BR112017005523B1 - TOPIC OCULAR SUSPENSION FORMULATION AND USE THEREOF - Google Patents

Abstract

OCULAR FORMULATIONS FOR DRUG RELEASE AND PROTECTION OF THE ANTERIOR SEGMENT OF THE EYE. The present application relates to topical formulations comprising Compound-I or its free base, and a second active agent selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof, for treating ocular neovascularization. The present application also relates to pharmaceutical compositions comprising particles of Compound-I or its free base, and suspension formulations comprising particle compositions of Compound-I or its free base.

Description

RELATED REQUEST

[01]This application claims priority to, and the benefit of, U.S.S.N. 62/051,794, filed on September 17, 2014, the contents of which are incorporated by reference in their entirety.

ORDER FIELD

[02] Embodiments disclosed herein are generally directed to the topical administration of a combination of a first active agent, for example, a pharmaceutical compound or a salt thereof, and a second active agent, to treat ocular diseases or conditions. Disclosed embodiments include ocular formulations comprising a first active agent or a salt thereof and a second active agent, where the formulation is a solution or a suspension. The solution or suspension may further comprise a solubilizing agent, and is suitable for delivering the first active agent, or a salt thereof, to the posterior segment of the eye while protecting the anterior segment of a subject's eye.

FUNDAMENTALS

[03]Treatment of diseases or disorders of the posterior segment of the eye with topically applied active agents was not effective because of inefficient delivery of the active agent to the target site. The vast majority of topical drugs penetrate through the cornea. However, the cornea is not equally permeable to all topically applied active agents, as the basic structure of the cornea dictates the relative penetration of the active agent. Effectively, the greatest barrier to penetration of the active agent is the corneal epithelium, which is rich in cellular membranes and is, therefore, more susceptible to penetration by lipophilic agents. On the contrary, since the corneal stroma is basically made up of water, active agents pass through more easily if they are hydrophilic. The endothelium represents a monolayer that, once again, is lipophilic. Active agents that are lipophilic or amphiphilic, in that they can behave as charged or uncharged, penetrate the cornea better. Similar to the cornea, the conjunctival epithelium and blood vessels within or beneath the conjunctival epithelium can be penetrated by the same type of lipophilic or biphasic agents. However, because of the nature of the lipophilic membranes in the conjunctiva and its inherent vasculature, most active agents typically do not penetrate through the conjunctiva and into the eye. Agents with limited penetration into vascular tissues in the conjunctival and subconjunctival regions are drained into the systemic circulation.

[04] Because of the limited permeability of many topical drops to the corneal and conjunctival barriers, a major disadvantage of topical drops may be the need for high concentrations of active agents in the topical formulation in order to obtain significant therapeutic drug levels in internal ocular tissues . Depending on the active agent, the molecule itself or high concentrations thereof, a topical formulation can be toxic to the anterior segment of the eye, including the conjunctiva, cornea and/or lens, causing various injuries to the ocular surface, such as corneal epithelial defects and erosions. .

[05]Ocular side effects observed following treatment with anti-EGFR drug therapies, e.g., anti-EGFR cancer therapies, have illuminated the essential role that the EGFR signaling pathway plays in maintaining and restoring epithelial health human cornea. Patients treated with anti-EGFR drug therapies may develop corneal changes such as epithelial degeneration and defects, ulceration, corneal epithelial thinning, corneal erosions and/or edema, keratitis as well as perforation while undergoing therapy or even after discontinuation of therapy. anti-EGFR. The important role of EGFR signaling in homeostasis and pathophysiology of the corneal epithelium is well established. EGFR activation is both necessary and sufficient for corneal epithelial migration, proliferation, and differentiation. Furthermore, EGFR is the primary mediator of wound healing during in vitro experiments with immortalized human corneal epithelial cells. Therefore, treating ocular diseases or disorders (e.g., diseases or disorders of the posterior segment of the eye) would benefit from administration of an agent that maintains EGFR activity in addition to administration of a formulation comprising an active agent (e.g., an active agent having toxic or anti-EGFR activity).

[06] The present application provides new formulations that overcome the problems encountered in the ocular delivery of existing topical therapeutic agents. The present application effects the combined effects of decreasing drug exposure to the cornea and anterior segment and protecting corneal and anterior segment tissues, while increasing posterior segment bioavailability. By reducing corneal exposure, protecting corneal tissues, and increasing posterior segment bioavailability, the formulation of the present application improves ocular tolerability and increases the therapeutic index of the active agent.

summary

[07] The present application relates to pharmaceutical formulations in the form of a solution and/or a suspension, which reduce exposure to a first active agent in the anterior segment of the eye, for example the ocular surface comprised of the cornea and conjunctiva, and protect the eye, for example the anterior segment of the eye, by maintaining EGFR activity. The pharmaceutical formulations of the application increase the bioavailability of the first active agent in the posterior segment of the eye, for example in the central choroid and/or central retina, and protect and/or improve the health of the anterior segment of the eye.

[08] The present application provides a formulation comprising a first active agent, and optionally a second active agent, in the form of a solution or a suspension, with superior characteristics compared to a composition formed as a gel. The present application establishes that the first active agent and/or the second active agent can be formulated together as a solution and/or a suspension. Increased levels of the first active agent in the anterior segment of the eye limit the ocular tolerability of topical drops containing the first active agent and may cause corneal epithelial defects and erosions. The presence of the second active agent may prevent damage that may be caused by exposure to the first active agent, treat any damage that may be caused by exposure to the first active agent, and/or improve the overall health of the ocular surface, specifically the corneal epithelium. . The second active agent may be formulated together with the first active agent, or formulated as a separate formulation that is administered in combination with the formulation comprising the first active agent.

[09] The formulations of the present application reduce exposure of the first active agent to the anterior segment of the eye, such as the corneal or conjunctival surface, protect from and/or repair damage to the anterior segment of the eye, such as the corneal or conjunctival surface, and maintain adequate concentrations of a first active agent necessary to bind the relevant receptors in target tissues and confer a therapeutic effect on the posterior segment of the eye, such as the choroid and retina.

[10] The present application relates to formulations and methods useful for treating pathological conditions that arise from or are exacerbated by ocular angiogenesis, neovascularization, and/or vascular leakage, for example, in diabetic retinopathy (including background diabetic retinopathy, retinopathy proliferative diabetic, and diabetic macular edema); age-related macular degeneration (AMD) (including neovascular (wet/exudative) AMD, dry AMD, and Geographic Atrophy); pathological choroidal neovascularization (CNV) and vascular leakage of any mechanism (e.g., high myopia, trauma, sickle cell anemia; ocular histoplasmosis, angioid lines, traumatic choroidal rupture, optic nerve drusen, or some retinal dystrophies); pathological retinal neovascularization and vascular leakage of any mechanism (e.g., sickle cell retinopathy, Eales disease, ocular ischemic syndrome, carotid-cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic occlusive arteriolitis, birdshot retinochoroidopathy, retinal vasculitis , sarcoidosis, or toxoplasmosis); uveitis; retinal vein occlusion (central or branch); eye trauma; surgery-induced edema; surgery-induced neovascularization; cystoid macular edema; ocular ischemia; retinopathy of prematurity; coat disease; sickle cell retinopathy; and/or neovascular glaucoma. In one embodiment, the pathological state is AMD. In one embodiment, the pathological states arise from or are exacerbated by ocular angiogenesis and/or neovascularization.

[11] The present application also relates to formulations and methods useful for preventing and/or treating disruptions of the corneal epithelium associated with a disease, including a systemic disease (e.g., cancer, diabetes, etc.) and an ocular disease, or with a side effect of a locally or systemically administered drug (e.g., anti-EGFR agents or EGFR inhibitors). The formulation of the present application has at least one anti-angiogenic agent, anti-inflammatory agent, or anti-vascular permeability agent for use in treating angiogenic ocular disorders and an EGFR modulator (e.g., an agent that maintains the activity of or active EGFR) for use in preventing and/or treating disruptions of the corneal epithelium.

[12] According to embodiments of the application, the first active agent is an anti-angiogenic kinase inhibitor and the second active agent is a kinase modulator (e.g., activator). In one embodiment, the first active agent inhibits a kinase that is different from the kinase that is modulated (e.g., activity maintained or activated) by the second active agent. Examples of some kinase inhibitors that can be used to produce beneficial therapeutic results include inhibitors of receptor tyrosine kinases, for example, without limitation, VEGFR, FGFR, Tie-2, and Ephrin kinase receptors. Examples of kinase modulators that can be used to produce beneficial therapeutic results include ErbB receptor tyrosine kinase activators, e.g., without limitation, EGFR1/ErbB1/HER1, ErbB2/HER2/Neu, ErbB3/HER3, and ErbB4/HER . In one embodiment, the ErbB receptor tyrosine kinase is EGFR1. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In another embodiment, the second active agent is nicotinic acid or nicotinamide. In another embodiment, the second active agent is vitamin K.

[13] In some embodiments, a second active agent of the present application reduces or alleviates EGFR inhibition in the anterior segment of the eye caused by high concentrations of a first active agent of the present application (e.g., transient high concentrations of the first agent active after administration of the first active agent to the anterior segment of the eye).

[14] Embodiments of the present application provide an ophthalmic formulation for treating ocular neovascularization comprising a first active agent of Formula I: or a pharmaceutically acceptable salt thereof; a second active agent wherein the second active agent is an EGFR modulator (e.g., activator), such as nicotinic acid, nicotinamide, or vitamin K, or a combination thereof; and pharmaceutically acceptable excipients; the first active agent or pharmaceutically acceptable salt is present at about 0.02% to about 1.2% w/v such that the formulation forms a solution or suspension, and wherein: X 1 is O or S; R1 is H, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C(O)(C1C10 alkyl), (CH2)t(C6-C10 aryl), (CH2)t(4- to 10-membered heterocyclic) , C(O)(CH2)t(aryl C6-C10), or C(O)(CH2)t (5- to 10-membered heterocyclic), where: t is an integer from 0 to 5; the alkyl group optionally includes 1 or 2 hetero moieties selected from O, S and N(R6) with the proviso that two O atoms, two S atoms, or one O atom and one S atom are not directly bonded together. to the other; the aryl and heterocyclic groups are optionally fused with a C6-C10 aryl group, a C5-C8 saturated cyclic group, or a 5- to 10-membered heterocyclic group; 1 or 2 carbon atoms in the preceding heterocyclic moieties are optionally replaced with an oxo moiety (=O) or an oxygen anion; the (CH2)t moieties optionally include a carbon-carbon double or triple bond when t is an integer from 2 to 5; and the preceding R1 groups, except H, are optionally substituted with 1 to 3 R4 groups; R2 is H; R3is (CH2)t(aryl C6-C10), where: t is an integer from 0 to 5; the aryl group is optionally fused with a C6-C10 aryl group, a C5-C8 saturated cyclic group, or a 5- to 10-membered heterocyclic group; the (CH2)t moieties optionally include a carbon-carbon double or triple bond when t is an integer from 2 to 5; and the preceding R3 groups are optionally substituted with 1 to 5 R4 groups; each R4 is independently selected from C1-C10 alkyl, C2C10 alkenyl, C2-C10 alkynyl, halo, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, OR5, C(O)R5, C(O)OR5, NR6C(O)R5 , NR6C(O)OR5, OC(O)R5, NR6SO2R5, SO2NR5R6, C(O)NR5R6, NR5R6, S(O)jR7 where j is an integer from 0 to 2, SO3H, NR5(CR6 R7)tOR6, (CH2)t(C6-C10 aryl), SO2(CH2)t(C6-C10 aryl), S(CH2)t(C6-C10 aryl), O(CH2)t(C6-C10 aryl), (CH2) t(heterocyclic from 5 to 10 members), and (CR6 R7)mOR6, where: m is an integer from 1 to 5; t is an integer from 0 to 5; the alkyl group optionally includes 1 or 2 hetero moieties selected from O, S and N(R6) with the proviso that two O atoms, two S atoms, or one O atom and one S atom are not directly bonded together. to the other; the aryl and heterocyclic groups are optionally fused with a C6-C10 aryl group, a C5-C8 saturated cyclic group, or a 5- to 10-membered heterocyclic group; 1 or 2 carbon atoms in the preceding heterocyclic moieties are optionally replaced with an oxo moiety (=O) or an oxygen anion; and the alkyl, aryl and heterocyclic moieties of the preceding R4 groups are optionally substituted with 1 to 3 substituents independently selected from halo, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, NR6SO2R5, SO2NR5 R6, C(O)R5, C(O) OR5, OC(O)R5, NR6C(O)R5, C(O)NR5R6 , NR5R6, (CR6R7)mOR6 where m is an integer from 1 to 5, OR5, and the substituents listed in the definition of R5 ; and R5, R6, and R7 are all independently H or C1-C6 alkyl.

[15] In one embodiment, R3 is (CH2)t(C6-C10 aryl), wherein t is an integer from 1 to 3 and R3 is optionally substituted with 1 to 4 R4 groups.

[16] In another embodiment, R3 is benzyl, optionally substituted with 1 to 4 independently selected halo and C1-C4 alkyl substituents. In another embodiment, R3 is benzyl substituted with 1 to 4 substituents independently selected from methyl, fluoro, chloro and bromine.

[17] In one embodiment, R1 is (CH2)t (5- to 10-membered heterocyclic), wherein t is an integer from 0 to 5, optionally substituted with 1 or 2 independently selected C1-C4 alkyl substituents , hydroxy and hydroxymethyl.

[18] The present application provides heterocyclic portion of the R1 group in Formula I selected from morpholino, pyrrolidinyl, imidazolyl, piperazinyl, piperidinyl, and 2,5-diaza-bicyclo[2.2.1]hept-2-yl, the variable t of R1 group ranges from 2 to 5, and the R1 group is optionally substituted with one or more hydroxy groups.

[19] For example, the heterocyclic portion of the R1 group in Formula I of the present application is pyrrolidine.

[20] In another embodiment of the present application, the first active agent is:

[21] In another embodiment of the present application, the first active agent is a hydrochloride salt of the compound of Formula II, that is, Compound-I:

[22] In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In another embodiment, the second active agent is vitamin K.

[23] Embodiments of the present application provide formulations comprising about 0.005% to about 5.0% w/v of the first active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, e.g., Compound-I . In some embodiments, the concentration of Compound-I or its free base (Formula II) in the formulations is about 0.005% to about 0.01%, about 0.01% to about 0.05%, about from 0.05% to about 0.1%, about 0.1% to about 0.2%, about 0.2% to about 0.3%, about 0.3% to about 0.4%, about 0.4% to about 0.5%, about 0.5% to about 0.6%, about 0.6% to about 0.7%, about 0 7% to about 0.8%, about 0.8% to about 0.9%, about 0.9% to about 1.0%, about 1.1 to about 2.0 %, about 2.1 to about 3.0%, about 3.1 to about 4.0%, or about 4.1 to about 5.0% w/v for topical administration. In some embodiments, the concentration of Compound-I or its free base (Formula II) in the formulations is about 0.1% to about 1.2%, about 0.2% to about 1.2% , about 0.3% to about 1.2%, about 0.4% to about 1.2%, 0.1% to about 1.1%, about 0.2% to about 1.1%, about 0.3% to about 1.1%, about 0.4% to about 1.1%, 0.1% to about 1.0%, about 0.2 % to about 1.0%, about 0.3% to about 1.0%, about 0.4% to about 1.0%, 0.1% to about 0.8%, about from 0.2% to about 0.8%, about 0.3% to about 0.8%, about 0.4% to about 0.8%, 0.1% to about 0, 6%, about 0.2% to about 0.6%, about 0.3% to about 0.6%, about 0.4% to about 0.6%, 0.1% to about 0.5%, about 0.2% to about 0.5%, about 0.3% to about 0.5%, about 0.4% to about 0.5%, 0 .1% to about 0.4%, about 0.2% to about 0.4%, about 0.3% to about 0.4% w/v for topical administration. In some embodiments the formulations include about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3 0.0%, about 4.0%, or about 5.0% w/v of Compound-I or its free base (Formula II).

[24] In some embodiments, the present application provides a solution of a first active agent (e.g., Compound-I) and a second active agent (e.g., nicotinic acid, nicotinamide, or vitamin K, or a combination thereof ), which includes one or more solubilizing agents.

[25] In some embodiments, the formulation comprises about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4 %, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0% , about 3.0%, about 4.0%, or about 5.0% w/v of Formula I or II, or a pharmaceutically acceptable salt thereof, e.g., Compound-I, and a second active agent and a solubilizing agent.

[26] In some embodiments, the solubilizing agent in the formulation may be cyclodextrin, for example, 2-hydroxypropyl-ß-cyclodextrin, methyl-ß-cyclodextrin, randomly methylated ß-cyclodextrin, ethylated ß-cyclodextrin, triacetyl-ß- cyclodextrin, peracetylated ß-cyclodextrin, carboxymethyl-ß-cyclodextrin, hydroxyethyl-ß-cyclodextrin, 2-hydroxy-3-(trimethylammonium)propyl-ß-cyclodextrin, glycosyl-ß-cyclodextrin, maltosyl-ß-cyclodextrin, sulfobutyl ether-ß -cyclodextrin, ß—branched cyclodextrin, hydroxypropyl—Y—cyclodextrin, Y—randomly methylated cyclodextrin, trimethyl—Y—cyclodextrin, or a combination thereof.

[27] In one embodiment, the solubilizing agent in the formulation is 2-hydroxypropyl-ß-cyclodextrin or ß-cyclodextrin sulfobutyl ether.

[28] In one embodiment, the formulation may further comprise one or more benzalkonium chloride (BAK), sodium chloride, and a pH adjusting agent.

[29] In additional embodiments, the formulation comprises about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4 %, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0% , about 3.0%, about 4.0%, or about 5.0% w/v of a first active agent or a pharmaceutically acceptable salt thereof, and a buffer, for example, tromethamine. In one embodiment, the formulation comprises about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3.0%, about 4.0%, or about 5.0% w/v of the first active agent or a pharmaceutically acceptable salt thereof, and about 0.3% to about 1.0% w/v of tromethamine, and optionally further comprises about 0.005% w/v of benzalkonium chloride (BAK).

[30] In some embodiments, the second active agent is an EGFR modulator (e.g., activator). For example, the second active agent includes, but is not limited to, nicotinic acid (or niacin/vitamin B3), nicotinamide, and vitamin K, or a combination thereof. “Vitamin K,” as defined herein, includes one or more members of the vitamin K family and prodrugs thereof, whether naturally occurring or synthetic. The vitamin K family is comprised of vitamin K1 (also known as phylloquinone, phytomenadione, or phytonadione), vitamin K2, and any vitamin K2 homologues. Vitamin K2 homologues are called menoquinones and are characterized by the number of isoprenoid residues in their side chains. Synthetic vitamin K includes but is not limited to vitamin K3 (i.e. menadione), vitamin K4, vitamin K5. In one embodiment, the second active agent is nicotinic acid and/or nicotinamide. In another embodiment, the second active agent is nicotinic acid. In another embodiment, the second active agent is nicotinamide. In yet another embodiment, the second active agent is vitamin K. In another embodiment, the second active agent is vitamin K1. In yet another embodiment, the second active agent is vitamin K2. In yet another embodiment, the second active agent is a vitamin K2 homologue. In yet another embodiment, the second active agent is a synthetic vitamin K (e.g., vitamin K3, vitamin K4, or vitamin K5). In one embodiment, the second active agent is vitamin K3 (i.e., menadione).

[31] In some embodiments, the concentration of the second active agent in the formulations is about 0.00001% to about 5.0% w/v for topical administration. In some embodiments, the concentration of the second active agent in the formulations is about 0.00001% to about 1.0%, about 0.00001% to about 0.1%, about 0.00001% to about 0.01%, about 0.00001% to about 0.001%, about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001% w/v for topical administration. In some embodiments, the concentration of the second active agent in the formulations is about 0.00001% to about 0.0001%, 0.000012% to about 0.0001%, 0.000014% to about 0. 0001%, 0.000016% to about 0.0001%, 0.000018% to about 0.0001%, 0.00002% to about 0.0001%, 0.00003% to about 0.0001% , 0.00004 % to about 0.0001 %, 0.00005 % to about 0.0001 %, 0.00006 % to about 0.0001 %, 0.00007 % to about 0.0001 %, 0 .00008% to about 0.0001%, 0.00009% to about 0.0001%, 0.000016% to about 0.00009%, 0.000018% to about 0.00009%, 0.00002 % to about 0.00009 %, 0.00003 % to about 0.00009 %, 0.00004 % to about 0.00009 %, 0.00005 % to about 0.00009 %, 0.00006 % to about 0.00009%, 0.00007% to about 0.00009%, or 0.00008% to about 0.00009% w/v for topical administration. In some embodiments, the formulations include about 0.00001%, 0.00002%, 0.00003%, 0.00004%, 0.00005%, 0.00006%, 0.00007%, 0.00008% , 0.000081 %, 0.000082 %, 0.000083 %, 0.000084 %, 0.000085 %, 0.000086 %, 0.000087 %, 0.000088 %, or 0.000089 % w/v of second active agent, or a pharmaceutically acceptable salt thereof.

[32] In some embodiments, the concentration of the second active agent in the formulations is about 0.5 µM, about 0.6 µM, about 0.7 µM, about 0.8 µM, about 0. 9 µM, about 1 µM, about 2 µM, about 3 µM, about 4 µM, about 5 µM, about 6 µM, about 7 µM, about 8 µM, or about 9 µM. In some embodiments, the concentration of the second active agent is about 1 µM.

[33] The present application provides a formulation having a pH value of about 4.5 to about 7.5 at or below 40 °C. In some embodiments, the pH value of the formulation is between about pH 5.0 to about 7.0. In one embodiment, the pH value of the formulation is about 6.0 at or below 40 °C.

[34] In another embodiment, the present application provides the use of a formulation comprising a first active agent (for example, Formula I or Compound-I or its free base (Formula II)), and a second active agent ( for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof), for the manufacture of a medicament to access the posterior segment of the eye and/or to treat and/or ameliorate a posterior segment disease or a vasculopathic disease or inflammatory of the eye, as described herein, for example, diabetic retinopathy (including background diabetic retinopathy, proliferative diabetic retinopathy and diabetic macular edema); age-related macular degeneration (AMD) (including neovascular (wet/exudative) AMD, dry AMD, and Geographic Atrophy); pathological choroidal neovascularization (CNV) of any mechanism (e.g., high myopia, trauma, sickle cell anemia; ocular histoplasmosis, angioid lines, traumatic choroidal rupture, optic nerve drusen, and some retinal dystrophies); pathologic retinal neovascularization of any mechanism (e.g., sickle cell retinopathy, Eales disease, ocular ischemic syndrome, carotid-cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic occlusive arteriolitis; birdshot retinochoroidopathy, retinal vasculitis, sarcoidosis, and toxoplasmosis); uveitis; retinal vein occlusion (central or branch); eye trauma; surgery-induced edema; surgery-induced neovascularization; cystoid macular edema; ocular ischemia; retinopathy of prematurity; coat disease; sickle cell retinopathy and/or neovascular glaucoma. In one embodiment, the eye disease is AMD. In one embodiment, diseases of the eye arise from or are exacerbated by ocular angiogenesis and/or neovascularization.

[35] In another embodiment, the present application relates to a formulation for use in the manufacture of a medicament suitable for accessing the posterior segment of the eye and/or for treating and/or ameliorating a disease of the posterior segment or a vasculopathic or inflammatory disease of the eye. In one embodiment, the formulation comprises a first active agent and a second active agent. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the first active agent is a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is a compound of Formula II or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is Compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof. The formulation may further comprise one or more pharmaceutically acceptable excipients.

[36] In another embodiment, the present application relates to a formulation for use in the manufacture of a medicament to treat and/or improve a symptom of an ocular disease or disorder (e.g., a posterior segment or ocular disease). vasculopathic or inflammatory disease of the eye). In one embodiment, the formulation comprises a first active agent and a second active agent. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the first active agent is a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is a compound of Formula II or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is Compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof. The formulation may further comprise one or more pharmaceutically acceptable excipients.

[37] In yet another embodiment, the present application relates to a formulation for use in a method for accessing the posterior segment of the eye and/or for treating and/or improving a disease of the posterior segment of the eye. In one embodiment, the formulation comprises a first active agent and a second active agent. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the first active agent is a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is a compound of Formula II or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is Compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In one embodiment, the second active agent is selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof. The formulation may further comprise one or more pharmaceutically acceptable excipients.

[38] In yet another embodiment, the present application relates to a formulation for use in a method for treating and/or improving a symptom of an eye disease or disorder (for example, a vasculopathic or inflammatory disease of the eye). posterior segment of the eye). In one embodiment, the formulation comprises a first active agent and a second active agent. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the first active agent is a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is a compound of Formula II or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is Compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof. The formulation may further comprise one or more pharmaceutically acceptable excipients.

[39] In another embodiment, the present application relates to a combinational therapy for accessing the posterior segment of the eye and/or for treating and/or improving a disease of the posterior segment of the eye, wherein the therapy comprises administering a first active agent and a second active agent. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the first active agent is a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is a compound of Formula II or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is Compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof. In one embodiment, the first active agent is administered simultaneously with the second active agent. In another embodiment, the first active agent is administered prior to administration of the second active agent. In another embodiment, the first active agent is administered after administration of the second active agent.

[40] In another embodiment, the present application relates to a combinational therapy for treating and/or improving a symptom of an ocular disease or disorder (e.g., a vasculopathic or inflammatory disease of the posterior segment of the eye), comprising administering a first active agent and a second active agent. In one embodiment, the first active agent is a VEGFR inhibitor. In one embodiment, the first active agent is a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is a compound of Formula II or a pharmaceutically acceptable salt thereof. In another embodiment, the first active agent is Compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., activator). In another embodiment, the second active agent is selected from nicotinic acid, nicotinamide, and vitamin K, and a combination thereof. In one embodiment, the first active agent is administered simultaneously with the second active agent. In another embodiment, the first active agent is administered prior to administration of the second active agent. In another embodiment, the first active agent is administered after administration of the second active agent.

[41] In some embodiments, the exposure time of the first active agent (e.g., Compound-I) and the second active agent is between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, the exposure time of the first active agent (e.g., Compound-I) and the second active agent is more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). . In some embodiments, the dosage regimen involves several courses of topical ocular administration of a formulation comprising the first active agent (e.g., Compound-I) and a second active agent to a subject for between 1 and 90 days or longer. than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) or for longer than 90 days. For example, the dosage regimen involves once daily, twice daily, three times daily, or four times daily administration of the formulation for between 1 and 90 days or for more than 90 days (e.g., 4 months , 6 months, 8 months, or 12 months) or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves once, twice, three times, or four times administration of the formulation on alternate days (i.e., on day 1, 3, 5, 7, etc.) for up to 90 days. For example, the dosage regimen involves administering once on day 1, once, or twice on day 2 through day 90. For example, the dosage regimen involves administering once, twice, three times, or four times on day 1, followed by once daily for 2 to 90 days. For example, the dosage regimen involves administering once, twice, thrice, four times on day 1, followed by once, twice, thrice, or four times on alternate days (i.e., on day 1, 3 , 5, 7, etc.) for up to 90 days. For example, a dosage regimen involves administering once or twice a day for 1, 2, 3, 4, or 5 consecutive days. For twice or thrice daily dosing regimen, subjects receive a topical ocular dose of a first active agent (e.g., Compound-I) and a formulation of the second active agent on days 1 and 4 approximately 4. 6, or 8 hours apart. In another embodiment, subjects receive topical ocular doses of a first active agent (e.g., Compound-I) and a formulation of the second active agent approximately 4, 6, or 8 hours apart for four consecutive days. In some embodiments, subjects receive one or two topical ocular doses of a first active agent (e.g., Compound-I) and a formulation of the second active agent per day for 5 consecutive days. In still other embodiments, subjects receive one or two topical ocular doses of a first active agent (e.g., Compound-I) and a formulation of the second active agent for 5 to 90 consecutive days. In some embodiments, subjects receive one or two topical ocular doses of a first active agent (e.g., Compound-I) and a formulation of the second active agent for at least 25 consecutive days. In one embodiment, subjects receive one or two topical ocular doses for at least 90 consecutive days or more.

[42] For example, a formulation comprising about 1 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g. 4 months, 6 months, 8 months, or 12 months) or for more than 90 days (e.g. 4 months, 6 months, 8 months, or 12 months) . In some embodiments a formulation comprising about 1 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 10 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above. In some embodiments, the formulation of the present application is administered QD or BID. In some embodiments, the formulation of the present application is administered QD, BID, TID, or QID when administered in low doses (e.g., 1 mg/mL, 2 mg/mL, 3 mg/mL, 4 mg/mL, or 5 mg/mL), and QD or BID at high doses (e.g., 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, or 10 mg/mL).

[43] In some embodiments, the formulation of a first active agent (e.g., Formula II or Compound-I) and a second active agent is administered to one eye or both eyes of a subject. For example, about 0.2% to about 1.0% (w/v) of the compound of Formula II or about 0.1% to 1.2% (w/v) of Compound-I and a second active agent comprising the formulation of the present application is administered once daily (QD), twice daily (BID), three times daily (TID), or four times daily (QID) to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, the compound of Formula II or Compound-I is complexed with a complexing agent, e.g., cyclodextrin (e.g., hydroxypropyl-ß-cyclodextrin (HP-ß-CD, KLEPTOSE® HPB) (%)) in a ratio of about 1:8, wherein about 2% to 13% (w/v) cyclodextrin (e.g., Kleptose® HPB (%)) is added to the formulation. The formulation may further comprise about 0.1% to about 0.2% buffer, for example, 10 mM phosphate buffer. The desired osmolality of the formulation can be about 200 to about 300 mOsm, obtained by adding sufficient amount to obtain the osmolality with a salt, for example, sodium chloride. The pH of the formulation can be about 6.0.

[44] The present application relates to a pharmaceutical composition comprising particles of an active agent of the present application (e.g., a first active agent (e.g., Formula II or Compound-I) and/or a second active agent), or a pharmaceutically acceptable salt thereof, wherein the particles have an average diameter between 100 nm and 100 µm.

[45] The present application relates to a suspension formulation comprising a pharmaceutical composition, wherein the pharmaceutical composition comprises particles of an active agent of the present application (e.g., a first active agent (e.g., Formula II or Compound- I) and/or a second active agent), or a pharmaceutically acceptable salt thereof, as described herein.

Brief Description of the Drawings

[46] Figure 1 is a series of immunoblots of EGFR phosphorylation in immortalized corneal epithelial cells (hTCEpi cells) treated with varying concentrations of a first active agent of the present application or a control (AG1478, an EGFR kinase inhibitor) and of EGF (top panels: immunoblotting of total EGFR, middle panels: immunoblotting showing phosphorylation of EGFR tyrosine 1068, bottom panels: immunoblotting showing phosphorylation of EGFR tyrosine 1045).

