less than about 0.25% w/w. Thus, in some embodiments, the sub-therapeutic amount is an amount of about 0.0002, 0.0004, 0.0006, 0.0008, 0.001, 0.002, 0.004, 0.006, 0.008, 0.01, 0.02, 0.04, 0.06, 0.08, 0.1, 0.15, 0.20, or 0.25% (w/w).
The compositions provided herein may consist essentially of bimatoprost, a beta adrenergic antagonist, buffering agent, a tonicity agent, a salt, a thickening agent and a preservative. Where the compositions provided herein may consist essentially of bimatoprost, a beta adrenergic antagonist, buffering agent, a tonicity agent, a salt, a thickening agent and a preservative, the composition consists of bimatoprost and a beta adrenergic antagonist, and any appropriate buffering agent, tonicity agent, salt, thickening agent and preservative. In some embodiments, the beta adrenergic antagonist is present in a sub-therapeutic amount. Non-limiting examples of a beta adrenergic compound suitable for the compositions and methods according to the embodiments of the present invention are betaxolol, levobunolol, or metipranolol. Thus, in some embodiments, the composition consists essentially of bimatoprost, betaxolol, a buffering agent, a tonicity agent, a salt, a preservative and a thickening agent. In some further embodiments, the betaxolol is present in a sub-therapeutic amount. In further embodiments, the sub-therapeutic amount is an amount less than about 0.25% w/w.
In other embodiments, the composition consists essentially of bimatoprost, levobunolol, a buffering agent, a tonicity agent, a salt, a preservative and a thickening agent. In further embodiments, the levobunolol is present in a sub-therapeutic amount. In some further embodiment, the sub-therapeutic amount is an amount less than about 0.25% w/w.
In some embodiments, the composition consists essentially of bimatoprost, metipranolol, a buffering agent, a tonicity agent, a salt, a preservative and a thickening agent. In further embodiments, the metipranolol is present in a sub-therapeutic amount. In some further embodiment, the sub-therapeutic amount is an amount less than about 0.3% w/w.
Tables 1-12 describe various examples of combinations of effective amounts of the prostaglandin agent (e.g., bimatoprost, travoprost, latanoprost) and the vasoconstrictor agent (e.g., timolol, betaxolol, levobunolol, metipranolol, tetrahydrozoline hydrochloride, brimonidine etc.). In particular, Table 1 provides 342 different combination embodiments of bimatoprost, as shown in the first column labeled “Bimatoprost % w/w”, and timolol, as shown in the first row labeled “Timolol % w/w.” Specific concentrations of bimatoprost and timolol for each of the combination embodiments described in Table 1 and numbered from 1 to 19 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 2 provides 342 different combination embodiments of bimatoprost, as shown in the first column labeled “Bimatoprost % w/w”, and betaxolol as shown in the first row labeled “Betaxolol % w/w.” Specific concentrations of bimatoprost and betaxolol for each of the combination embodiments described in Table 2 and numbered from 343 to 361 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 3 provides 342 different combination embodiments of bimatoprost, as shown in the first column labeled “Bimatoprost % w/w”, and levobunolol as shown in the first row labeled “Levobunolol % w/w.” Specific concentrations of bimatoprost and levobunolol for each of the combination embodiments described in Table 3 and numbered from 685 to 703 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 4 provides 266 different combination embodiments of bimatoprost, as shown in the first column labeled “Bimatoprost % w/w”, and metipranolol as shown in the first row labeled “Metipranolol % w/w.” Specific concentrations of bimatoprost and metipranolol for each of the combination embodiments described in Table 4 and numbered from 1027 to 1045 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 5 provides 270 different combination embodiments of travoprost, as shown in the first column labeled “Travoprost % w/w”, and timolol, as shown in the first row labeled “Timolol % w/w.” Specific concentrations of travoprost and timolol for each of the combination embodiments described in Table 5 and numbered from 1293 to 1307 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 6 provides 270 different combination embodiments of travoprost, as shown in the first column labeled “Travoprost % w/w”, and betaxolol as shown in the first row labeled “Betaxolol % w/w.” Specific concentrations of travoprost and betaxolol for each of the combination embodiments described in Table 6 and numbered from 1563 to 1577 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 7 provides 270 different combination embodiments of travoprost, as shown in the first column labeled “Travoprost % w/w”, and levobunolol as shown in the first row labeled “Levobunolol % w/w.” Specific concentrations of travoprost and levobunolol for each of the combination embodiments described in Table 7 and numbered from 1833 to 1847 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 8 provides 210 different combination embodiments of travoprost, as shown in the first column labeled “Travoprost % w/w”, and metipranolol as shown in the first row labeled “Metipranolol % w/w.” Specific concentrations of travoprost and metipranolol for each of the combination embodiments described in Table 8 and numbered from 2103 to 2117 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 9 provides 270 different combination embodiments of latanoprost, as shown in the first column labeled “Latanoprost % w/w”, and timolol, as shown in the first row labeled “Timolol % w/w.” Specific concentrations of latanoprost and timolol for each of the combination embodiments described in Table 9 and numbered from 2313 to 2327 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 10 provides 270 different combination embodiments of latanoprost, as shown in the first column labeled “Latanoprost % w/w”, and betaxolol as shown in the first row labeled “Betaxolol % w/w.” Specific concentrations of latanoprost and betaxolol for each of the combination embodiments described in Table 10 and numbered from 2583 to 2597 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 11 provides 270 different combination embodiments of latanoprost, as shown in the first column labeled “Latanoprost % w/w”, and levobunolol as shown in the first row labeled “Levobunolol % w/w.” Specific concentrations of latanoprost and levobunolol for each of the combination embodiments described in Table 11 and numbered from 2853 to 2867 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.
Table 12 provides 210 different combination embodiments of latanoprost, as shown in the first column labeled “Latanoprost % w/w”, and metipranolol as shown in the first row labeled “Metipranolol % w/w.” Specific concentrations of latanoprost and metipranolol for each of the combination embodiments described in Table 12 and numbered from 3123 to 3137 are shown, respectively, in the cells in the first row, which correspond to the numbered cell.







TABLE 1





Effective Amounts of Bimatoprost and Timolol
















Bimatoprost
Timolol % w/w

















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080
0.0100





0.001
1
20
39
58
77
96
115
134
153
172


0.002
2
21
40
59
78
97
116
135
154
173


0.003
3
22
41
60
79
98
117
136
155
174


0.004
4
23
42
61
80
99
118
137
156
175


0.005
5
24
43
62
81
100
119
138
157
176


0.006
6
25
44
63
82
101
120
139
158
177


0.007
7
26
45
64
83
102
121
140
159
178


0.008
8
27
46
65
84
103
122
141
160
179


0.009
9
28
47
66
85
104
123
142
161
180


0.010
10
29
48
67
86
105
124
143
162
181


0.020
11
30
49
68
87
106
125
144
163
182


0.030
12
31
50
69
88
107
126
145
164
183


0.040
13
32
51
70
89
108
127
146
165
184


0.050
14
33
52
71
90
109
128
147
166
185


0.060
15
34
53
72
91
110
129
148
167
186


0.070
16
35
54
73
92
111
130
149
168
187


0.080
17
36
55
74
93
112
131
150
169
188


0.090
18
37
56
75
94
113
132
151
170
189


0.100
19
38
57
76
95
114
133
152
171
190













Bimatoprost
Timolol % w/w

















% w/w
0.0200
0.0400
0.0600
0.0800
0.10
0.15
0.20
0.25







0.001
191
210
229
248
267
286
305
324



0.002
192
211
230
249
268
287
306
325



0.003
193
212
231
250
269
288
307
326



0.004
194
213
232
251
270
289
308
327



0.005
195
214
233
252
271
290
309
328



0.006
196
215
234
253
272
291
310
329



0.007
197
216
235
254
273
292
311
330



0.008
198
217
236
255
274
293
312
331



0.009
199
218
237
256
275
294
313
332



0.010
200
219
238
257
276
295
314
333



0.020
201
220
239
258
277
296
315
334



0.030
202
221
240
259
278
297
316
335



0.040
203
222
241
260
279
298
317
336



0.050
204
223
242
261
280
299
318
337



0.060
205
224
243
262
281
300
319
338



0.070
206
225
244
263
282
301
320
339



0.080
207
226
245
264
283
302
321
340



0.090
208
227
246
265
284
303
322
341



0.100
209
228
247
266
285
304
323
342

















TABLE 2





Effective Amounts of Bimatoprost and Betaxolol
















Bimatoprost
Betaxolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.001
343
362
381
400
419
438
457
476
495