[47] Figure 2 is a series of immunoblots of EGFR phosphorylation in immortalized corneal epithelial cells (hTCEpi cells) treated with varying concentrations of a first active agent of the present application alone, or after treatment with vitamin K3 (50 µM), nicotinic acid (10 µM), or nicotinamide (10 µM) (first row: immunoblotting of total EGFR, second row: immunoblotting showing phosphorylation of EGFR tyrosine 1068, third row: immunoblotting showing phosphorylation of EGFR tyrosine 1045).

[48] Figure 3A is a series of microphotographs showing the migration/proliferation of immortalized corneal epithelial cells (hTCEpi cells) treated for 30 minutes with the indicated concentrations of a first active agent of the present application or a control (AG1478, an inhibitor of EGFR kinase), followed by 16-hour treatment with the first active agent or control along with the indicated concentrations of EGF or VEGF. Figure 3B is a series of bar graphs quantifying the migration/proliferation of hTCEpi cells in Figure 3A.

[49] Figure 4A is a series of microphotographs showing the migration/proliferation of immortalized corneal epithelial cells (hTCEpi cells) treated with the indicated concentrations of vitamin K3, menadione, for 4 hours, and then supplemented with varying concentrations of a first agent active ingredient of this application, followed by treatment with vitamin K3, the first active agent together with EGF. Figure 4B is a series of bar graphs quantifying the migration/proliferation of hTCEpi cells in Figure 4A.

[50] Figure 5A is a series of images of epithelial wounds at the time of initial wounding (0 h) and post-wounding of corneas treated with vehicle, the indicated concentrations of a compound of Formula I or II, or AG1478. Figures 5B and 5C are a series of bar graphs quantifying wound healing at 16 hours (Figure 5B) or 24 hours (Figure 5C) after wounding of corneas treated with vehicle, the indicated concentrations of a compound of Formula I or II, or AG1478. Figure 5D is a graph showing the time course wound healing of corneas treated with vehicle, the indicated concentrations of a compound of Formula I or II, or AG1478.

[51] Figure 6A is a series of images of epithelial wounds at the time of initial wounding (0 h) and post-wounding of corneas treated with vehicle, menadione, a compound of Formula I or II, or menadione in combination with a compound of Formula I or II. Figure 6B is a series of bar graphs quantifying wound healing at 16 hours or 24 hours after wounding of corneas treated with vehicle, menadione, a compound of Formula I or II, or menadione in combination with a compound of Formula I. or II. Figure 6C is a graph showing the time course wound healing of corneas treated with vehicle, menadione, a compound of Formula I or II, or menadione in combination with a compound of Formula I or II.

[52] Figure 7A is a series of immunoblots showing total VEGFR2, phosphorylated VEGFR2, and α-tubulin (as loading control) in human retinal endothelial cells treated with the indicated concentrations of menadione for 4 hours, followed by treatment of 30 minutes with varying concentrations of a compound of Formula I or II (1 nM, 10 nM, 100 nM, or 1 µM), and then 10 ng/ml of VEGF. Figure 7B is a graph quantifying the growth of human retinal endothelial cells following treatment with the indicated concentrations of menadione for 4 hours, followed by 30 minute treatment with varying concentrations of a compound of Formula I or II (0.1 nM , 1 nM, 10 nM, or 100 nM), and then 10 ng/ml VEGF overnight.

[53] Figure 8 is a series of immunoblots showing total EGFR2, phosphorylated EGFR, and α-Tubulin (as loading control) in hTCEpi cells treated with the indicated concentrations of menadione for 4 hours, followed by incubation with 8.0 nM EGF over the indicated time periods.

[54] Figure 9A shows the particle size distribution comprising 5% of a compound of Formula II and 1% of Pluronic F-127 produced using small grinding media for a long period of time at a high cylinder speed. Figure 9B shows the particle size distribution comprising 5% of a compound of Formula II and 1% of Pluronic F-127 produced using large grinding media for a short period of time at a low cylinder speed. Figure 9C shows the size distribution of particles comprising 3% Compound-I, 0.6% Tris HCl, and 2% glycerol produced without milling. Figure 9D shows the size distribution of particles comprising 0.4% of a compound of Formula II, 10 µM menadione, 0.08% Pluronic F-127, and 2.5% glycerol after storage at 40° C for 7 days.

[55] Figure 10 shows the size distribution of particles comprising 5% menadione and 1% HPMC.

[56] Figure 11 shows the particle size distribution comprising 5% of a compound of Formula II and 1% of Pluronic F-127 produced using large grinding media for a short period of time at a low cylinder speed. Detailed Description

[57] The materials, compounds, compositions, articles, and methods described herein can be more easily understood by reference to the following detailed description of specific aspects of the disclosed subject matter and the Examples included therein. Before the present materials, compounds, compositions, articles, devices, and methods are disclosed and described, it should be understood that the aspects described below are not limited to specific methods or specific reagents, as such may vary. It should also be understood that the terminology used here is for the purpose of describing particular aspects only and is not intended to be limiting.

[58]Also, throughout this descriptive report, several publications are referenced. The disclosures of these publications in their entirety are hereby incorporated by reference into this application so as to more fully describe the prior art to which the disclosed subject matter belongs. The disclosed references are also individually and specifically incorporated by reference herein to the material contained therein that is discussed in the sentence in which the reference is recounted.

[59] The present application provides compositions or formulations containing a first active agent and/or a second active agent for use in treating ocular disorders caused by endothelial cell proliferation, enhanced vascular permeability, inflammation, angiogenesis, or neovascularization, and a second active agent for use in preventing and/or treating damage caused to the anterior segment of the eye by the first active agent, a systemic disease and/or an ocular disease.

[60] The present application also relates to a combination of a first active agent, for example, a compound of Formula I or II, and a second active agent, for example, nicotinic acid, nicotinamide, vitamin K, or a combination of these. In one embodiment, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, and a second active agent or a pharmaceutically acceptable salt thereof, are administered simultaneously. Alternatively, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered prior to administration of a second active agent, or a pharmaceutically acceptable salt thereof. In another embodiment, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered after administration of a second active agent, or a pharmaceutically acceptable salt thereof.

[61] The formulations of the application are useful in preventing or inhibiting neovascularization and vascular leakage associated with ocular disorders while preventing or inhibiting corneal diseases. In some cases, the application formulations cause regression of neovascularization. Briefly, within the context of the present application, the first active agents should be understood to be any molecule, synthetic or naturally occurring, that acts to inhibit vascular growth, reduce vascular permeability, and/or decrease inflammation.

[62] The formulations of the application are also useful in preventing and/or treating disruptions of the corneal epithelium caused by systemic diseases (e.g., cancer, diabetes, etc.), ocular diseases, or a side effect of a locally or systemically administered drug. (e.g., anti-EGFR agents, or compounds of Formula I or II having anti-EGFR activity). Briefly, within the context of the present application, second active agents should be understood to be any molecule, synthetic or naturally occurring, that acts to protect from and/or repair corneal edema, ulceration or any other corneal abnormality. In particular, the present application provides formulations comprising a first active agent and a second active agent, each in a therapeutically effective amount.

General Settings

[63] In this specification and in the claims that follow, reference will be made to various terms, which should be defined as having the following meanings. All percentages, ratios and proportions herein are by weight unless otherwise specified. All temperatures are in degrees Celsius (°C) unless otherwise specified.

[64]By “pharmaceutically acceptable” is meant a material that is not biologically or otherwise undesirable, that is, the material can be administered to an individual along with the relevant active compound without causing clinically unacceptable biological effects or interacting in a manner deleterious with any of the other components of the pharmaceutical composition or formulation in which it is contained.

[65]A percentage by weight of a component, unless specifically stated otherwise, is based on the total weight of the formulation or composition in which the component is included.

[66] By “effective amount” as used herein means “an amount of one or more of the disclosed compounds, effective in dosages and for periods of time necessary to obtain the therapeutic or desired result”. An effective amount may vary according to factors known in the art, such as the state of the disease, age, sex, and weight of the human or animal being treated. Although particular dosage regimens may be described in the examples herein, one skilled in the art will appreciate that the dosage regimen can be altered to provide optimal therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionately reduced as indicated by the demands of the therapeutic situation. Furthermore, the formulations of this disclosure can be administered as frequently as necessary to obtain a therapeutic amount.

[67]The effective amount or effective dose in a human can be determined from that in an animal (e.g., an experimental animal). For example, the effective dose in a human can be calculated based on the conversion shown in the table below.

[68]Conversion of animal doses to human equivalent doses (HEDs) using the exponent 0.67 for body surface area

[69] “Excipient” is used herein to include any other compound that may be contained in or combined with one or more of the disclosed inhibitors that is not a therapeutically or biologically active compound. As such, an excipient must be pharmaceutically or biologically acceptable or relevant (e.g., an excipient should generally be non-toxic to the subject). “Excipient” includes a single such compound and is also intended to include a plurality of excipients. For the purposes of the present application, the terms “excipient” and “carrier” are used interchangeably throughout the description of the present application and said terms are defined herein as, “ingredients that are used in the practice of formulating a safe and effective".

[70]As used here, by a “subject” is meant an individual. Thus, the “subject” may include domestic animals (e.g., cats, dogs, etc.), farmed animals (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratory animals (e.g., mouse, rabbit, rat, guinea pig, etc.), and birds. “Subject” may also include a primate or a human.

[71]By “reduce” or other forms of the word, such as “reducing” or “reduction”, is meant the diminution of an event or characteristic (e.g., vascular leak, or tissue swelling). It is understood that this is typically in relation to some standard or expected value, in other words, it is relative, but that it is not always necessary that the standard or relative value be referred to.

[72] The term “treat” or other forms of the word such as “treated” or “treatment” is used herein to mean that administration of a compound or formulation of the present application alleviates a disease or disorder in a host and/or reduces, inhibits, or eliminates a particular feature or event associated with a disorder (e.g., vascular stroke or corneal ulceration).

[73] As the methods of the present application are directed to preventing disorders, it is understood that the term “prevent” does not require that the disease state be completely prevented. Preferably, as used herein, the term prevention refers to the ability of the skilled artisan to identify a population that is susceptible to disorders, such that administration of the compounds of the present application can occur before the onset of a disease. The term does not imply that the disease state is completely avoided.

[74] The term “improvement” or other forms of the word such as “improve” is used herein to mean that administration of a therapeutic agent of the present application alleviates one or more symptoms of a disease or disorder in a host and/or reduces, inhibits, or eliminates a particular symptom associated with the disease or disorder before and/or after administration of the therapeutic agent.

[75] The disclosed first active agent compounds affect vascular leakage or pathological neovascularization by inhibiting a receptor tyrosine kinase. The disclosed second active agent compounds affect disruption of the corneal epithelium by modulating (e.g., activating) a receptor tyrosine kinase.

[76]Throughout the description and claims of this specification, the word “comprise” and other forms of the word, such as “comprising” and “comprises”, means including but not limited to, and is not intended to exclude, e.g. , other additives, or components.

[77] As used in the description and the appended claims, the singular forms “a”, “an”, and “the”, “a” include plural referents unless the context clearly dictates otherwise.

[78]“Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes examples where the event or circumstance occurs and examples where it does not occur.

[79]The term “about” refers to any minute change in the concentration or amount of a therapeutic agent (e.g., a first active agent or a second active agent) that does not change the effectiveness of the agent in preparing a formulation and in the treatment of a disease or disorder. For example, without being limiting, the concentration of a therapeutic agent would be effective if the concentration were varied from 0.005% to 5.0% (e.g., ±0.0005%). The term "about" with respect to the concentration range of the therapeutic/active agents of the present application, e.g., first active agent or second active agent, also refers to any variation from an established amount or range that was an amount or effective range.

[80]Ranges can be expressed here as “about” a particular value, and/or “about” another particular value. When such a range is expressed, another aspect includes that particular value and/or the other particular value. Similarly, when values are expressed as approximations, through the use of the antecedent “about”, the particular value is understood to form another aspect. It is further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are several values disclosed herein, and that each value is also disclosed herein as “about” this particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that when a value is disclosed, then “less than or equal to” value, “greater than or equal to value”, and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “10” is disclosed, then “less than or equal to 10” as well as “greater than or equal to 10” are also disclosed. It is also understood that throughout the application data is provided in several different formats and that this data represents endpoints and starting points and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as are between 10 and 15. It is also understood that each unit between two particular units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[81] The term “halo”, as used herein, unless otherwise indicated, includes fluoro, chlorine, bromine or iodine. Preferred halo groups are fluoro, chloro and bromine.

[82] The term “alkyl”, as used herein, unless otherwise indicated, includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specific number of carbon atoms. For example, C1-6 alkyl is intended to include C1, C2, C3, C4, C5, and C6 alkyl groups. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl, and n-hexyl. In certain embodiments, a straight-chain or branched-chain alkyl has six or fewer carbon atoms in its main chain (e.g., C1-C6 for straight chain, C3-C6 for branched chain), and in another form of embodiment, a straight-chain or branched-chain alkyl has four or fewer carbon atoms. Likewise, cycloalkyls have three to eight carbon atoms in their ring structure, and in other embodiments, cycloalkyls have five or six carbons in the ring structure.

[83]Alkyl can be substituted by substituting hydrogen on one or more carbons of the hydrocarbon main chain. Such substituents may include, for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonate, phosphinate, cyano, amino (including alkylamino , dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano , azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. Cycloalkyls may be further substituted, for example, with the substituents described above and additional substituents such as alkyl, alkenyl, and alkynyl. An “alkylaryl” or “aralkyl” moiety is an alkyl substituted with an aryl (e.g., phenylmethyl (benzyl)).

[84] The term “alkenyl”, as used herein, unless otherwise indicated, includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double bond. For example, the term “alkenyl” includes straight-chain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched-chain alkenyl groups, cycloalkenyl groups (e.g., alicyclic ) (e.g., cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloeptenyl, cyclooctenyl), cycloalkenyl groups substituted by alkyl or alkenyl, and alkenyl groups substituted by cycloalkyl or cycloalkenyl. In certain embodiments, a straight-chain or branched-chain alkenyl group has six or fewer carbon atoms in its main chain (e.g., C2-C6 for straight chain, C3-C6 for branched chain). Likewise, cycloalkenyl groups can have from three to eight carbon atoms in their ring structure, and in some embodiments, cycloalkenyl groups have five or six carbons in the ring structure. The term “C2-C6” includes alkenyl groups containing two to six carbon atoms. The term “C3-C6” includes alkenyl groups containing three to six carbon atoms. Alkenyl can be substituted by substituting hydrogen on one or more carbon atoms of the hydrocarbon backbone. Such substituents may include, for example, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonate, phosphinate, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[85] The term “alkynyl”, as used herein, unless otherwise indicated, includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one triple bond. For example, "alkynyl" includes straight-chain alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexinyl, heptynyl, octinyl, nonynyl, decynyl), branched-chain alkynyl groups, and alkynyl groups substituted by cycloalkyl or cycloalkenyl. In certain embodiments, a straight-chain or branched-chain alkynyl group has six or fewer carbon atoms in its main chain (e.g., C2-C6 for straight chain, C3-C6 for branched chain). The term “C2C6” includes alkynyl groups containing two to six carbon atoms. The term “C3-C6” includes alkynyl groups containing three to six carbon atoms. Alkynyl can be substituted by substituting hydrogen on one or more carbon atoms of the hydrocarbon backbone. Such substituents may include, for example, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonate, phosphinate, cyano, amino (including alkylamino , dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano , azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[86] The term “alkoxy”, as used herein, unless otherwise indicated, includes O-alkyl groups where “alkyl” is as defined above.

[87]The term “aryl,” as used herein, unless otherwise indicated, includes “unconjugated,” or single-ring, 5- and 6-membered aromatic groups, as well as “conjugated,” or multicyclic systems with at least minus one aromatic ring. Examples of aryl groups include benzene, phenyl, and the like. Furthermore, the term "aryl" includes multicyclic aryl groups, e.g., tricyclic, bicyclic, e.g., naphthalene.

[88]Aryl groups having heteroatoms in the ring structure may be referred to as “aromatic heterocycles”, “aryl heterocycles”, “heterocycles”, “heteroaryls” or “heteroaromatics”.

[89] The aryl or heteroaryl may be substituted at one or more positions of the ring with such substituents as described above, such as, for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonate, phosphinate, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido) , amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. Aryl groups can also be fused or bridged with alicyclic or heterocyclic rings, which are non-aromatic, to form a multicyclic system (e.g., tetralin, methylenedioxyphenyl).

[90] The term “4- to 10-membered heterocyclic,” as used herein, unless otherwise indicated, includes aromatic and non-aromatic heterocyclic groups containing one or more heteroatoms all selected from O, S, and N, wherein each heterocyclic group has 4 to 10 atoms in its ring system. Non-aromatic heterocyclic groups include groups having only 4 atoms in their ring system, but heterocyclic aromatic groups must have at least 5 atoms in their ring system. An example of a 4-membered heterocyclic group is azetidinyl (azetidine derivative). An example of a 5-membered heterocyclic group is thiazolyl and an example of a 10-membered heterocyclic group is quinolinyl.

[91]Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl and quinolizinyl.

[92] Examples of heterocyclic aromatic groups are pyridinyl, imidazolilla, pyrimidinilla, pyrazolilla, triazolilla, pyrazinyl, tetrazolilla, tienilla, isoxazolilla, thiazolilla, oxazolilla, isotiazolilla, pyrolylin, quinolinilla, isoquinolinyl, indolylla, benzimidadozolilla, benzimidadozolilla benzofuranila, cinolinila, Indazolila , indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl.

[93] The preceding groups, as derivatives of the compounds listed above, can be C-linked or N-linked where possible. For example, a pyrrole-derived group may be pyrrole-1-yl (N-linked) or pyrrole-3-yl (C-linked). The “4- to 10-membered heterocyclic” portion can be substituted.

[94] The phrase “pharmaceutically acceptable salt(s)”, as used herein, unless otherwise indicated, includes salts of acidic or basic groups that may be present in the compounds of the first active agents (e.g., Formula I or II) or second active agents. Compounds of Formula I or II and second active agents that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds of Formula I or II and second active agents are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as such as the salts of hydrochloride, hydrobromide, iodide, nitrate, sulfate, bisulfate, phosphate, phosphate acid, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentilinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)].

[95] Those compounds of the first active agents (e.g., Formula I or II) and second active agents that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include the alkali metal or alkaline earth metal salts and particularly the sodium and potassium salts.

[96] In some embodiments, the salt is an acid addition salt, for example, HCl salt.

[97] Certain compounds of Formula I or II and certain second active agents may have asymmetric centers and therefore exist in different enantiomeric forms. This application relates to the use of all optical isomers and stereoisomers of compounds of Formula I or II and mixtures thereof and second active agents and mixtures thereof. Compounds of Formula I or II and second active agents may also exist as E/Z geometric isomers or tautomers. This application relates to the use of all such geometric isomers and tautomers and mixtures thereof.

[98] The subject application also includes isotopically labeled compounds, and pharmaceutically acceptable salts thereof, which are identical to those reported in Formula I or II, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O, 17O, 35S, 18F, and 36Cl , respectively. Compounds of Formula I or II, conjugates thereof, and pharmaceutically acceptable salts of said compounds or said conjugates containing the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this application. Certain isotopically labeled compounds of the present application (e.g., compounds of Formula I or II), for example those in which radioactive isotopes such as 3H and 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon-14, i.e., 14C isotopes are particularly preferred for their ease of preparation and detectability. Furthermore, substitution with heavier isotopes such as deuterium, i.e., 2H, may provide certain therapeutic advantages that result from greater metabolic stability, e.g. increased in vivo half-life or reduced dosage requirements, and consequently may be preferred. in some circumstances. Isotopically labeled compounds of Formula I or II of this application and esters or lipid conjugates thereof generally can be prepared by carrying out the procedures disclosed in the schemes and/or in the Examples and Preparations below, substituting a readily available isotopically labeled reagent in place of a non-isotopically labeled reagent.

[99] This application also covers pharmaceutical formulations containing derivatives of compounds of Formula I or II, or pharmaceutically acceptable salts thereof and the second active agents, or pharmaceutically acceptable salts thereof. Compounds of Formula I or II, or pharmaceutically acceptable salts thereof, and second active agents, or pharmaceutically acceptable salts thereof, having free amino or amide groups can be converted into conjugated derivatives, in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three or four) amino acid residues is covalently linked through an amide or ester linkage to a free amino group of compounds of Formula I or II, or pharmaceutically acceptable salts thereof, or second agents active ingredients or pharmaceutically acceptable salts thereof. Amino acid residues include, but are not limited to, the 20 naturally occurring amino acids commonly designated by three-letter symbols and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, citrulline homocysteine, homoserine, ornithine and methionine sulfone.

[100]Additional types of derivatives are also covered. Amide and ester moieties may incorporate groups including but not limited to ether, amine and carboxylic acid functionalities. Free hydroxy groups can be derived using groups including but not limited to hemisuccinates, phosphate esters, dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlined in D. Fleisher, et al., Advanced Drug Delivery Reviews (1996) 19, 115. Carbamate conjugates Hydroxy and amino groups are also included, as are conjugates of carbonate and sulfate esters of hydroxy groups. The derivatization of hydroxy groups such as (acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may be an alkyl ester, optionally substituted with groups including but not limited to ether, amine and carboxylic acid functionalities, or where the acyl group is an amino acid ester as described above are also covered. Derivatives of this type are described in R. P. Robinson et al., J. Medicinal Chemistry (1996) 39, 10.

[101]The term “kinase” refers to any enzyme that catalyzes the addition of phosphate groups to a protein residue; for example, serine and threonine kinases catalyze the addition of phosphate groups to serine and threonine residues.

[102]The terms “VEGFR kinase”, and “VEGFR”, refer to any of the vascular endothelial growth factor receptors.

[103] The terms “VEGF signaling” and “VEGF cascade” refer to both the upstream and downstream components of the VEGF signaling cascade.

[104]The terms “ErbB kinase”, and “ErbB receptor”, refer to any member of the ErbB family of receptor tyrosine kinases including EGFR (ErbB1 or HER1), HER2/c-neu (ErbB2), HER3 (ErbB3) and HER4 (ErbB4).

[105] The terms “EGF signaling” and “EGF cascade” refer to both the upstream and downstream components of the EGF signaling cascade.

[106] The term “pharmaceutically acceptable” refers to the fact that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not harmful to the recipient thereof.

[107] The terms “administration of a compound” or “administering a compound” refer to the action of providing a compound of the application or pharmaceutical formulation to the subject in need of treatment.

[108]The term “vasculostasis” refers to the maintenance of homeostatic vascular functioning leading to normal physiological functioning.

[109]The term “vasculostatic agents” refers to agents that seek to treat conditions in which vasculostasis is compromised by preventing the loss of or restoring or maintaining vasculostasis.

[110] In the present application “composition” and “formulation” are used interchangeably and refer to the conventional understanding, as known in the art, of a composition or formulation.

[111]The present application relates to an ophthalmic formulation. In some embodiments, the ophthalmic formulation of the present application is a gel formulation or a semi-gel formulation, or both.

[112] “Gel” according to the present application is a semisolid dosage form of the present application, containing suspended particles. A semisolid is not pourable; it does not flow or conform to its container at room temperature. A semisolid does not flow at low shear stress and generally exhibits plastic flow behavior. A colloidal dispersion is a system in which particles of colloidal size (i.e. typically between 1 nm and 1 µm) are distributed uniformly throughout a liquid.

[113] In some embodiments, “gel” is a semisolid system consisting of suspensions of small inorganic particles or organic molecules interpenetrated by a liquid. “Gels” are classified as single-phase or dual-phase systems. “Gels” also consist of a mesophase, or intermediate state of matter between a liquid and a solid that represents a partially ordered structure, which is the state for the active agents in the “Gel Drop” of the present embodiments. A dual-phase gel consists of a network of small separate particles. In a dual-phase system, the gel mass is sometimes referred to as magma (e.g., Bentonite Magma) if the particle size of the suspended material is large. Both gels and magmas are thixotropic, forming semisolids at rest and becoming liquids upon stirring. Semisolid formulations must be shaken before administration to ensure homogeneity and must be labeled as such (see Suspensions). Single-phase gels consist of organic macromolecules uniformly distributed throughout a liquid in such a manner that no apparent boundary exists between the dispersed macromolecules and the liquid. Single-phase gels can also consist of low molecular weight (LMW) organic molecules where the component responsible for gelation is the actual active ingredient. These so-called “LMW Hydrogels” are different from traditional water gelators such as synthetic polymers, polysaccharides, and high molecular weight proteins. High molecular weight gelators are highly ordered and unidirectional due to hydrogen bonding whereas the forces governing LMW hydrogels are basically nondirectional (hydrophobic) van der Waals force interactions. In practice, LMW hydrogels are observed as highly anisotropic structures (typically fibrillar) that propagate throughout the liquid producing a physically branched or entangled network. Gels can be thus unordered to slightly ordered showing some birefringence, liquid crystal character. Gels are administered topically or, after shaking, in the form of a hydrogel such as an ophthalmic drop.

[114] The semisolid “gel” according to the present application is a semisolid by USP definitions and literature referenced therein. The apparent viscosity of the semisolid formulation increases with concentration. The clinical dosage concentration of the present formulation ranges from a low concentration of <1 mg/mL (0.1%) to a high concentration of <6 mg/mL (0.6%). Low concentration doses are less viscous and fall under the category of a “solution”, whereas higher concentrations are more viscous and fit the definition of a gel.

[115] “Jelly” according to the present application is a class of gels, which are semisolid systems consisting of suspensions composed of small inorganic particles or large organic molecules interpenetrated by a liquid, in which the structural coherent matrix contains a high portion of liquid, usually water.

[116] “Solution” according to the present application is a homogeneous, clear liquid dosage form that contains one or more chemical substances dissolved in a solvent or mixture of mutually miscible solvents. A solution is a liquid preparation containing one or more chemical substances dissolved in a suitable solvent or mixture of mutually miscible solvents. Because molecules of a drug substance in solution are uniformly dispersed, the use of solutions as dosage forms generally provides assurance of uniform dosage during administration and good accuracy when the solution is diluted or otherwise mixed.

[117] “Liquid” according to the present application is a dosage form consisting of a pure chemical product in its liquid state. A liquid is pourable; it flows and conforms to its container at room temperature. Liquids exhibit Newtonian or pseudoplastic flow behavior.

[118] “Suspension” according to the present application is a liquid dosage form that contains solid particles dispersed in a liquid vehicle.

[119]The compounds of the first active agents (e.g., Formula I or II) and the second active agents are formulated into therapeutic formulations as natural or saline forms. Pharmaceutically acceptable non-toxic salts include base addition salts (formed with free carboxyl groups or other anionic groups) that are derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases such as isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, procaine, and the like. Such salts are formed as acid addition salts with any free cationic groups and are generally formed with inorganic acids such as, for example, hydrochloric, sulfuric, or phosphoric acids, or organic acids such as acetic, citric, p-toluenesulfonic, methanesulfonic, oxalic, tartaric, mandelic acid, and the like. Salts of the application include amine salts formed by protonation of an amino group with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and the like. Salts of the application also include amine salts formed by protonation of an amino group with suitable organic acids, such as p-toluenesulfonic acid, acetic acid, and the like. Additional excipients that are considered for use in the practice of the present application are those available to those of ordinary skill in the art, e.g., those found in United States Pharmacopeia Vol. XXII and National Formulary Vol. XVII, United States Pharmacopeia Convention, Inc. ., Rockville, Md. (1989), the relevant contents of which are incorporated herein by reference. Furthermore, polymorphs of the compounds of the application are included in the present application.

[120] Embodiments of the present application provide an ophthalmic composition or formulation for treating ocular neovascularization with a first active agent of Formula I: or a pharmaceutically acceptable salt thereof; a second active agent or a pharmaceutically acceptable salt thereof, wherein the second active agent is nicotinic acid, nicotinamide, or vitamin K, or a combination thereof; and pharmaceutically acceptable excipients; the first active agent or pharmaceutically acceptable salt is present at about 0.02% to about 1.2% w/v, wherein: X 1 is O or S; R1 is H, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C(O)(C1C10 alkyl), (CH2)t(C6-C10 aryl), (CH2)t(4- to 10-membered heterocyclic) , C(O)(CH2)t(aryl C6-C10), or C(O)(CH2)t (5- to 10-membered heterocyclic), where: t is an integer from 0 to 5; the alkyl group optionally includes 1 or 2 hetero moieties selected from O, S and N(R6) with the proviso that two O atoms, two S atoms, or one O atom and one S atom are not directly bonded together. to the other; the aryl and heterocyclic groups are optionally fused with a C6-C10 aryl group, a C5-C8 saturated cyclic group, or a 5- to 10-membered heterocyclic group; 1 or 2 carbon atoms in the preceding heterocyclic moieties are optionally replaced with an oxo moiety (=O) or an oxygen anion; the (CH2)t moieties optionally include a carbon-carbon double or triple bond when t is an integer from 2 to 5; and the preceding R1 groups, except H, are optionally substituted with 1 to 3 R4 groups; R2 is H; R3is (CH2)t(aryl C6-C10), where: t is an integer from 0 to 5; the aryl group is optionally fused with a C6-C10 aryl group, a C5-C8 saturated cyclic group, or a 5- to 10-membered heterocyclic group; the (CH2)t moieties optionally include a carbon-carbon double or triple bond when t is an integer from 2 to 5; and each R4 is independently selected from C1-C10 alkyl, C2C10 alkenyl, C2-C10 alkynyl, halo, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, OR5, C(O)R5, C(O)OR5, NR6C(O) R5, NR6C(O)OR5, OC(O)R5, NR6SO2R5, SO2NR5R6, C(O)NR5R6, NR5R6, S(O)jR7 where j is an integer from 0 to 2, SO3H, NR5(CR6 R7)tOR6 , (CH2)t(C6-C10 aryl), SO2(CH2)t(C6-C10 aryl), S(CH2)t(C6-C10 aryl), O(CH2)t(C6-C10 aryl), (CH2 )t(heterocyclic from 5 to 10 members), and (CR6 R7)mOR6, where: m is an integer from 1 to 5; t is an integer from 0 to 5; the alkyl group optionally includes 1 or 2 hetero moieties selected from O, S and N(R6) with the proviso that two O atoms, two S atoms, or one O atom and one S atom are not directly bonded together. to the other; the aryl and heterocyclic groups are optionally fused with a C6-C10 aryl group, a C5-C8 saturated cyclic group, or a 5- to 10-membered heterocyclic group; 1 or 2 carbon atoms in the preceding heterocyclic moieties are optionally replaced with an oxo moiety (=O) or an oxygen anion; and the alkyl, aryl and heterocyclic moieties of the preceding R4 groups are optionally substituted with 1 to 3 substituents independently selected from halo, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, NR6SO2R5, SO2NR5 R6, C(O)R5, C(O) OR5, OC(O)R5, NR6C(O)R5, C(O)NR5R6 , NR5R6, (CR6R7)mOR6 where m is an integer from 1 to 5, OR5, and the substituents listed in the definition of R5 ; and R5, R6, and R7 are all independently H or C1-C6 alkyl.