0.002
344
363
382
401
420
439
458
477
496


0.003
345
364
383
402
421
440
459
478
497


0.004
346
365
384
403
422
441
460
479
498


0.005
347
366
385
404
423
442
461
480
499


0.006
348
367
386
405
424
443
462
481
500


0.007
349
368
387
406
425
444
463
482
501


0.008
350
369
388
407
426
445
464
483
502


0.009
351
370
389
408
427
446
465
484
503


0.010
352
371
390
409
428
447
466
485
504


0.020
353
372
391
410
429
448
467
486
505


0.030
354
373
392
411
430
449
468
487
506


0.040
355
374
393
412
431
450
469
488
507


0.050
356
375
394
413
432
451
470
489
508


0.060
357
376
395
414
433
452
471
490
509


0.070
358
377
396
415
434
453
472
491
510


0.080
359
378
397
416
435
454
473
492
511


0.090
360
379
398
417
436
455
474
493
512


0.100
361
380
399
418
437
456
475
494
513













Bimatoprost
Betaxolol % w/w


















% w/w
0.0100
0.0200
0.0400
0.0600
0.0800
0.10
0.15
0.20
0.25







0.001
514
533
552
571
590
609
628
647
666



0.002
515
534
553
572
591
610
629
648
667



0.003
516
535
554
573
592
611
630
649
668



0.004
517
536
555
574
593
612
631
650
669



0.005
518
537
556
575
594
613
632
651
670



0.006
519
538
557
576
595
614
633
652
671



0.007
520
539
558
577
596
615
634
653
672



0.008
521
540
559
578
597
616
635
654
673



0.009
522
541
560
579
598
617
636
655
674



0.010
523
542
561
580
599
618
637
656
675



0.020
524
543
562
581
600
619
638
657
676



0.030
525
544
563
582
601
620
639
658
677



0.040
526
545
564
583
602
621
640
659
678



0.050
527
546
565
584
603
622
641
660
679



0.060
528
547
566
585
604
623
642
661
680



0.070
529
548
567
586
605
624
643
662
681



0.080
530
549
568
587
606
625
644
663
682



0.090
531
550
569
588
607
626
645
664
683



0.100
532
551
570
589
608
627
646
665
684

















TABLE 3





Effective Amounts of Bimatoprost and Levobunolol
















Bimatoprost
Levobunolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.001
685
704
723
742
761
780
799
818
837


0.002
686
705
724
743
762
781
800
819
838


0.003
687
706
725
744
763
782
801
820
839


0.004
688
707
726
745
764
783
802
821
840


0.005
689
708
727
746
765
784
803
822
841


0.006
690
709
728
747
766
785
804
823
842


0.007
691
710
729
748
767
786
805
824
843


0.008
692
711
730
749
768
787
806
825
844


0.009
693
712
731
750
769
788
807
826
845


0.010
694
713
732
751
770
789
808
827
846


0.020
695
714
733
752
771
790
809
828
847


0.030
696
715
734
753
772
791
810
829
848


0.040
697
716
735
754
773
792
811
830
849


0.050
698
717
736
755
774
793
812
831
850


0.060
699
718
737
756
775
794
813
832
851


0.070
700
719
738
757
776
795
814
833
852


0.080
701
720
739
758
777
796
815
834
853


0.090
702
721
740
759
778
797
816
835
854


0.100
703
722
741
760
779
798
817
836
855













Bimatoprost
Levobunolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.001
856
875
894
913
932
951
970
989
1008



0.002
857
876
895
914
933
952
971
990
1009



0.003
858
877
896
915
934
953
972
991
1010



0.004
859
878
897
916
935
954
973
992
1011



0.005
860
879
898
917
936
955
974
993
1012



0.006
861
880
899
918
937
956
975
994
1013



0.007
862
881
900
919
938
957
976
995
1014



0.008
863
882
901
920
939
958
977
996
1015



0.009
864
883
902
921
940
959
978
997
1016



0.010
865
884
903
922
941
960
979
998
1017



0.020
866
885
904
923
942
961
980
999
1018



0.030
867
886
905
924
943
962
981
1000
1019



0.040
868
887
906
925
944
963
982
1001
1020



0.050
869
888
907
926
945
964
983
1002
1021



0.060
870
889
908
927
946
965
984
1003
1022



0.070
871
890
909
928
947
966
985
1004
1023



0.080
872
891
910
929
948
967
986
1005
1024



0.090
873
892
911
930
949
968
987
1006
1025



0.100
874
893
912
931
950
969
988
1007
1026

















TABLE 4







Effective Amounts of Bimatoprost and Metipranolol








Bimatoprost
Metipranolol % w/w





















% w/w
0.0030
0.0040
0.0060
0.0080
0.01
0.03
0.04
0.06
0.08
0.10
0.15
0.20
0.25
0.30





0.001
1027
1046
1065
1084
1103
1122
1141
1160
1179
1198
1217
1236
1255
1274


0.002
1028
1047
1066
1085
1104
1123
1142
1161
1180
1199
1218
1237
1256
1275


0.003
1029
1048
1067
1086
1105
1124
1143
1162
1181
1200
1219
1238
1257
1276


0.004
1030
1049
1068
1087
1106
1125
1144
1163
1182
1201
1220
1239
1258
1277


0.005
1031
1050
1069
1088
1107
1126
1145
1164
1183
1202
1221
1240
1259
1278


0.006
1032
1051
1070
1089
1108
1127
1146
1165
1184
1203
1222
1241
1260
1279


0.007
1033
1052
1071
1090
1109
1128
1147
1166
1185
1204
1223
1242
1261
1280


0.008
1034
1053
1072
1091
1110
1129
1148
1167
1186
1205
1224
1243
1262
1281


0.009
1035
1054
1073
1092
1111
1130
1149
1168
1187
1206
1225
1244
1263
1282


0.010
1036
1055
1074
1093
1112
1131
1150
1169
1188
1207
1226
1245
1264
1283


0.020
1037
1056
1075
1094
1113
1132
1151
1170
1189
1208
1227
1246
1265
1284


0.030
1038
1057
1076
1095
1114
1133
1152
1171
1190
1209
1228
1247
1266
1285


0.040
1039
1058
1077
1096
1115
1134
1153
1172
1191
1210
1229
1248
1267
1286


0.050
1040
1059
1078
1097
1116
1135
1154
1173
1192
1211
1230
1249
1268
1287


0.060
1041
1060
1079
1098
1117
1136
1155
1174
1193
1212
1231
1250
1269
1288


0.070
1042
1061
1080
1099
1118
1137
1156
1175
1194
1213
1232
1251
1270
1289


0.080
1043
1062
1081
1100
1119
1138
1157
1176
1195
1214
1233
1252
1271
1290


0.090
1044
1063
1082
1101
1120
1139
1158
1177
1196
1215
1234
1253
1272
1291


0.100
1045
1064
1083
1102
1121
1140
1159
1178
1197
1216
1235
1254
1273
1292
















TABLE 5





Effective Amounts of Travoprost and Timolol
















Travoprost
Timolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.0002
1293
1308
1323
1338
1353
1368
1383
1398
1413


0.0004
1294
1309
1324
1339
1354
1369
1384
1399
1414


0.0006
1295
1310
1325
1340
1355
1370
1385
1400
1415


0.0008
1296
1311
1326
1341
1356
1371
1386
1401
1416


0.0010
1297
1312
1327
1342
1357
1372
1387
1402
1417


0.0020
1298
1313
1328
1343
1358
1373
1388
1403
1418


0.0040
1299
1314
1329
1344
1359
1374
1389
1404
1419


0.0060
1300
1315
1330
1345
1360
1375
1390
1405
1420


0.0080
1301
1316
1331
1346
1361
1376
1391
1406
1421


0.0100
1302
1317
1332
1347
1362
1377
1392
1407
1422


0.0200
1303
1318
1333
1348
1363
1378
1393
1408
1423


0.0400
1304
1319
1334
1349
1364
1379
1394
1409
1424


0.0600
1305
1320
1335
1350
1365
1380
1395
1410
1425


0.0800
1306
1321
1336
1351
1366
1381
1396
1411
1426


0.1000
1307
1322
1337
1352
1367
1382
1397
1412
1427













Travoprost
Timolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.0002
1428
1443
1458
1473
1488
1503
1518
1533
1548



0.0004
1429
1444
1459
1474
1489
1504
1519
1534
1549



0.0006
1430
1445
1460
1475
1490
1505
1520
1535
1550



0.0008
1431
1446
1461
1476
1491
1506
1521
1536
1551



0.0010
1432
1447
1462
1477
1492
1507
1522
1537
1552



0.0020
1433
1448
1463
1478
1493
1508
1523
1538
1553



0.0040
1434
1449
1464
1479
1494
1509
1524
1539
1554



0.0060
1435
1450
1465
1480
1495
1510
1525
1540
1555



0.0080
1436
1451
1466
1481
1496
1511
1526
1541
1556



0.0100
1437
1452
1467
1482
1497
1512
1527
1542
1557



0.0200
1438
1453
1468
1483
1498
1513
1528
1543
1558



0.0400
1439
1454
1469
1484
1499
1514
1529
1544
1559



0.0600
1440
1455
1470
1485
1500
1515
1530
1545
1560



0.0800
1441
1456
1471
1486
1501
1516
1531
1546
1561



0.1000
1442
1457
1472
1487
1502
1517
1532
1547
1562

















TABLE 6





Effective Amounts of Travoprost and Betaxolol
















Travoprost
Betaxolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.0002
1563
1578
1593
1608
1623
1638
1653
1668
1683


0.0004
1564
1579
1594
1609
1624
1639
1654
1669
1684


0.0006
1565
1580
1595
1610
1625
1640
1655
1670
1685


0.0008
1566
1581
1596
1611
1626
1641
1656
1671
1686


0.0010
1567
1582
1597
1612
1627
1642
1657
1672
1687


0.0020
1568
1583
1598
1613
1628
1643
1658
1673
1688


0.0040
1569
1584
1599
1614
1629
1644
1659
1674
1689


0.0060
1570
1585
1600
1615
1630
1645
1660
1675
1690


0.0080
1571
1586
1601
1616
1631
1646
1661
1676
1691


0.0100
1572
1587
1602
1617
1632
1647
1662
1677
1692


0.0200
1573
1588
1603
1618
1633
1648
1663
1678
1693


0.0400
1574
1589
1604
1619
1634
1649
1664
1679
1694


0.0600
1575
1590
1605
1620
1635
1650
1665
1680
1695


0.0800
1576
1591
1606
1621
1636
1651
1666
1681
1696


0.1000
1577
1592
1607
1622
1637
1652
1667
1682
1697













Travoprost
Betaxolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.0002
1698
1713
1728
1743
1758
1773
1788
1803
1818