[121] In one embodiment, R3 is (CH2)t (C6-C10 aryl), where t is an integer from 1 to 3 and R3 is optionally substituted with 1 to 4 R4 groups.

[122] In another embodiment, R3 is benzyl, optionally substituted with 1 to 4 independently selected halo and C1-C4 alkyl substituents. In another embodiment, R3 is benzyl substituted with 1 to 4 substituents independently selected from methyl, fluoro, chloro and bromine.

[123] In one embodiment, R1 is (CH2)t (5- to 10-membered heterocyclic), wherein t is an integer from 0 to 5, optionally substituted with 1 or 2 independently selected C1-C4 alkyl substituents , hydroxy and hydroxymethyl.

[124] The present application provides heterocyclic portion of the R1 group in Formula I selected from morpholino, pyrrolidinyl, imidazolyl, piperazinyl, piperidinyl, and 2,5-diaza-bicyclo[2.2.1]hept-2-yl, the variable t of R1 group ranges from 2 to 5, and the R1 group is optionally substituted with one or more hydroxy groups.

[125] For example, the heterocyclic portion of the R1 group in Formula I of the present application is pyrrolidine.

[126] In other embodiments of the present application, the first active agent is:

[127] A compound of the present application is 3-[(4-bromo-2,6-difluorophenyl)methoxy]-5-[[[[4-(1-pyrrolidinyl)butyl]amino]carbonyl]amino]- hydrochloride 4- isothiazolcarboxamide, with the molecular formula: C2oH24BrF2N5O3S • HCl, molecular weight: 568.86 g/mol, and with the property that the molecule does not contain an asymmetric center and is not chiral. A compound of the present application is represented by Compound-I:

(Compound-I).

[128] Compound-I of the present application is an inhibitor of the tyrosine kinase activity of VEGFR-2, which blocks VEGF-stimulated auto-phosphorylation of this receptor as well as endothelial cell proliferation. It is selective (>500x) in relation to the concentration necessary to inhibit the epidermal growth factor receptor (EGFR) and insulin receptor (IR) tyrosine kinases. Compound-I is described in US Patent No. 6,235,764. In some embodiments, compounds of Formula I or II are inhibitors of VEGFR-2.

[129] The second active agents of the present application are EGFR modulators (e.g., activators) of EGFR tyrosine kinase activity.

[130] The first active agent and the second active agent can be administered together as part of the same formulation comprising the first active agent, the second active agent, and a pharmaceutical excipient. The first active agent and the second active agent can also be administered separately. In one embodiment, the first active agent and the second active agent are administered separately as two formulations, wherein one formulation comprises the first active agent and a pharmaceutical excipient, and the second formulation comprises the second active agent and a pharmaceutical excipient.

[131] In one embodiment, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, and the second active agent or a pharmaceutically acceptable salt thereof, are administered simultaneously. Alternatively, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered prior to administration of the second active agent, or a pharmaceutically acceptable salt thereof. In another embodiment, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered after administration of the second active agent, or a pharmaceutically acceptable salt thereof.

General Properties

[132] Compound-I of the present application has the characteristics as shown in table 1. Embodiments provide three formulations of Compound-I or its free base - the compound of Formula II. Table 1A: General Properties of Compound-I Drug Substance

[133] The composition of the Compound-I formulations is listed in table 1B. Formulation materials are listed in Table 1C. Table 1B: Compound-I Formulations: Gel Drop, suspension, and solution.

[134] The composition of the formulations of Compound-I and a second active are listed in table 1D. Formulation materials are listed in table 1E. Table 1D: Compound-I Formulations: Gel Drop, suspension, and solution.

Ophthalmic Solutions

[135] The present application provides formulations of a first active agent (e.g., Compound-I and/or its free base (compound of Formula II)) and/or a second active agent, formed as a solution with a viscosity similar to water . The solution includes pharmaceutically acceptable agents/excipients, for example, without limitation, cyclodextrin. The solution thus formed is a clear, colorless solution, suitable for topical administration to the eye.

[136] The solutions in this application reduce exposure to the anterior segment of the first active agent; thus, they allow increased concentration of the first active agent, for example, a compound of Formula I or II, in the solution and increased frequency of release, thus promoting maintained high concentration of the first active agent in the posterior segment of the eye.

[137] The solutions of the application comprise about 0.005% to about 5.0% w/v of the first active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, for example, Compound-I. In some embodiments, the concentration of Compound-I or its free base (Formula II) in the solutions is about 0.005% to about 0.01%, about 0.01% to about 0.05%, about from 0.05% to about 0.1%, about 0.1% to about 0.2%, about 0.2% to about 0.3%, about 0.3% to about 0.4%, about 0.4% to about 0.5%, about 0.5% to about 0.6%, about 0.6% to about 0.7%, about 0 7% to about 0.8%, about 0.8% to about 0.9%, about 0.9% to about 1.0%, about 1.0 to about 2.0 %, about 2.0 to about 3.0%, about 3.0 to about 4.0%, or about 4.0 to about 5.0% w/v for topical administration. In some embodiments, the concentration of Compound-I or its free base (Formula II) in the formulations is about 0.1% to about 1.2%, about 0.2% to about 1.2% , about 0.3% to about 1.2%, about 0.4% to about 1.2%, 0.1% to about 1.1%, about 0.2% to about 1.1%, about 0.3% to about 1.1%, about 0.4% to about 1.1%, 0.1% to about 1.0%, about 0.2 % to about 1.0%, about 0.3% to about 1.0%, about 0.4% to about 1.0%, 0.1% to about 0.8%, about from 0.2% to about 0.8%, about 0.3% to about 0.8%, about 0.4% to about 0.8%, 0.1% to about 0, 6%, about 0.2% to about 0.6%, about 0.3% to about 0.6%, about 0.4% to about 0.6%, 0.1% to about 0.5%, about 0.2% to about 0.5%, about 0.3% to about 0.5%, about 0.4% to about 0.5%, 0 .1% to about 0.4%, about 0.2% to about 0.4%, about 0.3% to about 0.4% w/v for topical administration. In some embodiments, the solutions include about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3.0%, about 4.0%, or about 5.0% w/v of Compound-I or its free base (Formula II).

[138] The solutions of the application may further comprise about 0.00001% to about 5.0% w/v of a second active agent, or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide, or vitamin K , or a combination of these. The order solutions may further comprise about 0.00001% to about 1.0%, about 0.00001% to about 0.1%, about 0.00001% to about 0.01%, about from 0.00001% to about 0.001%, about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001% w/v of a second active agent, or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In some embodiments, the concentration of the second active agent in the solutions is about 0.00001% to about 0.0001%, 0.000012% to about 0.0001%, 0.000014% to about 0. 0001%, 0.000016% to about 0.0001%, 0.000018% to about 0.0001%, 0.00002% to about 0.0001%, 0.00003% to about 0.0001% , 0.00004 % to about 0.0001 %, 0.00005 % to about 0.0001 %, 0.00006 % to about 0.0001 %, 0.00007 % to about 0.0001 %, 0 .00008% to about 0.0001%, 0.00009% to about 0.0001%, 0.000016% to about 0.00009%, 0.000018% to about 0.00009%, 0.00002 % to about 0.00009 %, 0.00003 % to about 0.00009 %, 0.00004 % to about 0.00009 %, 0.00005 % to about 0.00009 %, 0.00006 % to about 0.00009%, 0.00007% to about 0.00009%, 0.00008% to about 0.00009% w/v for topical administration. In some embodiments, solutions include about 0.00001%, 0.00002%, 0.00003%, 0.00004%, 0.00005%, 0.00006%, 0.00007%, 0.00008% , 0.000081 %, 0.000082 %, 0.000083 %, 0.000084 %, 0.000085 %, 0.000086 %, 0.000087 %, 0.000088 %, or 0.000089 % w/v of second active agent.

[139] Order solutions may further comprise about 0.5 µM, about 0.6 µM, about 0.7 µM, about 0.8 µM, about 0.9 µM, about 1 µM, about 2 µM, about 3 µM, about 4 µM, about 5 µM, about 6 µM, about 7 µM, about 8 µM, or about 9 µM of a second active agent, or a pharmaceutically available salt acceptable thereof, for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In some embodiments, the concentration of the second active agent is about 1 µM.

[140] In some embodiments, the formulation comprises cyclodextrin to improve the solubility of a first active agent (e.g., Compound-I). Cyclodextrin, an oligosaccharide composed of six to eight dextrose units linked through one or four bonds increases the solubility of active agents that have poor or low solubility in water or aqueous solutions (e.g., in PBS buffer). Cyclodextrins form hydrophilic complexes with hydrophobic active agents.

[141] One or more cyclodextrins can be used in the solution of the present application. Non-limiting examples of cyclodextrins for use in the formulation of the present application are, for example: 2-hydroxypropyl-ß-cyclodextrin, methyl-ß-cyclodextrin, randomly methylated ß-cyclodextrin, ethylated ß-cyclodextrin, triacetyl-ß-cyclodextrin, ß -peracetylated cyclodextrin, carboxymethyl-ß-cyclodextrin, hydroxyethyl-ß-cyclodextrin, 2-hydroxy-3-(trimethylammonium)propyl-ß-cyclodextrin, glycosyl-ß-cyclodextrin, maltosyl-ß-cyclodextrin, sulfobutyl ether-ß-cyclodextrin, ß-branched cyclodextrin, hydroxypropyl-Y-cyclodextrin, randomly methylated Y-cyclodextrin, trimethyl-Y-cyclodextrin, or a combination thereof.

[142] In some embodiments, the solution of the compound of Formula II or Compound-I comprising cyclodextrin is a clear and colorless solution and has a viscosity similar to water. In some embodiments, the present application provides a solution comprising Compound-I, one or more cyclodextrins, and a second active agent for topical application and is topically applied to the eye.

[143] The ophthalmic solution of the present application comprises cyclodextrin and pharmaceutical excipients chosen at or below the ideal concentrations for ophthalmic solution. The excipients of the present application are, for example, benzalkonium chloride (BAK) and NaCl. In some embodiments, the ophthalmic solution comprises about 0.001 to about 0.005% w/v benzalkonium chloride (BAK). The amount of BAK varies depending on the application needs.

[144] The ophthalmic solution comprises, for example, without being limiting, about 0.005% to 5.0% Compound-I or its free base, about 2 to about 25% cyclodextrin, for example, without being limiting , Hydroxypropyl-ß-cyclodextrin (HPßCD) or methylcyclodextrin (KLEPTOSE® HPB), and/or sulfobutyl ether-p-cyclodextrin (CAPTISOL®), about 0.1 to about 0.7% salt, e.g. without being limiting, NaCl, and/or about 0.005% of an antimicrobial agent, for example, without being limiting, Benzalkonium Chloride (BAK). The formulation comprises ratio of Compound-I or its free base to cyclodextrin of 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 , or between 1:10 and 1:20. In some embodiments, the ophthalmic solution comprising cyclodextrin further comprises tromethamine (also known as Tris, Tris(Hydroxymethyl)aminomethane, or Tris buffer). In some embodiments, the ophthalmic solution comprises about 0.05% to 1% Tris. In some embodiments, the ophthalmic solution comprises about 0.05% to 0.5% Tris. In some embodiments, the ophthalmic solution comprises about 0.05% to 0.2% Tris. In some embodiments, the ophthalmic solution comprises about 0.1% to 0.15% Tris. In some embodiments, the ophthalmic solution comprises about 1% Tris. In some embodiments, the ophthalmic solution further comprises about 0.005% to 5.0% second active agent.

[145] Ophthalmic solutions of the present embodiments include, for example, without limitation: about 0.3% to about 5.0% Compound-I (about 3 mg/mL to about 50.0 mg /mL), about 0.05% monobasic sodium phosphate monohydrate, about 2% glycerin; about 0.4% Compound-I, about 7% HPßCD, about 0.7% NaCl, about 0.005% BAK; about 0.4% Compound-I, about 4% HPßCD, about 0.7% NaCl, about 0.005% BAK; about 0.4% Compound-I, about 7% HPßCD, about 1% tromethamine, about 0.4% NaCl, about 0.005% BAK; and about 0.6% Compound-I, about 7% HPßCD, about 0.7% NaCl, about 0.005% BAK. For concentrations of Compound-I between about 0.005% to about 5.0%, cyclodextrin is present in a corresponding molar ratio. In some embodiments, the ophthalmic solution further comprises about 0.005% to 5.0% second active agent.

[146] Additional ophthalmic solutions include, for example, without limitation: about 0.4% compound of Formula II (free base), about 7.15% HPßCD, about 0.7% NaCl; about 0.1% compound of Formula II (free base), about 1.79% HPßCD, about 0.85% NaCl; about 0.2% compound of Formula II (free base), about 3.57% HPßCD, about 0.8% NaCl; about 0.6% compound of Formula II (free base), about 10.72% HPßCD, about 0.6% NaCl; about 0.4% compound of Formula II (free base), about 8.41% HPßCD, about 0.65% NaCl; about 0.4% Compound-I, about 10.51 HPßCD, about 0.65% NaCl; about 0.4% compound of Formula II (free base), about 10.51% HPßCD, about 0.15% NaCl, about 1.0% tromethamine (Tris); and/or about 0.1% compound of Formula II (free base), about 2.63% HPßCD, about 0.8% NaCl; about 0.6% Compound-I (as free base), about 15.77% HPßCD, about 0.37% NaCl. For concentrations of Formula II between about 0.005% to about 5.0%, cyclodextrin is present in a corresponding molar ratio. In some embodiments, the ophthalmic solution further comprises about 0.005% to 5.0% second active agent.

[147] In some embodiments, ophthalmic solutions include from about 1.0% to about 25% cyclodextrin. For example, without limitation, Compound-I formulations include about 2.0% to about 3.0% HPßCD, about 3.0% to about 5.0% HPßCD, about 5. 0% to about 10% HPßCD, or about 10% to about 25% HPßCD. In some embodiments, the ophthalmic solution further comprises about 0.005% to 5.0% second active agent.

[148] In additional embodiments, the ophthalmic solutions are formulated as, for example, without limitation: about 8.41% KLEPTOSE® HPB and about 0.142% phosphate; about 8.9% KLEPTOSE® HPB and about 0.142% phosphate; about 4.88% CAPTISOL® and about 0.142% phosphate; and/or about 4.88% CAPTISOL® and about 0.122% phosphate.

[149] In some embodiments, ophthalmic solutions comprising cyclodextrins are clear and colorless, and are extremely viscous, moderately viscous, or have a viscosity similar to water.

[150] In some embodiments, the ophthalmic solution of the application has a pH value of about 4.5 to about 7.5 to 40 °C or below.

[151] In some embodiments, the ophthalmic solution of the order has a pH value of about 5.0 to about 7.0 at or below 40 °C.

[152] For example, the ophthalmic solution of the application has a pH value of about 6.0 at or below 40 °C.

[153] The ophthalmic solutions of the present application may contain various commonly incorporated additives, such as buffering agents (e.g., phosphate buffers, borate buffers, citrate buffers, tartrate buffers, acetate buffers, amino acids, sodium acetate, sodium citrate and the like), tonicizing agents (e.g., saccharides such as sorbitol, glucose and mannitol, polyhydric alcohols such as glycerin, concentrated glycerin, PEG and Propylene glycol, salts such as sodium chloride, etc.), preservatives or antiseptics (e.g. Benzalkonium chloride, Benzathonium chloride, P-oxybenzoates such as Methyl p-oxybenzoate or Ethyl p-oxybenzoate, Benzyl alcohol, Phenethyl alcohol, Sorbic acid or its salt, Thimerosal, Chlorobutanol, etc. ), solubilization aids or particle stabilizing agents (e.g., water-soluble polymers such as polyvinylpyrrolidone, surfactants such as tyloxapol, polysorbates, poloxamer, etc.), pH modifiers (e.g., hydrochloric acid, acetic acid, phosphoric acid , Sodium hydroxide, Potassium hydroxide, Ammonium hydroxide and the like), thickening agents (e.g. HEC, Hydroxypropyl cellulose, Methyl cellulose, HPMC, Carboxymethyl cellulose and its salts), chelating agents (e.g. sodium edetate, citrate sodium, condensed sodium phosphate, etc.), and second active agent stabilizers (e.g., EDTA, propyl gallate, and a combination thereof).

[154] The ophthalmic solutions of the present application comprise cyclodextrin, and may further comprise additional excipients, for example, without being limiting, about 0.5% to about 3% surfactant and emulsifier, for example, without being limiting, polysorbate 80 or equivalent excipients thereof; about 0.05 to about 0.4% non-ionic liquid polymer of the alcoholic type of alkyl aryl polyether, for example, without being limiting tyloxapol; and/or about 0.05% to about 0.6% hydrophilic nonionic surfactant, for example, without limitation, poloxamer, such as poloxamer 407.

[155] In some embodiments, the ophthalmic solution comprises about 0.01 to about 0.5%, about 0.02 to about 0.5%, about 0.04 to about 0.5% %, about 0.06 to about 0.5%, about 0.08 to about 0.5%, about 0.08 to about 0.4%, about 0.08 to about 0 .3%, about 0.08 to about 0.2%, about 0.08 to about 0.18%, about 0.08 to about 0.16%, about 0.08 to about from 0.14%, or about 0.08 to about 0.12% EDTA. In some embodiments, the ophthalmic solution comprises about 0.04%, about 0.06%, about 0.08%, about 0.1%, about 0.12, about 0.14% , about 0.16%, about 0.18%, or about 0.2% EDTA. In some embodiments, the ophthalmic solution comprises about 0.1% EDTA.

[156] In some embodiments, the ophthalmic solution comprises about 0.001 to about 0.5%, about 0.002 to about 0.5%, about 0.005 to about 0.5%, about 0. 01 to about 0.5%, about 0.02 to about 0.5%, about 0.03 to about 0.5%, about 0.04 to about 0.5%, about 0.01 to about 0.4%, about 0.01 to about 0.3%, about 0.01 to about 0.2%, about 0.01 to about 0.1%, about 0.01 to about 0.08%, about 0.01 to about 0.06% propyl gallate. In some embodiments, the ophthalmic solution comprises about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06 %, about 0.07%, about 0.08%, about 0.09% propyl gallate. In some embodiments, the ophthalmic solution comprises about 0.05% propyl gallate. Concentration in Various Ocular Tissues - Released as an Ophthalmic Solution

[157] The ocular solution comprising cyclodextrin improves the bioavailability of the first active agents of the present application in the posterior segment of the eye. Without being bound by theory, in one embodiment, the formulation comprising cyclodextrin forms a clear, colorless solution, which reduces corneal exposure of the active agent, e.g., Compound-I exposure, by about 5 to 15 times compared to corneal exposure with an equimolar Gel Drop formulation.

[158] Without being bound by theory, in one embodiment, an ophthalmic solution comprising cyclodextrin increases the therapeutic index of Compound-I during topical ocular administration. During administration, the hydrophilic cyclodextrin-Compound-I complex is pharmacologically inert in the cornea. Without being bound by theory, in some embodiments, the cyclodextrin-Compound-I complex increases the corneal tolerability of Compound-I. Without being bound by theory, in some embodiments, spontaneous dissociation of cyclodextrin from Compound-I in the peripheral vasculature increases bioavailability in the target tissue, for example, in the choroid or retina.

[159] Unlike other Compound-I formulations, which in some embodiments contribute to corneal toxicity, the cyclodextrin-based ophthalmic solution comprising a similar concentration of Compound-I reduces corneal exposures and thereby increases the therapeutic index and corresponding benefits to patients. In one embodiment, the use of Compound-I cyclodextrin-based solution provides approximately 10x reduction in corneal exposure when compared to equimolar concentrations of Gel Drop. In some embodiments, the Compound-I cyclodextrin-based solution reduces the corneal exposure of Compound-I by 5x, 20x, 30x, 40x, or 50x. In one embodiment, 1 to 90 days or 3 to 9 months of topical ocular dosage of about 0.005% to about 5.0% of Compound-I as a cyclodextrin-based solution has no adverse or toxic effect on cornea, choroid, and/or retina. In yet another embodiment, 1 to 90 days of topical ocular dosage of about 0.6% to about 5.0% of Compound-I as a cyclodextrin-based solution has no adverse or toxic effect on the cornea. , choroid, and/or retina.

[160] Reduction in corneal exposure is correlated with increasing bioavailability and therapeutic index of the active agent in the posterior segment, for example, the retina or choroid, of the eye. For example, no toxic effects attributable to the first active agent or a suitable carrier are observed to the cornea or other parts of the eye when about 0.1% to about 5.0% Compound-I formulation comprising cyclodextrin is administered topically to the eye. eye for at least 30 days or more than 60 days.

[161] In one embodiment, when a formulation comprising about 0.4% (about 4 mg/mL) of Compound-I or its free base, and cyclodextrin, when administered topically to the eye, the central choroidal concentration is between about 0.2 µM to about 0.9 µM, central retinal concentration of the active agent is between about 0.02 µM to about 0.4 µM, aqueous humor concentration of the active agent is about 0.003 µM to about 0.009 µM, and corneal concentration of the active agent is between 6 µM and 40 µM. The cyclodextrin used in the formulation is, for example, without being a limiting example, KLEPTOSE® HPB or CAPTISOL®.

[162] In some embodiments, a cyclodextrin-based solution of Compound-I or its free base increases the bioavailability of the active agent in the central choroid and central retina, while reducing the concentration in the cornea. In some embodiments, topical release of Compound-I or its free base formulated in the presence of cyclodextrin reduces the corneal concentration by about 5 to about 15 times over the equimolar Gel Drop corneal concentration.

[163] Without being bound by theory, in some embodiments, the combined effects of decreasing corneal exposure to the drug so as to avoid poor ocular tolerability while increasing posterior segment bioavailability increase the therapeutic index and corresponding benefits to patients.

[164] In some embodiments, the exposure time of Compound-I and the second active agent is between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, the dosage regimen involves several courses of topical ocular administration of a formulation comprising Compound-I and a second active agent, wherein the second active agent is administered as a separate formulation or as part of the Compound-I formulation. I, to a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves once daily, twice daily, three times daily, or four times daily administration of the formulation for between 1 and 90 days or for more than 90 days (e.g., 4 months , 6 months, 8 months, or 12 months). For example, the dosage regimen involves administration once, twice, thrice, or four times of a compound of Formula I or II and a second active agent on alternate days (i.e., on day 1, 3, 5 , 7 etc.) for up to 90 days. For example, the dosage regimen involves administering once on day 1, once, or twice on day 2 through day 90. For example, the dosage regimen involves administering once, twice, three times, or four times on day 1, followed by once daily for 2 to 90 days. For example, the dosage regimen involves administering once, twice, thrice, four times on day 1, followed by once, twice, thrice, or four times on alternate days (i.e., on day 1, 3 , 5, 7, etc.) for up to 90 days. For example, a dosage regimen involves once a day or twice a day for 1, 2, 3, 4, or 5 consecutive days. For a twice- or three-times-daily dosing regimen, subjects receive a topical ocular dose of a Compound-I formulation and a second active agent on days 1 and 4 approximately 4, 6, or 8 hours apart. In another embodiment, subjects receive topical ocular doses of a Compound-I formulation and a second active agent approximately 4, 6, or 8 hours apart for four consecutive days. In some embodiments, subjects receive one or two doses of topical ocular dosage of the Compound-I formulation and a second active agent per day for 5 consecutive days. In still other embodiments, subjects receive one or two doses of a topical ocular dose of Compound-I and a formulation of the second active agent for 5 to 90 consecutive days. In some embodiments, subjects receive one or two doses of topical ocular dosage of the Compound-I formulation and a second active agent for at least 25 consecutive days. In one embodiment, subjects receive one or two topical ocular doses for at least 90 consecutive days or more. The second active agent can be administered separately or as part of the Compound-I formulation. When administered separately, the second active agent, or a pharmaceutical salt thereof, can be administered alone or as a formulation.

[165] For example, a formulation comprising about 1 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL BID of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL QD of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL BID of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL QD of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL BID of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL QD of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL BID of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL QD of a first active agent (e.g., Compound-I) and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 10 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above. In some embodiments, the formulation of the present application is administered QD, BID, TID, or QID when administered in low doses (e.g., 1 mg/mL, 2 mg/mL, 3 mg/mL, 4 mg/mL, or 5 mg/mL), and QD or BID at high doses (e.g., 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, or 10 mg/mL).

[166] In some embodiments, a 1 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 1 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 2 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 2 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 3 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 3 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 4 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 4 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 5 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 5 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 6 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 6 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 7 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 7 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, an 8 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, an 8 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 9 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 9 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 10 mg/mL BID of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 10 mg/mL QD of a formulation of first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days. or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above.

[167] The present application provides cyclodextrin-based solutions containing hydroxypropyl-beta-cyclodextrin (HP-ß-CD, KLEPTOSE® HPB) or CAPTISOL® that are well tolerated when administered topically for 30 to 90 days or for 4 to 6 months . In some embodiments, once or twice daily administration of about 0.005% to about 5.0% w/v of Compound-I or its free base and a second active agent in a solution containing about 1. 0% to about 25% of HP-ß-CD or CAPTISOL® is well tolerated by the subject.

[168] In some embodiments, the formulation of Formula II or Compound-I and a second active agent is administered to one eye or both eyes of a subject. For example, about 0.2% to about 1.0% (w/v) of the compound of Formula II or about 0.1% to 1.2% (w/v) of Compound-I formulation and A second active agent comprising the formulation of the present application is administered once daily (QD), twice daily (BID), three times daily (BID), or four times daily (QID) to one eye or both eyes. a subject's eyes for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, the Formula II or Compound-I formulation and a second active agent are administered to one eye or both eyes of a subject. For example, about 0.2% to about 1.0% (w/v) of the compound of Formula II or about 0.1% to 1.2% (w/v) of Compound-I formulation and a second active agent is administered once daily (QD), twice daily (BID), three times daily (BID), or four times daily (QID) to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months).

[169] In some embodiments, the compound of Formula II or Compound-I is complexed with a complexing agent, for example, cyclodextrin (e.g., Kleptose® HPB (%)) in a ratio of about 1:8, in that about 2% to 13% (w/v) of cyclodextrin (e.g., Kleptose® HPB (%)) is added to the formulation. The formulation may further comprise about 0.1% to about 0.2% buffer, for example, 10 mM phosphate buffer. The desired osmolality of the formulation is about 200 to about 300 mOsm, obtained by adding sufficient amount to obtain the osmolality with a salt, for example, sodium chloride. The pH of the formulation is about 6.0 at or below 40°C. The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above.

Ophthalmic Suspensions

[170] The present application provides suspensions of a first active agent (e.g., Compound-I) comprising the agent and pharmaceutically acceptable excipients. The present application also provides suspensions of a first active agent (e.g., Compound-I) and a second active agent comprising the first active agent, the second active agent and pharmaceutically acceptable excipients. For example, Compound-I suspensions and second active agent suspensions may include, without limitation, buffering agents, acids & bases, for example, without limitation, HCl and NaOH. In one embodiment, suspensions of Compound-I or its free base may include a buffering agent, for example, without limitation, tromethamine (Tris). In another embodiment, suspensions of Compound-I or its free base and a second active agent may include a buffering agent, for example, without limitation, tromethamine (Tris). A tromethamine-based suspension of the compound of Formula II or Compound-I and a second active agent is useful for topical administration to the eye.

[171] The suspensions of the application comprise about 0.005% to about 5.0% w/v of a first active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, for example, Compound-I. In some embodiments, the concentration of Compound-I or its free base (Formula II) in the suspensions is about 0.005% to about 0.01%, about 0.01% to about 0.05%, about from 0.05% to about 0.1%, about 0.1% to about 0.2%, about 0.2% to about 0.3%, about 0.3% to about 0.4%, about 0.4% to about 0.5%, about 0.5% to about 0.6%, about 0.6% to about 0.7%, about 0 7% to about 0.8%, about 0.8% to about 0.9%, about 0.9% to about 1.0%, about 1.0 to about 2.0 %, about 2.0 to about 3.0%, about 3.0 to about 4.0%, or about 4.0 to about 5.0% w/v for topical administration. In some embodiments, the concentration of Compound-I or its free base (Formula II) in the formulations is about 0.1% to about 1.2%, about 0.2% to about 1.2% , about 0.3% to about 1.2%, about 0.4% to about 1.2%, 0.1% to about 1.1%, about 0.2% to about 1.1%, about 0.3% to about 1.1%, about 0.4% to about 1.1%, 0.1% to about 1.0%, about 0.2 % to about 1.0%, about 0.3% to about 1.0%, about 0.4% to about 1.0%, 0.1% to about 0.8%, about from 0.2% to about 0.8%, about 0.3% to about 0.8%, about 0.4% to about 0.8%, 0.1% to about 0, 6%, about 0.2% to about 0.6%, about 0.3% to about 0.6%, about 0.4% to about 0.6%, 0.1% to about 0.5%, about 0.2% to about 0.5%, about 0.3% to about 0.5%, about 0.4% to about 0.5%, 0 .1% to about 0.4%, about 0.2% to about 0.4%, about 0.3% to about 0.4% w/v for topical administration. In some embodiments, the suspensions include about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3.0%, about 4.0%, or about 5.0% w/v of Compound-I or its free base (Formula II).

[172] Suspensions of the application may further comprise about 0.00001% to about 5.0% w/v of a second active agent, or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide, or vitamin K , or a combination of these. Order stays may further comprise about 0.00001% to about 1.0%, about 0.00001% to about 0.1%, about 0.00001% to about 0.01%, about from 0.00001% to about 0.001%, about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001% w/v of a second active agent, or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In some embodiments, the concentration of the second active agent in the suspensions is about 0.00001% to about 0.0001%, 0.000012% to about 0.0001%, 0.000014% to about 0. 0001%, 0.000016% to about 0.0001%, 0.000018% to about 0.0001%, 0.00002% to about 0.0001%, 0.00003% to about 0.0001% , 0.00004 % to about 0.0001 %, 0.00005 % to about 0.0001 %, 0.00006 % to about 0.0001 %, 0.00007 % to about 0.0001 %, 0 .00008% to about 0.0001%, 0.00009% to about 0.0001%, 0.000016% to about 0.00009%, 0.000018% to about 0.00009%, 0.00002 % to about 0.00009 %, 0.00003 % to about 0.00009 %, 0.00004 % to about 0.00009 %, 0.00005 % to about 0.00009 %, 0.00006 % to about 0.00009%, 0.00007% to about 0.00009%, or 0.00008% to about 0.00009% w/v for topical administration. In some embodiments, the suspensions include about 0.00001%, 0.00002%, 0.00003%, 0.00004%, 0.00005%, 0.00006%, 0.00007%, 0.00008% , 0.000081 %, 0.000082 %, 0.000083 %, 0.000084 %, 0.000085 %, 0.000086 %, 0.000087 %, 0.000088 %, or 0.000089 % w/v of second active agent.