0.0004
1699
1714
1729
1744
1759
1774
1789
1804
1819



0.0006
1700
1715
1730
1745
1760
1775
1790
1805
1820



0.0008
1701
1716
1731
1746
1761
1776
1791
1806
1821



0.0010
1702
1717
1732
1747
1762
1777
1792
1807
1822



0.0020
1703
1718
1733
1748
1763
1778
1793
1808
1823



0.0040
1704
1719
1734
1749
1764
1779
1794
1809
1824



0.0060
1705
1720
1735
1750
1765
1780
1795
1810
1825



0.0080
1706
1721
1736
1751
1766
1781
1796
1811
1826



0.0100
1707
1722
1737
1752
1767
1782
1797
1812
1827



0.0200
1708
1723
1738
1753
1768
1783
1798
1813
1828



0.0400
1709
1724
1739
1754
1769
1784
1799
1814
1829



0.0600
1710
1725
1740
1755
1770
1785
1800
1815
1830



0.0800
1711
1726
1741
1756
1771
1786
1801
1816
1831



0.1000
1712
1727
1742
1757
1772
1787
1802
1817
1832

















TABLE 7





Effective Amounts of Travoprost and Levobunolol
















Travoprost
Levobunolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.0002
1833
1848
1863
1878
1893
1908
1923
1938
1953


0.0004
1834
1849
1864
1879
1894
1909
1924
1939
1954


0.0006
1835
1850
1865
1880
1895
1910
1925
1940
1955


0.0008
1836
1851
1866
1881
1896
1911
1926
1941
1956


0.0010
1837
1852
1867
1882
1897
1912
1927
1942
1957


0.0020
1838
1853
1868
1883
1898
1913
1928
1943
1958


0.0040
1839
1854
1869
1884
1899
1914
1929
1944
1959


0.0060
1840
1855
1870
1885
1900
1915
1930
1945
1960


0.0080
1841
1856
1871
1886
1901
1916
1931
1946
1961


0.0100
1842
1857
1872
1887
1902
1917
1932
1947
1962


0.0200
1843
1858
1873
1888
1903
1918
1933
1948
1963


0.0400
1844
1859
1874
1889
1904
1919
1934
1949
1964


0.0600
1845
1860
1875
1890
1905
1920
1935
1950
1965


0.0800
1846
1861
1876
1891
1906
1921
1936
1951
1966


0.1000
1847
1862
1877
1892
1907
1922
1937
1952
1967













Travoprost
Levobunolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.0002
1968
1983
1998
2013
2028
2043
2058
2073
2088



0.0004
1969
1984
1999
2014
2029
2044
2059
2074
2089



0.0006
1970
1985
2000
2015
2030
2045
2060
2075
2090



0.0008
1971
1986
2001
2016
2031
2046
2061
2076
2091



0.0010
1972
1987
2002
2017
2032
2047
2062
2077
2092



0.0020
1973
1988
2003
2018
2033
2048
2063
2078
2093



0.0040
1974
1989
2004
2019
2034
2049
2064
2079
2094



0.0060
1975
1990
2005
2020
2035
2050
2065
2080
2095



0.0080
1976
1991
2006
2021
2036
2051
2066
2081
2096



0.0100
1977
1992
2007
2022
2037
2052
2067
2082
2097



0.0200
1978
1993
2008
2023
2038
2053
2068
2083
2098



0.0400
1979
1994
2009
2024
2039
2054
2069
2084
2099



0.0600
1980
1995
2010
2025
2040
2055
2070
2085
2100



0.0800
1981
1996
2011
2026
2041
2056
2071
2086
2101



0.1000
1982
1997
2012
2027
2042
2057
2072
2087
2102

















TABLE 8







Effective Amounts of Travoprost and Metipranolol








Travoprost %
Metipranolol % w/w





















w/w
0.0030
0.0040
0.0060
0.0080
0.01
0.03
0.04
0.06
0.08
0.10
0.15
0.20
0.25
0.30





0.0002
2103
2118
2133
2148
2163
2178
2193
2208
2223
2238
2253
2268
2283
2298


0.0004
2104
2119
2134
2149
2164
2179
2194
2209
2224
2239
2254
2269
2284
2299


0.0006
2105
2120
2135
2150
2165
2180
2195
2210
2225
2240
2255
2270
2285
2300


0.0008
2106
2121
2136
2151
2166
2181
2196
2211
2226
2241
2256
2271
2286
2301


0.0010
2107
2122
2137
2152
2167
2182
2197
2212
2227
2242
2257
2272
2287
2302


0.0020
2108
2123
2138
2153
2168
2183
2198
2213
2228
2243
2258
2273
2288
2303


0.0040
2109
2124
2139
2154
2169
2184
2199
2214
2229
2244
2259
2274
2289
2304


0.0060
2110
2125
2140
2155
2170
2185
2200
2215
2230
2245
2260
2275
2290
2305


0.0080
2111
2126
2141
2156
2171
2186
2201
2216
2231
2246
2261
2276
2291
2306


0.0100
2112
2127
2142
2157
2172
2187
2202
2217
2232
2247
2262
2277
2292
2307


0.0200
2113
2128
2143
2158
2173
2188
2203
2218
2233
2248
2263
2278
2293
2308


0.0400
2114
2129
2144
2159
2174
2189
2204
2219
2234
2249
2264
2279
2294
2309


0.0600
2115
2130
2145
2160
2175
2190
2205
2220
2235
2250
2265
2280
2295
2310


0.0800
2116
2131
2146
2161
2176
2191
2206
2221
2236
2251
2266
2281
2296
2311


0.1000
2117
2132
2147
2162
2177
2192
2207
2222
2237
2252
2267
2282
2297
2312
















TABLE 9





Effective Amounts of Latanoprost and Timolol
















Latanoprost
Timolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.0003
2313
2328
2343
2358
2373
2388
2403
2418
2433


0.0005
2314
2329
2344
2359
2374
2389
2404
2419
2434


0.0007
2315
2330
2345
2360
2375
2390
2405
2420
2435


0.0009
2316
2331
2346
2361
2376
2391
2406
2421
2436


0.0010
2317
2332
2347
2362
2377
2392
2407
2422
2437


0.0030
2318
2333
2348
2363
2378
2393
2408
2423
2438


0.0050
2319
2334
2349
2364
2379
2394
2409
2424
2439


0.0070
2320
2335
2350
2365
2380
2395
2410
2425
2440


0.0090
2321
2336
2351
2366
2381
2396
2411
2426
2441


0.0100
2322
2337
2352
2367
2382
2397
2412
2427
2442


0.0300
2323
2338
2353
2368
2383
2398
2413
2428
2443


0.0500
2324
2339
2354
2369
2384
2399
2414
2429
2444


0.0700
2325
2340
2355
2370
2385
2400
2415
2430
2445


0.0900
2326
2341
2356
2371
2386
2401
2416
2431
2446


0.1000
2327
2342
2357
2372
2387
2402
2417
2432
2447













Latanoprost
Timolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.0003
2448
2463
2478
2493
2508
2523
2538
2553
2568



0.0005
2449
2464
2479
2494
2509
2524
2539
2554
2569



0.0007
2450
2465
2480
2495
2510
2525
2540
2555
2570



0.0009
2451
2466
2481
2496
2511
2526
2541
2556
2571



0.0010
2452
2467
2482
2497
2512
2527
2542
2557
2572



0.0030
2453
2468
2483
2498
2513
2528
2543
2558
2573



0.0050
2454
2469
2484
2499
2514
2529
2544
2559
2574



0.0070
2455
2470
2485
2500
2515
2530
2545
2560
2575



0.0090
2456
2471
2486
2501
2516
2531
2546
2561
2576



0.0100
2457
2472
2487
2502
2517
2532
2547
2562
2577



0.0300
2458
2473
2488
2503
2518
2533
2548
2563
2578



0.0500
2459
2474
2489
2504
2519
2534
2549
2564
2579



0.0700
2460
2475
2490
2505
2520
2535
2550
2565
2580



0.0900
2461
2476
2491
2506
2521
2536
2551
2566
2581



0.1000
2462
2477
2492
2507
2522
2537
2552
2567
2582

















TABLE 10





Effective Amounts of Latanoprost and Betaxolol
















Latanoprost
Betaxolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.0003
2583
2598
2613
2628
2643
2658
2673
2688
2703


0.0005
2584
2599
2614
2629
2644
2659
2674
2689
2704


0.0007
2585
2600
2615
2630
2645
2660
2675
2690
2705


0.0009
2586
2601
2616
2631
2646
2661
2676
2691
2706


0.0010
2587
2602
2617
2632
2647
2662
2677
2692
2707


0.0030
2588
2603
2618
2633
2648
2663
2678
2693
2708


0.0050
2589
2604
2619
2634
2649
2664
2679
2694
2709


0.0070
2590
2605
2620
2635
2650
2665
2680
2695
2710


0.0090
2591
2606
2621
2636
2651
2666
2681
2696
2711


0.0100
2592
2607
2622
2637
2652
2667
2682
2697
2712


0.0300
2593
2608
2623
2638
2653
2668
2683
2698
2713


0.0500
2594
2609
2624
2639
2654
2669
2684
2699
2714


0.0700
2595
2610
2625
2640
2655
2670
2685
2700
2715


0.0900
2596
2611
2626
2641
2656
2671
2686
2701
2716


0.1000
2597
2612
2627
2642
2657
2672
2687
2702
2717













Latanoprost
Betaxolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.0003
2718
2733
2748
2763
2778
2793
2808
2823
2838