[173] Order suspensions may further comprise about 0.5 µM, about 0.6 µM, about 0.7 µM, about 0.8 µM, about 0.9 µM, about 1 µM, about 2 µM, about 3 µM, about 4 µM, about 5 µM, about 6 µM, about 7 µM, about 8 µM, or about 9 µM of a second active agent, or a pharmaceutically available salt acceptable thereof, for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In some embodiments, the concentration of the second active agent is about 1 µM.

[174] Ophthalmic suspensions may contain various commonly incorporated additives, such as buffering agents (e.g., phosphate buffers, borate buffers, citrate buffers, tartrate buffers, acetate buffers, amino acids, sodium acetate, sodium citrate and the like), tonicizing agents (e.g., saccharides such as sorbitol, glucose and mannitol, polyhydric alcohols such as glycerin, concentrated glycerin, PEG and propylene glycol, salts such as sodium chloride), preservatives or antiseptics (e.g. , benzalkonium chloride, benzathiconium chloride, P-oxybenzoates such as methyl p-oxybenzoate or ethyl p-oxybenzoate, Benzyl alcohol, phenethyl alcohol, Sorbic acid or its salt, Thimerosal, Chlorobutanol and the like), solubilizing aids or agents particle stabilizers (e.g., cyclodextrins and their water-soluble polymers, derivatives such as polyvinylpyrrolidone, surfactants such as tyloxapol, polysorbates, poloxamer), pH modifiers (e.g., hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide, potassium hydroxide, ammonium hydroxide and the like), thickening agents (e.g. HEC, hydroxypropyl cellulose, methyl cellulose, HPMC, carboxymethyl cellulose and their salts), chelating agents (e.g. sodium edetate, sodium citrate, sodium phosphate condensed sodium etc.), and second active agent stabilizers (e.g., EDTA, propyl gallate, and a combination thereof).

[175] The ophthalmic suspension of the present application comprises pharmaceutical excipients chosen at or below ideal concentrations for ophthalmic solution. Excipients of the present application include, for example, without limitation, sodium phosphate monohydrate, glycerin, and benzalkonium chloride (BAK).

[176] In some embodiments, the ophthalmic suspension comprises about 0.01 to about 0.5%, about 0.02 to about 0.5%, about 0.04 to about 0.5% %, about 0.06 to about 0.5%, about 0.08 to about 0.5%, about 0.08 to about 0.4%, about 0.08 to about 0 .3%, about 0.08 to about 0.2%, about 0.08 to about 0.18%, about 0.08 to about 0.16%, about 0.08 to about from 0.14%, or about 0.08 to about 0.12% EDTA. In some embodiments, the ophthalmic suspension comprises about 0.04%, about 0.06%, about 0.08%, about 0.1%, about 0.12, about 0.14% , about 0.16%, about 0.18%, or about 0.2% EDTA. In some embodiments, the ophthalmic suspension comprises about 0.1% EDTA.

[177] In some embodiments, the ophthalmic suspension comprises about 0.001 to about 0.5%, about 0.002 to about 0.5%, about 0.005 to about 0.5%, about 0. 01 to about 0.5%, about 0.02 to about 0.5%, about 0.03 to about 0.5%, about 0.04 to about 0.5%, about 0.01 to about 0.4%, about 0.01 to about 0.3%, about 0.01 to about 0.2%, about 0.01 to about 0.1%, about 0.01 to about 0.08%, about 0.01 to about 0.06% propyl gallate. In some embodiments, the ophthalmic suspension comprises about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06 %, about 0.07%, about 0.08%, about 0.09% propyl gallate. In some embodiments, the ophthalmic suspension comprises about 0.05% propyl gallate.

[178] In some embodiments, the ophthalmic suspension comprises about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0. 4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0 %, about 3.0%, about 4.0%, or about 5.0% w/v of Formula I or II, or a pharmaceutically acceptable salt thereof, e.g., Compound-I, and a second agent active, and may also comprise tromethamine (i.e., Tris). In some embodiments, the ophthalmic suspension comprises about 0.05% to 1% Tris. In some embodiments, the ophthalmic suspension comprises about 0.2% to 1.0% Tris. In some embodiments, the ophthalmic suspension comprises about 0.4% to 0.8% Tris. The tromethamine-based suspension of Compound-I or its free base and/or a second active agent may comprise additional buffers and excipients, for example, without limitation, phosphate buffer. The suspensions may further comprise one or more surfactants and emulsifiers, for example, without limitation, polysorbate 80 or equivalent excipients thereof; one or more non-ionic liquid polymers of the alcoholic type of alkyl aryl polyether, for example, without being limiting tyloxapol; and/or one or more hydrophilic nonionic surfactants, for example, without limitation, poloxamer, such as poloxamer 407.

[179] The present application provides suspensions of the agents of the present application formulated in the presence of excipients such as, without limitation, Povidone, polysorbate 80 (PS80), polyethylene glycol (PEG) 400, tyloxapol, poloxamer, glycerin, and BAK in a buffer by Tris.

[180] In one embodiment the suspension of Compound-I or its free base comprises about 0.1 to 0.5% phosphate buffer. In another embodiment, the suspension of Compound-I or its free base and the second active agent comprises about 0.1 to 0.5% phosphate buffer. In some embodiments, the pH of the tromethamine-based suspension is between pH 4 and 7, for example, pH 6.0. In some embodiments, suspensions prepared in Tris further comprise from about 0.5% to about 2% polysorbate 80; about 0.05 to about 0.2% tyloxapol; and/or about 0.05% to about 0.4% poloxamer 407.

[181] In some such embodiments, the suspensions of the application further comprise from about 0.01 to about 1%, or from about 1 to about 2.0% w/v of glycerin. In a specific embodiment, the suspensions comprise about 2% w/v glycerin.

[182] In some embodiments, the suspensions of the application further comprise about 0.001 to about 0.005% w/v benzalkonium chloride (BAK). The amount of BAK can be varied depending on any adverse effects observed. BAK can be harmful to cells on the ocular surface, and therefore the amount in the formulation can be varied to obtain an optimal level of ocular penetration of Compound-I, without compromising the integrity of the ocular cell layer and increased toxicity.

[183] In some embodiments, the suspension optionally comprises buffers. Buffers when used, for example, may be basic sodium monophosphate, phosphoric acid and Tris buffer. The concentration of Compound-I in suspension is about 0.005% to about 5.0% w/v. The suspension prepared without additional buffer further comprises about 0.005% BAK and about 2% glycerin and has a pH of 6.0. In another embodiment, the suspension prepared without additional buffer comprises about 1% polysorbate 80, about 0.1% tyloxapol, about 0.2% poloxamer 407, about 0.005% BAK, about 2 .0% glycerin, and with a pH of 6.0.

[184] In suspensions prepared in phosphoric acid/Tris, the suspension comprises about 0.14% phosphoric acid, about 0.2% Tris base, about 1.0% polysorbate 80, about 0.005% BAK, approximately 2.0% glycerin and with a pH of 6.0. In one embodiment, the suspension further comprises about 0.2% tyloxapol. The pH of the suspension varies between about pH 6.0 and 7.2.

[185] Suspensions prepared in tromethamine (Tris) alone comprise about 1% polysorbate 80, about 0.1% tyloxapol, about 0.2% poloxamer 407, about 0.6% Tris, about of 0.005% BAK, and about 2.0% glycerin with pH 6.0. In another embodiment, a suspension prepared in Tris comprises about 1% Tris, about 0.45% NaCl, about 0.025% EDTA, about 0.2% HPMC, about 0. 1% polysorbate 80, approximately 0.005% BAK, with a pH of 6.0. In these suspensions 1 N HCl and/or 1 N NaOH are used for titration to the appropriate pH.

[186]The suspension of the order comprises about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3 0.0%, about 4.0%, or about 5.0% of an active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, e.g., Compound-I, and about 0.01% to about 0.05%, about 0.05 to about 0.09%, or about 0.09 to about 0.2% w/v sodium phosphate monobasic monohydrate and/or about 0.3 % to about 1.0% Tris. Alternatively, the suspension of the order comprises about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0 .5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3, 0%, about 4.0%, or about 5.0% of an active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, e.g., Compound-I, about 0.005%, about 0. 05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7 %, about 0.8%, about 0.9%, about 1.0%, about 2.0%, about 3.0%, about 4.0%, or about 5.0 % of a second active agent, and about 0.01% to about 0.05%, about 0.05 to about 0.09%, or about 0.09 to about 0.2% w/w v sodium phosphate monobasic monohydrate and/or about 0.3% to about 1.0% Tris.

[187] In a specific embodiment, the suspension comprises about 0.14% or about 0.2% w/v Tris buffer. In additional embodiments, suspensions are prepared at about 0.6% Tris or about 1.0% Tris. Other equivalent buffer systems well known in the art are also used in the suspensions of the present application. In one embodiment, the compound of Formula II or Compound-I is formulated as about 0.4% active agent, about 5% Cremophor RH40, about 2.0% glycerin, and about 0.005% from BAK. In another embodiment, the compound of Formula II or Compound-I is formulated as about 0.4% active agent, about 0.005% to about 5.0% second active agent, about 5% Cremophor RH40, about 2.0% glycerin, and about 0.005% BAK.

[188] In some embodiments, the order suspension has a pH value of about 4.0 to about 7.5 at or below 40 °C.

[189] In some embodiments, the order suspension has a pH value of about 5.0 to about 7.0 at or below 40 °C.

[190]For example, the order suspension has a pH value of about 6.0 at or below 40°C.

[191] In some embodiments, the first active agent may be formulated as a solution in accordance with the embodiments described herein, and the second active agent may be formulated as a suspension in accordance with the embodiments described herein.

[192] In other embodiments, the first active agent may be formulated as a suspension in accordance with the embodiments described herein, and the second active agent may be formulated as a solution in accordance with the embodiments described herein. Concentration in Various Ocular Tissues - Released as an Ophthalmic Suspension

[193] In some embodiments, a suspension of Compound-I or its free base provides similar concentration of the first active agent in the central choroid and central retina compared to the concentration of the first active agent released in the form of Gel Drop (discussed infra ).

[194] In some embodiments, suspension of Compound-I based on Tris or its free base with or without a second active agent increases the bioavailability of the first active agent in the central choroid and central retina, while reducing the concentration in the cornea. and preventing and/or treating corneal tears and/or diseases (e.g., corneal edema, ulceration, abnormalities, etc.). In some embodiments, topical delivery of Compound-I or its free base formulated in Tris base reduces the corneal concentration of Compound-I by about 5 to 10x, 10 to 20x, 20 to 30x, 30 to 40x, or about 50 to 100x compared to the equimolar corneal concentration of Compound-I or its free base released as a Gel Drop.

[195]The combined effects of decreasing corneal drug exposure so as to avoid poor ocular tolerability while maintaining or increasing posterior segment bioavailability so as to increase receptor tyrosine kinase (RTK) inhibition, e.g., VEGFR, significantly increases the therapeutic index and corresponding benefits to patients. Further prevention and/or treatment of disruption to the anterior surface of the eye with a second active agent, such as an EGFR modulator (e.g., activator), further improves the therapeutic index and corresponding benefits to patients.

[196] Once or twice daily administration of about 0.005% to about 5.0% w/v of Compound-I suspension of the present application for 30 to 90 days or 4 to 6 months is well tolerated in the eye.

Gel Drop

[197] In some embodiments the ophthalmic composition or formulation of the present application is formulated as a Gel Drop. The Gel Drop formulation includes no more than about 0.05% sodium phosphate monobasic monohydrate to provide the necessary buffering capacity and free flowing, filterable formulations at about 0.005% to about 2.0% Compound-I and/or at about 0.005% to about 5% of a second active agent without the need for surfactant additives.

[198] The Gel Drop formulation of the application comprises about 0.005% to about 2.0% w/v of the first active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, for example, Compound-I. The concentration of Compound-I or its free base (Formula II) in the Gel Drops can be about 0.005% to about 0.01%, about 0.01% to about 0.05%, about 0. 05% to about 0.1%, about 0.1% to about 0.2%, about 0.2% to about 0.3%, about 0.3% to about 0.4 %, about 0.4% to about 0.5%, about 0.5% to about 0.6%, about 0.6% to about 0.7%, about 0.7% to about 0.8%, about 0.8% to about 0.9%, about 0.9% to about 1.0%, or about 1.0% to about 2.0% w/v for topical administration. In some embodiments, the Gel Drops include about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4% , about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, or about 2% w/w v of Compound-I or its free base (Formula II).

[199] The Gel Drop formulation of the application may further comprise from about 0.00001% to about 5.0% w/v of the second active agent, or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide or vitamin K, or a combination of these. The Gel Drop formulation of the application may further comprise about 0.00001% to about 1.0%, about 0.00001% to about 0.1%, about 0.00001% to about 0. 01%, about 0.00001% to about 0.001%, about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001% w/v of the second active agent , or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide or vitamin K, or a combination thereof. The concentration of the second active agent in the Gel Drops can be about 0.00001% to about 0.0001%, 0.000012% to about 0.0001%, 0.000014% to about 0.0001%, 0.000016% to about 0.0001%, 0.000018% to about 0.0001%, 0.00002% to about 0.0001%, 0.00003% to about 0.0001%, 0. 00004% to about 0.0001%, 0.00005% to about 0.0001%, 0.00006% to about 0.0001%, 0.00007% to about 0.0001%, 0.00008% to about 0.0001%, 0.00009% to about 0.0001%, 0.000016% to about 0.00009%, 0.000018% to about 0.00009%, 0.00002% to about from 0.00009%, 0.00003% to about 0.00009%, 0.00004% to about 0.00009%, 0.00005% to about 0.00009%, 0.00006% to about 0 .00009%, 0.00007% to about 0.00009%, or 0.00008% to about 0.00009% w/v for topical administration. In some embodiments, the Gel Drops include about 0.00001%, 0.00002%, 0.00003%, 0.00004%, 0.00005%, 0.00006%, 0.00007%, 0. 00008%, 0.000081%, 0.000082%, 0.000083%, 0.000084%, 0.000085%, 0.000086%, 0.000087%, 0.000088%, or 0.000089% w/ v of the second active agent.

[200] The Gel Drop formulation of the application may further comprise about 0.5 µM, about 0.6 µM, about 0.7 µM, about 0.8 µM, about 0.9 µM, about of 1 µM, about 2 µM, about 3 µM, about 4 µM, about 5 µM, about 6 µM, about 7 µM, about 8 µM, or about 9 µM of a second active agent, or a pharmaceutically acceptable salt thereof, for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In some embodiments, the concentration of the second active agent is about 1 µM.

[201] In some embodiments, the Gel Drop ophthalmic compositions of the present application include glycerin as a tonicity agent. Some application embodiments provide ophthalmic composition including mannitol. The content of glycerin or mannitol in an amount to prevent any changes in the solubility of Compound-I, and at a level of about 2.0 to about 2.5% glycerin provides an osmolality of about 225 to about 300 mOsm/kg depending on phosphate concentration. In additional embodiments, glycerin is about 2% and phosphate is about 0.05% of the ophthalmic gel drop composition. The glycerin and phosphate concentrations of the present application are in such an amount that the tonicity level of the ophthalmic composition is about 240 mOsm/kg.

[202] The ophthalmic Gel Drop composition of the present application may also include Benzalkonium Chloride (BAK). In some embodiments, the BAK content is about 0.005%, sufficient to preserve the ophthalmic composition against microbial contamination. In some embodiments of the present application, BAK is not necessary for use of the ophthalmic composition in a sterile, single-use product.

[203] In some embodiments, the Compound-I ophthalmic Gel Drop formulation includes: about 0.005% to about 2.0% Compound-I or its free base, about 0.05% phosphate sodium, about 2% glycerin as the tonicity-adjusting agent, about 0.005% BAK as a preservative, water (purified, i.e., distilled, or deionized) as a carrier, and sodium hydroxide to adjust the tonicity. pH to 6.0. In one embodiment, no other excipients are added.

[204] The Gel Drop of the application comprises about 0.005% to about 2.0% of the active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, e.g., Compound-I, and about 0.01 % to about 0.05%, about 0.05 to about 0.09%, or about 0.09 to about 0.2% w/v monobasic sodium phosphate monohydrate. In a specific embodiment, the Gel Drop comprises about 0.05%, about 0.05 to 0.2%, or about 0.2% w/v of monobasic sodium phosphate monohydrate buffer. Other equivalent buffer systems well known in the art are also used in the Gel Drop of the present application. In one embodiment, Compound-I or its free base is formulated as about 0.4% to about 2.0% first active agent, about 5% Cremophor RH40, about 2.0% glycerin, and about 0.005% BAK.

[205] In one embodiment, the Compound-I Gel Drop includes about 0.3% to about 2.0% (3 to 20 mg/mL) of Compound-I, about 0.05% to about 0.2% sodium phosphate, and about 2% glycerin. The pH of the composition is between pH 5.0 to 7.0.

[206] The ophthalmic Gel Drop formulation of Compound-I and a second active agent includes: about 0.005% to about 2.0% Compound-I or its free base, about 0.00001% to about 5% (e.g., about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001%) second active agent, about 0.05% sodium phosphate , about 2% glycerin as the tonicity-adjusting agent, about 0.005% BAK as a preservative, water (purified, i.e., distilled, or deionized) as a carrier, and sodium hydroxide to adjust the pH to 6.0. In one embodiment, no other excipients are added.

[207] The Gel Drop of the application comprises about 0.005% to about 2.0% of the active agent of Formula I or II, or a pharmaceutically acceptable salt thereof, for example, Compound-I, about 0.00001% to about 5% (e.g., about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001%) second active agent, and about 0.01% to about 0.05%, about 0.05 to about 0.09%, or about 0.09 to about 0.2% w/v sodium phosphate monobasic monohydrate. In a specific embodiment, the Gel Drop comprises about 0.05%, about 0.05 to 0.2%, or about 0.2% w/v of monobasic sodium phosphate monohydrate buffer. Other equivalent buffer systems well known in the art are also used in the Gel Drop of the present application. In one embodiment, Compound-I or its free base is formulated as about 0.4% to about 2.0% first active agent, about 0.005% to about 5.0% second active agent. , about 5% Cremophor RH40, about 2.0% glycerin, and about 0.005% BAK.

[208] In one embodiment, the Compound-I Gel Drop includes about 0.3% to about 2.0% (3 to 20 mg/mL) of Compound-I, about 0.005% to about of 5.0% second active agent, about 0.05% to about 0.2% sodium phosphate, and about 2% glycerin. The pH of the composition is between pH 5.0 to 7.0.

[209] The present application provides Gel Drops of the agents (for example, the first active agent and the second active agent) of the present application formulated in the presence of excipients such as, without being limiting example, Povidone, polysorbate 80 (PS80), polyethylene glycol (PEG) 400, tyloxapol, poloxamer, glycerin, and BAK in a phosphate buffer.

Ophthalmic Drops

[210] Disclosed here is a formulation comprising a first active agent, for example, a compound of Formula I or II, and/or a second active agent, for example, nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. , such as ophthalmic drops, a form of drug delivery that is pharmaceutically acceptable to patients, convenient, safe, with an onset of action of several minutes. A standard eye drop used in therapy under US federal regulatory practice is sterile, has a pH of about 6.0 to 7.4, and, if used more than once, contains a preservative but has a Limited shelf life once opened, usually one month. If eye drops are packaged in a sterile, single-use, single-dose dispenser, the preservative may be omitted.

[211] An ophthalmic drop formulation method comprises the purest forms of the disclosed compound of Formula I or II (e.g., greater than 99% purity) and/or the second active agent, and the compound and/or the second active agent are mixed with buffer and tonicity adjusters to adjust for physiological pH and osmolarity. Examples of buffering agents for maintaining or adjusting pH include, but are not limited to, acetate buffers, citrate buffers, phosphate buffers, and borate buffers. Examples of tonicity adjusters are sodium chloride, mannitol and glycerin. In some embodiments, other pharmaceutically acceptable ingredients are also added.

[212] The formulated solution is then aliquoted into a plurality of separate sterile disposable cartridges, each of which is suitable for unit dosage, or a single cartridge for unit dosage. Such a single disposable cartridge is, for example, a conical or cylindrical specific volume dispenser, with a container having compressible side walls in a direction radial to a longitudinal axis so as to dispense the contents of the container therefrom at one end of the container.

[213] The present application provides ophthalmic eye drop solutions/suspensions packaged in multiple-dose or single-dose form, for example, as a plastic bottle with a dropper. In multiple dose form formulations, preservatives are necessary to prevent microbial contamination after opening the container. Suitable preservatives include, but are not limited to: benzalkonium chloride, thimerosal, chlorobutanol, methyl paraben, propyl paraben, phenylethyl alcohol, disodium edetate, sorbic acid, polyquaternium-1, or other agents known to those skilled in the art, and all of which are considered for use in the present application. Such preservatives are typically used at a level of 0.001 to about 1.0% weight/volume.

[214] Without wishing to be bound by theory, the formulation of the present application in an ophthalmic drop provides a pulse input of the drug. The route by which Compound-I gains access to the posterior segment is not by direct diffusion through the cornea with subsequent diffusion through the aqueous humor, vitreous humor, retina and finally, the choroid. Preferably, Compound-I obtains notable bioavailability following topical instillation using a circumferential route around, rather than across, the globe.

[215] In certain clinical conditions, ophthalmic drop solutions/suspensions can be formulated with other pharmaceutical agents, in order to mitigate the irritation of the other ingredient and facilitate the clinical response. Such agents include, but are not limited to, a vasoconstrictor such as phenylephrine, oxymetazoline, naphthazoline or tetrahydrozoline; a mast cell stabilizer such as olopatadine; an antihistamine such as azelastine; an antibiotic such as tetracycline; a steroidal anti-inflammatory drug such as betamethasone; a non-steroidal anti-inflammatory drug such as diclofenac; an immunomodulator such as imiquimod or interferons; and antiviral agents such as valacyclovir, cidofovir and trifluridine. Doses used for the purposes described above vary, but are in an amount effective to suppress discomfort, itching, irritation, or pain in the eye. When the compositions are dosed topically, the “pharmaceutically effective amount” of a compound of Formula I or II may generally be in a concentration range of 0.05 mg/mL to about 10 mg/mL and the “pharmaceutically effective amount” of the second active agent can generally be in a concentration range of 0.05 mg/mL to about 10 mg/mL, with 1 to 4 drops of the composition administered as a unit dose 1 to 4 times per day. The most common method of ocular drug delivery is instillation of drops into the cornea (i.e., “ophthalmic drops”).

[216] A key need is that the formulation is sterile and produced in a sterile environment. An ideal compound disclosed for use in ophthalmic solutions/suspensions should be soluble and/or miscible in aqueous media at normal ocular pH and tonicity. Furthermore, the disclosed compounds must be stable, non-toxic, long-lasting, and potent enough to act against dilution of drug concentration by blinking and tearing.

Dosage Forms

[217] The formulation of the present application may be suitable for ophthalmic use. In one embodiment, the formulation is a solution. The solution of the present application may be a clear, colorless, sterile, isotonic, buffered aqueous free-flowing liquid preparation. The drug product (e.g., the first active agent and/or the second active agent) has a pH of approximately 6.0 and can be stored at +5 °C. The drug product may be supplied in a container closure system consisting of a semi-transparent ophthalmic dispensing bottle with a dropper tip and cap.

[218] In some embodiments, the clinical concentration of Compound-I ophthalmic solution or suspension is equal to or less than about 0.1 mg/mL, equal to or less than about 0.2 mg/mL, mL, about 0.2 to about 1.0 mg/mL, about 0.3 to about 1.0 mg/mL, about 0.4 to about 1.0 mg/mL, about 0 .5 to about 1.0 mg/mL, about 0.6 to about 1.0 mg/mL, about 0.7 to about 1.0 mg/mL, about 0.8 to about 1.0 mg/mL, about 0.9 to about 1.0 mg/mL, about 1.0 to about 2.0 mg/mL, about 2.0 to about 3.0 mg/mL mL, about 3.0 to about 4.0 mg/mL, about 4.0 to about 5.0 mg/mL, about 5.0 to about 6.0 mg/mL, about 5 .0 to about 10.0 mg/mL, about 10 to about 20 mg/mL, about 20 to about 30 mg/mL, about 30 to about 40 mg/mL, or about 40 to about about 50 mg/mL.

[219] In other embodiments, clinical concentrations of the ophthalmic solution or suspension of Compound-I and the second active agent are independently equal to or less than about 0.1 mg/mL, equal to or less than about from 0.2 mg/mL, about 0.2 to about 1.0 mg/mL, about 0.3 to about 1.0 mg/mL, about 0.4 to about 1.0 mg /mL, about 0.5 to about 1.0 mg/mL, about 0.6 to about 1.0 mg/mL, about 0.7 to about 1.0 mg/mL, about 0.8 to about 1.0 mg/mL, about 0.9 to about 1.0 mg/mL, about 1.0 to about 2.0 mg/mL, about 2.0 to about from 3.0 mg/mL, about 3.0 to about 4.0 mg/mL, about 4.0 to about 5.0 mg/mL, about 5.0 to about 6.0 mg /mL, about 5.0 to about 10.0 mg/mL, about 10 to about 20 mg/mL, about 20 to about 30 mg/mL, about 30 to about 40 mg/mL , or about 40 to about 50 mg/mL.

[220] In one embodiment of the present application the concentration of the compound of Formula I or II is about 0.005% to about 5.0% (about 0.5 to about 50 mg/mL). A desired pharmacological activity (or concentration) of the formulation of the present application against pathological choroidal and retinal neovascularization is obtained following ocular administration of formulations containing about 0.005%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0 9%, about 1.0%, about 2.0%, about 3.0%, about 4.0%, or about 5.0% w/v of Compound-I. The present application establishes that following topical ocular administration at an optimal dose (e.g., between about 0.005% and about 5.0%) the pharmacologically active concentration is obtained and maintained in the central choroidal target tissue. In one embodiment the first active agent (Formula II or Compound-I) is formulated as about 0.005 to about 5.0% w/v concentration, the second active agent (nicotinic acid, nicotinamide, or vitamin K, or a combination thereof) is formulated as about 0.005 to about 5.0% w/v concentration, and the combination is dosed once or twice daily per eye for more than 60 consecutive days. Plasma concentrations observed following topical administration are substantially below the level expected to produce systemic toxicity. Table 2: In vitro summary of pharmacodynamic properties for Compound-I

[221] In some embodiments, Compound-I exhibits potent inhibition of tyrosine kinase activity for several pro-angiogenic growth factor receptors, with IC50 of less than about 100 nM (see Table 3). Compound-I also blocks high affinity VEGF receptors, e.g. VEGFR-1/Flt-1, but with lower potency (with IC50 of about 122 nM (69.41 ng/mL)). Table 3: In Vitro Inhibition of Tyrosine Kinases Using a Titration Curve. of 10 points (257 nM to 5000 nM) for Compound-I

[222]While inhibition of VEGFR appears to be essential for reducing vascular permeability and preventing further neovascular growth, simultaneous inhibition of VEGF signaling with inhibition of other growth factor signaling pathways (e.g., PDGF and angiopoietins/Tie2) can be linked to unique therapeutic effects. The therapeutic effects of broader inhibition of signaling pathways may contribute to the regression of newly established pathological vessels in the posterior segment of the eye.

[223] In some embodiments, about 300 nM (about 170.67 ng/mL) of Compound-I inhibits VEGFR-2 kinase function (see Table 4). Substantial blockade of a similar set of proangiogenic growth factor receptors, including FGFRs1-3, Tie-2, and EphB-4 is also observed. An unexpected finding is that approximately 300 nM concentration of Compound-I inhibits VEGFR-2 kinase function, which falls within the typical range found in the central choroid and retina following five days of topical ocular release. Table 4: In Vitro Inhibition of Tyrosine Kinases by 300 nM (170.67 ng/mL) of Compound-I Table 5: In vitro Inhibition of Tyrosine Kinases by 1 µM (568.9 ng/mL) of Compound-I Drug Substance and Drug Product Overview

[224] Drug product: ophthalmic formulations of Compound-I and/or second active agent for clinical studies are manufactured in dosage concentrations between 0.05% to 1.0% Compound-I and about 0.00001% to about of 5% (e.g., about 0.00001% to about 0.0002%, or about 0.00001% to about 0.0001%) of the second active agent. In some embodiments, the dosage of Compound-I in the formulation is 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.8%, or 1.0%. In some embodiments, the dosage of the second active agent in the formulation is 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.8%, 1.0%, 2.0%, 3.0%, 4.0% or 5.0%. Ophthalmic formulations of Compound-I and/or second active agent (solutions or suspensions) are for single, daily, topical administration to the eye in a clinical setting. In addition to the first and second active ingredients, in some embodiments the medicated product may further contain about 0.005% BAK as a preservative, purified water as a carrier, and is pH adjusted with sodium hydroxide to pH 6.0.

Sodium Phosphate Based Gel Drop

[225] The ophthalmic benefits of Compound-I in a sodium phosphate-based formulation (listed in table 6) result from the self-gelling properties of the API in tampons, such as sodium phosphate. The spontaneous formation of thixotropic, self-forming Compound-I gel from a clear solution is formed by increasing the concentration of the first active agent in sodium phosphate. Once the concentration of the first active agent in the phosphate buffer reaches a super-saturated state, insoluble Compound-I particulates are observed within the gel.

[226] The current state of the art predicts that applying a gel with increased viscosity to the surface of the eye would increase the residence time on the cornea. The increased residence time on the cornea in turn facilitates ocular absorption of the drug. As a result, intraocular drug concentrations of viscous gels would be increased compared to non-viscous formulations, such as water-like solutions. One way to increase viscosity is to use various viscosity enhancing excipients, for example, carboxymethylcellulose, which in effect achieves increased intraocular absorption of different drug substances following topical ocular administration. The present application provides a thixotropic Compound-I gel and/or a second active agent formed in the absence of any viscosity enhancing excipients. For example, when Compound-I or Compound-I and a second active agent are dissolved in a simple buffer, such as sodium phosphate, a thixotropic gel is formed. The thixotropic gel, which is formed without any viscosity enhancing excipients, is formulated as a Gel Drop.

[227] The present application provides dose-dependent and dose-frequency-dependent release of Compound-I to tissues of the posterior segment of the eye.

[228] The Gel Drop formulations of the present application (listed in table 6) differ from each other in several aspects, such as the first active concentration, sodium phosphate concentration, presence or absence of tonicity (glycerin) or preserving agents ( benzalkonium chloride/BAK), solubilizing surfactants (polysorbate 80, tyloxapol, and/or poloxamer), and pH.

Tromethamine-Based Suspension

[229] The present application provides a suspension of Compound-I and/or a second active agent in a tromethamine-based formulation. In some embodiments, the suspension of Compound-I and/or the second active agent in a tromethamine-based formulation has equal to or more than 95% of the first active drug substance in an insoluble form. This characteristic is distinguishable from the soluble or semi-soluble state of Compound-I in the Gel Drop (gel, which is not an entirely soluble state as the concentration of the first active agent increases) or in a formulation based in Cyclodextrin. Compound-I formulations based on tromethamine show increased turbidity with increased concentration of the first active agent. Administering a topical drop of Compound-I suspension and/or second active agent to the eye, which is a combination of soluble and insoluble first active agent components, is beneficial with respect to both safety/tolerability and efficacy.

[230] The present application provides Compound-I in tromethamine-based suspension, delivered in concentrations to target tissues between 10 and 1000x the cellular IC50 for the various pro-angiogenic RTKs. See, for example, Table 7.

[231] The corneal safety and tolerability of topical Compound-I is a direct consequence of the amount of the first soluble (as opposed to insoluble) active agent applied to the corneal surface, and the concentration in the resulting corneal tissue. In some embodiments, subjects receiving topical ocular administration of the tromethamine-based suspension are able to tolerate up to the highest level of concentration of the first active agent in the formulation when compared to equimolar formulations of the phosphate-based Gel Drop. of sodium. The corneal safety and tolerability of topical Compound-I is also a consequence of administration of a second active agent, e.g., nicotinic acid, nicotinamide, or vitamin K, or a combination thereof, which is a modulator (e.g., activator) of EGFR that prevents or treats corneal tears or diseases caused by EGFR inhibition.

[232] The present application provides ocular bioavailability of Compound-I in the posterior segment during administration of a tromethamine-based suspension. The ocular bioavailability of Compound-I in the posterior segment is directly proportional to the total amount of drug, Compound-I, administered (insoluble plus soluble, see Table 7). Although insoluble drug particulates are not readily available to anterior segment tissues; the inherent and unique physicochemical properties of Compound-I allow both insoluble and soluble components to gain entry into posterior segment tissues such as the choroid and retina. Consequently, even higher drug concentrations than those obtained with Gel Drop formulations containing equivalent amounts of the first active agent are obtained with the tromethamine-based suspension. Thus, the tromethamine-based suspension provides: a) improved corneal tolerability and b) maintained or increased bioavailability to the posterior segment, particularly the choroid, the primary target tissue for treating neovascular (wet) AMD. Additionally, a second active agent, e.g., nicotinic acid, nicotinamide, or vitamin K, or a combination thereof, that is a modulator (e.g., activator) of EGFR that prevents or treats corneal tears or diseases caused by EGFR inhibition , prevents and/or treats corneal tears potentially associated with the administration of Compound-I, thereby increasing the therapeutic index of Compound-I.

Cyclodextrin-based solution

[233] Cyclodextrins, which are cyclic oligosaccharides composed of six to eight dextrose units (a-, ß-, and Y—CDs) linked through one to four bonds, are well known for their ability to act as a solubilizing agent for relatively insoluble drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol., 36: 30 - 42 (2008).

[234] In some embodiments, 2-hydroxypropyl-ß-cyclodextrin (HP- ß-CD, also known as KLEPTOSE® HPB) in a molar ratio equal to or greater than 1:6 or Sulfobutyl ether-ß-cyclodextrin ( SBE-ß-CD, also known as CAPTISOL®) in a ratio equal to or greater than 1:2 in the proposed clinical formulation, Compound-I or its free base and an ophthalmic solution of the second active agent, provide solubility that satisfies concentrations clinical dose range of 0.1 to 1.2% of Compound-I.

[235] In some embodiments, solutions of Compound-I based on cyclodextrin or its free base and/or a second active agent not only have improved solubility of the first active agent in a uniform solution, but, during topical ocular administration , also have a novel and previously unobserved feature of significantly increased therapeutic index of the first active agent in the posterior segment of the eye. Solutions of Compound-I and/or the second active agent of the present application reduce exposure to the anterior segment of Compound-I, thereby increasing the concentration of the first active in the solution and increasing the frequency of its release in order to maintain concentrations of the high posterior segment. Both of these beneficial features are related to the known property of cyclodextrin to form hydrophilic complexes with hydrophobic drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol., 36: 30 - 42 (2008). Administration of the second active agent as a combination with the first active agent prevents corneal tears or diseases caused by EGFR inhibition by systemic diseases (e.g., cancer, diabetes), ocular diseases, or administration of the first active agent, e.g., a compound of Formula I or II, thereby increasing the therapeutic index of the first active agent, e.g., Compounds of Formula I or II.

[236] When formulated with Compound-I or its free base and/or a second active agent, cyclodextrin can form a colorless, clear solution that exhibits viscosity similar to water. Following topical ocular administration, the Compound-I/cyclodextrin complex appears to be pharmacologically inactive and metabolically inert. The Compound-I/cyclodextrin complex confers corneal tolerability until the cyclodextrin spontaneously dissociates from the first active agent, thus making the high concentration of Compound-I available at its intended site of action in the posterior segment of the eye, e.g., choroid and retina. .

[237] In some embodiments, cyclodextrin-based Compound-I solutions reduce Compound-I corneal exposures compared to Gel Drop formulations at similar drug concentrations. The use of cyclodextrin-based Compound-I solutions provides approximately 10x reduction in corneal concentrations when compared to dosing with equimolar Gel Drop formulations. In some embodiments, after 20 to 30 days of topical ocular dosing of about 0.2 to 2.0%, e.g., about 0.6%, Compound-I as a cyclodextrin-based solution, no unfavorable findings are attributed to the test article or vehicle. The present application provides higher concentrations of Compound-I within posterior segment target tissues, such as the central choroid and central retina, when the cyclodextrin-based solution of Compound-I is topically applied. In some embodiments, the combined effects of decreasing corneal drug exposure so as to avoid poor ocular tolerability, while increasing posterior segment bioavailability so as to increase RTK inhibition, significantly increases the therapeutic index and benefit(s). corresponding to treated subjects. In other embodiments, the combined effects of decreasing corneal drug exposure so as to avoid poor ocular tolerability and prevention or treatment of corneal disruption or disease by the second active agent, while increasing posterior segment bioavailability so as to increase inhibition of RTK can significantly increase the therapeutic index and corresponding benefit(s) to the treated subjects.

[238] The present application provides expansion of the therapeutic window for both suspension-based formulations (see Example 3) and cyclodextrin formulations of Compound-I due to significantly reduced exposure (about 10 to 100X or 1 to 2 log). Reduced exposure improves corneal safety/tolerability, which allows higher concentrations or dosing frequency of Compound-I to be administered topically. The higher concentration allows Compound-I to obtain higher target tissue concentration behind the eye, which improves the therapeutic efficacy of Compound-I.

[239] In some embodiments, topical ocular dosing of ophthalmic gel drops is associated with high corneal tissue exposure (>100uM) and corresponding unfavorable observations in the anterior segment, such as discomfort, corneal and conjunctival inflammation, erosion and/or or corneal epithelial thinning and degeneration. In contrast, repeated topical ocular dosing of Compound-I ophthalmic solution produces corneal exposure that is approximately 5 to 10 times lower than an equimolar dose of ophthalmic gel drops, and is free from unfavorable clinical or histopathological findings. Topical ocular dosing with Compound-I ophthalmic solution also achieves equal or higher therapeutic target exposure in the central choroid compared to an equimolar dose of the ophthalmic gel drop. Preferably, the combination of decreased corneal exposure and corresponding improved ocular tolerability, while simultaneously maintaining or promoting drug release to posterior segment target tissues, together with improved physicochemical stability, provides greater benefit to subjects compared to the Gel Drop ophthalmic formulation. . 1 to 5 Day PK Results with Topical Ocular Compound-I in Cyclodextrin-Based Solutions

[240] The present application provides ocular pharmacokinetics of various formulations and dose regimens of Compound-I following topical ocular administration. Three dosage concentrations in nine (9) different topical ocular formulations of Compound-I are used for dosing once daily (q.d.) or twice daily (b.i.d.) for 1, 2, 3, 4, or 5 consecutive days. Subjects each receive approximately 30 µL bilateral topical ocular dose of one of three (3) Compound-I formulations, or vehicle formulation, using a positive displacement pipette.

[241] The composition of each Compound-I formulation is described in table 8A. All doses were administered within ±1 hour of the scheduled dose time. On day 1, Groups 1, 2, 4 to 6, 8, 10, 11, 13, 15, and 17 receive one dose (q.d.) for one (1) or four (4) days. On days 1 through 4, Groups 3, 7, 9, 12, 14, and 16 receive b.i.d. dosing. approximately 8 hours apart at 7:00 AM and 3:00 PM for four (4) days. Some subjects receive b.i.d dosing of vehicle-only formulations for five (5) consecutive days.

[242] In some embodiments, ocular sampling is performed at about 0.5, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about of 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20 , about 21, about 22, about 23, or about 24 hours post-dose relative to the day 1 dose. Samples of aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid are collected to monitor the effects of the treatment. Samples of aqueous humor, cornea, central retina, and central choroid are evaluated.

[243] Table 8A-C lists PK results from 1 to 5 days with topical ocular Compound-I in Cyclodextrin-based Solutions. Table 8A: Ocular Formulations

[244] Table 8B lists average Compound-I concentrations in aqueous humor, retina, choroid, and cornea (LLOQ: Lower Limit of Quantification; the LLOQ is the lowest analyte concentration that can be quantified with acceptable precision and accuracy ). Table 8B LLOQ of the central retina = 0.0181 µM LLOQ of the peripheral retina = 0.00873 µM (Grps 1 to 8); LLOQ = 0.00898 µM (Grps 12 to 16) LLOQ of the central choroid = 0.175 µM LLOQ of the peripheral choroid = 0.0349 µM (Grps 1 to 8); LLOQ = 0.0359 µM (Grp 12 to 16) Corneal LLOQ = 0.0181 µM (Grp 1 to 5); LLOQ = 0.0453 µM (Grp 6 to 8,10 to 13,15,16A17); LLOQ = 0.0873 µM (Grp 4,9,16B) N/A = Not applicable; Samples not evaluated by study protocol. *Average based on n=1.

[245] Table 8C lists summary mean ocular tissue concentrations of Compound-I in aqueous humor, central and peripheral retina, central and peripheral choroid, and cornea for Groups 1 to 10. Any values <LLOQ were excluded from calculations statistics. When all values are <LLOQ for a given point in time, <LLOQ are reported as the average. Table 8C LLOQ OF AH = 0.000903 µM LLOQ of the central retina = 0.0181 µM LLOQ of the peripheral retina = 0.00873 µM (Grps 1 to 8); LLOQ = 0.00898 µM (Grps 12 - 16) LLOQ of the central choroid = 0.175 µM LLOQ of the peripheral choroid = 0.0349 µM (Grps 1 to 8); LLOQ = 0.0359 µM (Grp 12 to 16) Corneal LLOQ = 0.0181 µM (Grp 1 to 5); LLOQ = 0.0453 µM (Grp 6 to 8,10 to 13,15,16A17); LLOQ = 0.0873 µM (Grp 4,9,16B) N/A = Not applicable; Samples not evaluated by study protocol. *Average based on n=1. 5-day PK results with topical ocular Compound-I in cyclodextrin-based solutions

[246] The present application provides ocular pharmacokinetics of various dose regimens of Compound-I topical ocular solutions containing hydroxypropyl-ß-cyclodextrin (“HDßCD”) following ocular dose administration. Topical ocular solutions other than Compound-I are administered once daily (q.d.) or twice daily (b.i.d.) for 4 or 5 consecutive days. Subjects each receive a bilateral topical ocular dose of 30 µL of one of four dosage concentrations of Compound-I.

[247] All doses were administered within ± 1 hour of the scheduled dose time, except for some subjects receiving it on day 1. Ocular sampling after Compound-I administration is performed one hour after the first daily dose on day 5 to subjects, except in some, where ocular sampling is performed 24 hours after the first daily dose on day 4.

[248] Samples of aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid are collected. Samples of the cornea, central retina, and central choroid are evaluated; Aqueous humor, peripheral retina, and peripheral choroid samples are not evaluated.

[249] Table 9 (AB) lists the 5-day PK results with topical ocular Compound-I in cyclodextrin-based solutions. Table 9A: Ocular Formulations

[250] Table 9B lists a summary of average ocular tissue concentrations of Compound-I in the central retina, central choroid, and cornea. Any values <LLOQ were excluded from statistical calculations. When all values were <LLOQ for a given time point, <LLOQ was reported as the average. Table 9B: Average concentrations of Compound-I in the retina, choroid, and cornea. Central retinal LLOQ = 0.0218 µM Central choroidal LLOQ = 0.174 µM Corneal LLOQ = 0.0174 µM *Average based on n=1 Compound-I Concentrations (in µM) in various ocular fluids and tissues

[251] In some embodiments, the concentration of the first active agent in various tissues and fluids of the eye is measured during topical ocular administration of a solution of about 0.4% (about 4 mg/mL) of Compound- I and cyclodextrin. The average concentration of Compound-I is measured in the central choroid, central retina, aqueous humor, and cornea. Compound-I is in a solution (0.4% or 4 mg/mL) with 8.41% KLEPTOSE® and 0.142% phosphate buffer; 8.9% KLEPTOSE® HPB and 0.142% phosphate; 4.88% CAPTISOL® and 0.142% phosphate; or 4.88% CAPTISOL® and 0.122% phosphate. See Table 10A-B.

[252] In some embodiments, during topical ocular administration of a solution of about 0.4% (about 4 mg/mL) of Compound-I and cyclodextrin, the concentration of Compound-I in the central choroid is between about 0.2 µM and about 0.8 µM. The concentration of Compound-I in the central retina is between about 0.05 µM to about 0.15 µM. In some embodiments, during topical ocular administration of a solution of about 0.4% (about 4 mg/mL) of Compound-I and cyclodextrin, the concentration of Compound-I in the aqueous humor is between about 0.003 µM to about 0.008 µM. And the corneal concentration of Compound-I is about 6.0 µM to about 40 µM. KLEPTOSE® HPB or CAPTISOL® is used in Compound-I solution administered topically to the eye.

[253] In some embodiments, average ocular tissue concentrations of Compound-I following twice-daily topical dosing with 0.3% Compound-I ophthalmic gel drop formulations with and without benzylalkonium chloride are highest in the cornea with between about 200 µM to about 350 µM in the cornea, between about 2.0 µM to about 5.0 µM in the peripheral choroid, between about 0.2 µM to about 0.7 µM in the central choroid, between about 0.05 µM to about 0.5 µM in the peripheral retina, and between about 0.01 µM to about 0.05 µM in the aqueous humor.

[254] In some embodiments, Tris suspension-based formulations of Compound-I are well tolerated, without any corneal findings, and with only a few sporadic incidences of mild conjunctivitis. In some embodiments, mean ocular tissue concentrations of Compound-I, evaluated at 1 hour ± 15 minutes after the first daily topical ocular dose on day 30 for twice daily topical dosing with Tris-based suspensions of Compound -I at 0.3% with and without benzylalkonium chloride, are highest in the cornea, for example, between about 2.00 µM to about 4.0 µM. The concentration in the peripheral choroid of the same dose is between about 0.7 µM to about 1.5 µM; the concentration in the central choroid is between about 0.3 µM to about 0.4 µM; the concentration in the peripheral retina is between about 0.08 µM to about 0.09 µM); the concentration in the central retina is between about 0.04 µM to about 0.07 µM; and the concentration in the aqueous humor is about 0.001 µM to about 0.002 µM.

[255] The present application provides cyclodextrin-based solutions (e.g., solutions comprising hydroxypropyl-beta-cyclodextrin (HP-ß-CD, KLEPTOSE® HPB)) of Compound-I that were well tolerated when administered topically for up to 30 days , twice daily in about 0.1% Compound-I (in a solution with about 2.0% to about 2.5% HP-ß-CD), twice daily in about 0 .2% Compound-I (in a solution with about 4.0% to about 4.5% HP-ß-CD), once or twice daily in about 0.4% Compound- I (in a solution with about 8.0% to about 8.5% HP-ß-CD), and once or twice daily in about 0.6% Compound-I (in solution with up to about 14% HP-ß-CD) in subjects. Furthermore, in additional embodiments, cyclodextrin-based solutions of about 0.4% w/v of Compound-I in Kleptose® HPB, Kleptose® HP, or Captisol® are well tolerated when dosed twice daily for up to 24 days.

[256] The present application provides dose-limiting corneal toxicity observed with Compound-I ophthalmic Gel Drop formulations. In some embodiments, the ophthalmic gel drop renders about five times to about fifteen times the highest corneal concentrations of Compound-I compared to the cyclodextrin-based solution, and about fifty times to about a hundred times the concentrations higher corneal levels of Compound-I compared to Tris-based suspensions. Tris-based suspensions of Compound-I and cyclodextrin-based solutions of the present application are well tolerated with no evidence of overt ocular toxicity. In some embodiments, administration once or twice daily for at least 30 days of about 0.005% to about 5.0% w/v of a cyclodextrin-based solution or a Tris-based suspension of Compound-I is well tolerated in subjects. The present application provides higher central choroidal concentrations of Compound-I using cyclodextrin-based solutions compared to equimolar doses of Tris-based gels and/or formulations. Table 10A: Average concentration of Compound-I in µM in various ocular fluids and tissues

[257] Table 11 shows the central corneal and choroidal concentrations of Compound-I formulations. Table 11:

Phase I Protocol for Dose Escalation Study in Patients With Neovascular AMD

[258] The present application provides a Phase I study involving a twelve-week, open-to-the-public, dose-escalation, multicenter trial to evaluate the safety, tolerability, and pharmacokinetics following topical ocular administration of Compound-I in patients with neovascular age-related macular degeneration (AMD). Up to 60 patients in total are treated once to twice daily with topical ocular dosing of Compound-I ophthalmic solution for three months, where three dose-escalation monotherapy arms and one adjunctive therapy arms using a single intravitreal injection of Lucentis ® plus the maximally tolerated monotherapeutic dose are planned (15 patients per treatment arm). Patients meeting pre-specified vision and CNV lesion criteria confirmed by an independent reading center are allowed to simultaneously discontinue topical ocular dosing and receive standard of care treatment.

[259] The present application provides 3 dosage concentrations, ranging from 0.1% to 1.0% (w/v) (as Compound-I) of ophthalmic solution for clinical studies. The concentrations are about 0.1%, about 0.3%, about 0.6%, and about 1.0% (w/v) Compound-I HCl. Formulation Preparation

[260] Non-limiting examples of formulations of the present application are outlined in table 12. Table 12: Overview of product compositions tested in product screening studies a Molar ratio of Compound-I: cyclodextrin. b KLEPTOSE® HPB c CAPTISOL®

In-Cell EGFR Tyrosine Phosphorylation Assay to Determine the EGFR Activity of Compounds of Formula I or II

[261] An EGFR tyrosine assay in corneal epithelial cells is conducted to determine whether higher concentrations of EGF can overcome inhibition of EGFR kinase activity. Cells are serum starved and then pretreated with different concentrations of a compound of Formula I or II, e.g., Compound-I or a control followed by EGF treatment. The cells are then harvested and immunoblotted to determine the concentration of phosphorylated EGFR and total EGFR which was used to determine receptor activity (IC50).

In-Cell EGFR Tyrosine Phosphorylation Assay to Determine the EGFR Activity of a Combination of Compounds of Formula I or II, and a Second Active Agent

[262] An EGFR tyrosine assay in corneal epithelial cells is conducted to determine whether Vitamin K or nicotinic acid/nicotinamide can overcome EGFR inhibition and whether there is an increase in EGFR activity. Cells are serum starved and then pretreated with a different concentration of a compound of Formula I or II, e.g., Compound-I and saturating concentrations of vitamin K or nicotinic acid/nicotinamide followed by treatment with EGF. Cells are harvested and immunoblotted for determination of the concentration of phosphorylated EGFR (tyrosine 1068 and tyrosine 1045) and total EGFR which was used to determine receptor activity (IC50).

Determination of Effects of Varying Concentrations of Compounds of Formula I or II and EGF on Cell Migration/Proliferation in Cells (In Vitro Wound Healing)

[263] Determination of cell migration/proliferation in corneal epithelial cells is conducted to determine cell migration/proliferation in the presence of varying concentrations of a compound of Formula I or II and EGF. Cells are plated with silicone plugs. The cells are then serum starved and pretreated with varying concentrations of a compound of Formula I or II or a control. The silicone plugs are then removed to create the acellular area where the cells are treated with EGF. Cell migration is quantified from microphotographs.

Determination of Cell Migration/Proliferation in Cells Treated with Compounds of Formula I or II and/or a Second Active Agent

[264] The determination of cell migration/proliferation in corneal epithelial cells is conducted to determine cell migration/proliferation in the presence of varying concentrations of a compound of Formula I or II, vitamin K, or nicotinic acid/nicotinamide. Cells are plated with silicone plugs. The cells are then serum starved and pretreated with varying concentrations of a compound of Formula I or II or a control. The plugs are removed and the cells are treated with 1) EGF, 2) EGF and vitamin K, or 3) EGF and nicotinic acid/nicotinamide. Cell migration is quantified from microphotographs.

Determination of the Effects of Compounds of Formula I or II on Basal and EGF-Mediated Corneal Wound Healing In Vivo

[265] Determination of the effects of a compound of Formula I or II on basal and ligand stimulated rates of corneal wound healing was determined in mice. Corneas from C57/Bl mice are injured and then pretreated with a compound of Formula I or II followed by addition of EGF. Wound size is monitored by fluorescein staining and fluorescent photography and quantified.

Determination of the Effects of Compounds of Formula I or II and a Second Active Agent on Basal Corneal Wound Healing and Mediated by EGF In Vivo

[266] Determination of the effects of a compound of Formula I or II and/or a second active agent on corneal wound healing was determined in mice. Corneas from C57/Bl mice are injured and then pretreated with a compound of Formula I or II, followed by addition of EGF, vitamin K, or nicotinic acid/nicotinamide. The size and closure of the wound are monitored.

Treatment Doses

[267] The formulation of the present application is effective in treating (i.e., stabilizing or regressing the lesion) or preventing choroidal and retinal neovascularization (NV) in the eye of a mammalian subject. Compound-I of the present application, at a specific dose, inhibits a receptor tyrosine kinase and the second active agent at a specific dose, inhibits an ErbB receptor tyrosine kinase. In some embodiments, the Compound-I formulation, at a specific dose, inhibits receptor tyrosine kinases including, VEGFR, FGFRs, Tie2, and EphB-4. The inhibition of several RTKs by the formulation of the present application, at a specific dose, simultaneously has a synergistic effect, and is effective in treating or regressing NV in the posterior segment of the eye. In some embodiments, the second active agent modulates (e.g., activates) directly or indirectly an ErB receptor tyrosine kinase including, EGFR, HER2, HER3 and Erb4. The activation of several ErB receptor tyrosine kinases by the formulation of the present application, at a specific dose, simultaneously can have a synergistic effect, and is effective in preventing or treating corneal tears or diseases.

[268] In one embodiment, the present application provides a method of treating (i.e., lesion stabilization or regression) or preventing choroidal and retinal neovascularization (NV) in the eye by administering to the subject in need thereof a therapeutically effective amount of a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, and a second active agent or a pharmaceutically acceptable salt thereof, wherein a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, and the second active agent, or a pharmaceutically acceptable salt thereof, are administered simultaneously. Alternatively, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered prior to administration of a second active agent, or a pharmaceutically acceptable salt thereof. In another embodiment, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered after administration of a second active agent, or a pharmaceutically acceptable salt thereof. In another embodiment, the present application provides a method of treating (i.e., stabilizing or regressing the lesion) or preventing choroidal and retinal neovascularization (NV) in the eye by administering to the subject in need thereof a therapeutically effective amount of a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, before administering a therapeutically effective dose of a formulation described herein.

[269] In another embodiment, the present application provides a method of treating (i.e., stabilizing or regressing the lesion) or preventing choroidal and retinal neovascularization (NV) in the eye and preventing or treating a corneal tear or disease by administering if the subject in need thereof is provided with a therapeutically effective amount of a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, and a second active agent or a pharmaceutically acceptable salt thereof, wherein a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, and the second active agent, or a pharmaceutically acceptable salt thereof, are administered simultaneously. Alternatively, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered prior to administration of a second active agent, or a pharmaceutically acceptable salt thereof. In another embodiment, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is administered after administration of a second active agent, or a pharmaceutically acceptable salt thereof. In another embodiment, the present application provides a method of treating (i.e., stabilizing or regressing the lesion) or preventing choroidal and retinal neovascularization (NV) in the eye and preventing or treating a corneal tear or disease by administering to the subject in need thereof a therapeutically effective amount of a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, before administering a therapeutically effective dose of a formulation described herein.

[270] In another embodiment, the formulation of the present application is effective in treating NV and treating and/or preventing corneal disruption or disease caused by systemic disease, an ocular disease or administration of a compound of Formula I or II when administered once, twice, three, and four times daily by topical ocular delivery of about 0.005% to about 5.0% (about 0.05 to about 50 mg/mL) of Compound-I and a second active agent . The formulation of Compound-I or its free base (Formula II) and a second active agent, for the treatment or regression of NV and the treatment and/or regression of corneal disruption or disease, is a solution comprising a second active agent, a compound of Formula I or II, and cyclodextrin or in a suspension comprising Tris. The solution or suspension when released to a subject exposed to atmospheric oxygen to induce oxygen-induced retinopathy (OIR) or NV, for example, is capable of effectively reducing the average area of preretinal NV per retina with little or no disruption or disease. corneas. Prevention or treatment of NV by Compound-I formulation and/or suspension is achieved through inhibition of several receptor tyrosine kinases (RTKs), including VEGFR-2. Prevention or treatment of corneal tears or disease by a second active agent is achieved through modulation (e.g., activation) of EGFR.

[271] Any of the disclosed diseases or conditions described herein can be treated or prevented by obtaining target tissue concentrations of about 200 nM to about 2 µM of the disclosed compounds or pharmaceutically acceptable salts, formulation and/or suspension thereof. One embodiment of this application relates to a method for treating pathological angiogenesis in the posterior segment of the eye by obtaining target tissue concentration of about 200 nM to about 2 µM of the disclosed compounds or pharmaceutically acceptable salts, and/or formulation thereof. . Another iteration of this embodiment relates to obtaining target tissue concentration of about 300 nM to about 2 µM of one or more of the disclosed compounds or pharmaceutically acceptable salts, and/or formulation thereof.

[272] In one embodiment of the present application about 0.2 to about 1.0% (about 2 to about 10 mg/mL) of Compound-I formulated as a solution or suspension, during administration, can effectively inhibit VEGFR-2 kinase function and provide substantial blockade of a set of pro-angiogenic growth factor receptors, including FGFRs1-3, Tie-2, and EphB-4. The concentration of 2 to 10 mg/mL of Compound-I in the formulation provides pharmacologically effective concentrations of drug to the central choroid and retina following 1 to 5 days of topical ocular release.

[273] In some embodiments, the exposure time of Compound-I is between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, the dosage regimen involves several courses of topical ocular administration of a formulation comprising Compound-I to a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves once daily, twice daily, three times daily, or four times daily administration of the formulation for between 1 and 90 days or for more than 90 days (e.g., 4 months , 6 months, 8 months, or 12 months). For example, the dosage regimen involves administering the formulation once, twice, thrice, or four times every other day (i.e., on day 1, 3, 5, 7, etc.) for up to 90 days. . For example, the dosage regimen involves administering once on day 1, once, or twice on day 2 through day 90. For example, the dosage regimen involves administering once, twice, three times, or four times on day 1, followed by once daily for 2 to 90 days. For example, the dosage regimen involves administering once, twice, three times, four times on day 1, followed by once, twice, three times, or four times every other day (i.e., on day 1). , 3, 5, 7 etc.) for up to 90 days. For example, a dosage regimen involves once a day or twice a day for 1, 2, 3, 4, or 5 consecutive days. For twice or three times daily dosing regimen, subjects receive topical ocular dosing of a Compound-I formulation on days 1 and 4 approximately 4, 6, or 8 hours apart. In another embodiment, subjects receive topical ocular doses of a Compound-I formulation approximately 4, 6, or 8 hours apart for four consecutive days. In some embodiments, subjects receive one or two doses of a topical ocular dose of Compound-I formulation per day for 5 consecutive days. In still other embodiments, subjects receive one or two doses of a topical ocular dose of Compound-I formulation for 5 to 90 consecutive days. In some embodiments, subjects receive one or two doses of a topical ocular dose of Compound-I formulation for at least 25 consecutive days. In one embodiment, subjects receive one or two topical ocular doses for at least 90 consecutive days or more.

[274] In some embodiments, the present application provides a formulation of Compound-I or its free base and/or a second active agent administered topically to the anterior segment of the subject's eye to treat AMD, pathological CNV, and/or NV pathological. For example, the formulation is administered to a subject's eye 1, 2, 3, or 4 times per day. In specific embodiments, the formulation is administered to a subject's eye 2 or 3 times per day. For example, the formulation is administered to one eye or both eyes of a subject. For example, about 1 mg/ml of a first active agent comprising the formulation of the current disclosure is administered twice daily (BID) to one eye or both eyes of a subject. In some embodiments, about 1 mg/mL once daily (QD) or BID, about 2 mg/mL QD or BID, about 3 mg/mL QD or BID, about 4 mg/mL QD or BID, about 5 mg/mL QD or BID, about 6 mg/mL QD or BID, about 7 mg/mL QD or BID, about 8 mg/mL QD or BID, about 9 mg/mL QD or BID, or about 10 mg/mL QD or BID is administered to one eye or both eyes of a subject.

[275] For example, a formulation comprising about 1 mg/mL BID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for longer than 90 days (e.g. 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL QD of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 1 mg/mL TID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a formulation comprising about 1 mg/mL QID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL BID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL QD of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL TID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 2 mg/mL QID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL BID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL QD of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL TID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 3 mg/mL QID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL BID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL QD of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 4 mg/mL TID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a formulation comprising about 4 mg/mL QID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL BID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL QD of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 5 mg/mL TID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a formulation comprising about 5 mg/mL QID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL BID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL QD of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL TID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 6 mg/mL QID of Compound-I and/or a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days. days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 7 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 8 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL BID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL TID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 9 mg/mL QID of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a formulation comprising about 10 mg/mL QD of a first active agent (e.g., Compound-I) and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (for example, 4 months, 6 months, 8 months, or 12 months). The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above. In some embodiments, the formulation of the present application is administered QD, BID, TID, or QID when administered in low doses (e.g., 1 mg/mL, 2 mg/mL, 3 mg/mL, 4 mg/mL, or 5 mg/mL), and QD or BID at high doses (e.g., 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, or 10 mg/mL).