0.0005
2719
2734
2749
2764
2779
2794
2809
2824
2839



0.0007
2720
2735
2750
2765
2780
2795
2810
2825
2840



0.0009
2721
2736
2751
2766
2781
2796
2811
2826
2841



0.0010
2722
2737
2752
2767
2782
2797
2812
2827
2842



0.0030
2723
2738
2753
2768
2783
2798
2813
2828
2843



0.0050
2724
2739
2754
2769
2784
2799
2814
2829
2844



0.0070
2725
2740
2755
2770
2785
2800
2815
2830
2845



0.0090
2726
2741
2756
2771
2786
2801
2816
2831
2846



0.0100
2727
2742
2757
2772
2787
2802
2817
2832
2847



0.0300
2728
2743
2758
2773
2788
2803
2818
2833
2848



0.0500
2729
2744
2759
2774
2789
2804
2819
2834
2849



0.0700
2730
2745
2760
2775
2790
2805
2820
2835
2850



0.0900
2731
2746
2761
2776
2791
2806
2821
2836
2851



0.1000
2732
2747
2762
2777
2792
2807
2822
2837
2852

















TABLE 11





Effective Amounts of Latanoprost and Levobunolol
















Latanoprost
Levobunolol % w/w
















% w/w
0.0002
0.0004
0.0006
0.0008
0.0010
0.0020
0.0040
0.0060
0.0080





0.0003
2853
2868
2883
2898
2913
2928
2943
2958
2973


0.0005
2854
2869
2884
2899
2914
2929
2944
2959
2974


0.0007
2855
2870
2885
2900
2915
2930
2945
2960
2975


0.0009
2856
2871
2886
2901
2916
2931
2946
2961
2976


0.0010
2857
2872
2887
2902
2917
2932
2947
2962
2977


0.0030
2858
2873
2888
2903
2918
2933
2948
2963
2978


0.0050
2859
2874
2889
2904
2919
2934
2949
2964
2979


0.0070
2860
2875
2890
2905
2920
2935
2950
2965
2980


0.0090
2861
2876
2891
2906
2921
2936
2951
2966
2981


0.0100
2862
2877
2892
2907
2922
2937
2952
2967
2982


0.0300
2863
2878
2893
2908
2923
2938
2953
2968
2983


0.0500
2864
2879
2894
2909
2924
2939
2954
2969
2984


0.0700
2865
2880
2895
2910
2925
2940
2955
2970
2985


0.0900
2866
2881
2896
2911
2926
2941
2956
2971
2986


0.1000
2867
2882
2897
2912
2927
2942
2957
2972
2987













Latanoprost
Levobunolol % w/w


















% w/w
0.01
0.02
0.04
0.06
0.08
0.10
0.15
0.20
0.25







0.0003
2988
3003
3018
3033
3048
3063
3078
3093
3108



0.0005
2989
3004
3019
3034
3049
3064
3079
3094
3109



0.0007
2990
3005
3020
3035
3050
3065
3080
3095
3110



0.0009
2991
3006
3021
3036
3051
3066
3081
3096
3111



0.0010
2992
3007
3022
3037
3052
3067
3082
3097
3112



0.0030
2993
3008
3023
3038
3053
3068
3083
3098
3113



0.0050
2994
3009
3024
3039
3054
3069
3084
3099
3114



0.0070
2995
3010
3025
3040
3055
3070
3085
3100
3115



0.0090
2996
3011
3026
3041
3056
3071
3086
3101
3116



0.0100
2997
3012
3027
3042
3057
3072
3087
3102
3117



0.0300
2998
3013
3028
3043
3058
3073
3088
3103
3118



0.0500
2999
3014
3029
3044
3059
3074
3089
3104
3119



0.0700
3000
3015
3030
3045
3060
3075
3090
3105
3120



0.0900
3001
3016
3031
3046
3061
3076
3091
3106
3121



0.1000
3002
3017
3032
3047
3062
3077
3092
3107
3122

















TABLE 12







Effective Amounts of Latanoprost and Metipranolol








Latanoprost
Metipranolol % w/w





















% w/w
0.0030
0.0040
0.0060
0.0080
0.01
0.03
0.04
0.06
0.08
0.10
0.15
0.20
0.25
0.30





0.0003
3123
3138
3153
3168
3183
3198
3213
3228
3243
3258
3273
3288
3303
3318


0.0005
3124
3139
3154
3169
3184
3199
3214
3229
3244
3259
3274
3289
3304
3319


0.0007
3125
3140
3155
3170
3185
3200
3215
3230
3245
3260
3275
3290
3305
3320


0.0009
3126
3141
3156
3171
3186
3201
3216
3231
3246
3261
3276
3291
3306
3321


0.0010
3127
3142
3157
3172
3187
3202
3217
3232
3247
3262
3277
3292
3307
3322


0.0030
3128
3143
3158
3173
3188
3203
3218
3233
3248
3263
3278
3293
3308
3323


0.0050
3129
3144
3159
3174
3189
3204
3219
3234
3249
3264
3279
3294
3309
3324


0.0070
3130
3145
3160
3175
3190
3205
3220
3235
3250
3265
3280
3295
3310
3325


0.0090
3131
3146
3161
3176
3191
3206
3221
3236
3251
3266
3281
3296
3311
3326


0.0100
3132
3147
3162
3177
3192
3207
3222
3237
3252
3267
3282
3297
3312
3327


0.0300
3133
3148
3163
3178
3193
3208
3223
3238
3253
3268
3283
3298
3313
3328


0.0500
3134
3149
3164
3179
3194
3209
3224
3239
3254
3269
3284
3299
3314
3329


0.0700
3135
3150
3165
3180
3195
3210
3225
3240
3255
3270
3285
3300
3315
3330


0.0900
3136
3151
3166
3181
3196
3211
3226
3241
3256
3271
3286
3301
3316
3331


0.1000
3137
3152
3167
3182
3197
3212
3227
3242
3257
3272
3287
3302
3317
3332









II. METHODS OF TREATMENT
Methods of treating an ophthalmic disease are provided, including methods of treating glaucoma. In particular, the methods according to the embodiments of the present invention are useful in decreasing IOP without causing conjunctival hyperemia. Some embodiments of the methods provided herein include applying an ophthalmic formulation described herein to the region on or around the eye, which can treat ophthalmic diseases by sustained administration of an effective amount of an active pharmaceutical ingredient (e.g., a prostaglandin agent) and a sub-therapeutic amount of an active pharmaceutical ingredient (e.g., a vasoconstrictor agent), as described herein, to the ophthalmic tissue (i.e. conjunctiva, lacrimal tissue or cornea). The methods provided herein further include administering any appropriate ophthalmically acceptable excipient.
In another aspect, a method of reducing increased intraocular pressure (IOP) in a subject in need thereof is provided. The method includes administering to the subject a therapeutically effective amount of an ophthalmic pharmaceutical formulation including a prostaglandin agent and a vasoconstrictor agent as provided herein. In some embodiments, the subject is in need of treatment for glaucoma. In other embodiments, the subject displays conjunctival hyperemia.
In some embodiments, the prostaglandin agent is present in a therapeutically effective amount and the vasoconstrictor agent is present in a sub-therapeutic amount. In other embodiments, the prostaglandin agent is bimatoprost and the vasoconstrictor agent is timolol. In some embodiments, the ophthalmic pharmaceutical formulation further includes a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative. In other embodiments, the ophthalmic pharmaceutical formulation consists essentially of bimatoprost, timolol, a buffering agent, a tonicity agent, a salt, a thickening agent and a preservative. In some embodiments, the bimatoprost is present in a therapeutically effective amount and the timolol is present in a sub-therapeutic amount.
III. EXAMPLES
Embodiments of the present invention are further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope thereof. On the contrary, it is to be clearly understood that resort may be made to various other embodiments, modifications and equivalents, which, after reading the description provided herein, may suggest themselves to those skilled in the art without departing from the spirit of the invention.
Bimatoprost 0.03% (Lumigan®), given once daily, is a highly efficacious intraocular pressure (IOP) lowering drug (Chen J et al., Optometry Pract 2002; 3:95-102; Cohen J S et al., Surv Ophthalmol 2004; 49:S45-S52; Christiansen G A et al., Ophthalmology 2004; 111:1658-1662; DuBiner H et al., Surv Ophthalmol 2001; 45(Suppl 4):S353-S360; Eisenberg D L, Toris C B, Camras C B, Surv Ophthalmol 2002; 47:S105-S115; Gandolfi S A, Cimino L., Ophthalmology 2003; 110:609-614; Higginbotham E J, Schuman J S, Goldberg I, et al., Arch Ophthalmol 2002; 120:1286-1293; Noecker R S et al., Am J Ophthalmol 2003; 135:55-63; Parrish R K, Palmberg P, Sheu W-P., Am J Ophthalmol 2003; 135:688-703; Williams R D., Adv Ther 2002; 19:275-281). It has a very high systemic safety margin in studies conducted in laboratory animals (Woodward D F, Phelps R L, Krauss A H-P, et al., Cardiovasc Drug Rev 2004; 22:103-120). The most common ocular side effect associated with bimatoprost in humans is conjunctival hyperemia. This effect is usually mild and diminishes over time (Abelson M B et al., Adv Ther 2003; 20:1-13; Cohen J S et al., Surv Ophthalmol 2004; 49:S45-S52; DuBiner H et al., Surv Ophthalmol 2001; 45(Suppl 4):S353-S360; Eisenberg D L, Toris C B, Camras C B, Surv Ophthalmol 2002; 47:S105-S115; Higginbotham E J, Schuman J S, Goldberg I, et al., Arch Ophthalmol 2002; 120:1286-1293; Noecker R S et al., Am J Ophthalmol 2003; 135:55-63; Parrish R K, Palmberg P, Sheu W-P., Am J Ophthalmol 2003; 135:688-703; Stewart W C et al., Am J Ophthalmol 2003; 135:314-320).
Latanoprost (Xalatan®), a prostaglandin FP receptor agonist prodrug, is a well-established IOP lowering medication that has been used as a comparator for ocular hypotensive and conjunctival hyperemia effects (DuBiner H et al., Surv Ophthalmol 2001; 45(Suppl 4):S353-S360; Eisenberg D L, Toris C B, Camras C B, Surv Ophthalmol 2002; 47:S105-S115; Gandolfi S A, Cimino L., Ophthalmology 2003; 110:609-614; Noecker R S et al., Am J Ophthalmol 2003; 135:55-63; Parrish R K, Palmberg P, Sheu W-P., Am J Ophthalmol 2003; 135:688-703; Stewart W C et al., Am J Ophthalmol 2003; 135:314-320). Bimatoprost and latanoprost exhibit different pharmacological profiles and stimulate different receptor populations, bimatoprost-sensitive and prostanoid FP receptors, respectively (Chen J et al., Br J Pharmacol 2005; 144:493-501; Spada C S, Krauss A H-P, Woodward D F, et al., Exp Eye Res 2005; 80:135-145; Woodward D F, Krauss A H-P, Chen J, et al., Surv. Ophthalmol 2001; 45:S337-S345; Woodward D F, Krauss A H-P, Chen J, et al., J Pharmacol Exp Ther 2003; 305:772-785). Nevertheless, bimatoprost and prostanoid FP agonists have similarities with respect to endothelium-dependent vasodilatation in blood vessels (Astin M, Stjernschantz J, Selén G., Exp Eye Res 1994; 59:401-408; Astin M, Stjernschantz J., Eur J Pharmacol 1997; 340:195-201; Astin M, Stjernschantz J., Curr Eye Res 1997; 16:886-890; Astin M., J Ocul Pharmacol 1998; 14:119-128; Chen J. et al., Br J Pharmacol 1995; 116:3035-3041; Chen J, Woodward D F. Adv Exp Med Biol 2002; 507:331-336; Chen J et al., Br J Pharmacol 2005; 144:493-501; Stewart W C et al., Am J Ophthalmol 2003; 135:314-320) and stimulation of intracellular calcium signaling, but in different cell populations of the cat iris sphincter (Spada C S, Krauss A H-P, Woodward D F, et al., Exp Eye Res 2005; 80:135-145).
Vasorelaxation produced by the stimulation of endothelial muscarinic receptors and mediated by an endothelium-derived substance was originally discovered by Furchgott and Zawadzki (Furchgott R F, Zawadzki J V., Nature 1980; 288:373-376). This substance was identified as nitric oxide (NO) by numerous studies in the mid-1980s. In endothelial cells, increases in intracellular calcium activate endothelial nitric oxide synthase (eNOS) and lead to the generation of nitric oxide, an important regulator of ocular blood flow (Albrecht E W et al., J Pathol 2003; 1999:8-17; Koss M C., Eur J Pharmacol 1999; 374:161-174; Schmetterer L, Polak K, Prog Retin Eye Res. 2001; 20:823-847). All isoforms of NOS have been identified in eyes, including ocular surface cells (Kim J C et al., J Korean Med Sci 2002; 17:389-394; Schmetterer L, Polak K, Prog Retin Eye Res. 2001; 20:823-847). The endothelial NOS isoform, also referred to as NOS-3, has often been reported to have a protective role in a number of disease states and inflammatory processes (Albrecht E W et al., J Pathol 2003; 1999:8-17), but under certain conditions eNOS may also contribute to inflammatory responses (Cirino G, Fiorucci S, Sessa W C., Trends Pharmacol Sci 2003; 24:91-95).
Studies in patients have addressed the long-term safety of bimatoprost and the time course of bimatoprost-induced conjunctival hyperemia (Abelson M B et al., Adv Ther 2003; 20:1-13; Cohen J S et al., Surv Ophthalmol 2004; 49:S45-S52; Higginbotham E J, Schuman J S, Goldberg I, et al., Arch Ophthalmol 2002; 120:1286-1293). More recent studies investigated the inflammatory potential of bimatoprost using human conjunctival biopsies and cells (Guenoun J-M et al., Invest Ophthalmol Vis Sci 2005; 46:2444-2450; Leal B C et al., Am J Ophthalmol 2004; 138:310-313; Mroz M, Abelson M B et al., Invest Ophthalmol Vis Sci 2003; 44:ARVOE-Abstract 4417). However, information from humans is limited, so animal models have been used to investigate safety issues, the pharmacological mechanism of hyperemia, and the possible involvement of ocular surface inflammation. The ocular surface safety of topical bimatoprost treatment in animals was established in multiple-dose studies performed for the development of bimatoprost (Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275_Lumigan.htm). Other preclinical studies showed that hyperemic effects of bimatoprost on the vasculature occur by a non-inflammatory, nitric oxide-mediated vasodilatation.
Clinical Hyperemia
A month-long, multicenter, open-label study evaluated the onset and progression of conjunctival hyperemia associated with bimatoprost 0.03% once daily during 6 office visits (Abelson M B et al., Adv Ther 2003; 20:1-13). The 39 patients enrolled in the study had bilateral open-angle glaucoma or ocular hypertension and had not been treated previously with bimatoprost. An overall mean hyperemia score was calculated for each visit by averaging three vessel-bed scores obtained by slit-lamp biomicroscopy. The frequency and severity of hyperemia peaked approximately 1 day after the first instillation of bimatoprost and decreased consistently throughout the study, returning to near baseline levels by day 28 (Abelson M B et al., Adv Ther 2003; 20:1-13; FIG. 1). The hyperemia was considered not clinically significant.
The long-term safety of bimatoprost was compared to that of timolol in patients with glaucoma or ocular hypertension (Higginbotham E J, Schuman J S, Goldberg I, et al., Arch Ophthalmol 2002; 120:1286-1293). In two identical, multicenter, randomized, double-blind, 1-year clinical trials, patients were treated with bimatoprost 0.03% once daily (n=474), bimatoprost 0.03% twice daily (n=483), or timolol maleate 0.5% twice daily (n=241). Laser-flare photometry measurements (a common measure of intraocular inflammation) were taken in a subset of 310 patients (124 in the bimatoprost q.d. group, 123 in the bimatoprost b.i.d. group, and 63 in the timolol group). There were no significant differences among the treatment groups in mean laser-flare photometry readings or in mean changes from baseline. In addition, no increase in flare readings was observed in those patients who developed conjunctival hyperemia during the trial. The results indicate that bimatoprost-induced conjunctival hyperemia is not associated with intraocular inflammation in patients treated for one year. Bimatoprost was also shown to be safe and well-tolerated in the study extension to 2 years (Cohen J S et al., Surv Ophthalmol 2004; 49:S45-S52).
In a prospective interventional study, histological signs of inflammation were evaluated in conjunctival biopsies from patients with bimatoprost-associated conjunctival hyperemia who were scheduled to undergo cataract surgery (Leal B C et al., Am J Ophthalmol 2004; 138:310-313; FIG. 2). Patients with primary open-angle glaucoma were treated with bimatoprost 0.03% q.d. for 2 to 4 weeks. Nine of 13 eyes treated with bimatoprost developed conjunctival hyperemia and only those eyes were included in the study. The control eyes (n=6) were untreated and had no history of ocular disease other than cataracts. Results showed that signs of inflammation were no more frequent in conjunctival specimens from untreated control patients than in those from bimatoprost-treated patients with trace to moderate hyperemia. Vascular congestion was observed in 83% (5/6) of eyes in the control group and 78% (7/9) of eyes in the bimatoprost treatment group. Signs of inflammatory cell margination or infiltration were present in 33% (2/6) of control eyes compared with 22% (2/9) of bimatoprost-treated eyes and were not related to treatment. The findings of this study demonstrated that bimatoprost-related hyperemia was not associated with histological evidence of inflammation. In another study, bulbar conjunctival biopsies taken at day 7 and impression cytology specimens from patients treated with bimatoprost 0.03% q.d. for 60 days also showed no evidence of inflammation (Mroz M, Abelson M B et al., Invest Ophthalmol Vis Sci 2003; 44:ARVOE-Abstract 4417).
An in vitro study of the expression of various inflammation-associated markers in human cultured conjunctiva-derived epithelial cells was conducted for bimatoprost, along with the prostaglandin FP agonists: latanoprost and travoprost (Guenoun J-M et al., Invest Ophthalmol Vis Sci 2005; 46:2444-2450). None of the drugs appeared to induce direct stimulation of inflammatory pathways involving adhesion molecules or class II antigens. The toxicity observed was mild and primarily related to the concentration of the preservative benzalkonium chloride (BAK). These results are consistent with those obtained from the in-life studies and suggest that conjunctival hyperemia is most likely not an inflammatory response to bimatoprost. Furthermore, it appears too early after onset of treatment to be the inflammatory consequence of long-term treatment.
Pharmacology in Animal Models
Nitric oxide is a predominant second messenger, with a possible compensatory role for potassium channels, in prostaglandin F2a (PGF2a)-induced relaxation of rabbit endothelium-intact jugular veins (Chen J. et al., Br J Pharmacol 1995; 116:3035-3041). The vasorelaxation produced by PGF2a and latanoprost free acid in rabbit submental veins was also found to be mediated by NO and, in addition, by sensory nerves (Astin M, Stjernschantz J., Eur J Pharmacol 1997; 340:195-201). In rabbits, NO and sensory nerves were reported to have roles in PGF2a-induced ocular hyperemia as determined by NOS inhibition and sensory denervation (Astin M, Stjernschantz J, Selén G., Exp Eye Res 1994; 59:401-408; Astin M, Stjernschantz J., Curr Eye Res 1997; 16:886-890). The effects of PGF2a and its analogs, which did not include latanoprost, on ocular surface hyperemia (OSH) in dog eyes correlated with the vasorelaxation in endothelium-intact rabbit precontracted jugular veins (Chen J, Woodward D F. Adv Exp Med Biol 2002; 507:331-336). Taken together, these studies suggest that the pharmacological mechanism of conjunctival hyperemia elicited by bimatoprost and latanoprost may be elucidated by comparing their responses in the isolated jugular vein and conscious dog eyes and using NOS inhibition to determine NO involvement in vasorelaxation and OSH. A methodological difference between these in vitro tissue studies and the in vivo and human in-life studies is that a cyclooxygenase inhibitor such as indomethacin is generally used in the isolated tissue bioassay experiments. Thus, the possibility that products of arachidonic acid cyclooxygenation could be generated de novo and contribute to the vasorelaxation was examined in the rabbit jugular vein preparation.