[276] In another embodiment, a BID of 1 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for longer than 90 days (e.g. 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 1 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 2 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 2 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 3 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 3 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments a BID of 4 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g. example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 4 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 5 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 5 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 6 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 6 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 7 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 7 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 8 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, an 8 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 9 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 9 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a BID of 10 mg/mL of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 10 mg/mL QD of Compound-I formulation and a second active agent is administered to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days ( for example, 4 months, 6 months, 8 months, or 12 months). The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above.

[277] The present application provides formulations as shown in table 13 for administering to one eye or both eyes of a subject. Table 13

[0278] In some embodiments, the formulation of Formula II or Compound-I is administered to one eye or both eyes of a subject. For example, about 0.2% to about 1.0% (w/v) of the compound of Formula II or about 0.1% to 1.2% (w/v) of Compound-I comprising the formulation of the current disclosure is administered once daily (QD) or twice daily (BID) to one eye or both eyes of a subject for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, the compound of Formula II or Compound-I is complexed with a complexing agent, e.g., cyclodextrin (e.g., Kleptose® HPB (%)) in a ratio of about 1:8, wherein about 2% to 13% (w/v) cyclodextrin (e.g. Kleptose® HPB (%)) is added to the formulation. The formulation further comprises about 0.1% to about 0.2% buffer, for example, 10 mM phosphate buffer. The desired osmolality of the formulation is about 200 to about 300 mOsm, obtained by adding sufficient amount to obtain the osmolality with a salt, for example, sodium chloride. The pH of the formulation is about 6.0 at or below 40°C. The dosage regimen for between 1 and 90 days or for more than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) may be any of the regimens involving consecutive or alternating days described in the paragraph above. In some embodiments, the compound of Formula II or Compound-I formulation further comprises a second active agent. In some embodiments, the compound of Formula II or Compound-I formulation is administered with a second active agent.

[279] The second active agent, e.g., nicotinic acid, nicotinamide, or vitamin K or a combination thereof, at a specific dose modulates (e.g., activates) directly or indirectly, ErbB receptor tyrosine kinase (RTK) including, EGFR , HER2, HER3, and HER4. Modulation (e.g., activation) of one or more ErbB RTKs by the formulation of the present application, at a specific, simultaneous dose, has a synergistic effect, and is effective in preventing or treating corneal tears or diseases, (e.g., corneal ulcers, epithelial corneal defects, keratitis, etc.) in the anterior segment of the eye.

[280] The methods of the present application are combined with standard of care, including but not limited to laser treatment and treatment with injectable anti-neovascular agents.

Particle Compositions and Formulations Understanding Particle Compositions

[281] The present application relates to a pharmaceutical composition comprising particles of an active agent of the present application (e.g., a first active agent (e.g., Formula II or Compound-I) and/or a second active agent), or a pharmaceutically acceptable salt thereof, wherein the particles have an average diameter between 100 nm and 100 µm. In some embodiments, the particles have an average diameter between 20 µm and 90 µm. In some embodiments, the particles have an average diameter between 20 µm and 80 µm. In some embodiments, the particles have an average diameter between 20 µm and 70 µm. In some embodiments, the particles have an average diameter between 30 µm and 70 µm. In some embodiments, the particles have an average diameter between 30 µm and 60 µm. In some embodiments, the particles have an average diameter between 30 µm and 50 µm. In some embodiments, the particles have an average diameter between 30 µm and 40 µm. In some embodiments, the particles have an average diameter between 50 µm and 70 µm. In some embodiments, the particles have an average diameter between 50 µm and 60 µm. In some embodiments, the particles have an average diameter of at least 30 µm. In some embodiments, the particles have an average diameter of about 30 µm, about 35 µm, about 40 µm, about 45 µm, about 50 µm, about 55 µm, about 60 µm, about 65 µm, or about 70 µm. In some embodiments, the particles have an average diameter of about 30 µm, about 35 µm, about 50 µm, or about 60 µm. In some embodiments, the particles have an average diameter between 100 nm and 8 µm. In some embodiments, the particles have an average diameter between 100 nm and 200 nm. In some embodiments, the particles have an average diameter of no more than 150 nm. In some embodiments, the particles have an average diameter of about 150 nm, about 140 nm, about 130 nm, about 120 nm, about 110 nm, or about 100 nm. In other embodiments, the particles have an average diameter between 1 µm and 5 µm. In some embodiments, the particles have an average diameter between 2 µm and 4 µm. In some embodiments, the particles have an average diameter of about 1 µm, about 2 µm, about 3 µm, about 4 µm, or about 5 µm. In some embodiments, the particles have an average diameter of about 3 µm.

[282] In some embodiments, at least 90% of the particles have a diameter of 70 µm or less. In some embodiments, at least 90% of the particles have a diameter of 60 µm or less. In some embodiments, at least 90% of the particles have a diameter of 10 µm or less. In some embodiments, at least 90% of the particles have a diameter of 9 µm or less. In some embodiments, at least 90% of the particles have a diameter of 8 µm or less. In some embodiments, at least 90% of the particles have a diameter of 7 µm or less. In some embodiments, at least 90% of the particles have a diameter of 6 µm or less. In some embodiments, at least 90% of the particles have a diameter of 5 µm or less. In some embodiments, at least 90% of the particles have a diameter of 4 µm or less. In some embodiments, at least 90% of the particles have a diameter of 300 nm or less. In some embodiments, at least 90% of the particles have a diameter of 200 nm or less.

[283] In some embodiments, the pharmaceutical composition comprises particles of a first active agent (e.g., a first active agent (e.g., Formula II or Compound-I)) and particles of a second active agent (e.g., nicotinic acid, nicotinamide, vitamin K, or a combination of these). In some embodiments, the pharmaceutical composition comprises particles of a first active agent (e.g., a first active agent (e.g., Formula II or Compound-I)) and particles of vitamin K (e.g., menadione).

[284] In some embodiments, a first active agent (e.g., Formula II or Compound-I) and a second active agent (e.g., nicotinic acid, nicotinamide, vitamin K, or a combination thereof) are milled together in roller mill to form particles comprising the first active agent and the second active agent. In some embodiments, the first active agent and the second active agent are milled in a roller mill separately, and the particles comprising the first active agent and the particles comprising the second active agent are then mixed or milled together in a roller mill. In some embodiments, the first active agent or the second active agent is roller mill milled first, and the particles are then added to the other active agent for additional roller mill milling.

[285] In some embodiments, the pharmaceutical composition comprising particles of an active agent of the present application (for example, a first active agent or a second active agent) further comprises one or more excipients. The excipient may be selected from any suitable excipient known in the art, for example, for preparing an ophthalmic formulation. In some embodiments, the excipient is selected from Polysorbate (Tween) 80, Poloxamer (Pluronic) F-127, Hypromellose (Hydroxypropyl Methylcellulose or HPMC), Povidone (PVP K-29/32 or K-30), and Tyloxapol. and a combination of these. In some embodiments, the excipient is selected from HPMC, Tween 80, Pluronic F-127, and Tyloxapol, and a combination thereof.

[286] In some embodiments, particles comprising a first active agent (e.g., Formula II or Compound-I) and particles comprising a second active agent (e.g., nicotinic acid, nicotinamide, vitamin K, or a combination of these) comprise the same excipient(s).

[287] In some embodiments, the pharmaceutical composition comprising particles of an active agent of the present application (for example, a first active agent or a second active agent) further comprises a surfactant, such as benzalkonium chloride (BAC).

[288] In some embodiments, the pharmaceutical composition comprising particles of an active agent of the present application (e.g., a first active agent or a second active agent) further comprises an excipient (e.g., to enhance the bioavailability of the active agent ). In some embodiments, the excipient is hydroxyethyl cellulose (HEC). In some embodiments, the HEC is present in an amount of about 0.1% to about 1%, about 0.1% to about 0.9%, about 0.1% to about 0. .8%, about 0.1% to about 0.7%, about 0.1% to about 0.6%, about 0.1% to about 0.5%, about 0. 1% to about 0.4%, or about 0.1% to about 0.3%. In some embodiments, HEC is present in an amount of about 0.2% or about 0.3%.

[289] In some embodiments, the particles of the present application comprise Formula II (i.e., free base of Compound-I). In other embodiments, the particles of the present application comprise Compound-I.

[290] In some embodiments, the particles of the present application are prepared by grinding in a roller mill. Factors that may affect particle size include, but are not limited to, the use of the free base or a salt of the active agent (e.g., Compound-I vs. Formula II), the addition of an excipient, the roller speed during the grinding, the size of the grinding media, and the duration of grinding.

[291] In some embodiments, the particles of the present application are prepared without grinding in a roller mill.

[292] In some embodiments, particles of an active agent of the present application (e.g., a first active agent (e.g., Formula II or Compound-I) and/or a second active agent) are sterilized. In some embodiments, sterilization is conducted with gamma irradiation. In some embodiments, particles of an active agent of the present application (e.g., a first active agent (e.g., Formula II or Compound-I) and/or a second active agent) are stable after sterilization (e.g. , remaining substantially pure of any impurities or degradation products as a result of sterilization).

[293] The present application relates to a suspension formulation comprising a pharmaceutical composition, wherein the pharmaceutical composition comprises particles of an active agent of the present application (e.g., a first active agent (e.g., Formula II or Compound- I) and/or a second active agent), or a pharmaceutically acceptable salt thereof, as described herein.

[294] In some embodiments, the suspension formulation comprises particles of a first active agent (e.g., Formula II or Compound-I), wherein the first active agent is at a concentration of about 0.1 mg / mL to about 10 mg/mL. In some embodiments, the first active agent is at a concentration of about 0.2 mg/mL to about 10 mg/mL, about 0.5 mg/mL to about 10 mg/mL, about 1 mg/mL to about 10 mg/mL, about 2 mg/mL to about 9 mg/mL, about 2 mg/mL to about 8 mg/mL, about 3 mg/mL to about 8 mg /mL, about 3 mg/mL to about 7 mg/mL, about 3 mg/mL to about 6 mg/mL, about 4 mg/mL to about 6 mg/mL, or about 4 mg /mL to about 5 mg/mL of a first active agent (e.g., Formula II or Compound-I). In some embodiments, the first active agent is at a concentration of about 0.1 mg/mL, 0.3 mg/mL, 0.5 mg/mL, about 1 mg/mL, about 2 mg/mL, about 2 mg/mL, mL, about 3 mg/mL, about 4 mg/mL, about 6 mg/mL, or about 10 mg/mL. In some embodiments, the first active agent is at a concentration of about 1 mg/mL to about 4 mg/mL or about 2 mg/mL to about 4 mg/mL. In some embodiments, the first active agent is at a concentration of about 2 mg/mL or about 4 mg/mL.

[295] In some embodiments, the suspension formulation comprising particles of a first active agent (e.g., Formula II or Compound-I) further comprises a second active agent (e.g., nicotinic acid, nicotinamide, vitamin K, or a combination of these). In some embodiments, the second active agent is vitamin K (e.g., menadione). In some embodiments, the second active agent is present in an amount of less than 10 µM. In some embodiments, the second active agent is present in an amount of about 0.5 µM, about 0.6 µM, about 0.7 µM, about 0.8 µM, about 0.9 µM , about 1 µM, about 2 µM, about 3 µM, about 4 µM, about 5 µM, about 6 µM, about 7 µM, about 8 µM, or about 9 µM. In some embodiments, the second active agent is present in an amount of about 1 µM.

[296] In some embodiments, the suspension formulation comprising particles of a first active agent (e.g., Formula II or Compound-I) further comprises an excipient selected from Polysorbate (Tween) 80, Poloxamer (Pluronic) F-127 , Hypromellose (Hydroxypropyl Methylcellulose or HPMC), Povidone (PVP K-29/32 or K-30), and Tyloxapol, and a combination of these. In some embodiments, the excipient is Pluronic F-127, Tween 80, HPMC, or Tyloxapol, or a combination thereof. In some embodiments, the excipient is present at a concentration of about 0.01% to about 0.2%, about 0.01% to about 0.15%, about 0.01% to about from 0.12%, about 0.01% to about 0.1%, about 0.01% to about 0.09%, about 0.02% to about 0.09%, about 0.03% to about 0.09%, about 0.04% to about 0.09%, or about 0.04% to about 0.08%. In some embodiments, the suspension formulation comprises about 0.08% or about 0.04% Pluronic F-127. In some embodiments, the suspension formulation comprises about 0.08% HPMC. In other embodiments, the suspension formulation comprises about 0.04% tyloxapol.

[297] In some embodiments, the suspension formulation comprising particles of a first active agent (e.g., Formula II or Compound-I) further comprises a buffering agent. In some embodiments, the buffering agent is selected from phosphate buffers, borate buffers, citrate buffers, tartrate buffers, acetate buffers, amino acids, sodium acetate, sodium citrate, Tris buffers, and the like. In some embodiments, the buffering agent is Tris. In some embodiments, the buffering agent is present at a concentration of about 0.1% to about 2%, about 0.2% to about 1.8%, about 0.3% to about 1.6%, about 0.4% to about 1.4%, about 0.4% to about 1.2%, about 0.4% to about 1%, about 0.4 % to about 0.8%, about 0.4% to about 0.7%, or about 0.5% to about 0.7%. In some embodiments, the suspension formulation comprises about 0.6% Tris.

[298] In some embodiments, the suspension formulation comprising particles of a first active agent (e.g., Formula II or Compound-I) further comprises an osmolality adjusting reagent. In some embodiments, the osmolality adjustment reagent is glycerol. In some embodiments, the osmolality adjusting reagent is present in an amount of about 1% to about 10%, about 2% to about 10%, about 3% to about 10%, about 4% to about 10%, about 5% to about 10%, about 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, or about 2% to about 3%. In some embodiments, the suspension formulation comprises about 2% or about 2.5% glycerol.

[299] In some embodiments, the suspension formulation comprising particles of a first active agent (e.g., Formula II or Compound-I) has a pH of less than 7.5. In some embodiments the pH is between about 6.0 to about 7.0. In some embodiments the pH is about 6.0 or about 7.0.

[300] In some embodiments, the suspension formulation comprising particles of a first active agent (e.g., Formula II or Compound-I) further comprises hydroxyethyl cellulose (HEC). In some embodiments, the HEC is present in an amount of about 0.1% to about 1%, about 0.1% to about 0.9%, about 0.1% to about 0. .8%, about 0.1% to about 0.7%, about 0.1% to about 0.6%, about 0.1% to about 0.5%, about 0. 1% to about 0.4%, or about 0.1% to about 0.3%. In some embodiments, HEC is present in an amount of about 0.2% or about 0.3%.

[301] One of a first active agent, a second active agent, and one or more excipients described in the present application may be present at any concentration or level described herein in combination with the remainder of the first active agent, the second active agent, and one or more excipients described in the present application at any concentration or level described herein.

[302] In some embodiments, the formulation of the present application comprises a first active agent at about 0.1% to about 1.2% (or any intermediate range as described herein), Tris at about 0.4% % to about 0.8% (or any intermediate range as described herein), glycerol from about 2% to about 4% (or any intermediate range as described herein), HEC from about 0.1% to about 0.3% (or any intermediate range as described herein), and HPMC at about 0.04% to about 0.09% (or any intermediate range as described herein). In some embodiments, the first active agent is Formula II or Compound-I. In some embodiments, the formulation of the present application comprises Formula II or Compound-I at about 0.4%, Tris at about 0.6%, glycerol at about 2%, HEC at about 0.2%. , and HPMC at around 0.08%. In some embodiments, the formulation comprises particles of the first active agent wherein the particles have an average diameter between 30 µm and 60 µm. In some embodiments, the formulation comprises particles of the first active agent wherein the particles have an average diameter between 50 µm and 60 µm. In some embodiments, the formulation comprises particles of the first active agent wherein the particles have an average diameter of about 50 µm or 60 µm. In some embodiments, the formulation has a pH of less than 7.0. In some embodiments, the formulation has a pH of about 6.

[303] In some embodiments, the formulation of the present application comprises a first agent active at about 0.1% to about 1.2% (or any intermediate range as described herein), a second agent active at about 0.00001% to about 0.0001% (or any intermediate range as described herein), Tris at about 0.4% to about 0.8% (or any intermediate range as described herein), glycerol at about 2% to about 4% (or any intermediate range as described here), HEC at about 0.1% to about 0.3% (or any intermediate range as described herein), and HPMC at about 0.04 % to about 0.09% (or any intermediate range as described here). In some embodiments, the first active agent is Formula II or Compound-I. In some embodiments, the second active agent is vitamin K3 (e.g., menadione). In some embodiments, the formulation of the present application comprises Formula II or Compound-I at about 0.4%, vitamin K3 at about 0.000086%, Tris at about 0.6%, glycerol at about 2%. %, HEC at about 0.2%, and HPMC at about 0.08%. In some embodiments, the formulation comprises particles of the first active agent wherein the particles have an average diameter between 30 µm and 60 µm. In some embodiments, the formulation comprises particles of the first active agent wherein the particles have an average diameter between 50 µm and 60 µm. In some embodiments, the formulation comprises particles of the first active agent wherein the particles have an average diameter of about 50 µm or 60 µm. In some embodiments, the particle also comprises the second active agent. In some embodiments, the formulation has a pH of less than 7.0. In some embodiments, the formulation has a pH of about 6.

Indications and Treatment Methods

[304] Methods for treating diseases or conditions of the eye are disclosed. The disclosed methods relate to the treatment, prevention, or control of ocular neovascularization (NV), or treating a disease or condition that is related to the onset of NV by administering to a subject one or more of the disclosed compounds (e.g., a first active agent (e.g., a compound of Formulas I or II), and optionally a second active agent), and formulations thereof.

[305] One aspect of the disclosed method relates to treating or preventing NV by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent ( for example, vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof. One embodiment of this aspect relates to a method of treating NV by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and optionally b) one or more compatible carriers or excipients.

[306] The disclosed methods relate to preventing or controlling pathological ocular neovascularization (NV), or treating a disease or condition that is related to the onset of NV by administering to a subject one or more of the disclosed compounds of Formulas I or II, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and formulations thereof.

[307] Current embodiments provide for the use of a formulation of Compound-I or its free base (Formula II), and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof ), for the manufacture of a medicament to treat a subject with a vasculopathic or inflammatory disease of the posterior segment of the eye. These include, for example, diabetic retinopathy (including background diabetic retinopathy, proliferative diabetic retinopathy and diabetic macular edema); age-related macular degeneration (AMD) (including neovascular (wet/exudative) AMD, dry AMD, and Geographic Atrophy); pathological choroidal neovascularization (CNV) of any mechanism (i.e. high myopia, trauma, sickle cell anemia; ocular histoplasmosis, angioid lines, traumatic choroidal rupture, optic nerve drusen, and some retinal dystrophies); pathologic retinal neovascularization of any mechanism (i.e., sickle cell retinopathy, Eales disease, ocular ischemic syndrome, carotid-cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic occlusive arteriolitis; birdshot retinochoroidopathy, retinal vasculitis, sarcoidosis, or toxoplasmosis); uveitis; retinal vein occlusion (central or branch); eye trauma; surgery-induced edema; surgery-induced neovascularization; cystoid macular edema; ocular ischemia; retinopathy of prematurity; coat disease; sickle cell retinopathy and/or neovascular glaucoma. In one embodiment, the eye disease is AMD. In one embodiment, diseases of the eye arise from or are exacerbated by ocular angiogenesis and/or neovascularization.

[308] In one aspect of the present application the formulation is used in the treatment of age-related macular degeneration (AMD) (including neovascular (wet/exudative) AMD, dry AMD, and Geographic Atrophy). The solutions or suspensions are used in the treatment of neovascular (exudative or wet) AMD. In another embodiment, the solutions or suspensions are used to treat dry AMD. In yet another embodiment, solutions or suspensions are used to treat Geographic Atrophy.

[309] The formulation of the present application prevents, delays, or treats the onset of pathological choroidal neovascularization (CNV) from any mechanism (i.e. high myopia, trauma, sickle cell anemia; ocular histoplasmosis, angioid lines, traumatic choroidal rupture, nerve drusen optic, and some retinal dystrophies) in subjects.

[310] The formulation of the present application delays the onset, prevents the progression, or treats the formation of pathological choroidal neovascularization (CNV) beneath the neurosensory retina. The formulation of the present application is effective in treating CNV.

[311] One aspect of this method relates to treating or preventing ocular neovascularization by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts thereof, and optionally a second active agent. (e.g. vitamin K, nicotinic acid, or nicotinamide, or a combination of these). One embodiment of this aspect relates to a method of treating ocular edema and neovascularization by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and optionally b) one or more compatible carriers or excipients.

[312] The disclosed methods also relate to preventing or controlling ocular edema or treating a disease or condition that is related to the onset of ocular edema by administering to a subject one or more of the disclosed compounds of Formulas I or II and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof).

[313] One aspect of this method relates to treating or preventing ocular edema by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts thereof, and optionally a second active agent. (e.g. vitamin K, nicotinic acid, or nicotinamide, or a combination of these). One embodiment of this aspect relates to a method of treating ocular edema by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and optionally b) one or more compatible carriers or excipients.

[314] Another disclosed method relates to preventing or controlling retinal edema or retinal neovascularization or treating a disease or condition that is related to the onset of retinal edema or retinal neovascularization by administering to a subject one or more of the disclosed compounds of Formulas I or II and optionally a second active agent (for example, vitamin K, nicotinic acid, or nicotinamide, or a combination thereof). One aspect of this method relates to treating or preventing retinal edema or retinal neovascularization by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts thereof, and optionally a second active agent. (e.g. vitamin K, nicotinic acid, or nicotinamide, or a combination of these). One embodiment of this aspect relates to a method of treating retinal edema or retinal neovascularization by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts , and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and optionally b) one or more compatible carriers or excipients.

[315] Another embodiment of this aspect relates to a method for delaying or preventing the progression of non-proliferative retinopathy to proliferative retinopathy by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and optionally b) one or more carriers or compatible excipients.

[316] One aspect of the disclosed methods relates to diseases that are a direct or indirect result of diabetes, inter alia, diabetic macular edema and diabetic retinopathy. The diabetic ocular vasculature becomes unstable over time leading to conditions such as non-proliferative retinopathy, macular edema, and proliferative retinopathy. As fluid leaks into the center of the macula, the part of the eye where sharp, straight-ahead vision occurs, the buildup of fluid and associated protein begins to deposit on or under the macula. This results in swelling that causes the subject's central vision to gradually become distorted. This condition is referred to as “macular edema”. Another condition that can occur is non-proliferative retinopathy in which vascular changes, such as microaneurysms, outside the macular region of the eye can be seen. During proliferative DR, pathological new blood vessels grow from the retina into the vitreous body, where these abnormal vessels can alter retinal morphology in the macula, and/or hemorrhage into the vitreous humor and obscure the visual axis.

[317] Another disclosed method relates to the treatment, prevention or control of diabetic retinopathy or treatment of a disease or condition that is related to the onset of diabetic retinopathy by administering to a subject one or more of the disclosed compounds of Formulas I or II , and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof).

[318] One aspect of the disclosed method relates to treating or preventing diabetic retinopathy by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts thereof, and optionally a second agent active ingredient (e.g. vitamin K, nicotinic acid, or nicotinamide, or a combination of these). One embodiment of this aspect relates to a method of treating diabetic retinopathy by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and optionally b) one or more compatible carriers or excipients.

[319]Diabetic proliferative retinopathy is characterized by neovascularization. New blood vessels are fragile and are susceptible to bleeding. The result is scarring of the retina, as well as occlusion or complete blocking of the path of light through the eye due to the abnormal formation of new blood vessels. Typically subjects having diabetic macular edema are suffering from the non-proliferative stage of diabetic retinopathy; however, it is not uncommon for subjects to begin to manifest macular edema only at the beginning of the proliferative stage.

[320] Another disclosed method relates to preventing or controlling diabetic macular edema or treating a disease or condition that is related to the onset of diabetic macular edema by administering to a subject one or more of the disclosed compounds of Formulas I or II and optionally a second active agent (for example, vitamin K, nicotinic acid, or nicotinamide, or a combination thereof).

[321] One aspect of this method relates to treating or preventing diabetic macular edema by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent. (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), or formulations thereof. One embodiment of this aspect relates to a method of treating diabetic macular edema by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and optionally a second active agent (e.g., vitamin K, nicotinic acid, or nicotinamide, or a combination thereof), and/or formulations thereof, and b) one or more compatible carriers or excipients.

[322] Another aspect of the disclosed method relates to the treatment or prevention of NV and treatment and/or prevention of corneal tears or diseases by administering to a subject an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, a second active agent, and/or formulations thereof. One embodiment of this aspect relates to a method of treating NV and treating and/or preventing corneal tears or diseases by administering to a subject a composition of: a) an effective amount of one or more of the disclosed compounds of Formulas I or II, or pharmaceutically acceptable salts, and/or formulations thereof, b) a second active agent, and optionally c) one or more compatible carriers or excipients. In one embodiment, the corneal disruption or disease is caused by a compound of Formulas I or II.

Kits

[323] Kits of the disclosed compounds and compositions for drug release in a human, mammal, or cell are also disclosed. Kits may comprise one or more packaged unit doses of a composition comprising one or more compounds of Formulas I or II and one or more packaged unit doses of a second active agent to be delivered into a human, mammal, or cell. Unit dosage ampoules or multiple dose containers, in which the compounds of Formulas I or II or the second active agent to be released are packaged prior to use, may comprise a hermetically sealed container enclosing a quantity of the active agent or pharmaceutically acceptable salt , or formulation thereof, suitable for a pharmaceutically effective dose thereof, or multiples of an effective dose. Compounds can be packaged as a sterile formulation, and the hermetically sealed container is designed to preserve the sterility of the formulation until use.

[324] The kit of the present application has a single-use ophthalmic drop dispensing bottle for dispensing ophthalmic formulation. In an alternative embodiment, the kit of the present application has a multiple-use eye drop dispensing bottle. The multiple dose dispensing bottle has appropriate amount of anti-infective agent and/or preservative, for example without being limited to, 0.005% BAK. The ophthalmic dispenser of the present application has a top and a lid. The container of the present application has a semi-transparent LDPE ophthalmic dispensing bottle with an LDPE dropper tip and HDPE cap. The container may be of other type and shape as required and/or as used in the art.

[325] The following examples are illustrative, but not limiting, of the methods and compositions of the present application. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in therapy and which are obvious to those skilled in the art are within the spirit and scope of the embodiments.

General Methodology roller mill

[326]The horizontal roller mill consists of multiple motor-driven rollers contained within a metal housing. Individual containers placed between the rollers will rotate at an rpm determined by the speed of the rollers and the diameter of the container. The drug slurry consisting of API, and optionally stabilizers, water, and/or grinding media is added to the container before being placed between the rollers. The media used are granules of various sizes, for example from 800 microns to 3000 microns, and can be made with Yttria Zirconium. After grinding, the dispersion is separated from the media by transferring the contents to a centrifuge tube insert fitted with a mesh sieve. After centrifugation, for example at approximately 300 x G for approximately 5 minutes, the dispersion is collected below the mesh (which retained the media). Optical Microscopy (OM)

[327] The morphology and particle size distribution of compositions can be evaluated by optical microscopy, and photomicrographs of the particles can be taken, for example, using an Olympus BX51 system equipped with a 100x oil immersion objective (magnification 1000x). A calibration bar (from 1 µm to 100 µm) can be placed as a comparator on each photomicrograph. Particle Size Distribution (PSD)

[328] Particle size distribution can be analyzed using laser diffraction light scattering, for example with a Horiba LA-950 V2. Generic assumptions are made on fit conditions and the refractive index value. Distributions are based on volume. The sample density is adjusted to a generic percentage transmission range in the blue LED light source. A small sample cell (filled with water) can be used instead of the flow through cell, to minimize the amount of sample.

Calculation

[329] The following equation was used to determine the volume of diluent (50:50 methanol: water) required in order to prepare samples of the choroid, retina, and cornea at a specified tissue concentration. Where: Tissue Conc = Desired tissue concentration (mg/ml) Tissue Mass = Tissue mass (mg) Diluent Vol = Volume of diluent (50:50 methanol:water) (ml) Tissue Vol = Tissue volume (ml) , assuming a density of 1.0 g/ml

[330] To calculate the concentration in ng/g (ng of drug/g of tissue), the following equation was used: Where: Conc.ng/g = Calculated concentration (ng of drug/g of tissue) Conc.ng/ml = Calculated concentration (ng of drug/ml of homogenate) Vol.Total = Total volume of tissue homogenate (ml) MassTissue = Fabric mass (g)

[331]To then calculate the drug concentration in tissue* in µM (µmol of drug/volume of tissue), the following equation was used, assuming a tissue density of 1 g/ml: Where: Conc.µM = Calculated concentration (µmol of drug/volume of tissue) Conc.ng/g = Calculated concentration (ng of drug/g of tissue) MW = Molecular weight (g/mol) *Tissue = choroid, retina, or cornea

[332]To calculate the drug concentration in fluid** in µM (µmol of drug/volume of fluid), the following equation was used: Where: Conc.µM = Calculated concentration (µmol of drug/volume of fluid) Conc.ng/ml = Calculated concentration (ng of drug/ml of fluid) MW = Molecular weight (g/mol) **Fluid = plasma

[333]Note: Any sample concentrations <LLOQ within the body of this report have been dropped or excluded for the calculation of reported statistical values (mean, standard deviation, and percent coefficient of variation). Draize Ocular Irritation Counting System for cornea, iris, and connective tissue Draize et al. (1944) Methods for the study of irritation and toxicity of substances applied topically to the skim and mucous membranes Counts of 0 are assigned to each parameter if the cornea, iris, or conjunctiva are normal.

EXAMPLES

[334] Compound-I is a potent and selective small molecule inhibitor of VEGFR-2, along with other proangiogenic RTKs such as FGF receptors (FGFR-1 to 3), Tie-2, and ephrin B4 receptor (EPHB-4). Compound-I has been shown to inhibit the phosphorylation of specific RTKs, endothelial cell proliferation, and pathological angiogenesis following systemic administration in murine cornea and rat plaque growth models, as well as the growth of human tumor xenografts in mice. athymic. With respect to potential ophthalmic indications, the Examples of the present application described below demonstrated that topical ocular delivery of Compound-I provided significant inhibition of pathological retinal and choroidal neovascularization in clinically relevant rodent models. A summary of these data follows.

[335] The following studies were conducted to measure the effect of the disclosed compounds on vascular leakage and neovascularization of retinal tissue.