Effects of Bimatoprost and Latanoprost Free Acid on Rabbit Isolated Jugular Veins
The pharmacological mechanism of ocular surface hyperemia elicited by bimatoprost and latanoprost was investigated using the rabbit jugular vein preparation as a quantitative in vitro model (Chen J. et al., Br J Pharmacol 1995; 116:3035-3041). In these studies, tissues were suspended in jacketed organ baths containing Krebs buffer with 1 μM indomethacin (endothelium-intact and -denuded) or in its absence (endothelium-intact) and precontracted with 3 μM histamine. Vasorelaxant and contractile activity were determined as a percentage (%) of the control tone elicited by histamine. Statistical comparisons for the vasorelaxant activity consisted of testing for significance of difference at each of the concentrations tested using the Student'-s t-test for unpaired samples (indomethacin vs no indomethacin) and paired samples (L-NAME vs D-NAME). Differences were considered statistically significant for P-values≦0.05.
Bimatoprost and latanoprost free acid produced endothelium-dependent vasorelaxant effects that are consistent with and extend previously reported data (Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609; Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275_Lumigan.htm). Bimatoprost produced a weak vasorelaxant response (EC50=3,981 nM) at high concentrations in endothelium-intact jugular veins and was inactive in the endothelium-denuded preparation (Chen J et al., Br J Pharmacol 2005; 144:493-501; FIG. 3A). This response profile is consistent with a wide separation between its IOP lowering and ocular surface hyperemia effects. Bimatoprost produced vasorelaxant responses at the 10−6 M to 10−4 M concentrations (FIGS. 3A, 5A), which encompassed concentrations (10−6-10−5 molar equivalent) found in non-human primate ocular surface tissues after single or multiple topical bimatoprost dosing (Woodward D F, Krauss A H-P, Chen J, et al., J Pharmacol Exp Ther 2003; 305:772-785; Woodward D F, Phelps R L, Krauss A H-P, et al., Cardiovasc Drug Rev 2004; 22:103-120). The weak vasorelaxant response to bimatoprost in the rabbit jugular vein was predictive for the findings of mean mild conjunctival hyperemia in human subjects, although the hyperemia severity may vary according to the tissue concentrations achieved.
Latanoprost free acid produced complex concentration-response curves of maximal vasorelaxation at 10−7 M and a reversal of the relaxant effects at higher concentrations of 10−6 M and 10−5 M in the endothelium-intact preparation and only contractions at the 10−5 M concentration in endothelium-denuded veins (Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609; Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275 Lumigan.htm; FIG. 3B). The response to latanoprost free acid in the jugular veins was interesting in that it produced vasorelaxation at the lower concentrations tested and this response reversed at higher concentrations. This effect, particularly the reversal at high concentrations, may contribute to a decreased ability of latanoprost to elicit ocular surface hyperemia compared to bimatoprost and correlates with its mild conjunctival hyperemia response in humans (Eisenberg D L, Toris C B, Camras C B, Surv Ophthalmol 2002; 47:S105-S115; Noecker R S et al., Am J Ophthalmol 2003; 135:55-63; Parrish R K, Palmberg P, Sheu W-P., Am J Ophthalmol 2003; 135:688-703; Stewart W C et al., Am J Ophthalmol 2003; 135:314-320). The vasoconstrictor effects of latanoprost free acid in both the endothelium-intact and -denuded veins suggested the possible involvement of contractile prostanoid TP receptors. However, TP stimulation does not explain reversal of the vasorelaxant response to latanoprost free acid at pharmacological concentrations up to 10−6 M, since the TP antagonist SQ 29548 used in these studies is highly effective yet did not block the reversal of the vasorelaxant response. Moreover, latanoprost free acid did not contract the endothelium-denuded preparation over the concentration range of 10−10 to 10−6 M. Therefore, reversal of the vasorelaxant response to latanoprost free acid in the rabbit jugular vein may involve an endothelial FP receptor or even perhaps a previously unrecognized receptor that causes endothelium-induced vasoconstriction. The increased variability in latanoprost free acid responses at the higher concentrations was likely due to the opposing vasorelaxant and vasoconstrictor effects.
These different activity profiles for latanoprost free acid and bimatoprost in the rabbit jugular vein provide further pharmacological evidence that these agents are recognized by different receptor populations. Latanoprost free acid is a well-known potent and selective prostanoid FP receptor agonist, while bimatoprost is a putative prostamide receptor agonist. Studies using isolated tissue preparations and DNA synthesis in mouse cultured fibroblasts have shown that species-, tissue-, or preparation-related factors, partial agonism, and metabolism all do not provide acceptable explanations of the different activity profiles of bimatoprost and prostanoid FP receptor agonists (Chen J et al., Br J Pharmacol 2005; 144:493-501). The results from the rabbit jugular vein model add support to the contention that bimatoprost is a prostamide and produces its effects by stimulating bimatoprost-sensitive receptors as opposed to prostanoid FP receptors.
Indomethacin is a cyclooxygenase inhibitor that is routinely used in prostanoid receptor assays (Chen J et al., Current protocols in pharmacology. New York: John Wiley & Sons, Inc., 2001; 4.18.1-4.18.41). Studies of intact and viable isolated tissues, such as the rabbit jejunum (Ferreira S H, Herman A G, Vane J R., Br J Pharmacol 1976; 56:469-477) and human bronchial muscle (Hage-Legrand I, Cerrina J, Raffestin B et al., J Pharmacol Exp Ther 1986; 239:536-541), found the basal output of prostaglandins to be 8-23 pg/mg of tissue at baseline or over 30 min collection periods. In the presence of 1.7 and 2.79 μM indomethacin, the amounts of prostaglandins generated in isolated tissues become negligible (Botting J H, Salzmann R., Br J Pharmacol 1974; 50:119-124; Haye-Legrand I, Cerrina J, Raffestin B et al., J Pharmacol Exp Ther 1986; 239:536-541). The 1 μM concentration of indomethacin in the tissue bath, in our experience, is sufficient to prevent biosynthesis of prostanoids within isolated vascular tissue preparations (Alster P, Wennmalm Å., Eur J Pharmacol 1983; 86:441-446; Chen J et al., Current protocols in pharmacology. New York: John Wiley & Sons, Inc., 2001; 4.18.1-4.18.41). At substantially higher concentrations, indomethacin has been reported to lose selectivity for prostanoid biosynthesis inhibition (Aboulafia J et al., Br J Pharmacol 1976; 58:223-228; Goodfriend T L, Simpson R U., Br J Pharmacol 1981; 72:247-255; Northover B J, Br J Pharmacol 1971; 41:540-551; Sawdy R et al., Br J Pharmacol 1998; 125:1212-1217). The rabbit jugular vein studies were conducted under experimental conditions of intact isolated tissues continually bathed in buffer containing 1 μM indomethacin. Results of these studies are consistent with and extend previously reported findings (Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275_Lumigan.htm). The contractile response to histamine in all of the tissues examined without indomethacin was superimposed with spontaneous spike activity, indicating a contribution of prostaglandin production to the contractions. In the presence of indomethacin, the histamine-induced contractions were well-maintained, stable, and showed minimal spontaneous activity. Endothelium-intact tissues incubated with and without 1 μM indomethacin showed no statistically significant differences in effects between the means and at each of the concentrations tested for bimatoprost and latanoprost free acid (Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275 Lumigan.htm; FIG. 4). These findings suggest that responses to these agents in the rabbit isolated jugular vein are mediated by endothelial located receptors and not by de novo biosynthesis of prostaglandins. Consequently, these in vitro responses can be compared with those observed in the in vivo and in-life studies.
The involvement of nitric oxide in bimatoprost- and latanoprost free acid-induced relaxation of the rabbit jugular vein was determined using L-NAME (L-NG-nitroarginine methyl ester), a potent enantiomerically specific competitive inhibitor of NOS in cell types that include vascular endothelial cells (Koss M C., Eur J Pharmacol 1999; 374:161-174; Rees D D et al., Br J Pharmacol 1990; 101:746-752), and its inactive isomer D-NAME. In the endothelium-intact histamine precontracted tissues, 100 μM L-NAME significantly inhibited the vasorelaxant responses to bimatoprost and latanoprost free acid, compared to their respective responses in the 100 μM D-NAME control tissues, P≦0.05, paired t-test (FIG. 5). This result indicates that NO is an important contributor to vasorelaxation produced by bimatoprost and latanoprost free acid in the rabbit jugular vein and is consistent with previous findings for PGF2a. (Chen J. et al., Br J Pharmacol 1995; 116:3035-3041). L-NAME at 100 μM was less effective in inhibiting the responses to PGF2a. (Chen J. et al., Br J Pharmacol 1995; 116:3035-3041) compared to bimatoprost and latanoprost free acid, which may reflect the selectivity of these agents for their respective receptors.
Effects of Nitric Oxide Synthase Inhibition on Conjunctival Hyperemia in Dog Eye
The ocular surface hyperemic effects of bimatoprost, latanoprost (isopropyl ester), or prostaglandin E2 (PGE2) were determined in eyes of conscious dogs treated with topical L-NAME or D-NAME. The dog is suitable for OSH studies since it responds with mild conjunctival hyperemia to bimatoprost and latanoprost, which is consistent with the hyperemic responses observed in humans. The rabbit eye is less predictive for conjunctival hyperemia since latanoprost has no significant acute effects on blood flow in the conjunctiva at a topical dose of 10 μg (0.03%, based on a 30 μl volume) (Astin M, Stjernschantz J, Selén G., Exp Eye Res 1994; 59:401-408; Stjernschantz J et al., Prog Retin Eye Res 2000; 19:459-496). In vivo experiments consisted of visually grading ocular surface hyperemia (OSH) in conscious Beagle dogs (males or females). All test articles were administered in a masked fashion. The test compound was applied bilaterally as a single drop at time (t)=0 in the one day study. L-NAME 1% (w/v) was topically applied to one eye, while its inactive isomer D-NAME 1% (w/v) was given to the contralateral eye, at 90, 60, 30, 5 min pre-dose, and 1, 2, 3, and 4 h post-dose for a total of 8 drops (approximately 2.8 mg) each per animal. OSH grading was semi-quantitative and assessed according to a 5-point scoring scale used for clinical evaluations: 0=none; 0.5=trace; 1=mild; 2=moderate; and 3=severe. The statistical comparisons for OSH in the dog eyes consisted of testing for significance of mean differences from baseline at each time point using the Wilcoxon signed-rank test for paired observations. Differences were considered statistically significant for P-values≦0.05.
L-NAME and D-NAME did not produce conjunctival hyperemia or any adverse ocular effects at baseline (time 0). Our studies using direct evaluation of L-NAME on conjunctival hyperemia in dogs indicated the involvement of NO in OSH elicited by bimatoprost and latanoprost (Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609; FIG. 6A, 6B). The hyperemia effects elicited by bimatoprost 0.1% and latanoprost 0.005% were blocked in eyes treated with L-NAME, as shown by responses that were not different from the respective paired baseline values at 0-6 h of the study. In D-NAME treated control dog eyes, bimatoprost and latanoprost elicited trace to mild ocular surface hyperemic responses that were significantly different compared to baseline (P≦0.05). The inhibition by L-NAME in dog eyes and the in vitro rabbit jugular vein model for OSH suggests that NO is the predominant mediator of OSH for bimatoprost. L-NAME treatment also inhibited latanoprost-induced conjunctival hyperemia in dog eyes but, in this case, the D-NAME treated eyes were unaffected. This finding may possibly reflect a partial role for NO in latanoprost-induced OSH, since the vasorelaxant effects in rabbit submental veins were shown to be mediated by NO, calcitonin gene-related peptide, substance P or another tachykinin released from perivascular sensory nerves (Astin M, Stjernschantz J., Eur J Pharmacol 1997; 340:195-201; Stewart W C et al., Am J Ophthalmol 2003; 135:314-320).
In these studies, PGE2 0.01% served as a comparator. It produced OSH of mild to moderate severity; its effects were significantly different from those at the baseline (P≦0.05) in both the D-NAME- and L-NAME-treated eyes (Chen J et al., Invest Ophthalmol Vis Sci 2004; 45:ARVO E-Abstract 2609; FIG. 6C). The finding that L-NAME had no effect on the observed conjunctival hyperemia was predicted, since PGE2 does not require an intact vascular endothelium to exert its relaxant response (Chen J. et al., Br J Pharmacol 1995; 116:3035-3041). These results showed that the nitric oxide synthase inhibitor L-NAME inhibits the in vivo acute OSH response to bimatoprost or latanoprost, but has no effect on PGE2-induced OSH.
In-Life Observation and Histopathological Assessment
The ocular surface safety following bimatoprost treatment was addressed using rabbits, dogs, and non-human primates in studies of one month to one year duration. Results of the studies were submitted to worldwide regulatory authorities for the approval of bimatoprost as an antiglaucoma drug (Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www fda.gov/cder/foi/nda/2001/21275 Lumigan.htm). The possible involvement of ocular surface inflammation in animal eyes treated with subclinical, clinical, and exaggerated doses of bimatoprost and controls were evaluated by in-life observations and histopathological assessment. The in-life ocular evaluations consisted of clinical observations, gross ocular observations, ophthalmoscopy, and slit lamp biomicroscopy. Histopathological assessments were conducted by three board-certified veterinary pathologists, each applying somewhat different criteria and terminology, in separate studies. The animals comprised of those scheduled for terminal sacrifice and similar numbers of males and females were used in each group. As part of a comprehensive assessment of the eye, tissues from the upper and lower eyelids with associated palpebral conjunctiva were excised for histopathological evaluation. Ocular tissues were preserved in 10% formalin, processed to paraffin, sectioned at 5 microns, and stained using H&E. The tissues were sectioned in an anatomically consistent manner for all animals in all groups. One section was evaluated for each designated site.
Rabbit Studies
The rabbit is the species most frequently used in eye irritation tests. New Zealand White (albino) rabbits were assigned to 4 groups (n=16 per group) for a one month study (Table 13). The animals received four times daily (q.i.d.) topical applications of vehicle or bimatoprost (0.001%, 0.01%, 0.1%) in the left eye, while the right eye was untreated.







TABLE 13







Incidence of minimal-to-mild mononuclear cell infiltration in palpebral


conjunctiva of albino rabbits (n = 16 in each group) after daily application


of bimatoprost, q.i.d. for one month









Groups












1
2
3
4









Eyes

















Right
Left
Right
Left
Right
Left
Right



Left
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated


Treatment
Vehicle
Control
0.001%
control
0.01%
Control
0.1%
control


















Upper
1
3
4
3
6
4
8
6


Eyelid


Lower
15
12
12
10
13
15
14
14


Eyelid





Incidence: number of animals with histopathologic findings


q.i.d. = four times daily; Study 2968-58. Adapted from ref. 31.






Dutch-Belted (pigmented) rabbits were assigned to 2 groups (n=19-20 per group) and received twice daily (b.i.d.) topical ocular applications of vehicle or bimatoprost 0.03% in the left eye for one month (Table 14). The right eye was untreated. The vehicle and drug solutions were preserved with 0.005% benzalkonium chloride (BAK).







TABLE 14







Incidence of mononuclear cell infiltration in palpebral conjunctiva of


pigmented rabbits after daily application of bimatoprost,


b.i.d. for one month









Groups










1 (n = 20)
2 (n = 19)









Eyes













Right

Right



Left
Untreated
Left
Untreated


Treatment
Vehicle
Control
Bimatoprost 0.03%
Control














Upper Eyelid






Minimal/mild
10
10
6
14


Lower Eyelid


Minimal/mild
16
15
12
13


Moderate
1
0
0
0





Incidence: number of animals with histopathologic findings;


n = number of animals per group.


b.i.d. = twice daily;


Study TX97032. Adapted from ref. 31.






In a six month study, Dutch-Belted (pigmented) rabbits were assigned to 4 groups (n=20 per group) (Table 15). The left eye received b.i.d. topical ocular applications of vehicle, once daily (q.d.) or b.i.d. bimatoprost 0.03%, or b.i.d. bimatoprost 0.1%. The right eye was untreated. The vehicle and drug solutions were preserved with 0.005% BAK.







TABLE 15







Mononuclear cell infiltration in palpebral conjunctiva of pigmented rabbits


(n = 20 per group) after daily application of bimatoprost, q.d. or b.i.d. for 6 months









Groups












1
2
3
4









Eyes
















Left
Right
Left
Right
Left
Right
Left
Right



Vehicle
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated


Treatment
b.i.d.
Control
0.03% q.d.
control
0.03.% b.i.d.
Control
0.1% b.i.d.
Control





Upper
N
N
N
N
N
N
N
N


Eyelid


Lower
N
N
N
N
N
N
N
N


Eyelid





N = Tissues within normal histological limits


q.d. = once daily;


b.i.d. - twice daily;


Study TX98004. Adapted from ref. 31.






Dog Study

Beagle dogs are recognized as appropriate for use in multiple-dose ocular irritation and safety studies. In a one month study, Beagle dogs were assigned to 4 groups (n=6 per group) (Table 16). The left eye was treated with q.i.d. topical ocular applications of vehicle or bimatoprost at 0.001%, 0.01%, or 0.1% doses and the right eye was untreated. All test solutions were non-preserved.







TABLE 16







Incidence of minimal-to-mild mononuclear cell infiltration in palpebral


conjunctiva of dogs (n = 6 per group) after daily application of bimatoprost,


q.i.d. for one month.









Groups












1
2
3
4









Eyes

















Right
Left
Right
Left
Right
Left
Right



Left
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated


Treatment
Vehicle
Control
0.001%
Control
0.01%
Control
0.1%
Control





Upper
5
5
4
4
3
5
5
3


Eyelid


Lower
5
5
3
4
5
5
6
6


Eyelid





Incidence: number of animals with histopathologic findings


q.i.d. = four times daily;


Study 3137-5. Adapted from ref. 31.






Non-Human Primate Study

Cynomolgus monkeys have been historically used in safety evaluation studies and are recommended by most regulatory agencies. In a 12 month study, Cynomolgus monkeys scheduled for terminal sacrifice were assigned to 4 groups (n=6-8 per group) (Table 17). The right eyes were treated with topical ocular applications of vehicle b.i.d., bimatoprost 0.03% q.d., bimatoprost 0.03% b.i.d., or bimatoprost 0.1% b.i.d. The left eye was not treated. All test solutions were preserved with 0.005% BAK.