EXAMPLE 1 Primary Pharmacodynamics

[336]Pharmacology of In Vitro Efficacy of Compound-I. Compound-I potently inhibits the tyrosine kinase activity of vascular endothelial growth factor receptor-2 (VEGF-2), as well as a selected subset of other proangiogenic RTKs, during several in vitro assays. Specifically, Compound-I blocked VEGF-stimulated VEGFR-2 phosphorylation in whole cells along with the proliferation of cultured endothelial cells. Compound-I inhibited the recombinant tyrosine kinase activity of VEGFR-2 and FGFR-2 with a 50% inhibitory concentration (IC50) of 10.55 nM (6 ng/ml) and 8.79 nM (5 ng/ml ), respectively; and inhibited VEGFR-2 autophosphorylation in intact cells with an IC50 = 5.27 nM (3 ng/ml). This inhibition was selective versus many other tyrosine kinases, for example, the IC50 of VEGFR-2 was approximately 500x and 1000x lower than those for the epidermal growth factor receptor (EGFR) and insulin receptor tyrosine kinases ( IR), respectively (see Table 2).

[337] When using a 10-point titration curve that ranged from 257 to 5000 nM (146 to 2845 ng/ml), Compound-I exhibited potent inhibition of tyrosine kinase activity for several proangiogenic growth factor receptors , as evidenced by an IC50 < 100 nM (56.89 ng/ml) (see Table 3). The IC50s for this select group of kinases were as follows: recombinant KDR (human isoform of VEGFR-2) = 1.27 nM (0.72 ng/ml), Tie-2 = 10.10 nM (5.75 ng/ml) ml), and FGFRs 1 to 3 = 8.50 nM (4.84 ng/ml), 3.08 nM (1.75 ng/ml), and 33.9 nM (19.29 ng/ml), respectively . The compound also blocked the other high-affinity VEGF receptor, VEGFR-1/Flt-1, but with lower potency: IC50 = 122 nM (69.41 ng/ml).

[338]While inhibition of VEGFR appears to be essential for reducing vascular permeability and preventing further neovascular growth, simultaneous inhibition of VEGF signaling with inhibition of other growth factor signaling pathways (e.g., PDGF and angiopoietins/Tie2 ) may be linked to unique therapeutic effects. The therapeutic effects of broader inhibition of signaling pathways may contribute to the regression of newly established pathological vessels in the posterior segment of the eye.

[339]300 nM (170.67 ng/ml) of Compound-I completely inhibited VEGFR-2 kinase function (see Table 4) and provided substantial blockade of a similar set of proangiogenic growth factor receptors, including FGFRs-1 to 3, Tie-2, and EphB-4. An unexpected finding was that the 300 nM concentration was able to completely inhibit VEGFR-2 kinase function. This concentration falls within the typical range found in the central choroid and retina following five days of topical ocular release in rabbits and dogs. Overview of Drug Substance and Drug Product

[340]Drug substance: The active pharmaceutical ingredient (API), Formula II Hydrochloride (Compound-I, CP-547,632 - 01), is a small molecule of a single polymorph. The API substance is consistently manufactured in purity that exceeds 99.7%. Any impurity in drug substance > 0.15% is appropriately qualified in toxicology studies and the current specification for new individual unknown impurities is adjusted to NMT 0.2%. The final drug substance and drug product are analyzed using standard methods.

[341] Drug Product: Compound-I ophthalmic formulations for clinical studies were manufactured at dosage concentrations between 0.05% and 1.0% (as Compound-I). The concentrations used for LPG batches are 0% (placebo), 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.8%, and 1.0% Compound-I. Compound-I ophthalmic formulations (solutions or suspensions) are used for once-daily topical administration to the eye in clinical testing. In addition to the active ingredient, the pharmaceutical product may also contain 0.005% BAK as a preservative, purified water as a vehicle, and is pH adjusted with sodium hydroxide to pH 6.0.

EXAMPLE 2 Sodium Phosphate Based Gel Drop

[342] The ophthalmic benefits of Compound-I in a sodium phosphate-based formulation (listed in table 6) result from the self-gelling properties of the API in buffers, such as sodium phosphate. The spontaneous formation of a self-forming, thixotropic gel of Compound-I from a clear solution was formed by increasing the concentration of API in sodium phosphate. This gel initially appeared clear and then demonstrated increased thickness/viscosity at higher API concentrations, as well as becoming increasingly more opaque, i.e., cloudy. Once the API concentration in the phosphate buffer reached the supersaturated state, insoluble Compound-I particulates were also observed within the gel.

[343] Applying a gel with increased viscosity to the surface of the eye increases the time it remains on the cornea. The increased residence time on the cornea in turn facilitates ocular absorption of the drug. As a result, intraocular drug concentrations of viscous gels increase compared to non-viscous formulations such as aqueous solutions. One way to increase viscosity is to use various viscosity enhancing excipients, for example, carboxymethyl cellulose, which in effect achieves increased intraocular absorption of different drug substances following topical ocular administration. In this study, however, a thixotropic gel of Compound-I was unexpectedly formed in the absence of any viscosity-enhancing excipient. for example, when Compound-I was dissolved in a simple buffer, such as sodium phosphate, a thixotropic gel was formed. The thixotropic gel, which was formed without any viscosity enhancing excipient, was formulated as a Gel Drop in this Example.

[344] Compound-I Gel Drop was applied to the eyes of Dutch girdle rabbits. Compound-I Gel Drops were administered to Dutch girdle rabbits for 4 or 5 consecutive days, dosing three times a day. Compound-I concentrations in target tissues were measured within 1 hour of the last administered dose. The release of Compound-I to posterior segment tissues was dose-dependent and dose-frequency dependent.

[345]The Gel Drop formulations (listed in table 6) differed in several aspects, such as API concentration, sodium phosphate concentration, presence or absence of tonicity agents (glycerin) or preservative (benzalkonium chloride/ BAK), solubilizing surfactants (polysorbate 80, tyloxapol, and/or poloxamer), and pH.

EXAMPLE 3 Tromethamine-based suspension

[346] Compound-I (about 1 mg/ml to about 10 mg/ml) in a tromethamine-based formulation formed a suspension. The suspension of Compound-I in a tromethamine-based formulation had >95% of the active drug substance in an insoluble form. This characteristic is distinguishable from the soluble or semi-soluble states of Compound-I in the Gel Drop (the Gel Drop (gel) is not a fully soluble state as the concentration of the active agent increases) or in a Cyclodextrin-based formulation. The tromethamine-based formulations of Compound-I showed increased turbidity with increasing active agent concentration. Administration of a topical drop of Compound-I suspension to the eye (which is a combination of soluble and insoluble active agent components) was expected to provide unique benefits relevant to both safety/tolerability and efficacy.

[347] The tromethamine-based suspension of Compound-I was administered to Dutch girdle rabbits for 4 or 5 consecutive days dosing three times a day. Ocular tissue and plasma concentrations of Compound-I were measured within 1 hour of the last administered dose. Compound-I in the tromethamine-based suspension delivered concentrations to target tissues between 10 and 1000x the cellular IC50 for the various pro-angiogenic RTKs. See Table 7.

[348] The corneal safety and tolerability of topical Compound-I was a direct consequence of the amount of soluble (as opposed to insoluble) active agent applied to the corneal surface, and the resulting concentration in corneal tissue. Animals that received topical ocular administration of the tromethamine-based suspension were able to tolerate up to a higher level of concentration of the active agent in the formulation, when compared to equimolar formulations of the Gel Drop based on sodium phosphate. Results obtained from both Dutch girdle rabbits and beagle dogs suggested that ocular side effects, such as discomfort and inflammation and in some cases, corneal thinning, were more consistently observed when the corneal concentration of Compound-I exceeded 100 µM.

[349] Administration of the tromethamine-based suspension had the unexpected effect on the ocular bioavailability of Compound-I in the posterior segment. The ocular bioavailability of Compound-I in the posterior segment was observed to be directly proportional to the total amount of drug administered (insoluble plus soluble, see Table 7). Although insoluble drug particulates were not readily available to anterior segment tissues; The inherent and unique physicochemical properties of Compound-I allowed both insoluble and soluble components to gain entry into posterior segment tissues such as the choroid and retina. Consequently, even higher drug concentrations than those obtained with Gel Drop formulations containing equivalent amounts of the active agent were obtained with the tromethamine-based suspension. Thus, the tromethamine-based suspension provided: a) improved corneal tolerability and b) increased bioavailability to the posterior segment, particularly to the choroid, the primary target tissue for treating neovascular (wet) AMD.

EXAMPLE 4 Cyclodextrin-based solution

[350] Cyclodextrins, which are cyclic oligosaccharides composed of six to eight dextrose units (a-, ß-, and Y—CDs) linked through one to four bonds, are well known for their ability to act as a solubilizer for relatively insoluble drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol., 36: 30 - 42 (2008).

[351] A clinical formulation of ophthalmic solution of Compound-I or its free base in 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD, KLEPTOSE® HPB) in a molar ratio equal to or greater than 1:6 or Ether sulfobutflic-ß-cyclodextrin (SBE-ß-CD, CAPTISOL®) in ß in a ratio equal to or greater than 1:2 provided solubility with clinical dose concentrations of 0.1 to 1.0% of Compound-I.

[352] Cyclodextrin-based solutions of Compound-I or its free base not only improved the solubility of the active agent in a uniform solution, but, during topical ocular administration, also had a new and previously unobserved therapeutic index feature of the compound. significantly increased active agent in the posterior segment of the eye. The solution reduced exposure to the anterior segment, thereby providing increased concentration of the active ingredient in the solution and increased release frequency, which maintained high concentrations in the posterior segment. Both of these beneficial characteristics are related to the known property of cyclodextrin to form hydrophilic complexes with hydrophobic drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol., 36: 30 - 42 (2008). When formulated with Compound-I or its free base, cyclodextrin formed a clear, colorless solution and exhibited viscosity like that of water. Following topical ocular administration, the compound-I/cyclodextrin complex appeared to be pharmacologically inactive and metabolically inert. The Compound-I/cyclodextrin complex conferred corneal tolerability until the cyclodextrin spontaneously dissociated from the active agent, thus making a high concentration of Compound-I available at its intended site of action in the posterior segment of the eye, e.g., choroid and retina.

[353] During topical ocular dosing studies lasting 1 to 30 days in Dutch girdle rabbits, cyclodextrin-based solutions of Compound-I demonstrated dramatically decreased corneal exposures compared to Gel Drop formulations (see Example 2). at similar drug concentrations. The use of cyclodextrin-based solutions of Compound-I provided an approximate 10x reduction in corneal concentrations when compared to dosing with equimolar Gel Drop formulations. Consequently, after 30 days of topical ocular dosing of 0.6% Compound-I as a cyclodextrin-based solution, none of the unexpected findings were attributed to the test article or vehicle. The cyclodextrin-based solution of Compound-I also achieved equal or significantly higher drug concentrations within posterior segment target tissues, such as the central choroid and central retina. The combined effects of decreasing corneal drug exposure to avoid poor ocular tolerability, while increasing posterior segment bioavailability to increase RTK inhibition, can significantly increase the therapeutic index and corresponding benefit(s). experienced by patients.

[354] For both suspension-based formulations (see Example 3) and cyclodextrin formulations, the therapeutic window is expanded due to significantly reduced exposure (10 to 100X or 1 to 2 log reduction). Reduced exposure improves corneal safety/tolerability, which allows higher concentrations or dosing frequency of Compound-I to be topically administered. The higher concentrations allow the drug to reach higher concentrations in the target fundus tissue, which improves the therapeutic efficacy of Compound-I.

[355] This study demonstrated that in rabbits and dogs, topical ocular dosing of ophthalmic gel drops was associated with high exposure of corneal tissue (>100 µM) and corresponding unwanted observations in the anterior segment, such as discomfort, corneal inflammation and conjunctival, corneal epithelial erosion and/or thinning and degeneration. In contrast, repeated topical ocular dosing of Compound-I ophthalmic solution produced corneal exposures that are approximately 5 to 10 times lower than an equimolar dose of ophthalmic gel drops, and are free from unwanted clinical or histopathological findings. Furthermore, topical ocular dosing with Compound-I ophthalmic solution achieved equal or higher target therapeutic exposure in the central choroid compared to an equimolar dose of the ophthalmic gel drop. Overall, the combination of decreased corneal exposure and corresponding improved ocular tolerability, while simultaneously maintaining or promoting drug release to posterior segment target tissues, along with improved physicochemical stability, will provide greater benefit to patients compared to the ophthalmic formulation of Gout. of Gel.

EXAMPLE 5 1 to 5 Day PK Results with Topical Ocular Compound-I in Cyclodextrin-Based Solutions

[356]Following topical ocular dose administration of various formulations and dosage regimens of Compound-I in Dutch girdle rabbits, ocular pharmacokinetics were investigated. Nine (9) different topical ocular formulations having three doses of Compound-I were administered once daily (q.d.) or twice daily (b.i.d.) for 1, 4, or 5 consecutive days. The study design (see Tables 10A-B) consisted of seventy-two (72) rabbits each receiving a 30 µL bilateral topical ocular dose of one of three (3) Compound-I formulations, or vehicle formulation, using a positive displacement pipette.

[357] The composition of each Compound-I formulation is described in table 8A. All doses were administered within ±1 hour of the scheduled dose time. On day 1, Groups 1, 2, 4 - 6, 8, 10, 11, 13, 15, and 17 received one dose (q.d.) for one (1) or four (4) days. On days 1 to 4, Groups 3, 7, 9, 12, 14, and 16 received b.i.d. dosing. approximately 8 hours apart at 7:00 AM and 3:00 PM for four (4) days. Animals in Groups 18 and 19 received bi.i.d dosing of vehicle-only formulations for five (5) consecutive days.

[358]Ocular sampling occurred for Group 1 at 0.5, 1, 2, 4, 8, or 24 hours post dose relative to the day 1 dose. Ocular sampling occurred for Groups 2 and 6 at 1 , 8, and 24 hours after the dose in relation to the morning dose of day 5. Ocular sampling for Groups 3, 7, 8, 11, and 13 occurred at 1 and 24 hours after the dose in relation to the morning dose of the day 5. Eye sampling for Groups 4, 9, 10, 12, 14, 15, 16, and 17 occurred at 1 hour post-dose relative to the morning dose on day 5. Group 5 eye sampling occurred at 0. 5, 1, 2, 4, 8, and 24 hours post dose relative to the day 1 dose. Animals in Groups 18 and 19 were followed for clinical observations only for five (5) days.

[359] Samples of aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid were collected. Aqueous humor, cornea, central retina, and central choroid samples were then evaluated. Peripheral retinal and peripheral choroidal samples were evaluated only for Groups 1 to 8, 12, 14, and 16 per study protocol.

[360] Samples of rabbit aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid were analyzed. Calibration curves were prepared in control matrix to determine the concentration of Compound-I in the various tissues.

EXAMPLE 6 PK Results on Day 5 with Topical Ocular Compound-I in Cyclodextrin-Based Solutions

[361] Following ocular dose administration of various dose regimens of Compound-I topical ocular solutions containing hydroxypropyl-ß-cyclodextrin (“HDßCD”) in Dutch girdle rabbits, ocular pharmacokinetics were calculated. Nine (9) different topical ocular solutions having four different doses of Compound-I were administered once daily (q.d.) or twice daily (b.i.d.) for 4 or 5 consecutive days. The study design consisted of forty (40) rabbits each receiving a 30 µL bilateral topical ocular dose of one of four (4) dosing concentrations of Compound-I, using a positive displacement pipette.

[362]All doses were administered within ±1 hour of the scheduled dose time, except on day 1 for Groups 1, 4, and 10. The first dose on day 1 was administered at 12:00 PM, and the second dose (for Groups 1 and 10) was administered approximately 4 hours later. This was due to the late arrival of the formulations. All other dosing for these groups was as scheduled. On day 1, Groups 4 to 8, and 11 to 12 received one dose (q.d.) for four (4) days. On days 1 through 4, Groups 1 through 3, and 9 through 10 received b.i.d. dosing. approximately 8 hours apart over four (4) days.

[363] Eye sampling occurred one hour after the first daily dose on day 5 for all groups except Groups 5b and 6b, where eye sampling occurred 24 hours after the first daily dose on day 4.

[364]Blood samples for plasma collection were obtained exactly before scheduled euthanasia for all animals. Samples of aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid were collected. Corneal, central retina, and central choroid samples from groups 1 to 12 were evaluated. aqueous humor, peripheral retina, and peripheral choroid samples were not evaluated.

EXAMPLE 7 Compound-I Concentrations (in µM) in Various Ocular Fluids and Tissues

[365] Compound-I ocular solution comprising cyclodextrin was prepared and tested on different groups of animals. During topical ocular administration of a 0.4% (4 mg/ml) solution of Compound-I and cyclodextrin, the concentration of the active agent was measured in various tissues and fluids of the eye. The average concentration of Compound-I was measured in the central choroid, central retina, aqueous humor, and cornea. Compound-I was in a solution (0.4% or 4 mg/ml) with 8.41% KLEPTOSE® and 0.142% phosphate buffer; 8.9% KLEPTOSE® HPB and 0.142% phosphate; 4.88% CAPTISOL® and 0.142% phosphate; or 4.88% CAPTISOL® and 0.122% phosphate. See Table 10A-B.

[366]The concentration in the central choroid of Compound-I was between 0.259 µM and 0.769 µM. See Table 10A. The central retinal concentration of Compound-I was between 0.0531 µM and 0.124 µM. See Table 10A. The aqueous humor concentration of Compound-I was between 0.00313 µM and 0.00656 µM. See Table 10A. And the corneal concentration of Compound-I was 6.49 µM to 30 µM. See Table 10A. The cyclodextrins used to prepare the solutions were KLEPTOSE® HPB or CAPTISOL®. See Table 10B.

EXAMPLE 8 Ocular Toxicology Studies

[367]Dose-limiting ocular toxicity was characterized by corneal and conjunctival findings in Dutch girdle rabbits and beagle dogs. These ocular findings from repeat dose toxicology studies with Compound-I ophthalmic gel drops were substantiated during clinical and histopathological ophthalmic evaluations and were limited to conjunctival hyperemia, chemosis, congestion, and suppuration, corneal opacification and epithelial erosion, and keratoconjunctivitis. None of the unwanted changes involving deeper structures of the eye (iris, lens, ciliary body, retina, choroid, sclera) or the optic nerve were observed. Retinal function was normal in all groups treated with the test article and vehicle during full-field electroretinograms performed in rabbits.

[368] The objectives for the exploratory ocular toxicology studies were to identify: a) a topical ocular formulation that was well tolerated and b) one that could achieve the targeted therapeutic concentrations of Compound-I in the central choroid.

[369] Mean ocular tissue concentrations of Compound-I following twice-daily topical dosing with 0.3% Compound-I gel drop ophthalmic formulations with and without benzylalkonium chloride were highest in the cornea (236 to 260 µM) >> peripheral choroid (2.79 to 4.10 µM), central choroid (0.340 to 0.496 µM), peripheral retina (0.150 to 0.309 µM) and aqueous humor (0.0197 to 0.0395 µM ) in Dutch girdle rabbits.

[370]Tris-based suspension formulations were well tolerated, where ophthalmic clinical examinations revealed a notable absence of corneal findings with only a few sporadic incidences of mild conjunctivitis in Dutch girdle rabbits. Furthermore, the eyes of animals that received 0.3% w/v Tris-based suspension of Compound-I twice daily for 30 days were considered normal during microscopic evaluations. Mean ocular tissue concentrations of Compound-I, evaluated at 1 hour ± 15 minutes after the first daily topical ocular dose on day 30 for twice daily topical dosing with 0.3% Tris-based Suspensions of Compound-I I with and without benzylalkonium chloride, were higher in the cornea (2.69 to 3.10 µM) >> peripheral choroid (0.781 to 1.21 µM), central choroid (0.303 to 0.319 µM), peripheral retina (0. 0819 to 0.0868 µM), central retina (0.0495 to 0.0592 µM), and aqueous humor (0.00127 to 0.00145 µM).

[371] Cyclodextrin-based solutions, using hydroxypropyl-beta-cyclodextrin (HP-ß-CD, KLEPTOSE® HPB), were well tolerated when administered topically for 30 days, twice daily at 0.1% Compound-I (2.1% HP-ß-CD), twice daily at 0.2% Compound-I (4.21% HP-ß-CD), once or twice daily at 0.4 % Compound-I (8.41% HP- ß-CD), and once or twice daily at 0.6% Compound-I (up to 12.62% HP- ß-CD) in rabbits Dutch belts. Furthermore, in a similar repeated dosing study, cyclodextrin-based solutions of 0.4% w/v of Compound-I in Kleptose® HPB, Kleptose® HP, or Captisol® were well tolerated when dosed twice daily for 24 days. No overt ocular toxicity related to Compound-I or vehicle treatment was discovered during clinical ophthalmic or microscopic examinations in each study.

[372]In the day 24 study, ocular tissue concentrations from eyes treated with cyclodextrin-based solutions of Compound-I were evaluated at 1 hour ± 15 minutes after the first daily topical ocular dose on day 24. high in the cornea (6.49 to 30 µM) >> centrally perforated choroid (0.212 to 0.769 µM) > centrally perforated retina (0.0531 to 0.124) > aqueous humor (0.002 - 0.007).

[373] In summary, dose-limiting corneal toxicity was observed with ophthalmic Compound-I Gel Drop formulations. The ophthalmic Gel Drop makes corneal Compound-I concentrations five times to fifteen times higher compared to the cyclodextrin-based solution, and corneal Compound-I concentrations fifty times to hundreds of times higher compared to suspensions. based on Tris. Tris-based suspensions of Compound-I and cyclodextrin-based solutions were well tolerated with no evidence of overt ocular toxicity. Dose levels that were well tolerated for the cyclodextrin-based solutions or Tris-based suspensions of Compound-I when administered once or twice daily ranged from about 0.005% to about 5.0% w/v. for at least 30 days. The cyclodextrin-based solutions also provided the highest Compound-I concentrations in the central choroid when using equimolar doses of the three formulations, and met or exceeded target therapeutic concentrations.

EXAMPLE 9 Phase I Protocol for Dose Escalation Study in Patients With Neovascular AMD

[374]The Phase I study is a one-week, open-label, dose-escalation, multicenter trial designed to evaluate the safety, tolerability, and pharmacokinetics of topical ocular administration of Compound-I in patients with degeneration. age-related macular neovascular (AMD). Up to 60 patients in total are treated once to twice daily with topical ocular dosing of Compound-I ophthalmic solution for three months, where three arms of dose escalation monotherapy and one arm of adjunctive therapy using a single intravitreal injection of Lucentis® plus the maximally tolerated monotherapy dose are planned (15 patients per treatment arm). Patients who meet pre-specified vision and CNV lesion criteria confirmed by an independent reading center are simultaneously allowed to discontinue topical ocular dosing and receive standard of care treatment.

[375] Compound-I ophthalmic solution for clinical studies is manufactured in at least 3 dosage concentrations, ranging from 0.1% to 1.0% (as Compound-I). The concentrations for the GLP lots are 0% (placebo), 0.1%, 0.3%, 0.6%, and 1.0% Compound-I HCl. Up to 2 to 3 times incremental doses (steps of approximately % log unit) are administered to subsequent groups.

EXAMPLE 10

[376]A homogeneous “non-gel”, “non-viscous” solution ophthalmic topical formulation that is both physically and chemically stable at drug concentrations of 0.1 to 1.0% (1 to 10 mg/ml) was prepared measuring the concentration of compound-I, concentration of cyclodextrin complexing agent, pH, and tonicity, the solubility and stability of Compound-I. An adequate buffering system prevents the pH from fluctuating in stability at concentrations less than 1 mg/ml. Both phosphate and Trometamol (Tris) have been evaluated as buffering agents. Sodium chloride was used to adjust tonicity.

[377]Product quality attributes are shown in table 14. Table 14: Product Quality Attributes

Formulation Preparation

[378] The formulations outlined in tables 12 and 13 were prepared using the general procedure listed.

[379] The formulation was prepared to volume with water for injection and stirred for 30 minutes at 500 rpm. The final pH was checked and adjusted with NaOH or HCl to the target range. Aliquots of approximately 5 ml are directly filtered into 5 ml semi-transparent LDPE bottles under continuous stirring at constant speed with the help of a Watson Marlow Pumpsil D probe, adapted to a Flexicon filler and connected to the 0.2 PVDF capsule filter. micron. Samples are stored at 2 to 8 °C until all sample preparation is complete. All samples will then be submitted to analytical for storage and testing.

EXAMPLE 11 EGFR tyrosine phosphorylation assay in corneal epithelial cells (hTCEpi cells) to determine the EGFR activity of compounds of Formulas I or II

[380] An EGFR tyrosine assay in corneal epithelial cells was performed to determine whether higher concentrations of EGF could overcome the inhibition of EGFR kinase activity by a compound of Formulas I or II.

[381] 35 mm dishes of hTCEpi cells were serum starved and then pretreated with different concentrations of a compound of Formulas I or II, e.g., Compound-I, or a control, AG1478 (a EGFR), for 30 minutes, followed by treatment with EGF (10, 50, 100 ng/ml) for 15 minutes. Six concentrations of a compound of Formulas I or II, e.g., 0.1 µM, 3 µM, 10 µM, 30 µM, 100 µM, or 1 µM AG1478 as a control, were used for each concentration of EGF (10, 50 , 100 ng/ml). Additional controls conducted were without any compounds of Formulas I or II or any EGF. All experiments were repeated three times. The cells were then harvested and immunostained for the determination of phosphorylated EGFR (tyrosine 1068 and tyrosine 1045) and total EGFR.

[382]EGFR phosphorylation serves as a readout of receptor activity. As shown in Fig. 1, high concentrations of a compound of Formulas I or II were able to inhibit ligand-stimulated EGFR tyrosine phosphorylation, and increasing concentrations of a compound of Formulas I or II decreased EGFR tyrosine phosphorylation in both the locals. Increasing EGF concentrations overcomes EGFR inhibition and corneal adversity due to the compound of Formulas I or II (Fig. 1). The specific EGFR inhibitor, AG1478, has an IC50 = 3 nM, and 1 µM AG1478 completely blocked EGFR phosphorylation. It is estimated that 100 µM of a compound of Formulas I or II inhibits EGFR phosphorylation by ~75 to 80%.

EXAMPLE 12 Assay of EGFR tyrosine phosphorylation in corneal epithelial cells (hTCEpi cells) with compounds of Formulas I or II, and a second active agent

[383] An EGFR tyrosine assay in corneal epithelial cells was performed to determine whether vitamin K, nicotinic acid, or nicotinamide can prevent compounds of Formulas I or II from inhibiting EGFR and whether there is an increase in EGFR activity.

[384]35 mm dishes of hTCEpi cells were serum starved and then pretreated with vitamin K (50 µM), nicotinic acid (10 µM), or nicotinamide (10 µM) for 4 hours, followed by treatment with different concentrations of a compound of Formulas I or II, for example, Compound-I, for 30 minutes. The cells were then incubated with EGF (50 ng/ml) for 15 minutes. Cells were harvested and cell lysates were immunoblotted to determine the concentration of phosphorylated EGFR (tyrosine 1068 and tyrosine 1045) and total EGFR. As shown in Fig. 2, vitamin K, nicotinic acid, or nicotinamide overcome corneal adversity mediated by the compound of Formulas I or II (EGFR inhibition). Furthermore, Fig. 2 shows that vitamin K3 (menadione) most effectively attenuated the compound of Formulas I or II-mediated inhibition of EGFR tyrosine phosphorylation.

EXAMPLE 13 Determination of cell migration/proliferation (in vitro wound healing) in corneal epithelial cells (hTCEpi cells): effects of compounds of Formulas I or II and EGF

[385] Determination of cell migration/proliferation in corneal epithelial cells was performed to determine cell migration/proliferation in the presence of varying concentrations of a compound of Formulas I or II and EGF.

[386] hTCEpi cells were plated with silicone plugs. The cells were then serum starved and pretreated with varying concentrations of a compound of Formulas I or II or a control compound, AG1478, for 30 minutes. The silicone plugs were then removed to create the acellular area. Cells were photographed and then treated with EGF (10, 50, or 100 ng/ml) or VEGF (10 ng/ml) together with a compound of Formulas I or II or AG1478 for 16 hours. The cells were photographed again and cell migration was quantified from the photomicrographs. All experiments were repeated three times.

[387] Four concentrations of a compound of Formulas I or II (0.3 µM, 10 µM, 30 µM) or 3.2 µM of AG1478 (an EGFR kinase inhibitor) were used for each concentration of EGF or VEGF. . Additional controls conducted did not include any compounds of Formulas I or II and no EGF.

[388] The data allows determination of whether a compound of Formulas I or II impedes in vitro measurements of corneal epithelial wound healing (cell migration and proliferation) and whether higher levels of EGF overcome inhibition of receptor tyrosine kinase. As shown in Fig. 3A and 3B, a compound of Formulas I or II caused a dose-dependent inhibition of EGFR-mediated hTCEpi cell in vitro “wound healing”.

EXAMPLE 14 Determination of cell migration/proliferation (in vitro wound healing) in corneal epithelial cells (hTCEpi cells): effects of compounds of Formulas I or II and a second active agent

[389] Determination of cell migration/proliferation in corneal epithelial cells was performed to determine cell migration/proliferation in the presence of varying concentrations of a compound of Formulas I or II alone or together with vitamin K, nicotinic acid, or nicotinamide .

[390] hTCEpi cells were plated with silicone plugs. The cells were then serum starved and pretreated with varying concentrations of the vitamin K3 analogue, menadione, for 4 hours and supplemented with varying concentrations of a compound of Formulas I or II for 30 minutes. The tampons were removed. The cells were photographed and then treated with various concentrations of vitamin K, EGF, and/or a compound of Formulas I or II. The cells were photographed again and cell migration was quantified from the photomicrographs. Additional controls conducted did not include any compounds of Formulas I or II and no EGF. All experiments were repeated three times.

[391] As shown in Figs. 4A and 4B, the inhibitory effects of a compound of Formulas I or II on in vitro wound healing can be reversed by the addition of 0.3 µM menadione. There was a statistically significant difference in wound healing in vitro when 0.3 µM of a compound of Formulas I or II was combined with 0.3 µM of menadione versus 0.3 µM of a compound of Formulas I or II alone.

EXAMPLE 15 Determination of basal and EGF-mediated in vivo corneal wound healing: effects of compounds of Formulas I or II

[392] Determination of the effects of a compound of Formulas I or II on basal and ligand-stimulated rates of corneal wound healing was determined in C57/Bl mice.

[393]The corneas of 8-week-old C57/Bl mice were wounded (1.5 mm superficial epithelial wound) and then pretreated with a compound of Formulas I or II (0, 1.0 µM, 10 µM , 30 µM, or 50 µM), followed by the addition of EGF (0, 10, 100 ng/ml). As a control, AG1478 (an EGFR kinase inhibitor) was used in the absence and presence of 100 ng/ml EGF. Wound size was monitored by fluorescein staining and fluorescent photography over the course of 40 hours. Wound closure was quantified using Image J software.