TABLE 17







Incidence of inflammation in palpebral conjunctiva of cynomolgus


monkeys after daily application of bimatoprost, q.d. or b.i.d. for 12 months.









Groups












1 (n = 6)
2 (n = 8)
3 (n = 6)
4 (n-6)









Eyes
















Left
Right
Left
Right
Left
Right
Left
Right



Untreated
Vehicle
Untreated
Bimatoprost
Untreated
Bimatoprost
Untreated
Bimatoprost


Treatment
control
b.i.d.
control
0.03% q.d.
control
0.03% b.i.d.
control
0.1% b.i.d.





Upper Eyelid










Chronic active
5
5
4
4
3
5
5
3


Lower Eyelid


Subacute


Chronic active





Incidence: number of animals with histopathologic findings;


n = number of animals per group.


q.d. = once daily;


b.i.d. = twice daily;


Study 6177-110. Adapted from ref. 31.






Summary of In-Life Observations and Histopathological Assessments

No compound-related findings of corneal toxicity, discomfort or irritation, or histopathological changes were observed in eyes of the animal species treated with multiple subclinical, clinical, and exaggerated doses of bimatoprost. Histopathological evaluations of eyes treated with the highest dose and frequency of application of bimatoprost in each study were compared to vehicle treatment. The results are summarized in Table 18. Only vehicle-treated eyes showed inflammatory changes above background stimulation in the palpebral conjunctiva, whereas no such changes were related to bimatoprost treatment. The presence of minimal to mild inflammatory infiltrates (background stimulation) in conjunctiva of untreated control eyes, vehicle-treated eyes, and bimatoprost-treated eyes was considered to be incidental and unrelated to treatment. In all the studies, bimatoprost did not increase the intensity or alter the characteristics of inflammatory processes in the conjunctiva.







TABLE 18







Summary of incidence of histopathologic findings above background


stimulation in the lower palpebral conjunctiva after topical


administration of bimatoprost















Incidence of





Highest dose &
inflammatory




Duration
frequency
infiltrates above


Species
Strain
months
tested
background














Rabbits
New Zealand
1
0.1% q.i.d.
0 (16)



White

Vehicle
0 (16)


Rabbits
Dutch-Belted
1
0.03% b.i.d.
0 (19)





Vehicle
1 (20)


Rabbits
Dutch-Belted
6
0.1% b.i.d.
1 (20)





Vehicle
0 (20)


Dogs
Beagle
1
0.1% q.i.d.
0 (6)





Vehicle
0 (6)


Monkeys
Cynomolgus
12
0.1% b.i.d.
0 (60)





Vehicle
1 (6)





Incidence: number of animals with histopathologic findings (per total number of animals) Adapted from ref. 31.






The in-life evaluations indicated no treatment-related ocular surface inflammation in animals treated with bimatoprost for periods of up to one year exposure. Minimal to mild mononuclear infiltrates were recognizable in routine sampling of the conjunctiva of untreated and treated laboratory animals. Such changes represent background stimulation of this mucosal barrier. Thus, across all studies and species, the results indicated that bimatoprost does not increase the intensity or alter the characteristics of inflammatory processes in the conjunctiva of animals. These findings are more in agreement with a protective role for eNOS rather than involvement in pro-inflammatory processes. The safety evaluation results for animals treated with bimatoprost correlate with the clinical assessments suggesting that bimatoprost-related conjunctival hyperemia occurs by a non-inflammatory mechanism (Abelson M B et al., Adv Ther 2003; 20:1-13; Guenoun J-M et al., Invest Ophthalmol Vis Sci 2005; 46:2444-2450; Higginbotham E J, Schuman J S, Goldberg I, et al., Arch Ophthalmol 2002; 120:1286-1293; Leal B C et al., Am J Ophthalmol 2004; 138:310-313; Lumigan (Bimatoprost) Ophthalmic Solution. Pharmacology Review of New Drug Application 21-275. FDA/Center for Drug Evaluation and Research. 2001; Part 1 and Part 2:1-107. Available online at http://www.fda.gov/cder/foi/nda/2001/21275_Lumigan.htm; Mroz M, Abelson M B et al., Invest Ophthalmol Vis Sci 2003; 44:ARVOE-Abstract 4417; Noecker R S et al., Am J Ophthalmol 2003; 135:55-63).
CONCLUSIONS
The results of pharmacological studies of OSH indicate that bimatoprost and latanoprost elicit conjunctival hyperemia by a common signaling mechanism that involves intracellular calcium and endothelial-derived nitric oxide. Extensive safety evaluation studies of bimatoprost at clinical and exaggerated doses in three species of laboratory animals found no evidence of treatment-related ocular surface inflammation. The results showed that bimatoprost has a very high safety margin and is well-tolerated in animals.
Clinical Studies with Bimatoprost in Combination with Timolol
Approximately 60% of patients being treated for glaucoma require two or more agents (adjuvant therapy) to control the intraocular pressure. For this reason, combination therapies, in which two pharmaceutically active agents are combined in the same formulation, have been developed to decrease complexity in the treatment regimen, improve patient compliance, and to minimize the exposure to excipients with possible toxicity such as benzalkonium chloride. Currently, the most commonly prescribed adjuvant therapy includes a beta-blocker and a prostaglandin analogue. For this reason, Allergan has formulated bimatoprost 0.03% together with timolol 0.5% (Combination). To date, four pivotal clinical trials have been conducted that compares the combination with monotherapy (bimatoprost 0.03% once a day given individually) Unexpectedly, there were substantial reductions in the degree of ocular surface hyperemia observed with the combination compared with monotherapy as reported as adverse events or treatment-related adverse events in the pooled data from the 018T and 021T studies as well as from the 026T study (table) suggesting that the timolol may have been responsible for the decrease in hyperemia.







TABLE 19







Phase 3 Studies: Incidence of Conjunctival hyperaemia Detected on


Biomicroscopy and Reported as a Treatment-Related AE for


Patients in the Combination Group










Combination
Bimatoprost












Biomicro
Rx-rel
Biomicro
Rx-rel



≧+1
AE
≧+1
AE















Pooled 192024-018T & 021T
15.6%a
22.7%a
30.9%
38.5%


(3 months)


Pooled 192024-018T & 021T
22.1%a
25.7%a
41.5%
43.4%


(3 months)


192024-504T (3 months)
20.4%
25.0%
21.1%
29.9%


192024-026T (3 weeks)
8.5%
19.3%
18.9%
27.8%





* Combination statistically significantly lower than Bimatoprost (p ≦ 0.024)






The mechanism of the effect is not certain; however, it is well-known that beta blockade can induce an unopposed alpha agonism leading to vasoconstriction and decrease ocular hyperemia. (see first two reference at the end) More recently, it has been shown that nitric oxide synthase is under beta-adrenergic control. This suggests that beta-blockade should decrease the production of nitric oxide resulting from the topical exposure of the prostaglandin analogue to the ocular surface reducing the severity of ocular surface hyperemia (the remainder of references at the end of the document).

## Claims

1. A composition comprising a prostaglandin agent and a vasoconstrictor agent, wherein said composition is an ophthalmic pharmaceutical formulation further comprising an ophthalmically acceptable excipient, and wherein said vasoconstrictor agent is present in a sub-therapeutic amount, and wherein said prostaglandin agent and said vasoconstrictor agent are present in a combined amount effective to treat an ophthalmic disease.

2. The composition of claim 1, wherein said ophthalmic pharmaceutical formulation is a gel formulation.

3. The composition of claim 1, wherein said ophthalmic pharmaceutical formulation is an aqueous solution.

4. The composition of claim 1, wherein said prostaglandin agent is bimatoprost.

5. The composition of claim 4, wherein said bimatoprost is present in an amount approximately equal to or less than about 0.1% w/w.

6. The composition of claim 4, wherein said bimatoprost is present in an amount of about 0.03% w/w.

7. The composition of claim 1, wherein said prostaglandin agent is travoprost.

8. The composition of claim 7, wherein said travoprost is present in an amount approximately equal to or less than about 0.1% w/w.

9. The composition of claim 7, wherein said travoprost is present in an amount of about 0.004% w/w.

10. The composition of claim 1, wherein said prostaglandin agent is latanoprost.

11. The composition of claim 10, wherein said latanoprost is present in an amount approximately equal to or less than about 0.1% w/w.

12. The composition of claim 10, wherein said latanoprost is present in an amount of about 0.005% w/w.

13. The composition of claim 1, wherein said vasoconstrictor agent is an alpha adrenergic agonist.

14. The composition of claim 1, wherein said vasoconstrictor agent is selected from the group consisting of befunolol, betaxolol, carteolol, levobunolol, metipranolol, timolol, brimonidine, tetrahydrozolone hydrochloride and mepindolol.

15. The composition of claim 14, wherein the vasoconstrictor agent is timolol.

16. The composition of claim 14, wherein the vasoconstrictor agent is befunolol.

17. The composition of claim 14, wherein the vasoconstrictor agent is betaxolol.

18. The composition of claim 14, wherein the vasoconstrictor agent is carteolol.

19. The composition of claim 14, wherein the vasoconstrictor agent is levobunolol.

20. The composition of claim 14, wherein the vasoconstrictor agent is metipranolol.

21. The composition of claim 14, wherein the vasoconstrictor agent is brimonidine.

22. The composition of claim 14, wherein the vasoconstrictor agent is tetrahydrozolone hydrochloride.

23. The composition of claim 14, wherein the vasoconstrictor agent is mepindolol.