[394] As shown in Figs. 5A, 5B, 5C, and 5D, in vivo corneal epithelial wound healing was reduced in the presence of a compound of Formulas I or II in a dose-dependent manner. Maximum inhibition of wound healing was observed with a compound concentration of Formulas I or II that was five times greater than that of AG1478. Pretreatment with a compound of Formulas I or II for 48 hours before wounding the cornea inhibited corneal epithelial wound healing at a lower concentration of a compound of Formulas I or II than without pretreatment.

EXAMPLE 16 Determination of basal and EGF-mediated corneal wound healing in vivo: effects of the compound of Formulas I or II and a second active agent

[395] Determination of the effects of a compound of Formulas I or II and/or a second active agent on corneal wound healing was determined in C57/Bl mice.

[396] The corneas of 8-week-old C57/Bl mice were pretreated with topical administration of vehicle, 0.3 µM menadione, 10 µM of a compound of Formulas I or II, 0.3 µM menadione and 10 µM of a compound of Formulas I or II 48 hours before the corneas were injured (1.5 mm superficial epithelial wound). Wound size was monitored by fluorescein staining and fluorescent photography. Wound closure was monitored over the course of 40 hours.

[397] As shown in Figs. 6A, 6B, and 6C, menadione (0.3 µM) reversed the effects of a compound of Formulas I or II (10 µM) in the pretreatment of corneal epithelial wound healing.

EXAMPLE 17 Effect of menadione on VEGFR inhibition by compounds of Formulas I or II

[398] Human retinal endothelial cells were treated with 0, 0.3, or 3 µM menadione for 4 hours, followed by 30 minutes of treatment with varying concentrations of a compound of Formulas I or II (1 nM, 10 nM, 100 nM, or 1 µM) in menadione. Cells were then stimulated with 0.5 nM (10 ng/ml) VEGF. The cells were lysed. Lysates were resolved by 7.5% SDS-PAGE, transferred to nitrocellulose, and immunoblotted for phosphorylated VEGFR2, total VEGFR2, or α-tubulin as a loading control. As shown in Fig. 7A, menadione did not change the IC50 of inhibition of VEGFR2 phosphorylation mediated by a compound of Formulas I or II.

EXAMPLE 18 Effect of menadione on the proliferation of primary human retinal microvascular endothelial cells

[399] Human retinal endothelial cells were pretreated with menadione for 4 hours, then with the indicated concentration of a compound of Formulas I or II for 30 minutes, and supplemented with 0.5 nM (10 ng/ml) VEGF for at night. Viable cells were quantified by the Alamar Blue Assay (Thermo Fisher). Data were plotted as the fold growth relative to cells treated with no VEGF or a compound of Formulas I or II, and are shown as the mean ± S.E.M. In Fig. 7B. As shown in Fig. 7B, menadione did not change the IC50 of VEGF-mediated inhibition of retinal endothelial cell proliferation mediated by a compound of Formulas I or II.

EXAMPLE 19 Determination of the mechanism by which menadione enhances EGFR Activity

[400] hTCEpi cells were pretreated with menadione (0, 0.3, or 3.0 µM) for 4 hours. The cells were then incubated with 8.0 nM (50 ng/ml) EGF for 0 to 3 hours. Cells were lysed and lysates were resolved by SDS-PAGE, and immunoblotted for phosphorylated EGFR (pY1068), total EGFR, or α-tubulin. As shown in Fig. 8, menadione treatment did not change basal EGFR phosphorylation or slow the rate of EGFR dephosphorylation. 0.3 mM menadione decreased the rate of EGFR degradation kinetics, indicating that menadione may maintain EGFR receptor signaling by decreasing the rate of receptor degradation kinetics or increasing the rate of new receptor synthesis.

EXAMPLE 20 - 1 Ocular pharmacokinetics of application formulations in Dutch girdle rabbits

[401] A non-GLP study was conducted to evaluate the ocular pharmacokinetics of a first active agent of the present application (e.g., Formula II or Compound-I), in suspension or solution, when administered once or twice daily, as a topical instillation, to both eyes of Dutch girdle rabbits for four days. The study design (Table 15 - 1) consisted of forty-five (45) Dutch girdle rabbits, each receiving a bilateral topical ocular dose of 30 µL. Administration was conducted according to the dosage regimen in table 15 - 1. Table 15 - 1: Study Design aA single dose will be administered on day 5. OQ - once a day; BID - twice a day

[402]Ocular sampling occurred one hour after the first daily dose on day 5 for all groups. Whole blood samples using K2EDTA as an anticoagulant were collected from all animals on day 5, 1 hour post dose. Whole blood was placed on ice until the samples were spun in a centrifuge to separate the plasma. Samples of aqueous humor, conjunctiva, cornea, central retina, peripheral retina, central choroid, and peripheral choroid were collected following necropsy. Samples were then frozen at -70°C or lower. All plasma, central retina, central choroid, and cornea samples from Groups 1 to 15 were evaluated.

[403]For all dose groups, ocular tissue concentrations of Formula II or Compound-I evaluated on day 5 were in descending order: highest in cornea >> central choroid > central retina (Table 15 - 2). Higher corneal concentrations of Formula II or Compound-I were observed in Group 10. Lower corneal concentrations of Formula II or Compound-I were observed in Group 8. In general, the highest concentrations in the central retina and central choroid were associated with higher corneal concentrations of Formula II or Compound-I. No advantage of other formulations over Formula II or Compound-I ophthalmic solutions was observed with respect to a reduction in corneal concentrations or an increase in central retinal or central choroid concentrations of Formula II or Compound-I. Table 15 – 2 *Average based on n = 3 LLOQ in Plasma = 0.00188 µM LLOQ in central Retina = 0.00751 µM LLOQ in central Choroid = 0.0563 µM LLOQ in Cornea = 0.0376 µM

[404] Gross ocular examinations using the Draize scale for counting ocular irritation were performed on all animals before the first dose on each day during the study. All formulations were well tolerated. Additionally, animals were observed for general health before the study, during dosing, and at necropsy. All animals assigned to the study were normal. The necropsy evaluations consisted of observations in addition to those already observed in the Draize Count sheets and Ophthalmological Examination forms. For all formulations, dosing observations were evaluated using the Draize ocular irritation counting system. No abnormal observations were noted except for the following: Before necropsy on day 5, Level 1 redness in both eyes was observed in one animal in Group I and one animal in Group M.

EXAMPLE 20-2 Ocular Pharmacokinetics of Order Formulations in Dutch Belted Rabbits

[405] A non-GLP study was conducted to evaluate the ocular pharmacokinetics of a first active agent of the present application (e.g., Formula II or Compound-I), in suspension or solution, when administered one, three or four times daily, as a topical instillation, into both eyes of Dutch Belted rabbits for four days. The study design (Table 16-1) consisted of thirty-nine (39) Dutch Belted rabbits, each receiving a 30 µL bilateral topical ocular dose. Administration was conducted according to the dosage regimen in Table 16-1. Table 16-1: Study Design aA single dose will be administered on day 5. QD - Once daily, TID - Three times daily, QID - Four times daily

[406]Ocular sampling occurred one hour after the first daily dose on day 5 for all groups. Whole blood samples using K2EDTA as an anticoagulant were collected from all animals on day 5, 1 hour post-dose. Whole blood was placed on ice until the samples were spun in a centrifuge to separate the plasma. Samples of aqueous humor, conjunctiva, cornea, central retina, peripheral retina, central choroid and peripheral choroid were collected after necropsy. Samples were then frozen at -70°C or lower. All plasma, central retina, central choroid and cornea samples were evaluated, Groups 1-13.

[407]For all dosage groups, ocular tissue concentrations of Formula II or Compound-I evaluated on day 5 were in descending order: highest: in cornea >> central choroid > central retina (Table 16-2). The highest corneal concentrations of Formula II or Compound-I were observed in Group 1. The lowest corneal concentrations of Formula II or Compound-I were observed in Group 3. In general, the highest central retinal and central choroidal concentrations High concentrations were associated with higher corneal concentrations of Formula II or Compound-I. No advantage of other formulations over the ophthalmic solution of Formula II or Compound-I was observed with respect to a reduction in corneal concentrations or an increase in central retinal or central choroidal concentrations of Formula II or Compound-I. Table 16-2 *Average based on n=3 Plasma LLOQ = 0.00188 µM Central retina LLOQ = 0.00751 µM Central choroid LLOQ = 0.0282 µM Cornea LLOQ = 0.0376 µM

[408] Gross ocular examinations using the Draize scale to score ocular irritation were performed on all animals before the first dose on each day during the study. All formulations were well tolerated. In addition, the animals were observed for general health pre-study, during dosing, and at necropsy. All animals assigned to study were normal. Necropsy evaluations consisted of observations beyond those already noted on Draize Score sheets and Ophthalmologic Examination forms. For all formulations, dosing observations were evaluated using the Draize ocular irritation scoring system. The following abnormal observations were observed throughout the duration of the study: animals in Groups 7, 9, 10, 11, 12 and 13 experienced increased blinking following dosing for several days of the study; one animal in Group 11 exhibited Secretion Level 1 on day 3.

EXAMPLE 20-3 Ocular pharmacokinetics of order formulations in Dutch Belted rabbits

[409] A non-GLP study was conducted to evaluate the ocular pharmacokinetics of a first active agent of the present application (e.g., Formula II or Compound-I), in suspension, when administered one, two or three times daily, as a topical instillation into both eyes of Dutch Belted rabbits for four days. The study design (Table 17-1) consisted of thirty-nine (39) Dutch Belted rabbits, each receiving a 30 µL bilateral topical ocular dose. Various suspension formulations of the present application were evaluated and compared against two control formulations: a) a cyclodextrin-based ophthalmic drop solution (Control Group 13, Table 17-1), and b) a pilot suspension formulation used in pharmacokinetic studies of previous rabbits, (Group 10, Table 171). A comparison of the formulation variables for each suspension listed in Table 17-1 is provided in Table 17-2. Formula II or Compound-I was used in the test suspensions, whereas the eye drop solution formulation is derived with Compound-I. Table 17-1: Study Design

[410]Ocular sampling occurred one hour after the first daily dose on day 5 for all groups. Whole blood samples using K2EDTA as an anticoagulant were collected from all animals on day 5, 1 hour post-dose. Whole blood was placed on ice until the samples were spun in a centrifuge to separate the plasma. Samples of aqueous humor, conjunctiva, cornea, central retina, peripheral retina, central choroid and peripheral choroid were collected after necropsy. Samples were then frozen at -70°C or lower. For all groups 1-13, plasma, central retina, central choroid, and cornea samples were evaluated.

[411]Ocular tissue concentrations of Formula II or Compound-I assessed on day 5 were in descending order: highest in cornea >> central choroid > central retina. Corneal concentrations of Formula II or Compound-I ranged from 1.5 to 14.7 µM with the highest concentrations being observed in Group 13 and lowest in Group 1. All suspension formulations, Groups 112, provided higher mean corneal concentrations. low when compared to the control eye drop solution, Group 13. These observations did not appear to be influenced by the presence or absence of Vitamin K3/menadione in the suspension formulation. Central choroidal concentrations ranged from <0.028 to 0.27 µM with the highest concentrations observed in Group 11 and lowest in Group 2. Central retinal concentrations ranged from 0.009 to 0.069 µM; Group 11 had the highest central retinal concentrations and Group 1 had the lowest central retinal concentrations. Plasma concentrations of Formula II or Compound-I were very low being approximately 10 times and 100 times lower than those in the central retina and central choroid, respectively. See, Table 17-3. Table 17-3 *Average based on n=3 Plasma LLOQ = 0.00188 µM Central retina LLOQ = 0.00751 µM Central choroid LLOQ = 0.0282 µM Cornea LLOQ = 0.0376 µM

[412] Additionally, Vitamin K3 concentrations were determined in plasma, central retina, central choroid and cornea samples for animals from Groups 8 and 9. A calibration curve prepared in control matrix was used to determine the concentration of Vitamin K3. See, Table 17-4. Table 17-4 *Average based on n=3 Plasma LLOQ = 0.291 µM Central retina LLOQ = 11.6 µM Central choroid LLOQ = 43.6 µM Cornea LLOQ = 2.91 µM

[413] Gross ocular examinations using the Draize scale to score ocular irritation were performed on all animals before the first dose on each day during the study. All formulations were well tolerated, with the exception that for one animal in Group 11 on day 4 of dosing, where conjunctival chemosis was observed, and for one animal in Group 12 on day 4 of dosing, where conjunctival redness was observed. Additionally, animals were observed for general health pre-study, during dosing, and at necropsy. All animals assigned to study were normal. Necropsy assessments consisted of observations beyond those already noted on Draize score sheets. For all formulations, dosing observations were evaluated using the Draize ocular irritation scoring system. The following abnormal observations were observed throughout the duration of the study: animals in Groups 4, 6, 7, 8, 9, and 11 experienced increased blinking following dosing for several days of the study; one animal in Group 4 exhibited Redness Level 1 in both eyes and Chemosis Level 1 in the left eye prior to necropsy on day 5; and one animal in Group 5 exhibited Chemosis Level 1 in the right eye prior to necropsy on day 5.

EXAMPLE 20-5 Ocular pharmacokinetics of order formulations in Dutch Belted rabbits

[414] A non-GLP study was conducted to evaluate the ocular pharmacokinetics of a first active agent of the present application (e.g., Formula II or Compound-I), in suspension, when administered once or twice daily, as a topical instillation, to both eyes of Dutch Belted rabbits for four days. The study design (Table 18-1) consisted of forty-nine (49) Dutch Belted rabbits, each receiving a 30 µL bilateral topical ocular dose. Various suspension formulations of the present application were evaluated and compared against a cyclodextrin-based ophthalmic drop solution (Control Groups 2 and 3, Table 18-1). A comparison of the formulation variables for each suspension listed in Table 18-1 is provided in Table 18-2. Formula II or Compound-I was used in the test suspensions, whereas the eye drop solution formulation is derived with Compound-I.

[415]Ocular sampling occurred one hour after the first daily dose on day 5 for all groups. Whole blood samples using K2EDTA as an anticoagulant were collected from all animals on day 5, 1 hour post-dose. Whole blood was placed on ice until the samples were spun in a centrifuge to separate the plasma. Samples of aqueous humor, conjunctiva, cornea, central retina, peripheral retina, central choroid and peripheral choroid were collected after necropsy. Samples were then frozen at -70°C or lower. For all groups 1-16, plasma, central retina, central choroid, and cornea samples were evaluated for PAN-90806.

[416]Ocular tissue concentrations of Formula II or Compound-I assessed on day 5 were in descending order: highest in cornea >> central choroid > central retina. Corneal concentrations of Formula II or Compound-I ranged from 3 to 34 µM with the highest concentrations being observed in Group 9 and lowest in Group 1. Central choroidal concentrations ranged from <0.056 to 0.24 µM with the highest concentrations observed in Groups 6 and 14 and lower in Group 1. Central retinal concentrations ranged from 0.02 to 0.14 µM; Group 6 had the highest central retinal concentrations and Group 1 had the lowest central retinal concentrations. Plasma concentrations of Formula II or Compound-I were very low being approximately 5 to 10 times and 100 times lower than those in the central retina and central choroid, respectively. See, Table 18-3. Table 18-3 Central retina LLOQ = 0.00751 µM Central choroid LLOQ = 0.0563 µM Cornea LLOQ = 0.0188 µM

[417] Gross ocular examinations using the Draize scale to score ocular irritation were performed on all animals before the first dose on each day during the study. All formulations were well tolerated. Additionally, animals were observed for general health pre-study, during dosing, and at necropsy. All animals assigned to study were normal. Necropsy assessments consisted of observations beyond those already noted on Draize score sheets. For all formulations, dosing observations were evaluated using the Draize eye irritation scoring system and are summarized below. No abnormal observations were noted.

EXAMPLE 20-4 Ocular pharmacokinetics of order formulations in Dutch Belted rabbits

[418] A non-GLP study was conducted to evaluate the ocular pharmacokinetics of a first active agent of the present application (e.g., Formula II or Compound-I), in suspension, when administered once or twice daily, as a topical instillation, to both eyes of Dutch Belted rabbits for four days. The study design (Table 19-1) consisted of forty-seven (47) Dutch Belted rabbits, each receiving a 30 µL bilateral topical ocular dose. Various suspension formulations of the present application were evaluated and compared against a cyclodextrin-based eye drop solution (Control Group 1, Table 19-1). A comparison of the formulation variables for each suspension listed in Table 19-1 is provided in Table 19-2. Formula II or Compound-I was used in the test suspensions as well as the eye drop solution. However, different batches of Compound-I were tested in certain suspensions (Groups 13, 14, and 15, Table 19-1). Table 19-1

[419]Ocular sampling occurred one hour after the first daily dose on day 5 for all groups. Whole blood samples using K2EDTA as an anticoagulant were collected from all animals on day 5, 1 hour post-dose. Whole blood was placed on ice until the samples were spun in a centrifuge to separate the plasma. Samples of aqueous humor, conjunctiva, cornea, central retina, peripheral retina, central choroid and peripheral choroid were collected after necropsy. Samples were then frozen at -70°C or lower. For all groups 1-15, plasma, central retina, central choroid, and cornea samples were evaluated. For Group 5 only, samples were analyzed for Vitamin K3.

[420]Ocular tissue concentrations of Formula II or Compound-I assessed on day 5, 1 hour after the first dose, were in descending order: highest in cornea >> central choroid > central retina. Corneal concentrations of Formula II or Compound-I ranged from 1 to 25 µM with the highest concentrations being observed in Group 12 and lowest in Group 7. Central choroidal concentrations ranged from 0.06 to 0.2 µM with the highest concentrations high observed in Groups 12 and 14 and lowest in Group 7. Central retinal concentrations ranged from 0.03 to 0.1 µM; Group 12 had the highest central retinal concentrations and Group 7 had the lowest central retinal concentrations. Plasma concentrations of Formula II or Compound-I were very low being approximately 10 times and 100 times lower than those in the central retina and central choroid, respectively. See, Table 19-3. Plasma LLOQ = 0.00188 µM Central retina LLOQ = 0.00751 µM Central choroid LLOQ = 0.0563 µM Cornea LLOQ = 0.0376 µM

[421] An analytical method for the analysis of Vitamin K3 was determined on samples of plasma, central retina, central choroid and cornea from Group 5 animals. A calibration curve prepared in control vitreous humor was used to determine the concentration of Vitamin K3 . Internal standard (IS) responses were inconsistent for the central choroidal and plasma samples. Thus, positive plasma results were likely due to reduced IS response. The Vitamin K3 response in plasma was similar to knowledge levels. Central choroidal samples had a positive Vitamin K3 response, but the reduced IS response made quantitative results suspicious. Central retinal and cornea samples had a normal IS response and no quantitative results for Vitamin K3. See, Table 19-4. Table 19-4 Plasma LLOQ = 0.0145 µM Central retina LLOQ = 0.581 µM Central choroid LLOQ = 2.18 µM Cornea LLOQ = 0.145 µM

[422] Gross ocular examinations using the Draize scale to score ocular irritation were performed on all animals before the first dose on each day during the study. In general, the formulations were well tolerated, with the exception of two animals in Group 6 on day 4 of dosing where conjunctival chemosis was observed, one animal in Group 8 on day 3 where conjunctival secretion was observed, one animal in Group 10 on day 5 of dosing, where chemosis and conjunctival discharge were observed, one animal in Group 13, and one animal in Group 14 on day 5 of dosing, where conjunctival chemosis was observed. Additionally, animals were observed for general health pre-study, during dosing, and at necropsy. All animals assigned to study were normal. Necropsy assessments consisted of observations beyond those already noted on Draize score sheets. For all formulations, dosing observations were evaluated using the Draize ocular irritation scoring system. The following clinical observations were noted throughout the study: One animal in Group 8 exhibited discharge in the left eye during dosing on day 3. Animals in Group 12 exhibited chemosis in both eyes at the time of necropsy.

EXAMPLE 21 Preparation of particles comprising Formula II

[423] Suspension formulations comprising Formula II particles were prepared by roller milling. Large particles were produced using larger grinding media (e.g., 3 mm in diameter) for a short period of time at low roller speed (Figure 9B). Nanoparticles were produced using smaller media (e.g., 0.8 mm in diameter) for a longer period of time at a high roller speed (Figure 9A). The two concentrations of Formula II (i.e., 0.4% and 0.2%) were diluted from a 5% API stock with a glycerol solution to obtain the appropriate tonicity. Improved cleaning precautions included autoclaving of product contact parts, filtration of excipient solutions, and transfer using prepackaged sterile supplies. Additionally, all transfers were within a laminar flow hood in a Clean Room area.

[424] Additional examples of particles comprising Formula II or Compound-I are shown in Tables 21 and 22. Table 21

EXAMPLE 22 Preparation of Compound-I/Tris Formulation

[425] An unground suspension of Compound-I was prepared. The formulation consisted of: Compound-I at 3%; 0.6% Tris-HCl pH 6; 2% glycerol. Compound-I was added to the solution directly and mixed without grinding and characterized (Figure 9C).

EXAMPLE 23 Particles comprising Formula II and Vitamin K3

[426]Addition of Vitamin K3 (menadione) to the suspension formulation may have a beneficial in vivo effect. Menadione was added to the formulation as a ground suspension. Menadione was initially milled exactly as Formula II was milled; a 5% suspension with Pluronic F-127 as the stabilizer. This suspension was then added to Formula II before milling (large and nanoparticulate formulations). The ground 5% suspensions (containing menadione suspension) were then diluted to 0.4% Formula II (in glycerol, as previously described).

[427] Menadione did not appear to affect any physical characteristics of the Formula II milled particles (Figure 9D). However, at the highest concentration of added menadione (10 µM), Vitamin K3 particles were detected and increased in size. Additionally, there was a small population of many large particles outside the range of the free base, which was presumably Vitamin K3. It was concluded that Vitamin K3 could not be distributed homogeneously like the other particles in the suspension, as they were growing at a faster rate than Formula II.

EXAMPLE 24 Preparation of particles comprising menadione

[428]Menadione/Formula II formulation may form larger crystals over time. An optimized formulation was needed to be able to add menadione as a suspension. Formulations containing 5% menadione were screened using a roller mill in the same manner as when screening Formula II formulations. An HPMC formulation produced a fine homogeneous suspension that demonstrated short-term stability at 40 °C (Figure 10).

EXAMPLE 25 Preparation of particles comprising Formula II

[429] The HPMC formulation for menadione was used with Formula II to provide a formulation in which menadione can be added as a suspension. It was found that an X-large particle size (30 to 40 microns) using HPMC can be milled in a shorter time than when using Pluronic F-127 as the stabilizer (Figures 11A and 11B). The suspension can then be diluted with a menadione/HPMC suspension.

EXAMPLE 25 Stability of menadione in formulations of this application

[430]The concentration of menadione was 20 µM in the stability study. Menadione was formulated as follows: 0.43% (w/v) Compound-I (unmilled, 50 µm), 0.6% Tris/HCl, pH 6, 2% Glycerol, 0.2 HEC %, 0.08% HPMC and 20 µM menadione. From this formulation, ten test formulations were prepared: 1. less glycerin (no N2) 2. control formulation; (none N2) 3. control formulation; Powdered N2 4. + 0.3% Na2 thiosulfate (pentahydrate), powdered N2* 5. + 0.1% Na2 EDTA, powdered N2 6. + 0.5% TPGS vitamin E, N2 powdered 7. + 0.05% propyl gallate, N2 powdered 8. + 0.02% BHT, N2 powdered 9. + menadione/HPb-cyclodextrin complex, N2 powdered 10. + antioxidant complex (#5 to # 9), N2 spray

[431] 7 mL of each formulation was added to a 40 mL amber vial (under amber lighting). Filtered nitrogen gas was slowly bubbled into the bottom of the flask until the generated foam began to come out of the top of the flask, which was then promptly capped. Menadione/HPb-Cyclodextrin Complex (#9) was prepared by mixing approximately equimolar concentrated solutions of cyclodextrin and menadione before addition (approximately 2 hours) to the formulation.

EXAMPLE 26 Methods for roller milling and particle characterization of the present application roller mill

[432]The horizontal roller mill (US Stoneware, model 755) consists of four 12” engine-driven rubber rollers contained within a metal casing. Individual vials placed between the rollers will rotate at an rpm determined by the speed of the rollers and the diameter of the vial. Drug suspension consisting of API, stabilizers, water and grinding media was added to the vial and the lid tightly sealed before placing between the rollers. The medium used was an extremely dense Yttria Zirconium bead ranging in diameter from 800 microns to 3000 microns. After grinding, the dispersion was separated from the medium by transferring the contents to an inserted centrifuge tube fitted with a mesh sieve. The small insert was placed in a centrifuge tube. The centrifuge was then run at approximately 300 x G for approximately 5 minutes. The dispersion collected below the mesh (which held the medium) in the tube.

Optical Microscopy (OM)

[433] Optical microscopy microphotographs of nanoparticles were taken using an Olympus BX51 system equipped with a 100x oil immersion objective (1000x magnification). A calibration bar (from 1 µm to 100 µm) was adjusted as a comparator on each photomicrograph. (The calibration bar effectively caters for larger sized particles at lower magnifications, too.)

Particle Size Distribution (PSD)

[434] Particle size distribution was analyzed using laser diffraction light scattering with a Horiba LA-950 V2. Generic assumptions were made on fitting conditions and the refractive index value. Distributions were based on volume. Sample density was adjusted to a generic range of percentage transmission in the blue LED light source. A small sample cell (filled with water) was used instead of flow through the cell, to minimize sample amount.

INCORPORATION BY REFERENCE

[435] The full disclosure of each of the patent documents and scientific articles referenced herein is incorporated by reference for all purposes. In the present application, the host document is identified with sufficient particularity and materials that are relevant to the disclosure are interpreted based on the context of the reference. Citation of publications and patent documents is not intended as an admission that any prior art is pertinent, nor does it constitute any admission as to the content or data thereof. The application having now been described by way of written description, those of skill in the art will recognize that the application may be practiced in a variety of embodiments and the foregoing description and examples are for purposes of illustration and not limitation of the claims that follow. .

EQUIVALENTS

[436]The request may be incorporated in other specific forms without departing from its spirit or essential characteristics. The foregoing embodiments are therefore considered in all respects illustrative rather than limiting the application described herein. The scope of the application is thus indicated by the appended claims rather than by the preceding description, and all changes that are within the meaning and range of equivalence of the claims are intended to be included therein.

Claims (16)

1. Topical ocular suspension formulation, CHARACTERIZED by the fact that it comprises: a. a first active agent of Formula II: or a pharmaceutically acceptable salt thereof; B. a second active agent, or a pharmaceutically acceptable salt thereof, wherein the second active agent is nicotinic acid, nicotinamide, or vitamin K, or a combination thereof; and c. a pharmaceutically acceptable excipient; wherein the first active agent or pharmaceutically acceptable salt thereof is present at 0.005% to 5.0% w/v. 2. Formulation according to claim 1, CHARACTERIZED by the fact that the first active agent or the pharmaceutically acceptable salt thereof is present in 0.1% to 2.0% w/v. 3. Formulation according to claim 1, CHARACTERIZED by the fact that the first active agent or the pharmaceutically acceptable salt thereof is present in 0.2% to 1.0% w/v. 4. Formulation according to any one of claims 1 to 3, CHARACTERIZED by the fact that the pharmaceutically acceptable salt of the first active agent is Compound I: 5. Formulation, according to any one of claims 1 to 4, CHARACTERIZED by the fact that it comprises 0.1%, 0.2%, 0.3%, or 0.4% w/v of the first active agent or of the pharmaceutically acceptable salt thereof. 6. Formulation according to any one of claims 1 to 5, CHARACTERIZED by the fact that the second active agent is vitamin K. 7. Formulation according to claim 1, CHARACTERIZED by the fact that it comprises particles of the first active agent or the pharmaceutically acceptable salt thereof, wherein the particles have an average diameter between 100 nm and 100 µm. 8. Formulation, according to claim 7, CHARACTERIZED by the fact that the particles have an average diameter between 30 µm and 60 µm, or between 1 µm and 5 µm. 9. Formulation, according to claim 7, CHARACTERIZED by the fact that the particles have an average diameter of a maximum of 150 nm. 10. Formulation according to claim 7, CHARACTERIZED by the fact that the particles have an average diameter of 2.85 µm to 3.15 µm, 28.5 µm to 31.5 µm, 33.25 µm to 36, 75 µm, or 47.5 µm to 52.5 µm. 11. Formulation according to claim 7, CHARACTERIZED by the fact that the second active agent is present in an amount of less than 10 µM, optionally wherein the second active agent is present in an amount of 0.95 µM to 1.05 µM. 12. Formulation, according to claim 7, CHARACTERIZED by the fact that it further comprises one or more excipients selected from Polysorbate (Tween) 80, Poloxamer (Pluronic) F-127, Hypromellose (Hydroxypropyl Methylcellulose or HPMC), Povidone ( PVP K-29/32 or K-30), and Tyloxapol, and a combination thereof. 13. Formulation according to claim 7, CHARACTERIZED by the fact that the first active agent is in a concentration of 0.1 mg/mL to 5 mg/mL, optionally 1 mg/mL to 4 mg/mL. 14. Formulation according to claim 1, CHARACTERIZED by the fact that it is for use in the treatment and/or improvement of a disease of the posterior segment of the eye selected from the group consisting of: diabetic retinopathy, age-related macular degeneration (AMD) , pathological choroidal neovascularization (CNV), pathological retinal neovascularization, uveitis, retinal vein occlusion, ocular trauma, surgery-induced edema, surgery-induced neovascularization, cystoid macular edema, ocular ischemia, retinopathy of prematurity, Coat's disease, sickle cell retinopathy , and neovascular glaucoma. 15. Formulation according to claim 7, CHARACTERIZED by the fact that it is for use in the treatment and/or improvement of a disease of the posterior segment of the eye selected from the group consisting of: diabetic retinopathy, age-related macular degeneration (AMD) , pathological choroidal neovascularization (CNV), pathological retinal neovascularization, uveitis, retinal vein occlusion, ocular trauma, surgery-induced edema, surgery-induced neovascularization, cystoid macular edema, ocular ischemia, retinopathy of prematurity, Coat's disease, sickle cell retinopathy , and neovascular glaucoma. 16. Use of the formulation, as defined in any one of claims 1 to 15, CHARACTERIZED by the fact that it is in the manufacture of a medicament to treat and/or improve a disease of the posterior segment of the eye selected from the group consisting of: diabetic retinopathy, age-related macular degeneration (AMD), pathological choroidal neovascularization (CNV), pathological retinal neovascularization, uveitis, retinal vein occlusion, ocular trauma, surgery-induced edema, surgery-induced neovascularization, cystoid macular edema, ocular ischemia, retinopathy of prematurity, Coat's disease, sickle cell retinopathy, and neovascular glaucoma.